U.S. patent application number 13/312291 was filed with the patent office on 2013-01-31 for methods of preparing substituted tetracyclines with transition metal-based chemistries.
This patent application is currently assigned to Trustees of Tufts College. The applicant listed for this patent is Darrell J. Koza, Mark L. Nelson, Glen Rennie. Invention is credited to Darrell J. Koza, Mark L. Nelson, Glen Rennie.
Application Number | 20130029943 13/312291 |
Document ID | / |
Family ID | 34865541 |
Filed Date | 2013-01-31 |
United States Patent
Application |
20130029943 |
Kind Code |
A1 |
Nelson; Mark L. ; et
al. |
January 31, 2013 |
Methods of Preparing Substituted Tetracyclines with Transition
Metal-Based Chemistries
Abstract
The present invention relates to novel chemistries which allow
for heretofore unobtainable substituted tetracycline compounds
which exhibit significant activity in tetracycline responsive
states. The methods disclosed herein utilize reactive
tetracycline-based precursor compounds, reactive organic
substituent precursors and transition metal catalysts under
conditions such that a tetracycline compound substituted with the
desired organic substituent is formed. In one embodiment of the
invention, a substituted tetracycline compound may be prepared by
combining a reactive tetracycline-based precursor compound such as
an arene tetracycline diazonium salt, and a reactive organic
substituent precursor, e.g., alkenes, substituted alkenes, vinyl
monomers, aromatics and heteroaromatics, in the presence of a
transition metal catalyst, such as palladium chloride, under
conditions such that a tetracycline compound substituted with the
organic substituent is formed. Such compounds may optionally act as
intermediates for making other compounds, e.g., hydrogenation of
unsaturated groups on the substituent.
Inventors: |
Nelson; Mark L.; (Norfolk,
MA) ; Rennie; Glen; (South Weymouth, MA) ;
Koza; Darrell J.; (Willimantic, CT) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Nelson; Mark L.
Rennie; Glen
Koza; Darrell J. |
Norfolk
South Weymouth
Willimantic |
MA
MA
CT |
US
US
US |
|
|
Assignee: |
Trustees of Tufts College
Medford
MA
|
Family ID: |
34865541 |
Appl. No.: |
13/312291 |
Filed: |
December 6, 2011 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11004559 |
Dec 3, 2004 |
8106225 |
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13312291 |
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10765233 |
Jan 26, 2004 |
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11004559 |
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10391923 |
Mar 18, 2003 |
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10765233 |
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09660598 |
Sep 13, 2000 |
6617318 |
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10391923 |
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60154701 |
Sep 14, 1999 |
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60232091 |
Sep 12, 2000 |
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60367050 |
Mar 21, 2002 |
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Current U.S.
Class: |
514/152 ;
552/203; 552/205 |
Current CPC
Class: |
C07D 295/185 20130101;
C07C 231/12 20130101; C07D 295/192 20130101; C07C 231/12 20130101;
C07C 237/26 20130101; C07C 237/26 20130101; C07C 2603/46 20170501;
A61P 31/04 20180101 |
Class at
Publication: |
514/152 ;
552/203; 552/205 |
International
Class: |
C07C 237/26 20060101
C07C237/26; A61P 31/04 20060101 A61P031/04; A61K 31/65 20060101
A61K031/65 |
Claims
1. A compound selected from the group consisting of:
[4S-(4.alpha.,12a.alpha.)]-9-[3'-(E)-propenoic
acid]-4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,5,10,12,12a-pent-
ahydroxy-6-methyl-1,1'-dioxo-2-naphthacenecarboxamide,
[4S-(4.alpha.,12a.alpha.)]-9-[3'-(E)-butylpropenoate]-4-(dimethylamino)-1-
,4,4a,5,5a,6,11,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-1,1'-dio-
xo-2-naphthacenecarboxamide,
[4S-(4.alpha.,12a.alpha.)]-9-[3'-(E)-butylpropenoate]-4,7-Bis(dimethylami-
no)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,1'-dioxo-2-n-
aphthacenecarboxamide,
[4S-(4.alpha.,12a.alpha.)]-9-[1'-(E)-(2'-phenyl)ethenyl]-4-(dimethylamino-
)-1,4,4a,5,5a,6,11,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-1,1'--
dioxo-2-naphthacenecarboxamide,
[4S-(4.alpha.,12a.alpha.)]-9-carboxy-4-(dimethylamino)-1,4,4a,5,5a,6,11,1-
2a-octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-1,1'-dioxo-2-naphthacenec-
arboxamide, 9-phenyl minocycline, 7,9-diphenyl sancycline,
7-ethylenyl sancycline,
[4S-(4.alpha.,12a.alpha.)]-7-[3'-(E)-butylpropenoate]-4-(dimethylamino)-1-
,4,4a,5,5a,6,11,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-1,1'-dio-
xo-2-naphthacenecarboxamide,
[4S-(4.alpha.,12a.alpha.)]-13-(4'-methylphenyl)-4-(dimethylamino)-1,4,4a,-
5,5a,6,11,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-methylene-1,1'-dioxo--
2-naphthacenecarboxamide,
[4S-(4.alpha.,12a.alpha.)]-13-(3'-carboxyphenyl)-4-(dimethylamino)-1,4,4a-
,5,5a,6,11,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-methylene-1,1'-dioxo-
-2-naphthacenecarboxamide, and
[4S-(4.alpha.,12a.alpha.)]-13-(4'-ethoxyphenyl)-4-(dimethylamino)-1,4,4a,-
5,5a,6,11,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-methylene-1,1'-dioxo--
2-naphthacenecarboxamide, or a pharmaceutically acceptable salt
thereof.
2. A compound selected from the group consisting of:
9-iodo-minocycline, 9-iodo-sancycline, 9-iodo-doxycycline,
7,9-diiodosancycline, and 7-iododoxycycline, or a pharmaceutically
acceptable salt thereof.
3. A 7-substituted tetracycline analog, wherein the substituent at
the 7 position is connected with a --C--C-- linkage, and wherein
the substituent comprises a --C.dbd.C-- bond adjacent to the
--C--C-- linkage.
4. The 7-substituted tetracycline analog of claim 3, wherein the
substituent is of the formula ##STR00030## wherein R.sub.2 and
R.sub.3 are each independently hydrogen, alkyl, alkenyl, alkynyl,
cycloalkyl, aryl, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl,
aryloxycarbonyl, amino, hydroxyl, cyano, alkoxy, aryloxy, or
carboxyl; or R.sub.2 and R.sub.3, taken together, form a
substituted or unsubstituted carbocyclic or heterocyclic ring
having 5 to 15 atoms in the ring.
5. The 7-substituted tetracycline analog of claim 4, wherein
R.sub.2 is hydrogen; R.sub.3 is ##STR00031## and R.sub.4 is
hydrogen, cyano, or C.sub.1-C.sub.5 alkoxy.
6. The 7-substituted tetracycline analog of claim 4, wherein
R.sub.2 and R.sub.3, taken together, form a substituted or
unsubstituted carbocyclic or heterocyclic ring having 5 to 15 atoms
in the ring.
7. The 7-substituted tetracycline analog of claim 5, wherein the
substituted or unsubstituted carbocyclic or heterocyclic ring is
selected from the group consisting of conjugated and unconjugated
aromatic ring systems.
8. A pharmaceutical composition comprising the compound of any one
of claims 1-7 and a pharmaceutically acceptable carrier.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. Ser. No.
11/004,559, filed on Dec. 3, 2004, allowed, which is a continuation
of U.S. Ser. No. 10/765,233, filed on Jan. 26, 2004, abandoned,
which is a continuation of U.S. Ser. No. 10/391,923, filed on Mar.
18, 2003, abandoned, which is a continuation-in-part of U.S. Ser.
No. 09/660,598, filed on Sep. 13, 2000, now U.S. Pat. No.
6,617,318, which claims priority to U.S. Ser. No. 60/232,091, filed
on Sep. 12, 2000 and to U.S. Ser. No. 60/154,701, filed on Sep. 14,
1999. U.S. Ser. No. 10/391,923 claims priority to U.S. Ser. No.
60/367,050, filed on Mar. 21, 2002.
BACKGROUND OF THE INVENTION
[0002] The development of the tetracycline antibiotics was the
direct result of a systematic screening of soil specimens collected
from many parts of the world for evidence of microorganisms capable
of producing bacteriocidal and/or bacteriostatic compositions. The
first of these novel compounds was introduced in 1948 under the
name chlortetracycline. Two years later oxytetracycline became
available. The detailed elucidation of the chemical structure of
these agents confirmed their similarity and furnished the
analytical basis for the production of a third member of this group
in 1952, tetracycline. By 1957, a new family of tetracycline
compositions characterized chemically by absence of the position 6
ring-attached OH group present in the earlier compositions was
prepared and became publicly available in 1967; and minocycline was
in use by 1972. Individual tetracycline-type agents are
structurally compared within Table I below, with reference made the
following structural formula:
TABLE-US-00001 TABLE I ##STR00001## Congener Substituent(s) At
Carbon Position Nos. Chlortetracycline --Cl (7) Oxytetracycline
--OH, --H (5) Demeclocycline --OH, --H; --Cl (6; 7) Methacycline
--OH, --H; .dbd.CH.sub.2 (5; 6) Doxycycline --OH, --H; --CH.sub.3,
--H (5; 6) Minocycline --H, --H; --N(CH.sub.3).sub.2 (6; 7)
[0003] More recent research efforts have focused on developing new
tetracycline antibiotic compositions effective under varying
therapeutic conditions and routes of administration; and for
developing new tetracycline analogues which might prove to be equal
or more effective then the originally introduced tetracycline
families beginning in 1948. Representative of such developments
include U.S. Pat. Nos. 3,957,980; 3,674,859; 2,980,584; 2,990,331;
3,062,717; 3,557,280; 4,018,889; 4,024,272; 4,126,680; 3,454,697;
and 3,165,531. It will be understood that these issued patents are
merely representative of the range of diversity of investigations
seeking tetracycline and tetracycline analogue compositions which
are pharmacologically active.
[0004] Historically, soon after their initial development and
introduction, the tetracyclines, regardless of specific formulation
or chemical structure, were found to be highly effective
pharmacologically against rickettsiae; a number of gram-positive
and gram-negative bacteria; and the agents responsible for
lymphogranuloma venereum, inclusion conjunctivitis, and
psittacosis. Hence, tetracyclines became known as "broad spectrum"
antibiotics. With the subsequent establishment of their in vitro
antimicrobial activity, effectiveness in experimental infections,
and pharmacological properties, the tetracyclines as a class
rapidly became widely used for therapeutic purposes. However, this
widespread use of tetracyclines for both major and minor illnesses
and diseases led directly to the emergence of resistance to these
antibiotics even among highly susceptible bacterial species both
commensal and pathogenic--as for example pneumococci and
Salmonella. The rise of tetracycline-resistant organisms has
resulted in a general decline in use of tetracyclines and
tetracycline analogue compositions as antibiotics of choice.
SUMMARY OF THE INVENTION
[0005] The present invention relates to novel chemistries which
allow for the production of substituted tetracycline compounds
including substituted tetracycline compounds which exhibit
significant antibacterial activity. The methods disclosed herein
utilize reactive tetracycline-based precursor compounds, reactive
organic substituent precursors and transition metals or transition
metal catalysts under conditions such that a tetracycline compound
substituted with the desired organic substituent is formed. In one
embodiment of the invention, a substituted tetracycline compound
may be prepared by combining a reactive tetracycline-based
precursor compound such as an arene tetracycline diazonium salt,
and a reactive organic substituent precursor, e.g., alkenes,
substituted alkenes, vinyl monomers, aromatics and heteroaromatics,
in the presence of a transition metal catalyst, such as palladium
chloride, under conditions such that a tetracycline compound
substituted with the organic substituent is formed. In another
embodiment, a substituted tetracycline compound may be prepared by
contacting a reactive tetracycline chemical complex comprising a
reactive tetracycline-based precursor compound and a transition
metal or transition metal catalyst forming a reactive chemical
intermediate with a reactive organic substituent precursor under
conditions such that a tetracycline compound substituted with the
organic substituent is formed.
[0006] The invention relates in another embodiment to reactive
tetracycline chemical complexes comprising a reactive
tetracycline-based precursor compound and a transition metal
catalyst forming a chemical intermediate, which can advantageously
be used in the methods of the invention.
[0007] In yet another embodiment substituted tetracycline analogs
are disclosed, wherein the substituent (denoted herein as "Z") at
the desired position, e.g., 7, 9, 13, is connected with a --C--C--
linkage, and wherein the substituent comprises an aromatic or
heteroaromatic moiety. The substituent may also comprise a
--C.dbd.C-- bond adjacent to the --C--C-- linkage, e.g.,
##STR00002##
wherein R.sub.2 and R.sub.3 are each independently hydrogen, alkyl,
alkenyl, alkynyl, cycloalkyl, aryl, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aryloxycarbonyl, amino, hydroxy, cyano, alkoxy,
aryloxy, carboxyl, alkoxycarbonyl, aryloxycarbonyl; or R.sub.2 and
R.sub.3, taken together, form a substituted or unsubstituted
carbocyclic or heterocyclic ring having 5 to 15 atoms in the
ring.
[0008] The methods and chemical intermediates disclosed herein
allow for novel substituted tetracycline-type compounds and
therapeutic methods and pharmaceutical compositions that comprise
such compounds.
[0009] The method of the invention includes providing Z
substituents, above, on the basic tetracycline ring structure
through a process involving forming a reactive intermediate
(comprising a tetracycline arenediazonium salt in a preferred
embodiment) at the desired position and adding a reactive compound,
e.g., a n-bond containing compound in the presence of a transition
metal catalyst to that position. The reactive intermediate may be
formed in situ. In an advantageous embodiment such substituents are
provided on the D ring of the basic tetracycline ring structure,
e.g., positions 7 and/or 9. In another advantageous embodiment,
such substitutions may be made at position 13. Such synthetic
schemes are heretofore new in this art and advantageously allow for
direct substitution of different and/or heretofore complex
substituent groups at desired positions.
[0010] Compounds of the invention are active against susceptible
microorganisms, including tetracycline-sensitive bacteria as well
as tetracycline-resistant bacteria. Particularly preferred
compounds of the invention exhibit 24-hr minimum inhibitory
concentration (MIC) values of about 10 .mu.g/mL or less, more
preferably about 1 .mu.g/mL or less, against tetracycline-resistant
E. coli, S. aureus and E. faecalis strains such as E. coli pHCM1,
S. aureus RN4250 and E. faecalis pMV158. Preferred compounds of the
invention also include those that exhibit such MIC values against
tetracycline-sensitive E. coli, S. aureus and E. faecalis strains
such as E. coli D31 m4, S. aureus RN450 and E. faecalis
ATCC9790.
[0011] The invention provides methods of treatment against
susceptible microorganisms such as bacteria, fungi, rickettsia,
parasites and the like, and diseases associated with such
microorganisms. These therapeutic methods in general comprise
administration of a therapeutically effective amount of one or more
compounds of the invention to a living subject that is suffering
from or susceptible to infection by a susceptible microorganism
such as bacteria, fungi, rickettsia and the like. Suitable subjects
for treatment include animals, particularly a mammal such as human,
or plants.
[0012] Pharmaceutical compositions comprising one or more compounds
of the invention and a suitable carrier are also provided.
DETAILED DESCRIPTION OF THE INVENTION
[0013] The present invention will be more fully illustrated by
reference to the definitions set forth below.
[0014] "Tetracycline" or "tetracycline-type" is intended to include
tetracycline and other tetracycline family members such as
oxytetracycline; chlortetracycline; demeclocycline; doxycycline;
chelocardin; minocycline; rolitetracycline; lymecycline;
sancycline; methacycline; apicycline; clomocycline; guamecycline;
meglucycline; mepylcycline; penimepicycline; pipacycline;
etamocycline; penimocycline, etc. as well as other tetracycline
compounds, such as 4-dedimethylamino tetracycline compounds, having
the characteristic naphthacene A-B-C-D ring structure noted in the
Background Of The Invention. Additionally, numbered tetracycline
ring positions as referred to herein are the same as designated in
the above structural formula.
[0015] "Reactive tetracycline-based precursor compound" or
"RT-based precursor compound" includes tetracyclines which have a
reactive position on the tetracycline ring structure, e.g., at 7, 9
or 13, such that substitution of the reactive tetracycline-based
precursor compound may be accomplished as disclosed herein to form
a substituted tetracycline compound. Examples of RT-based precursor
compounds include derivatives from art-recognized tetracycline
compound families. Without limitation, such tetracycline compound
families include minocycline, doxycycline and sancycline
compounds.
[0016] "Minocycline-based precursor compound" is intended to
include compounds having the core structure of minocycline, which
differs from the core structure of tetracycline by the presence of
a dimethylamino group at position 7, and the absence of methyl and
hydroxyl groups at position 6, and the absence of a hydroxyl group
at position 5. The core structure of minocycline-based precursor
compounds is shown below for the purposes of illustration:
##STR00003##
It should be understood that minocycline-based precursor compounds
can be substituted, unsubstituted or derivatized, e.g., at
positions other than positions 5 and 6. For example, other
positions in the core structure, e.g., position 8, can be
substituted or unsubstituted and others can be substituted or
derivatized, such as the 2-position amido group. Suitable
substituents include moieties such as hydrogen, alkyl, alkenyl,
alkynyl, cycloalkyl, aryl, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aryloxycarbonyl, amino, hydroxy, cyano, alkoxy,
aryloxy, carboxyl, carboxamido, carboxy ester, alkoxycarbonyl,
aryloxycarbonyl, carbocyclic or heterocyclic groups, and
combinations thereof. Other substituent groups will be recognized
by those of skill in the art. Further, R in the above formula can
represent a group other than methyl, e.g., lower alkyl such as
ethyl, propyl, etc. Reactive minocycline-based precursor compounds
include, without limitation, 9-diazonium minocycline-based
compounds, 9-iodo minocycline-based compounds, 9-bromo
minocycline-based compounds, and 9-chloro minocycline-based
compounds.
[0017] "Doxycycline-based precursor compound" is intended to
include compounds having the core structure of doxycycline, which
differs from the core structure of tetracycline by the substitution
of a hydrogen for a hydroxyl at position 6, and the substitution of
a hydroxyl for a hydrogen at position 5. The core structure of
doxycycline-based precursor compounds is shown below for the
purposes of illustration:
##STR00004##
It should be understood that doxycycline-based precursor compounds
can be substituted, unsubstituted or derivatized, e.g., at
positions 7, 8 and/or 9. For example, other positions in the core
structure, e.g., position 8, can be substituted or unsubstituted
and others can be substituted or derivatized, such as the
5-position hydroxyl group or the 2-position amido group. Suitable
substituents include moieties such as hydrogen, alkyl, alkenyl,
alkynyl, cycloalkyl, aryl, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aryloxycarbonyl, amino, hydroxy, cyano, alkoxy,
aryloxy, carboxyl, carboxamido, carboxy ester, alkoxycarbonyl,
aryloxycarbonyl, carbocyclic or heterocyclic groups, and
combinations thereof. Other substituent groups will be recognized
by those of skill in the art. Further, R in the above formula can
represent a group other than methyl, e.g., lower alkyl such as
ethyl, propyl, etc. Reactive doxycycline-based precursor compounds
include, without limitation, 7- and/or 9-diazonium doxycycline
compounds, 7- and/or 9-iodo doxycycline compounds, 7- and/or
9-bromo doxycycline compounds, and 7- and/or 9-chloro doxycycline
compounds.
[0018] "Sancycline-based precursor compound" is intended to include
compounds having the core structure of sancycline, which differs
from the core structure of tetracycline by the substitution of a
hydrogen for a methyl group and hydrogen for a hydroxyl at position
at position 6. The core structure of sancycline-based precursor
compounds is shown below for the purposes of illustration:
##STR00005##
It should be understood that sancycline-based precursor compounds
can be substituted, unsubstituted or derivatized, e.g., at
positions 7, 8 and/or 9. For example, other positions in the core
structure, e.g., position 8, can be substituted or unsubstituted
and others can be substituted or derivatized, such as the
2-position amido group. Suitable substituents include moieties such
as hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl,
alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aryloxycarbonyl,
amino, hydroxy, cyano, alkoxy, aryloxy, carboxyl, carboxamido,
carboxy ester, alkoxycarbonyl, aryloxycarbonyl, carbocyclic or
heterocyclic groups, and combinations thereof. Other substituent
groups will be recognized by those of skill in the art. Reactive
sancycline-based precursor compounds include, without limitation,
7- and/or 9-diazonium sancycline compounds, 7- and/or 9-iodo
sancycline compounds, 7- and/or 9-bromo sancycline compounds, and
7- and/or 9-chloro sancycline compounds.
[0019] In a preferred embodiment, the reactive tetracycline-based
precursor compound is an arene tetracycline diazonium salt, and
alternately iodo derivatized tetracycline compounds, or
tetracycline compounds that possess a double bond and are reactive
with boronic acid derivatives, e.g., at position 13. In one
embodiment, the reactive tetracycline-based precursor compound and
a transition metal catalyst form a reactive chemical intermediate
useful in making novel tetracyclines, through techniques known in
the art (see, for example, Hegedus, Transition Metals in the
Synthesis of Complex Organic Molecules, University Science Books,
Mill Valley, Calif., 1994, incorporated herein by reference). The
reactive chemical intermediate are preferably formed in situ with
the reactive organic substituent precursor.
[0020] "Transition metal catalyst" is an art-recognized term which
includes transition metals and catalysts comprising a transition
metal, e.g., including elements 21 through 29, 39 through 47, 57
through 79, and 89 on. Exemplary transition metal catalysts include
CuCl.sub.2, copper (I) triflate, copper thiophene chloride,
palladium (II) chloride, organopalladium catalysts such as
palladium acetate, Pd(PPh.sub.3).sub.4, Pd(AsPh.sub.3).sub.4,
Pd(dppf).sub.2Cl.sub.2, Pd(dppf).sub.2, PdCl.sub.2(PhCN).sub.2,
PdCl.sub.2 (Ph.sub.3P).sub.2, Pd.sub.2(dba).sub.3--CHCl.sub.3
("dba"=dibenzylacetone); and combinations thereof. Other transition
metal catalysts include those containing metals such as rhodium
(e.g. rhodium (II) acetate and Rh.sub.6(CO).sub.16), iron, iridium,
chromium, zirconium, and nickel. A skilled artisan will be able to
select the appropriate transition metal catalyst to perform the
desired reaction, based on the existing literature (see, for
example, Lipshutz, B. H. Org. React. 1992, 41:135, incorporated
herein by reference.)
[0021] "Reactive organic substituent precursor" includes organic
substituents having a reactive group that allows for addition to
the reactive tetracycline-based precursor compound as disclosed
herein. Preferably the reactive organic substituent precursor
comprises at least one reactive group. In an embodiment, the
reactive organic substituent precursor may include .pi.-bonded
species such as methylene compounds, aryl boronic acids, active
aromatic rings and unsubstituted and substituted olefins and
alkynes, nitriles, acetylenes, substituted acetylenes,
arylethylenes, styrenes, conjugated dienes, isoprenes, vinyl
ethers, .alpha.,.beta.-unsaturated aldehydes and ketones, aryl
vinyl and arylisoprenyl ketones, iodoalkenes and iodoarenes,
quinones, carbon monoxide, .alpha.,.beta.-unsaturated acids and
their derivatives.
[0022] "Reactive organic substituent precursors" also include
compounds (which may be formed in situ) which react with the
reactive intermediate to form a desired tetracycline analog. For
example, the reactive intermediate can be transmetallated to form a
wide variety analogs through reactions with other organometal
complexes such as tributyltin compounds (e.g., amino tributyl tin
compounds, aryl tributyltin compounds, alkynyl tributyl tin
compounds, and alkenyl tributyl tin compounds, etc.) and lithium
diorganocuprates (see for example, Kalanin, Synthesis, 1992, 413;
Sawamuru, Chem. Rev. 1992, 92:857; Negeishi, Acct. Chem. Res.,
1982, 15:340, incorporated herein by reference). Other precursors
include those suitable for transition metal catalyzed reactions
include compounds with bonds which are reactive with the transition
metal containing intermediates. Such precursors include, for
example, compounds with halogen groups, hydroxyl groups, triflate
groups, thiol groups, or amino groups. Intramolecular reactions are
also included wherein the reactive organic substituent precursor is
bonded or associated with the reactive chemical intermediate (see
Hegedus, supra).
[0023] The appropriate reaction conditions may include additional
reactive organic substituent precursors in combination with the
reactive organic substituent precursor, to form a particular
desired substituted tetracycline compound.
[0024] The term "in combination with" includes the addition of the
additional reactive organic substituent precursor simultaneously
with the other reactive organic substituent precursor, prior to the
other reactive organic substituent precursor, or after the other
reactive organic substituent precursor.
[0025] The term "additional reactive organic substituent precursor"
include compounds and reagents such that a reactive intermediate
can be reacted to form a desired substituted tetracycline compound.
One example of a reactive intermediate includes carboxylic acids of
tetracycline compounds which can be further derivatized by a
variety of methods known to those of ordinary skill in the art.
Examples of reactive organic substituent precursors include, but
are not limited to, amide precursors, ester precursors, anhydride
precursors, hydrazone precursors, imide precursors, ketone
precursors, acid halide precursors, or nitrile precursors.
[0026] The term "amide precursors" includes reagents and conditions
which can be reacted with an appropriate reactive intermediate of
the invention (such as, for example, a carboxylic acid), such that
an amide bond is formed. Examples of amide precursors include
primary and secondary amines (See, for example, Syn. 453 (1974) and
LaRock, Comprehensive Organic Transformations (VCH Publishers,
Inc.: New York, 1989), pp. 972-976).
[0027] The term "ester precursors" includes reagents and conditions
which can be reacted with an appropriate reactive intermediate of
the invention, such as, for example, a carboxylic acid, such that
an ester bond is formed. Examples of ester precursors include
alcohols. Other ester precursors are known in the art (See, for
example, Tetr. 36, 2409 (1980) and LaRock, supra, pp. 966-972).
[0028] The term "anhydride precursors" includes reagents and
conditions which can be reacted with an appropriate reactive
intermediate of the invention, such as, for example, a carboxylic
acid, such that an anhydride bond is formed. Examples of ester
precursors include carboxylic acids. Other anhydride precursors are
known in the art (See, for example, LaRock, supra, pp.
965-966).
[0029] The term "hydrazone precursors" includes reagents and
conditions which can be reacted with an appropriate reactive
intermediate of the invention, such as, for example, a carboxylic
acid, such that a hydrazone bond is formed. Examples of hydrazone
precursors include hydrazines (See, for example, Phillips, Org.
Reac. 10 (1959) 143-178).
[0030] The term "imide precursors" includes reagents and conditions
which can be reacted with an appropriate reactive intermediate of
the invention, such as, for example, a carboxylic acid, such that
an imide is formed. Examples of imide precursors include amides
(See, for example, Challis, in Zabicky The Chemistry of Amides,
(Wiley: New York, 1970), pp. 731-857.)
[0031] The term "ketone precursors" includes reagents and
conditions which can be reacted with an appropriate reactive
intermediate of the invention, such as, for example, a carboxylic
acid, such that a ketone is formed. Examples of ketone precursors
include organic lithium reagents and other precursors are known in
the art (See, for example, Jorgenson, Org. React. 1970, 18,
1-97).
[0032] The term "nitrile precursors" includes reagents and
conditions which can be reacted with an appropriate reactive
intermediate of the invention, such as, for example, a carboxylic
acid, such that a nitrile is formed. Examples of nitrile precursors
include ammonia and other precursors are known in the art (See, for
example, LaRock, supra, pp. 976-977).
[0033] Compounds of the invention include 7-substituted
tetracycline analogs, 9-substituted tetracycline analogs, and
13-substituted tetracycline analogs. These compounds may be
illustrated by the general formula
##STR00006##
wherein Z.sub.1, Z.sub.2, and Z.sub.3 are individually H or
##STR00007##
wherein R.sub.2 and R.sub.3 are each independently hydrogen, alkyl,
alkenyl, alkynyl, cycloalkyl, aryl, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aryloxycarbonyl, amino, hydroxy, cyano, alkoxy,
aryloxy, carboxyl, alkoxycarbonyl, aryloxycarbonyl; or R.sub.2 and
R.sub.3, taken together, form a substituted or unsubstituted
carbocyclic or heterocyclic ring having 5 to 15 atoms in the ring;
and R.sub.1 is H or OH.
[0034] In another embodiment R.sub.2 is hydrogen, and R.sub.3
is
##STR00008##
where R.sub.4 is hydrogen, cyano, or a C.sub.1-C.sub.5 alkoxy
group. In another embodiment R.sub.1 and R.sub.2, taken together,
form a substituted or unsubstituted carbocyclic or heterocyclic
ring having 5 to 15 atoms in the ring; the ring may be a conjugated
or unconjugated aromatic ring system, preferably C.sub.5 to
C.sub.8. Suitable substituents for Z include
##STR00009##
In a further embodiment, the invention also pertains to substituted
tetracycline compounds wherein the substituent at the desired
position (e.g., 7, 9, or 13) is connected with a --C--C-- linkage
and is a carbonyl moiety. Substituted tetracycline compounds with
carbonyl moieties include compounds with a carbonyl group (e.g.,
C.dbd.O or a derivative thereof) linked through a --C--C-- bond to
the carbon at the desired position of the tetracycline
compound.
[0035] The term "carbonyl moiety" also includes moieties which
comprise a C.dbd.O bond, such as amides, esters, anhydrides, acid
halides, carboxylic acids, and ketones. It also includes
thiocarbonyl moieties, hydrazone moieties, imide moieties (which
comprise C.dbd.NR' bonds), nitrile moieties, as well as moieties.
In certain embodiments, the C.dbd.O moiety is linked to the
tetracycline compound through a C--C bond to the carbon at the
desired position of the tetracycline compound.
[0036] Example of thiocarbonyl moieties include moieties which
comprise a C.dbd.S bond. In a further embodiment, tetracycline
compounds comprising a thiocarbonyl moiety include those with a
thiocarbonyl group linked through a --C--C-- bond to the carbon at
the desired position of the tetracycline compound.
[0037] Example of hydrazone moieties include compounds which
comprise C.dbd.N--N-- bonds. In a further embodiment, tetracycline
compounds comprising a hydrazone moiety include compounds with a
hydrazone moiety (e.g., C.dbd.N--N--) linked through a --C--C--
bond to the carbon at the desired position of the tetracycline
compound.
[0038] Example of imide moieties include compounds which comprise
C.dbd.N-bonds. In a further embodiment, tetracycline compounds
comprising an imide moiety include compounds with a imide moiety
(e.g., C.dbd.N--) linked through a --C--C-- bond to the carbon at
the desired position of the tetracycline compound.
[0039] The invention also provides in another aspect a method for
preparing substituted tetracycline compounds, desirably 7, 9 or
13-substituted compounds, and, in another aspect, tetracycline
compounds prepared by this method. The compounds can be prepared as
generally depicted in the Schemes set forth hereinbelow. In the
discussions of the Schemes, the various substituent groups are the
same as defined above; "R" includes R.sub.2 and R.sub.3. Also, for
purposes of exemplification only, doxycycline is depicted as the
"base" tetracycline compound, although it will be understood that a
wide variety of tetracycline compounds can be employed in the same
manner. For example, the base tetracycline compound substituted at
the 7-, 9- and/or 13-positions suitably may be oxytetracycline;
chlortetracycline; demeclocycline; doxycycline; chelocardin;
minocycline; rolitetracycline; lymecycline; sancycline;
methacycline; apicycline; clomocycline; guamecycline; meglucycline;
mepylcycline; penimepicycline; pipacycline; etamocycline;
penimocycline, semi-synthetic intermediates thereof, and the like,
such as 4-dedimethylaminotetracycline compounds.
[0040] Tetracycline compounds of the present invention may readily
be prepared by the methods outlined in accordance with the
following schemes. Scheme I refers to the preparation of
tetracycline compounds which may be prepared from a starting
compound of formula 1, a clinically useful tetracycline antibiotic
named doxycycline. It has been found that
6-(substituted)-5-hydroxy-6-deoxytetracyclines (1,
R.sub.2.dbd.CH.sub.3, doxycycline) or their mineral acid salts can
be dissolved in concentrated acids, e.g., H.sub.2SO.sub.4 as an
appropriate solvent, and reacted with nitrating reagents such as
sodium or potassium nitrate to produce 7 and 9-nitro tetracycline
derivatives (2, 3). These compounds are separated by a variety of
techniques, with the preferred method being preparative HPLC on
C.sub.18-reverse phase silica gel with a binary gradient system
comprising either phosphate-buffered ethylene diamine tetraacetic
acid, sodium salt (EDTA) with a methanol gradient or a gradient of
acetonitrile over 0.1% trifluoroacetic acid. These isolated
compounds are readily reduced to the amine functional group using
typical reducing reagents such as hydrogen with transition metal
catalysts, platinum oxide, palladium on carbon, or similar to
produce the 7-NH.sub.2 and 9-NH.sub.2 tetracyclines (4, 5) in good
yield. Alternatively, 7-NH.sub.2 tetracyclines (doxycycline) can be
prepared (such as detailed in U.S. Pat. No. 3,483,251, incorporated
herein by reference) via a reductive alkylation of 7-(N,N
dicarboxybenzyloxyhydrazino)tetracyclines.
[0041] Compounds possessing the anilino functional groups can
undergo a diazotization reaction with nitrous acid (HONO) or
organic agents such as butyl nitrite readily forming the diazonium
salts, (such as the hydrochloride or tetrafluoroborate salts) (6,
7) in nearly quantitative yield. This reactive tetracycline-based
precursor compound (6, 7) as a suitable diazonium salt form, can
now chemically complex with organopalladium catalysts and species
that results in carbon-carbon bond formation between the
tetracycline reactant intermediate and the reactive organic
substituent precursor of choice. Transition metal catalysts such as
CuCl.sub.2 (the Meerwin reaction) as well as palladium catalysts
such as palladium chloride, palladium acetate or other catalysts
mentioned above, with palladium acetate being preferred, are used
to produce the 7 and 9 substituted position derivatives of
tetracyclines. The reactions are typically run in polar solvents
such as DMSO, water, and alcohols with trace mineral acids (HCl,
0.1%) to react with substituted or unsubstituted aromatic or
heteroaromatic, alkyl, alkenyl, or alkynyl substructures producing
the desired substituted compounds. Non-polar solvents may also be
used in which to run the reactions.
[0042] It is known that transition metal halides, such as palladium
and copper halides, react with arenediazonium salts to form
complexes capable of further reactions. Transition metal halides as
catalysts facilitate carbon-carbon bond formation via a radical
oxidation-reduction addition of carbon substructures (double bonds
and other structures possessing n-bonds) to the electron deficient
nitrogen diazonium reactive group. For example, palladium catalyzed
carbon-carbon bond formation occurs readily when a suitable alkene
in the reacting system forms reactive coordination complexes. This
is followed by insertion into carbon sigma bonds to give a ternary
complex. Catalysts such as palladium are cycled and regenerated
via, for example, a .beta.-hydride elimination, thereby forming a
carbon-carbon covalent bond. Using these conditions, molecular
substructures possessing a .pi.-bond system, such as alkenes or
acrylic acid esters or any one of the many other compounds
possessing a double bond, are readily arylated with reactive
tetracycline-based precursor compounds, e.g., tetracycline
arenediazonium salts. Other transition metal catalyzed reactions
such as transmetallation and insertion reactions, e.g., of carbon
monoxide) are also contemplated (see, Hegedus, supra for examples
of transition metal catalyzed reactions).
[0043] Homogeneous catalysis of carbon-carbon bond formation is
possible using palladium complexes and suitable reactive species.
Tetracyclines, e.g., doxycycline or minocycline, are used to
generate the reactive diazonium functional group within the D ring
while the reactive addend is available from structurally diverse
chemical families.
[0044] Therefore, reactive tetracycline-based precursor compounds
such as tetracycline arenediazonium salts, i.e., having reactive
functional groups at, e.g., positions 7 and 9 of the tetracycline
molecule similarly can be reacted with alkenes, substituted
alkenes, vinyl monomers, aromatic and heteroaromatic reactive
groups (unsubstituted or substituted) in the presence of the
appropriate transition metal catalyst to produce 7-(substituted)
and 9-(substituted) tetracyclines (8, 9, Scheme I) in good yield.
7-position substituted tetracycline 9-diazonium salts, for example
(Scheme II), produced by the reaction sequence of minocycline (10)
nitration to the 9-NO.sub.2 derivative (11), followed by catalytic
reduction to the 9-NH.sub.2 derivative (12) followed by
diazotisation (13), may also be reacted with double bond compounds
such as olefins and reactive products and reagents producing
minocycline derivatives of formula (II) (14, Scheme II).
[0045] In an embodiment, reaction products of formulas I and II may
be further derivatized and reacted with reagents as depicted in
Schemes III-VII, thus acting as intermediates for making other
compounds not readily obtained otherwise. 9-alkenyl substituted
doxycyclines (8, 9) of formula I may undergo hydrogenation of the
9-alkenyl group with platinum or palladium catalysts on carbon
under low pressure hydrogen to form the 9-alkyl derivatives of
doxycycline (15, 16, Scheme III). Similarly, 9-alkenyl derivatives
of minocycline (14) may also be reduced to the alkyl derivatives
using catalytic hydrogenation methods as shown in Scheme IV
(17).
[0046] 7 or 9 derivatives of doxycycline of formula I (Schemes I
and III) may also react with carboxylic acids while dissolved in
strong acids such as anhydrous hydrogen fluoride or
methanesulphonic acid or trifluoromethanesulfonic acid to produce
the 5-ester derivatives of 7 and 9 substituted doxycyclines (18,
19, Scheme V).
[0047] 7 or 9 derivatives of doxycycline of formula I (Schemes I,
III and V) may also form Mannich base derivatives by the reaction
of the 7 or 9 derivatives with formaldehyde and an appropriate base
(pyrrolidine) to produce Mannich base addition products (20, 21,
Scheme VI).
[0048] 9 derivatives of minocycline of formula II (14, Scheme II)
may also form Mannich base derivatives by the reaction of the 7 or
9 derivative with formaldehyde and an appropriate base
(pyrrolidine) to produce the Mannich base addition products (22,
Scheme VII).
[0049] Tetracycline diazonium reactive functional groups generated
in Scheme I may also be reacted with carbon monoxide in alcohols in
the presence of transition metal catalysts such as palladium
acetate to produce 7 and 9-carboxylic acid derivatives (23, 24) in
good yield which readily esterified to produce 9 position
tetracycline esters (25, 26, Scheme VIII).
[0050] Minocycline diazonium reactive functional groups generated
in Schemes II may also be reacted with carbon monoxide in alcohols
in the presence of transition metal catalysts such as palladium
acetate to produce the 9-carboxylic acid derivative (2) in good
yield which is readily esterified to produce 9 position minocycline
carboxylic acid esters (28, Scheme IX).
[0051] Other reactions are possible with 7 and 9 aminotetracyclines
via a diazonium functional group. Tetracycline arene diazonium
salts also react with active methylene compounds such as esters of
acetoacetate, and derivatives thereof, active aromatic rings and
unsubstituted and substituted olefins, acetylenes, substituted
acetylenes, arylethylenes, styrenes, conjugated dienes, isoprenes,
vinyl ethers, .alpha.,.beta.-unsaturated aldehydes and ketones,
aryl vinyl and arylisoprenyl ketones, quinones, .alpha.,
.beta.-unsaturated acids and their derivatives. All of the multiple
bond compounds are readily coupled to arenediazonium salts, as well
as nucleophiles.
[0052] Position 7 and the 7 and 9 reactive tetracycline-based
precursor compounds (halogenated derivatives of tetracyclines as
shown in Scheme X) also produce 7 and 9 derivatives of
tetracyclines. Aromatic substitution reactions by iodination,
bromination or chlorination to produce the 7 and 9 halogen
derivatives of doxycycline (29, 30) or sancycline (31, 32) in good
yield by reactions described, e.g., by Hlavka, J. J., et al., J.
Am. Chem. Soc., 84, 1961, 1426-1430. Position 7 and 9 halogenated
derivatives of the tetracyclines may be further coupled with
iodoalkenes or iodoarenes in N-methylpyrrolidinone with transition
metal catalysts such as copper thiophene chloride or others to
produce the position 7 or 9 derivatives of doxycycline (33, or
position 7 or 9 derivatives of doxycycline (35, M) in good
yield.
[0053] Position 13-derivatives of tetracyclines may be prepared via
the reaction of phenylboronic acids with the exocyclic double bond
of methacycline (37) (Scheme XI) in alcohols such as methanol, in
the presence of palladium chloride or other transition metal
catalysts to produce the 13-phenyl derivatives of methacycline in
good yield (38).
[0054] The following synthetic schemes are illustrative of the
present invention: [0055] Scheme 17-(substituted)-6-methyl-6-deoxy
5-hydroxy tetracyclines and 9-(substituted)-6-methyl-6-deoxy
5-hydroxy tetracyclines [0056] Scheme II 9-(substituted)
minocyclines [0057] Scheme III 7-(alkyl
substituted)-6-methyl-6-deoxy 5-hydroxy tetracyclines and 9-(alkyl
substituted)-6-methyl-6-deoxy 5-hydroxy tetracyclines [0058] Scheme
IV 9-(alkyl substituted) minocyclines [0059] Scheme V 7-(alkyl or
aryl substituted)-6-methyl-6-deoxy 5-acyloxy tetracyclines and
9-(alkyl or aryl substituted)-6-methyl-6-deoxy 5-acyloxy
tetracyclines [0060] Scheme VI 7-(alkyl or aryl
substituted)-6-methyl-6-deoxy 5-hydroxy tetracyclines and 9-(alkyl
or aryl substituted)-6-methyl-6-deoxy 5-hydroxy 2-(carboxamido
substituted) tetracyclines [0061] Scheme VII 9-(alkyl
substituted)-2-(carboxamido substituted) minocyclines [0062] Scheme
VIII 7-(carboxy or carboxy ester)-6-methyl-6-deoxy 5-hydroxy
tetracyclines and 9-(carboxy or carboxy ester)-6-methyl-6-deoxy
5-hydroxy 2-(carboxamido substituted) tetracyclines [0063] Scheme
IX 9-(carboxy or carboxy ester) minocyclines [0064] Scheme X
7-(alkenyl or aryl)-6-methyl-6-deoxy 5-hydroxy tetracyclines and
9-(alkenyl or aryl)-6-methyl-6-deoxy 5-hydroxy tetracyclines, X
9-(alkenyl or aryl)-6-demethyl-6-deoxytetracyclines and 9-(alkenyl
or aryl)-6-demethyl-6-deoxytetracyclines [0065] Scheme XI
13-(substituted)-6-methylene-5-hydroxy-6-deoxytetracyclines
##STR00010##
##STR00011##
##STR00012##
##STR00013##
##STR00014##
##STR00015##
##STR00016##
##STR00017##
##STR00018##
##STR00019##
##STR00020##
[0066] Compounds of the invention are active against susceptible
microorganisms such as bacteria, fungi, rickettsia, parasites and
the like, and diseases associated with such microorganisms,
including tetracycline-sensitive bacteria as well as
tetracycline-resistant bacteria. Particularly preferred compounds
of the invention exhibit 24-hr minimum inhibitory concentration
(MIC) values of about 10 .mu.g/mL or less, more preferably about 1
.mu.g/mL or less, against tetracycline-resistant E. coli, S. aureus
and E. faecalis strains such as E. coli pHCM1, S. aureus RN4250 and
E. faecalis pMV158. Preferred compounds of the invention also
include those that exhibit such MIC values against
tetracycline-sensitive E. coli, S. aureus and E. faecalis strains
such as E. coli D31 m4, S. aureus RN450 and E. faecalis
ATCC9790.
[0067] As discussed above, the invention provides methods of
treatment against microorganism infections and associated diseases,
which methods in general will comprise administration of a
therapeutically effective amount of one or more compounds of the
invention to a subject, which may be an animal or plant, and
typically is a mammal, preferably a primate such as a human.
[0068] In therapeutic methods of the invention, one or more
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. 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.
[0069] At least many of the compounds of the invention suitably may
be administered to a subject in a protonated and water-soluble
form, e.g., as a pharmaceutically acceptable salt of an organic or
inorganic acid, e.g., hydrochloride, sulfate, hemi-sulfate,
phosphate, nitrate, acetate, oxalate, citrate, maleate, mesylate,
etc. Also, where an appropriate acidic group is present on a
compound of the invention, a pharmaceutically acceptable salt of an
organic or inorganic base can be employed such as an ammonium salt,
or salt of an organic amine, or a salt of an alkali metal or
alkaline earth metal such as a potassium, calcium or sodium
salt.
[0070] Therapeutic compounds can be administered to a subject in
accordance with the invention by any of a variety of routes.
Topical (including transdermal, buccal or sublingual), and
parenteral (including intraperitoneal, subcutaneous, intravenous,
intradermal or intramuscular injection) are generally
preferred.
[0071] For parenteral application, particularly suitable are
solutions, preferably oily or aqueous solutions as well as
suspensions, emulsions, or implants, including suppositories.
Therapeutic compounds will 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.
[0072] 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. Controlled release formulations of the
compounds are also contemplated. 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.
[0073] For topical applications, therapeutic compounds can be
suitably admixed in a pharmacologically inert topical carrier such
as a gel, a spray (e.g., aerosol), 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.
[0074] In addition to treatment of humans, 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.
[0075] 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.
[0076] In general, compounds of the invention for treatment can be
administered to a subject in dosages used in prior tetracycline
therapies. See, for example, the Physicians' Desk Reference. For
example, a suitable effective dose of one or more compounds of the
invention will be in the range of 0.01 to 100 milligrams per
kilogram of bodyweight of recipient per day, preferably in the
range of 0.1 to 50 milligrams per kilogram bodyweight of recipient
per day, more preferably in the range of 1 to 20 milligrams per
kilogram bodyweight 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.
[0077] It will also be understood that normal, conventionally known
precautions will be taken regarding the administration of
tetracyclines generally to ensure their efficacy under normal use
circumstances. Especially when employed for therapeutic treatment
of humans and animals in vivo, the practitioner should take all
sensible precautions to avoid conventionally known contradictions
and toxic effects. Thus, the conventionally recognized adverse
reactions of gastrointestinal distress and inflammations, the renal
toxicity, hypersensitivity reactions, changes in blood, and
impairment of absorption through aluminum, calcium, and magnesium
ions should be duly considered in the conventional manner.
Exemplification of the Invention
Biological Activity: Method for In Vitro Evaluation
[0078] Various compounds made in accordance with the invention were
evaluated for anti-bacterial activity in vitro as follows. The
minimum inhibitory concentration, the lowest concentration of drug
that inhibits bacterial growth at 18 hrs at their appropriate
temperature, is determined by the broth dilution method using
L-broth or Mueller-Hinton broth. The Mueller-Hinton broth was
cation-adjusted accordingly and all bacteriological methods were
performed as was described by Waitz, J. A., National Commission for
Clinical Laboratory Standards Document M7-A2, vol. 10, no. 8, pp.
13-20, 2.sup.nd edition, Villanova, Pa. (1990). The organisms
tested represent gram-positive and gram-negative bacterial species
that are susceptible to tetracyclines or are resistant to
tetracyclines due to the ability to efflux tetracyclines or which
confer resistance by ribosomal protection mechanisms. The clinical
strains used are either susceptible to tetracyclines or are
resistant to them by either drug efflux or ribosomal
protection.
TABLE-US-00002 TABLE I LEGEND FOR COMPOUNDS Compound Name =
Doxycycline
[4S-(4a,12a.alpha.)]-4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,5-
, 10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-2-
naphthacenecarboxamide Minocycline [4S-(4a,12a.alpha.)]-4,7-Bis
(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-
3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide A
[4S-(4a,12a.alpha.)]-9-(nitro)-4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-
octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-2-
naphthacenecarboxamide (9-nitro-6-deoxy-5-hydroxy tetracycline) B
[4S-(4a,12a.alpha.)]-9-(amino)-4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-
octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-2-
naphthacenecarboxamide (9-amino-6-deoxy-5-hydroxy tetracycline) C
[4S-(4a,12a.alpha.)]-9-(diazonium)-4-(dimethylamino)-1,4,4a,5,5a,6,11,12-
a- octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-2-
naphthacenecarboxamide (9-diazonium-6-deoxy-5-hydroxy tetracycline)
D [4S-(4a,12a.alpha.)]-9-[3'-(E)-propenoic acid]-4-(dimethylamino)-
1,4,4a,5,5a,6,11,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-
1,11-dioxo-2-naphthacenecarboxamide (9-[3'-(E)-ethylpropenoic
acid]-6-deoxy-5-hydroxy tetracycline) E
[4S-(4a,12a.alpha.)]-9-[3'-(E)-butylpropenoate]-4-(dimethylamino)-
1,4,4a,5,5a,6,11,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-
1,11-dioxo-2-naphthacenecarboxamide
(9-[3'-(E)-butylpropenoate]-6-deoxy-5-hydroxy tetracycline) F
[4S-(4a,12a.alpha.)]-9-[3'-(E)-butylpropenoate]-4,7-Bis
(dimethylamino)-
1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-
naphthacenecarboxamide 9-[3'-(E)-butylpropenoate] minocycline G
[4S-(4a,12a.alpha.)]-7-[4'-Cl-phenyl)]-4-(dimethylamino)-
1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-
naphthacenecarboxamide 7-(4'-Cl-phenyl) sancycline H
[4S-(4a,12a.alpha.)]-7-phenyl-9-phenyl-4-(dimethylamino)-
1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-
naphthacenecarboxamide 7,9-diphenyl sancycline I
[4S-(4a,12a.alpha.)]-13-(4'-methylphenyl)-4-(dimethylamino)-
1,4,4a,5,5a,6,11,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-
methylene-1,11-dioxo-2-naphthacenecarboxamide
13-(4'-methylphenyl)-6-deoxy-6-methylene-5-hydroxy tetracycline J
[4S-(4a,12a.alpha.)]-13-(3'-carboxyphenyl)-4-(dimethylamino)-
1,4,4a,5,5a,6,11,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-
methylene-1,11-dioxo-2-naphthacenecarboxamide
13-(3'-carboxyphenyl)-6-deoxy-6-methylene-5-hydroxy tetracycline K
[4S-(4a,12a.alpha.)]-13-(4'-ethoxyphenyl)-4-(dimethylamino)-
1,4,4a,5,5a,6,11,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-
methylene-1,11-dioxo-2-naphthacenecarboxamide
13-(4'-ethoxyphenyl)-6-deoxy-6-methylene-5-hydroxy tetracycline
TABLE-US-00003 TABLE II Antibacterial Activity of Transition Metal
Catalyzed Derivatives of Tetracyclines Doxy D E F G H I J K E. coli
0.78 25 6.25 >50 >50 >50 >50 >50 12.5 ML308-225
Tc.sup.s E. coli 25 >50 >50 >50 >50 >50 >50
>50 >50 D1-299 Tc.sup.r E. coli 50 >50 >50 >50
>50 >50 >50 >50 >50 D1-209 Tc.sup.r E. coli 1.56
>50 3.12 3.12 0.78 1.56 6.25 >50 12.5 D31m4 Tc.sup.s E. coli
D 31m4 25 >50 6.25 6.25 0.78 -- 25 >50 >50 pHCM1 Tc.sup.r
S. aureus <0.098 3.12 0.78 1.56 .ltoreq.0.098 1.56 .ltoreq.0.098
0.39 0.195 RN450 Tc.sup.s S. warnerii Tc.sup.r 50 >50 6.25 3.12
.ltoreq.0.098 0.78 12.5 >50 12.5 ATCC12715 S. aureus 25 >50
6.25 3.12 .ltoreq.0.098 0.78 12.5 >50 6.25 RN4250 Tc.sup.r S.
aureus 6.25 >50 0.39 3.12 0.195 0.78 6.25 >50 6.25 MRSA5
Tc.sup.r E. hirae 0.195 3.12 3.12 3.12 .ltoreq.0.098 0.78 0.39 3.12
0.39 ATCC9790 Tc.sup.s E. hirae 9790 6.25 12.5 6.25 3.12
.ltoreq.0.098 0.39 3.12 >50 6.25 with pMV158 Tc.sup.r E. hirae
9790 6.25 >50 6.25 3.12 .ltoreq.0.098 1.56 12.5 >50 3.12 with
pAM211 Tc.sup.r Tc.sup.s = tetracycline susceptible Tc.sup.r =
tetracycline resistant
EXPERIMENTAL
[0079] Compounds of the invention may be prepared as presented in
schemes I through IX, above, and/or as described below.
[0080] In scheme I, doxycycline is dissolved in cold concentrated
sulfuric acid and an equivalent of potassium nitrate is added. The
reaction temperature was maintained in the range 0 to 5.degree. C.
for a period of 1 to 3 hrs, producing 7 and
9-nitro-6-substituted-5-hydroxy tetracyclines of formula IV. These
intermediates, with suitable chemically reactive functionality, can
be reacted with a broad range of reducing agents such as PtO.sub.2
or hydrogen and palladium or platinum catalysts producing compounds
of general formula IV. The diazonium salts of the 7 and 9 amino
derivatives are produced by the action of nitrites (sodium nitrite,
butyl nitrite or equivalent) and the intermediate used without
further purification.
Example 1
[4S-(4.alpha.,12a.alpha.)]-9-(nitro)-4-(dimethylamino)-1,4,4a,5,5a,6,11,12-
a-octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-2-naphthaceneca-
rboxamide
[0081] To an ice cold solution of 1.0 g of doxycycline
hydrochloride in 10 mL of concentrated sulfuric acid was added
0.231 g of potassium nitrate. The reaction was stirred for 1 hr
under ambient atmosphere. The mixture was then poured over 150 g of
ice and the resulting solid was extracted with n-butanol and dried
to afford 0.9 g of the desired product as a yellow-green solid.
[0082] MS (FAB): m/z 490 (M+H).
[0083] .sup.1H NMR (CD.sub.3OD): .delta. 7.50 (d, 1H, J=8.07 Hz,
H-8); 6.86 (d, 1H, J=8.07 Hz, H-7); 4.44 (bs, 1H, H-4); 3.62 (dd,
1H, J=11.42; 8.35 Hz, H-5); 2.95 (bs, 6H, NMe.sub.2); 2.81 (d, 1H,
J=11.45 Hz, H-4a); 2.71 (dq, 1H, J=12.41; 6.5 Hz, H-6); 2.53 (dd,
1H, J=12.23; 8.20 Hz, H-5a); 1.51 (d, 3H, J=6.78 Hz, CH.sub.3).
Example 2
[4S-(4.alpha.,12a.alpha.)]-9-(amino)-4-(dimethylamino)-1,4,4a,5,5a,6,11,12-
a-octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-2-naphthaceneca-
rboxamide
[0084] Into a 200 mL hydrogenation bottle is added 1.0 g of product
from example 1, 40 mL of methanol, 1 mL of concentrated HCl, and
100 mg of 10% palladium on carbon. Using a hydrogenation apparatus,
the mixture is subjected to 30 psi of hydrogen for 3 hrs. The
catalyst is filtered off and the filtrate is dried to afford 0.9 g
of the dihydrochloride as a yellow solid.
[0085] MS (FAB): m/z 460 (M+H). .sup.1H NMR (CD.sub.3OD): .delta.
7.54 (d, 1H, J=8.08 Hz, H-8); 6.88 (d, 1H, J=8.08 Hz, H-7); 5.16
(dd, J=10.44; 7.94 Hz, H-5); 4.44 (bs, 1H, H-4); 3.74 (d, 1H,
J=2.07 Hz, H-4); 3.04 (bs, 6H, NMe.sub.2); 2.90 (dd, 1H, J=7.94;
2.07 Hz, H-4-a); 2.72 (dq, 1H, J=12.31; 6.56 Hz, H-6); 2.61 (dd,
1H, J=12.31; 10.44 Hz, H-5a); 2.54 (q, 2H, J=7.48 Hz, CH.sub.2--C);
1.44 (bs, 9H, CMe.sub.3); 1.29 (d, 3H, J=6.56 Hz, CH.sub.3); 1.20
(t, 3H, J=7.48 Hz, C--CH.sub.3).
Example 3
[4S-(4.alpha.,12a.alpha.)]-9-(diazonium)-4-(dimethylamino)-1,4,4a,5,5a,6,1-
1,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-2-naphthace-
necarboxamide
[0086] A 10 mL round bottom flask was charged with 100 mg of
product from example 2 and dissolved in 4 mL of 0.1 N methanolic
hydrochloric acid. The solution was cooled to 0.degree. C. and 35
.mu.l of butyl nitrite was added with stirring. After 1 hr, the
bright red reaction mixture was added dropwise to 100 mL of cold
anhydrous diethyl ether. The product was collected by filtration,
washed with ether, and dried in a vacuum dessicator to give 73 mg
of the diazonium salt as an orange solid.
[0087] MS (FAB): m/z 472 (M+H).
[0088] .sup.1H NMR (CD.sub.3OD): .delta. 7.55 (d, 1H, J=8.08 Hz,
H-8); 6.86 (d, 1H, J=8.08 Hz, H-7); 5.13 (dd, J=10.44; 7.94 Hz,
H-5); 4.41 (bs, 1H, H-4); 3.72 (d, 1H, J=2.07 Hz, H-4); 3.04 (bs,
6H, NCH.sub.3); 2.90 (dd, 1H, J=7.94; 2.07 Hz, H-4-a); 2.70 (dq,
1H, J=12.31; 6.56 Hz, H-6); 2.61 (dd, 1H, J=12.31; 10.44 Hz, H-5a);
2.2 (m, 6H, J=7.48 Hz, Acetyl); 1.44 (bs, 9H, C(CH.sub.3).sub.3);
1.29 (d, 3H, J=6.56 Hz, CH.sub.3); 1.20 (t, 3H, J=7.48 Hz,
C--CH.sub.3).
[0089] General Procedure for Olefination.
[0090] To a solution of 0.1 g of 9-diazonium compound in (wet or
dry) methanol is added 0.05 equivalents of palladium acetate. The
reaction mixture is stirred for 5 minutes at room temperature, and
2 equivalents of the desired olefin is added. Stirring is continued
for 18 hrs under ambient atmosphere or followed by HPLC. The
stirring may also be continued under N.sub.2 atmosphere. Upon
completion, the catalyst is filtered off and the filtrate dried to
give the crude product. The purified product is isolated by
preparative reverse-phase HPLC using methanol and phosphate buffer
gradient.
Example 4
[4S-(4.alpha.,12a.alpha.)]-9-[3'-(E)-propenoic
acid]-4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,5,10,12,12a-pent-
ahydroxy-6-methyl-1,1'-dioxo-2-naphthacenecarboxamide MS (FAB): m/z
515 (M+H)
Example 5
[4S-(4.alpha.,12a.alpha.)]-9-[1'-(E)-(2'-phenyl)ethenyl]4-(dimethylamino)--
1,4,4a,5,5a,6,11,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-1,11-di-
oxo-2-naphthacenecarboxamide MS (FAB): m/z 547 (M+H)
Example 6
[4S-(4.alpha.,12a.alpha.)]-7-[3'-(E)-butylpropenoate]-4-(dimethylamino)-1,-
4,4a,5,5a,6,11,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-1,1'-diox-
o-2-naphthacenecarboxamide
[0091] General Procedure for Arylation.
[0092] To a solution of 9-diazonium compound in methanol is added
0.10 equivalents of palladium acetate. The mixture is stirred at
room temperature for 5 minutes, and 2 equivalents of aryl boronic
acid is added. After 6 hrs, the catalyst is filtered off and the
filtrate is dried down. The crude product is purified by
preparative reverse-phase HPLC using a methanol phosphate buffer
gradient.
[0093] General Procedure for Carboxylation.
[0094] To a three neck round bottom flask equipped with two rubber
septa, a vacuum source, and a stirbar, is added 100 mg of diazonium
compound, 6.0 mg of palladium acetate, and 10 mL of anhydrous
dimethylformamide. The reaction vessel is evacuated, and CO is
passed through the mixture for 1 hr via a syringe. The mixture is
stirred for an additional 2 hr, then the solvent removed in vacuo
to yield the crude product. The title compound was isolated by
preparative C.sub.18 reverse-phase HPLC by using a binary solvent
gradient.
Example 7
[4S-(4.alpha.,12a.alpha.)]-9-(carboxy)-4-(dimethylamino)-1,4,4a,5,5a,6,11,-
12a-octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-2-naphthacene-
carboxamide MS (FAB): m/z 489 (M+H)
[0095] General Procedure for Hydrogenation.
[0096] The compound is prepared by dissolving 0.100 g of Example 4
into 10 mL of methanol, adding 0.1% concentrated HCl and 10 mg of
10% palladium on carbon. The mixture is hydrogenated in at 40 psi
in a Parr apparatus for 6 hrs at room temperature and monitored by
HPLC. The resulting crude product is chromatographed on C.sub.18
reverse-phase via semi-preparative binary solvent methods to give
the desired product.
[0097] General Procedure for 7 Position Olefination.
[0098] To a solution of 0.1 g of 7-diazonium compound, generated in
similar methods as described in Examples 1 and 2, in wet methanol
is added 0.05 equivalents of palladium acetate. The reaction
mixture is stirred for 5 minutes at room temperature, and 2
equivalents of the desired olefin are added. Stirring is continued
for 18 hrs under ambient atmosphere and followed by HPLC. Upon
completion, the catalyst is filtered through Celite and the
filtrate dried to give the crude product. The purified product is
isolated by preparative reverse-phase HPLC using methanol and
phosphate buffer gradient.
Example 8
9-phenyl minocycline
[4S-(4.alpha.,12a.alpha.)]-9-(phenyl)-4,7-Bis(dimethylamino)-1,4,4a,5,5a,6-
,11,12a-octahydro-3,5,10,12,12a-tetrahydroxy-6-methyl-1,11-dioxo-2-naphtha-
cenecarbox amide
[0099] The compound was prepared using 0.100 g of 9-amino
minocycline and reagents and conditions similar to those found in
Example 5. The reaction was stirred overnight under a nitrogen
atmosphere and the solvent removed in vacuo to produce 0.063 g of
the crude product. Chromatography using C.sub.18 reverse-phase
preparative methods and binary solvent systems followed by
extraction of the product into butanol and evaporation of the
product in vacuo, furnished 0.027 g of the desired product as a
yellow solid.
[0100] MS (FAB): m/z 571 (M+H).
Example 9
7-iododoxycycline
[0101] 30.0 mL of concentrated sulfuric acid was added to 1.00 g of
doxycycline hydrochloride hemihydrate with stirring and the
solution cooled to 0.degree. C. 0.973 g of N-iodosuccinimide was
added portionwise to the solution over one hr and the reaction
monitored by HPLC and TLC to ensure completion. The solution was
poured into 250 mL of ice water, extracted three times with
butanol, and the solvent removed under reduced pressure. The crude
residue was purified by preparative HPLC to provide 1.13 g (89%) of
the title compound as dark yellow crystals.
[0102] MS (FAB): m/z 587 (M+H)
[0103] .sup.1H NMR (Methanol d-4, 300 MHz) .delta. 7.94 (d, J=8.19
Hz, 1H), 6.78 (d, J=8.18 Hz, 1H), 4.13 (s, 1H), 3.53 (m, 3H), 2.85
(s, 7H), 2.66 (m, 4H), 2.41 (s, 1H), 1.49 (d, J=6.52 Hz, 3H), 0.95
(t, J=7.27 Hz, 2H).
EXAMPLES 10 and 11
7-iodosancycline and 7,9-diiodosancyline
[0104] 30.0 mL of concentrated sulfuric acid was added to 1.00 g of
sancycline hydrochloride hemihydrate with stirring and the solution
cooled to 0.degree. C. 1.09 g of N-iodosuccinimide was added
portionwise to the solution over one hr and the reaction mixture
monitored by HPLC and TLC. The reaction mixture was poured into 250
mL of ice water, extracted three times with n-butanol, and the
solvent removed under reduced pressure. The crude residue was
purified by preparative HPLC yielding 787 mg (61%) of
7-iodosancycline and 291 mg (22%) of 7,9-diiodosancycline as yellow
and dark yellow crystals respectively.
[0105] MS (FAB): m/z 587 (M+H) 7-iodosancycline
[0106] .sup.1H NMR (Methanol d-4, 300 MHz) .delta. 7.89 (d, J=8.86
Hz, 1H), 6.67 (d, 8.87 Hz, 1H), 3.56 (s, 1H), 3.03 (s, 2H), 2.84
(s, 6H), 2.46 (m, 2H), 1.63 (m, 4H) 0.95 (m, 2H).
[0107] MS (FAB): m/z 667 (M+H) 7,9-diiodosancycline
[0108] .sup.1H NMR (Methanol d-4, 300 MHz) .delta. 8.35 (s, 1H),
3.78 (s, 1H), 3.33 (s, 2H), 2.88 (s, 7H), 2.41 (m, 2H), 1.41 (m,
5H).
Example 12
General Coupling Procedure
7-4'-Cl-phenyl sancycline
[0109] 100 mg of 7-iodosancycline or 7-iodo doxycycline (0.18 mM)
and 4 mg of Pd(OAc).sub.2 is added to an argon degassed solution of
methanol followed by 200 .mu.l of 2 M Na.sub.2CO.sub.3. The
resultant solution was stirred for 10 minutes at room temperature.
4'-Cl-phenyl boronic acid (58 mg, 0.37 mM) was dissolved in 1 mL
methanol, added to the iodotetracycline and the reaction flask
degassed with argon 3 times. The reaction was stirred for 15
minutes at room temperature, then heated to reflux for 18 hrs. The
solution was cooled, filtered and the solvent removed under reduced
pressure. The crude product was purified by C18-reverse phase
chromatography to yield 23 mg of product as dark yellow
crystals.
[0110] MS (FAB): m/z (M+H) 525.1852
[0111] .sup.1H NMR (Methanol d.sub.4, 300 MHz) .delta. 7.35-7.44
(m, 4H), 7.21-7.24 (d, 1H), 6.85-6.88 (d, 1H), 3.55 (s, 1H), 2.88
(s, 6H), 2.47 (m, 2H) 1.52 (m, 2H)
Example 13
7,9-diphenyl sancycline
[0112] MS (FAB) m/z (M+H) 567.2545
[0113] .sup.1H NMR (Methanol d.sub.4, 300 MHz) .delta. 7.22-7.85
(m, 11H), 4.02 (M, 1H), 3.53 (s, 1H), 2.86 (br s, 6H), 2.41 (m,
2H), 1.52 (m, 2H)
Example 14
7-(4-fluorophenyl)sancycline
[0114] MS (FAB): m/z 509 (M+H)
[0115] .sup.1H NMR (Methanol d-4, 300 MHz) .delta. 7.41 (d, J=8.61
Hz, 1H), 7.30 (td, J=6.87, 2.16 Hz, 2H), 7.16 (td, J=6.84, 2.11 Hz,
2H), 6.89 (d, J=8.59 Hz, 1H) 3.56 (s, 2H), 2.91 (s, 7H), 1.52 (m,
4H), 0.95 (m, 2H).
Example 15
7-(4-nitrophenyl)sancycline
[0116] MS (FAB): m/z 536 (M+H)
[0117] .sup.1H NMR (Methanol d-4, 300 MHz) .delta. 8.28 (d, J=8.50,
2H), 7.52 (d, J=8.52, 2H), 7.42 (d, J=8.64, 1H), 6.93 (d, J=8.65,
1H), 3.51 (s, 2H), 6.73 (s, 7H), 1.50 (m, 5H), 0.92 (m, 2H).
Example 16
7-(2-pyridyl)doxycycline
[0118] MS (FAB): m/z 522 (M+H)
[0119] .sup.1H NMR (Methanol d-4, 300 MHz) .delta. 8.62 (s, 1H),
7.94 (m, 2H), 7.49 (m, 1H), 7.40 (m, 1H), 6.94 (m, 1H), 4.21 (s,
1H), 3.56 (m, 2H), 2.91 (s, 7H), 2.70 (m, 3H), 1.038 (s, 3H), 0.92
(m, 2H).
Example 17
7-ethenylsancycline
[0120] MS (FAB): m/z 471 (M+H)
[0121] .sup.1H NMR (Methanol d-4, 300 MHz) .delta. 7.65 (d, J=8.79
Hz, 1H), 6.80 (d, J=8.76 Hz, 1H), 5.56 (d, J=18.42 Hz, 1H), 5.25
(d, J=12.15 Hz, 1H), 3.84 (s, 1H), 3.19 (m, 2H), 2.98 (s, 6H), 2.82
(m, 1H), 2.32 (m, 2H), 0.92 (m, 1H).
Example 18
7-Sancycline Carboxylic Acid
##STR00021##
[0123] In the Parr apparatus were placed 9-iodosansycline
trifluoroacetic acid salt (7.5 g, 1 eq.), K.sub.2CO.sub.3 (6.3 g, 4
eq.), Pd(OAc).sub.2 (0.257 g, 0.1 eq.), H.sub.2O (500 mL) and DMF
(250 mL) (N. A. Bumagin at al. J. Organomet. Chem. 1988, 563). The
apparatus was closed, purged with CO then filled with CO under
20-100 psi. The reaction mixture was stirred at room temperature
for 4-5 days. The mixture was then acidified with TFA. The product
was concentrated in vacuo and purified by HPLC. The yield was
4.0-5.5 g of the product.
Example 19
7-Sancycline Butyl Ester
##STR00022##
[0125] A Parr reactor was loaded with 7-iodosancycline (0.654 g, 1
mmol, NaOAc (0.3 g, 3.6 mmol), Pd(dppf).sub.2, CH.sub.2Cl.sub.2
(100 mg) and n-BuOH (20 mL). The reaction mixture was degassed and
charged with CO (300 psi). It was then stirred at 70.degree. C. for
14 hours. The butanol was then evaporated and the product was
purified using HPLC. 170 mg of the product as a TFA salt was
obtained.
Example 20
7-Sancycline Methyl Ester
##STR00023##
[0127] A Parr reactor was loaded with 7-iodosancycline (0.654 g, 1
mmol), NaOAc (0.3 g, 3.6 mmol), Pd(dppf).sub.2, CH.sub.2Cl.sub.2
(100 mg) and MeOH (20 mL). The reaction mixture was degassed and
charged with CO (300 psi). It was then stirred at 70.degree. C. for
14 hours. The methanol was then evaporated and the product was
purified using HPLC. The product was obtained as a TFA salt.
Example 21
9-Sancycline Methyl Ester
##STR00024##
[0129] In the Parr apparatus were placed: 9-iodosancycline
trifluoroacetic acid salt (0.8 g, 1.17 mmol), NaOAc (0.64 g, 4
eq.), Pd(dppf).sub.2Cl.sub.2, and CH.sub.2Cl.sub.2 (48 mg, 5%). The
apparatus was closed, purged with CO then filled with CO under 400
psi. The reaction mixture was stirred for four hours at 70.degree.
C. It was then acidified with TFA and concentrated in vacuo. The
product was purified by HPLC as a single epimer.
Example 22
9-Minocycline Methyl Ester
##STR00025##
[0131] In the Parr apparatus were placed: 9-iodosancycline
trifluoroacetic acid salt (0.8 g, 1.17 mmol), NaOAc (0.64 g, 4
eq.), Pd(dppf).sub.2Cl.sub.2, and CH.sub.2Cl.sub.2 (48 mg, 5%). The
apparatus was closed, purged with CO, and then filled with CO under
450 psi. The reaction mixture was stirred for four hours at
80.degree. C. It was then acidified with TFA and concentrated in
vacuo. The product was purified by HPLC. A mixture of 3:1 epimers
was obtained. The yield was 188 mg of product.
Example 23
7-(Morpholin-4'-yl-ethane-1,2'-dione)-Sancycline
##STR00026##
[0133] In the Parr apparatus were placed 7-sancycline carboxylic
acid (0.5 g, 0.93 mmol), PdI.sub.2 (33 mg, 10% mol), PPh.sub.3
(48.5 mg, 20% mol) and morpholine (12 mL). The apparatus was
closed, purged with CO, and then filled with CO under 200 psi. The
reaction mixture was stirred for five hours at 70.degree. C. and
concentrated in vacuo. The product was purified by HPLC.
Example 24
7-(2'-N',N'-dimethylaminoethane-1',2'-dione)-Sancycline
##STR00027##
[0135] In the Parr apparatus were placed 7-sancycline carboxylic
acid (0.5 g, 0.93 mmol), Pd(PPh.sub.3).sub.2Cl.sub.2 (65 mg),
HNMe.sub.2, HCl (1.5 g), triethylamine (TEA) (6 mL) and dimethyl
formamide (DMF) (10 mL). The apparatus was closed, purged with CO,
and then filled with CO under 200 psi. The reaction mixture was
stirred for four hours at 70.degree. C., and concentrated in vacuo.
The product was purified by HPLC. The product yield was 100 mg. A
1:1 mixture of epimers was obtained.
Example 25
7-(N',N'-Dimethylbenzamide) Sancycline
##STR00028##
[0137] A mixture of 7-sancyclinecarboxylic acid TFA salt (200 mg,
0.35 mmol), dimethylamine hydrochloride (85 mg, 5 eq.), NEt.sub.3
(0.245 mL, 5 eq.), HBTU (0.4 g, 3 eq.) and DMF (1.5 mL) was stirred
at room temperature for 48 hours. The solvents were evaporated, the
residue was acidified with TFA, and the product was subsequently
isolated by HPLC.
Example 26
9-(Morpholin-4'-yl-methanone) Sancycline
##STR00029##
[0139] In the Parr apparatus were placed 7-sancycline carboxylic
acid (0.8 g, 1.17 mmol), Pd(PPh.sub.3).sub.2Cl.sub.2,
CH.sub.2Cl.sub.2 (43 mg, 5%) and morpholine (15 mL). The apparatus
was closed, purged with CO and then filled with CO under 150 psi.
The reaction mixture was stirred overnight at 70.degree. C.,
concentrated in vacuo, and acidified with TFA. The product was
purified by HPLC. The product yield was 146 mg.
Example 27
7-(2'-Pyridyl) Sancycline
[0140] To a DMF (20 mL) solution of 7-iodosancycline TFA salt (1
mmol), Pd(PPh.sub.3).sub.4 (0.1 mmol), and/or Pd(OAc).sub.2 (0.1
mmol) and CuI (0.1 mmol) were added and the reaction mixture was
purged with argon for 2 minutes. To this solution, 2-pyridyl
stannane (1 mmol) was added and the reaction mixture was stirred at
room temperature for 2-4 hours. Completion of the reaction was
monitored by HPLC/LCMS. The reaction mixture was then filtered
through celite and the filterate was concentrated to dryness. The
crude material was then purified by preparative HPLC.
EQUIVALENTS
[0141] 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 this invention
and are covered by the following claims. The contents of all
references, patents, and patent applications cited throughout this
application are hereby incorporated by reference.
* * * * *