U.S. patent application number 14/283734 was filed with the patent office on 2014-09-11 for pleuromutilin derivatives for the treatment of diseases mediated by microbes.
This patent application is currently assigned to NABRIVA THERAPEUTICS AG. The applicant listed for this patent is Nabriva Therapeutics AG. Invention is credited to Rudolf BADEGRUBER, Atchyuta Rama Chandra Murty BULUSU, Mathias FERENCIC, Werner HEILMAYER, Rosemarie MANG, Rodger NOVAK, Dirk B. STRICKMANN.
Application Number | 20140256731 14/283734 |
Document ID | / |
Family ID | 38668870 |
Filed Date | 2014-09-11 |
United States Patent
Application |
20140256731 |
Kind Code |
A1 |
MANG; Rosemarie ; et
al. |
September 11, 2014 |
PLEUROMUTILIN DERIVATIVES FOR THE TREATMENT OF DISEASES MEDIATED BY
MICROBES
Abstract
Pleuromutilin derivative compounds of the following formula, and
uses thereof for the treatment of diseases mediated by microbes,
are disclosed: ##STR00001##
Inventors: |
MANG; Rosemarie; (Vienna,
AT) ; HEILMAYER; Werner; (Zillingtal, AT) ;
BADEGRUBER; Rudolf; (Vienna, AT) ; STRICKMANN; Dirk
B.; (Vienna, AT) ; NOVAK; Rodger; (Vienna,
AT) ; FERENCIC; Mathias; (Vienna, AT) ;
BULUSU; Atchyuta Rama Chandra Murty; (Perchtoldsdorf,
AT) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Nabriva Therapeutics AG |
Wien |
|
AT |
|
|
Assignee: |
NABRIVA THERAPEUTICS AG
Wien
AT
|
Family ID: |
38668870 |
Appl. No.: |
14/283734 |
Filed: |
May 21, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13438160 |
Apr 3, 2012 |
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14283734 |
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13252732 |
Oct 4, 2011 |
8153689 |
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13438160 |
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12531839 |
Sep 17, 2009 |
8071643 |
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PCT/AT08/00097 |
Mar 19, 2008 |
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13252732 |
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Current U.S.
Class: |
514/239.5 ;
514/462; 514/511; 544/154; 549/342; 560/118; 560/125 |
Current CPC
Class: |
A61K 31/215 20130101;
C07C 2601/14 20170501; A61P 31/00 20180101; A61K 31/357 20130101;
A61P 17/10 20180101; C07C 323/61 20130101; C07C 2603/99 20170501;
A61P 31/06 20180101; C07D 317/72 20130101; C07D 295/096 20130101;
A61K 31/5375 20130101; C07C 323/52 20130101; A61P 31/04
20180101 |
Class at
Publication: |
514/239.5 ;
560/125; 514/511; 560/118; 544/154; 549/342; 514/462 |
International
Class: |
A61K 31/5375 20060101
A61K031/5375; A61K 31/357 20060101 A61K031/357; C07D 295/096
20060101 C07D295/096; C07D 317/72 20060101 C07D317/72; C07C 323/61
20060101 C07C323/61; A61K 31/215 20060101 A61K031/215 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 20, 2007 |
EP |
07450053.9 |
Claims
1. A compound of formula (I) ##STR00011## wherein n is 0 to 4; m is
0 or 1 with the proviso that the sulphur atom and R.sub.3 are in
vicinal position (if m=0 then R.sub.3 is in position 2', and if m=1
then R.sub.3 is on position 1'); R is ethyl or vinyl; R.sub.1 is
hydrogen or (C.sub.1-6)alkyl, R.sub.2 is hydrogen or
(C.sub.3-6)cycloalkyl, or unsubstituted (C.sub.1-6)alkyl, or
(C.sub.1-6)alkyl substituted by one or more of hydroxy; preferably
one or two, methoxy, halogen, (C.sub.3-6)cycloalkyl, or R.sub.1 and
R.sub.2 together with the nitrogen atom to which they are attached
form a 5 to 7 membered heterocyclic ring containing at least 1
nitrogen atom or 1 nitrogen and 1 additional heteroatome e.g.
selected from N or O, or R.sub.1 is hydroxy and R.sub.2 is formyl;
R.sub.3 is OH, OR.sub.4, a halogen atom, or with the proviso that
R.sub.3 is bound to 2'R.sub.3 represents --O--(CH.sub.2).sub.p--O--
with p is 2 or 3; R.sub.4 is unsubstituted (C.sub.1-6)alkyl or
(C.sub.3-6)cycloalkyl.
2. A compound of formula (II) ##STR00012## wherein n, R, R.sub.1,
R.sub.2 and R.sub.3 are as defined in claim 1.
3. A compound of formula (III) ##STR00013## wherein n, R, R.sub.1
and R.sub.2 are as defined in claim 1.
4. A compound of formula (TV) ##STR00014## wherein n, R.sub.1 and
R.sub.2 are as defined in claim 1.
5. A compound of formula (V) ##STR00015## wherein n, R.sub.1 and
R.sub.2 are as defined in claim 1.
6. A compound of formula (VI) ##STR00016## wherein n, R.sub.1 and
R.sub.2 are as defined in claim 1.
7. A compound according to one of the claims 1 to 6, selected from
the group consisting of
14-O-{[(1R,2R,4R)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
14-O-{[(1S,2S,4S)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
14-O-{[(1R,2R,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
14-O-{[(1S,2S,5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
14-O-{[(1R,2R,4S)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
and the (1S,2S,4R) diastereomer thereof
14-O-{[(1R,2R,5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
14-O-{[(1S,2S,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
14-O-{[(1R,2R,3R)-3-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
and the (1S,2S,3S) diastereomer thereof
14-O-{[(1R,2R,4R)-4-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin and the (1S,2S,4S) diastereomer thereof
14-O-{[(1R,2R,4R)-4-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin and the (1S,2S,4S) diastereomer thereof
14-O-{[(1R,2R,5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin and the (1S,2S,5R) diastereomer thereof
14-O-{[(1R,2R,5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin and the (1S,2S,5R) diastereomer thereof
14-O-{[(1R,2R,4S)-4-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin and the (1S,2S,4R) diastereomer thereof
14-O-{[(1R,2R,5R)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin and the (1S,2S,5S) diastereomer thereof
14-O-{[(1R,2R,3R)-3-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin and the (1S,2S,3S) diastereomer thereof
14-O-{[(1R,2R,3R)-3-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin and the (1S,2S,3S) diastereomer thereof
14-O-{[(11R,2R,4S)-4-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-
-acetyl}-mutilin and the (1S,2S,4R) diastereomer thereof
14-O-{[(1R,2R,5S)-5-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]--
acetyl}-mutilin and the (1S,2S,5R) diastereomer thereof
14-O-{[(1R,2R,3R/S)-3-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin and the (1S,2S,3R/S) diastereomer thereof
14-O-{[(1R,2R,5S)-2-Hydroxy-5-methylamino-cyclohexylsulfanyl]-acetyl}-mut-
ilin and the (1S,2S,5R) diastereomer thereof
14-O-{[(1R,2R,5S)-5-Allylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin and the (1S,2S,5R) diastereomer thereof
14-{[(1R,2R,5S)-2-Hydroxy-5-(2-methoxy-ethylamino)-cyclohexylsulfanyl]-ac-
etyl}-mutilin and the (1S,2S,5R) diastereomer thereof
14-O-{[(1R,2R,4R*)-2-Hydroxy-4-(2-hydroxy-ethylamino)-cyclohexylsulfanyl]-
-acetyl}-mutilin and the (1S,2S,4S*) diastereomer thereof
14-O-{[(1R,2R,4R*)-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-
-mutilin and the (1S,2S,4S*) diastereomer thereof
14-O-{[(1R,2R,4R*)-4-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-mutilin and the (1S,2S,4S*) diastereomer thereof
14-O-{[(1R,2R,5S*)-4-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-mutilin and the (1S,2S,5R*) diastereomer thereof
14-O-{[(1R,2R,4S*)-4-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-mutilin and the (1S,2S,4R*) diastereomer thereof
14-O-{[(1R,2R,5R*)-2-Hydroxy-5-morpholin-4-yl-cyclohexylsulfanyl]-acetyl}-
-mutilin and the (1S,2S,5S*) diastereomer thereof
14-O-{[(1R,2R,5S*)-2-Hydroxy-5-morpholin-4-yl-cyclohexylsulfanyl]-acetyl}-
-mutilin and the (1S,2S,5R*) diastereomer thereof
14-O-{[(1R,2R,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19,20-dih-
ydro-mutilin and the (1S,2S,5R) diastereomer thereof
14-O-{[(1R,2R,5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19,2-
0-dihydro-mutilin and the (1S,2S,5R) diastereomer thereof
14-O-{[(1R,2R,5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19,20-dih-
ydro-mutilin and the (1S,2S,5S) diastereomer thereof
14-O-{[(1R,2R)-4-Aminomethyl-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutili-
n and the (1S,2S) diastereomers thereof
14-O-{[5-Amino-2-chloro-cyclohexylsulfanyl]-acetyl}-mutilin
14-O-{[4-Amino-2-chloro-cyclohexylsulfanyl]-acetyl}-mutilin
14-O-[(4-Amino-1-hydroxy-cyclohexylmethylsulfanyl)-acetyl]-mutilin
14-O-{[(1R,2R)-2-Hydroxy-5-(3-methylamino-propyl)-cyclohexylsulfanyl]-ace-
tyl}-mutilin and the (1S,2S) diastereomer thereof
14-O-{[(1R,2R)-2-Hydroxy-4-(3-methylamino-propyl)-cyclohexylsulfanyl]-ace-
tyl}-mutilin and the (1S,2S) diastereomers thereof
14-O-{[(1R,2R)-5-(3-Amino-propyl)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-m-
utilin and the (1S,2S) diastereomers thereof
14-O-{[(1R,2R)-4-(3-Amino-propyl)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-m-
utilin and the (1S,2S) diastereomer thereof
14-O-{[(6R,8R)-8-Amino-1,4-dioxa-spiro[4.5]dec-6-ylsulfanyl]-acetyl}-muti-
lin and the (6S,8S) diastereomer thereof
14-O-{[4-Amino-2-methoxy-cyclohexylsulfanyl]-acetyl}-mutilin and
14-O-{[5-Amino-2-methoxy-cyclohexylsulfanyl]-acetyl}-mutilin
8. The compound according to any of claims 1 to 7 in the form of a
salt and/or solvate.
9. A compound according to any of claims 1 to 8 for use as a
pharmaceutical drug substance.
10. A method of treatment of diseases mediated by microbes which
comprises administering to a subject in need of such treatment an
effective amount of a compound of any one of claims 1 to 8.
11. A pharmaceutical drug composition comprising a compound of
anyone of claims 1 to 8, in association with at least one
pharmaceutical excipient.
12. A pharmaceutical drug composition according to claim 11,
further comprising another pharmaceutically active agent.
Description
[0001] The present invention relates to organic compounds, namely
pleuromutilins.
[0002] Pleuromutilin, a compound of formula A
##STR00002##
is a naturally occurring antibiotic, e.g. produced by the
basidomycetes Pleurotus mutilus and P. passeckerianus, see e.g. The
Merck Index, 13th edition, item 7617. A number of further
pleuromutilins having the principle ring structure of pleuromutilin
and being substituted at the hydroxy group have been developed,
e.g. as antimicrobials.
[0003] From WO 02/04414 A1 pleuromutilin derivatives, e.g.
14-O-[(Aminocyclohexan-2-yl (and -3-yl)-sulfanyl)-acetyl]-mutilins;
from WO 07/014409 A1 e.g. 14-O-[((Mono- or
dialkylamino)-cycloalkylsulfanyl)-acetyl]-mutilins and from WO
07/000004 A1 e.g.
[((Acyl-hydroxy-amino)-cycloalkylsulfanyl)-acetyl]-mutilins, are
known.
[0004] We have now found pleuromutilins with interesting activity
combined with an unexpected remarkable metabolic stability.
[0005] The pleuromutilin derivatives according to the invention are
compounds of formula (I)
##STR00003##
wherein n is 0 to 4; m is 0 or 1 with the proviso that the sulphur
atom and R.sub.3 are in vicinal position (if m=0 then R.sub.3 is in
position 2', and if m=1 then R.sub.3 is on position 1'); R is ethyl
or vinyl; R.sub.1 is hydrogen or (C.sub.1-6)alkyl, R.sub.2 is
hydrogen or [0006] (C.sub.3)cycloalkyl, or [0007] unsubstituted
(C.sub.1-6)alkyl, or [0008] (C.sub.1-6)alkyl substituted by one or
more of [0009] hydroxy, preferably one or two, [0010] methoxy,
[0011] halogen, [0012] (C.sub.3-6)cycloalkyl, or R.sub.1 and
R.sub.2 together with the nitrogen atom to which they are attached
form a 5 to 7 membered heterocyclic ring containing at least 1
nitrogen atom or 1 nitrogen and 1 additional heteroatom e. g.
selected from N or O, or R.sub.1 is hydroxy and R.sub.2 is formyl;
R.sub.3 is OH, OR.sub.4, a halogen atom, or [0013] with the proviso
that R.sub.3 is bound to 2' R.sub.3 represents
--O--(CH.sub.2).sub.p--O-- with p is 2 or 3; R.sub.4 is
unsubstituted (C.sub.1-6)alkyl or (C.sub.3-6)cycloalkyl.
[0014] Preferred compounds of the present invention are compounds
of formula (II)
##STR00004##
wherein n, R, R.sub.1, R.sub.2 and R.sub.3 are as defined
above.
[0015] More preferred compounds of the present invention are
compounds of formula (III)
##STR00005##
wherein n, R, R.sub.1 and R.sub.2 are as defined above.
[0016] Most preferred compounds of the present invention are [0017]
a compound of formula (IV)
[0017] ##STR00006## [0018] a compound of formula (V)
##STR00007##
[0018] and [0019] a compound of formula (VI)
##STR00008##
[0019] wherein n, R.sub.1 and R.sub.2 are as defined above.
[0020] Particularly preferred is a compound selected from the soup
consisting of [0021]
14-O-{[(1R,2R,4R)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
[0022]
14-O-{[(1S,2S,4S)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin [0023]
14-O-{[(1R,2R,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
[0024]
14-O-{[(1S,2S,5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin [0025]
14-O-{[(1R,2R,4S)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
and the (1S,2S,4R) diastereomer thereof [0026]
14-O-{[(1R,2R,5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
[0027]
14-O-{[(1S,2S,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin [0028]
14-O-{[(1R,2R,3R)-3-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
and the (1S,2S,3S) diastereomer thereof [0029]
14-O-{[(1R,2R,4R)-4-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin and the (1S,2S,4S) diastereomer thereof [0030]
14-O-{[(1R,2R,4R)-4-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin and the (1S,2S,4S) diastereomer thereof [0031]
14-O-{[(1R,2R,5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin and the (1S,2S,5R) diastereomer thereof [0032]
14-O-{[(1R,2R,5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin and the (1S,2S,5R) diastereomer thereof [0033]
14-O-{[(1R,2R,4S)-4-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin and the (1S,2S,4R) diastereomer thereof [0034]
14-O-{[(1R,2R,5R)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin and the (1S,2S,5S) diastereomer thereof [0035]
14-O-{[(1R,2R,3R)-3-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin and the (1S,2S,3S) diastereomer thereof [0036]
14-O-{[(1R,2R,3R)-3-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin and the (1S,2S,3S) diastereomer thereof [0037]
14-O-{[(1R,2R,4S)-4-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]--
acetyl}-mutilin and the (1S,2S,4R) diastereomer thereof [0038]
14-O-{[(1R,2R,5S)-5-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]--
acetyl}-mutilin and the (1S,2S,5R) diastereomer thereof [0039]
14-O-{[(1R,2R,3R/S)-3-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin and the (1S,2S,3R/S) diastereomer thereof [0040]
14-O-{[(1R,2R,5S)-2-Hydroxy-5-methylamino-cyclohexylsulfanyl]-acetyl}-mut-
ilin and the (1S,2S,5R) diastereomer thereof [0041]
14-O-{[(1R,2R,5S)-5-Allylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin and the (1S,2S,5R) diastereomer thereof [0042]
14-O-{[(1R,2R,5S)-2-Hydroxy-5-(2-methoxy-ethylamino)-cyclohexylsulfanyl]--
acetyl}-mutilin and the (1S,2S,5R) diastereomer thereof [0043]
14-O-{[(1R,2R,4R)-2-Hydroxy-4-(2-hydroxy-ethylamino)-cyclohexylsulfanyl]--
acetyl}-mutilin and the (1S,2S,4S*) diastereomer thereof [0044]
14-O-{[(1R,2R,4R*)-4-Cyclohexylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl-
}-mutilin and the (1S,2S,4S*) diastereomer thereof [0045]
14-O-{([(1R,2R,4R*)
4-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
and the (1S,2S,4S*) diastereomer thereof [0046]
14-O-{[(1R,2R,5S*)-4-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-mutilin and the (1S,2S,5R*) diastereomer thereof [0047]
14-O-{[(1R,2R,4S*)-4-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-mutilin and the (1S,2S,4R*) diastereomer thereof [0048]
14-O-{[(1R,2R,5R*)-2-Hydroxy-5-morpholin-4-yl-cyclohexylsulfanyl]-acetyl}-
-mutilin and the (1S,2S,5S*) diastereomer thereof [0049]
14-O-{[(1R,2R,5S)-2-Hydroxy-morpholin-4-yl-cyclohexylsulfanyl]-acetyl}-mu-
tilin and the (1S,2S,5R*) diastereomer thereof [0050]
14-O-{[(1R,2R,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19,20-dih-
ydro-mutilin and the (1S,2S,5R) diastereomer thereof [0051]
14-O-{[(1R,2R,5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19,2-
0-dihydro-mutilin and the (1S,2S,5R) diastereomer thereof [0052]
14-O-{[(1R,2R,5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19,20-dih-
ydro-mutilin and the (1S,2S,5S) diastereomer thereof [0053]
14-O-{[(1R,2R)-4-Aminomethyl-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutili-
n and the (1S,2S) diastereomers thereof [0054]
14-O-{[5-Amino-2-chloro-cyclohexylsulfanyl]-acetyl}-mutilin [0055]
14-O-{[4-Amino-2-cyclohexylsulfanyl]-acetyl}-mutilin [0056]
14-O-[(4-Amino-1-hydroxy-cyclohexylmethylsulfanyl)-acetyl]-mutilin
[0057]
14-O-{[(1R,2R)-2-Hydroxy-5-(3-methylamino-propyl)-cyclohexylsulfanyl]-ace-
tyl}-mutilin and the (1S,2S) diastereomers thereof [0058]
14-O-{[(1R,2R)-2-Hydroxy-4-(3-methylamino-propyl)-cyclohexylsulfanyl]-ace-
tyl}-mutilin and the (1S,2S) diasteromers thereof [0059]
14-O-{[(1R,2R)-5(3-Amino-propyl)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin and the (1S,2S) diastereomers thereof [0060]
14-O-{[(1R,2R)-4-(3-Amino-propyl)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-m-
utilin and the (1S,2S) diastereomer thereof [0061]
14-O-{[(6R,8R)-8-Amino-1,4-dioxa-spiro[4.5]dec-6-ylsulfanyl]-acetyl}-muti-
lin and the (6S,8S) diastereomer thereof [0062]
14-O-{[4-Amino-2-methoxy-cyclohexylsulfanyl]-acetyl}-mutilin and
[0063]
14-O-{[5-Amino-2-methoxy-cyclohexylsulfanyl]-acetyl}-mutilin.
[0064] A compound provided by the present invention is herein also
designated as "compound(s) of (according to) the present
invention". A compound of the present invention includes a compound
in any form, e.g. in free form, in the form of a salt, in the form
of a solvate and in the form of a salt and a solvate.
[0065] According to another aspect, the present invention provides
a compound of the present invention in the form of a salt and/or
solvate.
[0066] The salts include preferably pharmaceutically acceptable
salts, although pharmaceutically unacceptable salts are included,
e.g. for preparation/isolation/purification purposes.
[0067] A salt of a compound of the present invention includes a
base salt or an acid addition salt Pharmaceutically acceptable base
salts include ammonium salts such as trimethylammonium salt, alkali
metal salts such as those of sodium and potassium, alkaline earth
metal salts such as those of calcium and magnesium, and salts with
organic bases, including salts of primary, secondary and tertiary
amines, such as isopropylamine, diethylamine, ethanolamine,
trimethylamine, dicyclohexyl amine and N-methyl-D-glucamine,
preferably sodium salts. Acid addition salts include salts of a
compound of the present invention with an acid, e.g. hydrogen
fumaric acid, fumaric acid, tartaric acid, ethane-1,2-disulphonic
acid, maleic acid, naphthalin-1,5-sulphonic acid, acetic acid,
maleic acid, succinic acid, salicylic acid, azelaic acid,
2-[(2,6-dichlorophenyl)amino]benzene acetic acid, hydrochloric
acid, deuterochloric acid, preferably hydrochloric acid.
[0068] A compound of the present invention in free form may be
converted into a corresponding compound in the form of a salt, and
vice versa. A compound of the present invention in free form or in
the form of a salt and/or in the form of a solvate may be converted
into a corresponding compound in free form or in the form of a salt
in non-solvated form, and vice versa.
[0069] A compound of the present invention may exist in the form of
isomers and mixtures thereof e.g. optical isomers,
diastereoisomers, cis/trans conformers. A compound of the present
invention may e.g. contain asymmetric carbon atoms and may thus
exist in the form of enantiomers or diastereoisomers and mixtures
thereof, e.g. racemates or diastereomeric mixtures. Any asymmetric
carbon atom may be present in the (R)-, (S)- or
(R,S)-configuration, preferably in the (R)- or
(S)-configuration.
[0070] For example, in a compound of formula I the carbon atom of
the cycloalkyl ring which is attached to (CH.sub.2).sub.mS group,
the carbon atom of the cycloalkyl ring which is attached to the
R.sub.3 group, and the carbon atom of the cycloalkyl ring to which
the (CH.sub.2).sub.nN(R.sub.1R.sub.2) group is attached, all are
asymmetric carbon atoms. Substituents attached to such asymmetric
carbon atom may thus exist in (R) and (S) configuration, including
mixtures thereof. For example, if in a compound of formula I
R.sub.2 is substituted alkyl and that substituent is attached to a
carbon atom of the side chain of such alkyl, the carbon atom to
which such substituent is attached is an asymmetric carbon atom and
such substituent may be in the (R)- and (S)-configuration,
including mixtures thereof.
[0071] The configuration of substituents attached to asymmetric
carbon atoms of the mutilin-tricycles is preferably the same as in
natural pleuromutilin.
[0072] Isomeric mixtures may be separated as appropriate, e.g.
according e.g. analogously, to a method as conventional, to obtain
pure isomers. The present invention includes a compound of the
present invention in any isomeric form and in any isomeric mixture.
The present invention also includes tautomers of a compound of the
present invention, where tautomers can exist.
[0073] The compounds of the present invention exhibit
pharmacological activity and are therefore useful as
pharmaceuticals.
[0074] For example, the compounds of the present invention show
antimicrobial, e.g. antibacterial, activity against gram positive
bacteria, such as coagulase positive Staphylococci, e.g.
Staphylococcus aureus, coagulase negative Staphylococci, e.g.
Staphylococcus opidermidis, Staphylococcus haemolyticus, and
Streptococci. e.g. Streptococcus pyogenes, Streptococcus
pneumoniae, Enterococci, e.g. Enterococcus faccium and Listeria
monocytogenes and against gram negative bacteria such as Moraxella,
e.g. Moraxella catarrhalis, and Haemophilus, e.g. Haemophilus
influenzae, and Legionella, e.g. Legionella pneumophila,
Neisseriaceae, e.g. Neisseria gonorrhoeae, as well as against
Mycoplasms, Chlamydia and obligatory anaerobes, e.g. Bacteroides
fragilis, Clostridium difficile, Fusobacterium spp., and
Propionibacterium spp.
[0075] The in vitro activity against aerobic bacteria was
determined by Agar Dilution Test or Microdilution Test according to
the Clinical and Laboratory Standards Institute (CLSI, former
NCCLS) Document M7-A7 Vol. 26, No. 2: "Methods for dilution
Antimicrobial Susceptibility Tests for Bacteria that Grow
Aerobically--Approved Standard; Seventh Edition (2006)"; and the
test against anaerobic bacteria was performed according to the
Clinical and Laboratory Standards Institute (CLSI, former NCCLS),
Document, M11-A6, Vol. 24, No. 2: "Methods for Antimicrobial
Susceptibility Testing of Anaerobic Bacteria--Approved Standard;
Sixth Edition (2004)" and the in vivo activity was tested by the
septicaemia mouse model against Staphylococcus aureus.
[0076] Compounds of the present invention are therefore suitable
for the treatment and prevention of diseases which are mediated by
microbes, e.g. by bacteria. Diseases which may also be treated
include e.g. diseases mediated by Helicobacter, such as
Helicobacter pylori, and diseases mediated by Mycobacterium
tuberculosis. Diseases which may also be treated include in general
inflammatory diseases, where microbes are mediating said
inflammation, e.g. including acne.
[0077] In another aspect the present invention provides a compound
of the present invention for use as a pharmaceutical, preferably as
an antimicrobial, such as an antibiotic, e.g. and an
anti-anaerobic.
[0078] In another aspect the present invention provides a compound
of the present invention for use in acne treatment.
[0079] In a further aspect the present invention provides a
compound of the present invention for use in the preparation of a
medicament for the treatment of diseases, mediated by microbes,
such as bacterials, for example [0080] diseases mediated by
bacteria, e.g. selected from Staphylococci, Streptococci,
Enterococci; [0081] diseases mediated by bacteria, e.g. selected
from Moraxella, Haemophilus, Legionella, Neisseriaceae; [0082]
diseases mediated by Helicobacter, [0083] diseases mediated by
Mycobacterium tuberculosis; [0084] e.g. diseases mediated by
Mycoplasms, Chlamydia and obligatory anaerobes; and for the
treatment of acne.
[0085] In a further aspect the present invention provides a method
of treatment of diseases mediated by microbes which comprises
administering to a subject in need of such treatment an effective
amount of a compound of the present invention e.g. in the form of a
pharmaceutical composition.
[0086] In a further aspect the present invention provides a method
of treatment of acne which comprises administering to a subject in
need of such treatment an effective amount of a compound of the
present invention e.g. in the form of a pharmaceutical
composition.
[0087] Treatment includes treatment and prophylaxis.
[0088] For antimicrobial and acne treatment, the appropriate dosage
will, of course, vary depending upon, for example, the chemical
nature and the pharmacokinetic data of a compound of the present
invention employed, the individual host, the mode of administration
and the nature and severity of the conditions being treated.
However, in general, for satisfactory results in larger mammals,
for example humans, an indicated daily dosage is in the range from
about 0.5 mg to 3 g of a compound of the present invention
conveniently administered, for example, in divided doses up to four
times a day.
[0089] A compound of the present invention may be administered by
any conventional route, for example enterally, e.g. including
nasal, buccal, rectal, oral administration; parenterally, e.g.
including intravenous, intramuscular, subcutaneous administration;
or topically, e.g. including epicutaneous, intranasal,
intratracheal administration, e.g. in form of coated or uncoated
tablets, capsules, injectable solutions or suspensions, e.g. in the
form of ampoules, vials, in the form of creams, gels, pastes,
inhaler powder, foams, tinctures, lip sticks, drops, sprays, or in
the form of suppositories, e.g. in analogous manner to macrolides,
such as erythromycins, e.g. clarithromycin or azithromycin.
[0090] A compound of the present invention may be administered in
the form of a pharmaceutically acceptable salt, e.g. an acid
addition salt or a base addition salt, e.g. a metal salt. or in
free form, optionally in the form of a solvate. A compound of the
present invention in the form of a salt exhibits the same order of
activity as the compound in free form, optionally in the form of a
solvate.
[0091] A compound of the present invention may be used for
pharmaceutical treatment according to the present invention alone
or in combination with one or more other pharmaceutically active
agents. Such other pharmaceutically active agents include e.g.
other antibiotics and antiinflammatory agents, and, if a compound
of the present invention is used in the treatment of acne, other
pharmaceutically agents include furthermore agents which are active
against acne.
[0092] Combinations include fixed combinations, in which two or
more pharmaceutically active agents are in the same formulation;
kits, in which two or more pharmaceutically active agents in
separate formulations are sold in the same package, e.g. with
instruction for co-administration; and free combinations in which
the pharmaceutically active agents are packaged separately, but
instruction for simultaneous or sequential administration are
given.
[0093] In another aspect the present invention provides a
pharmaceutical composition comprising a compound of the present
invention in free form or in the form of a pharmaceutically
acceptable salt and/or in the form of a solvate in association with
at least one pharmaceutical, excipient, e.g. carrier or diluent,
e.g. including fillers, binders, disintegrators, flow conditioners,
lubricants, sugars and sweeteners, fragrances, preservatives,
stabilizes, wetting agents and/or emulsifiers, solubilizes, salts
for regulating osmotic pressure and/or buffers.
[0094] In another aspect the present invention provides a
pharmaceutical composition according to the present invention,
further comprising another pharmaceutically active agent.
[0095] Such pharmaceutical compositions may be manufactured
according, e.g. analogously, to a method as conventional, e.g. by
mixing, granulating, coating, dissolving or lyophilizing processes.
Unit dosage form may contain, for example, from about 0.5 mg to
about 2000 mg, such as 10 mg to about 500 mg.
[0096] The compounds of the present invention are additionally
suitable as veterinary agents, e.g. veterinary active compounds,
e.g. in the prophylaxis and in the treatment of microbial, e.g.
bacterial diseases, in animals, such as fowl, pigs and calves,
e.g., and for diluting fluids for artificial insemination and for
egg-dipping techniques.
[0097] In another aspect the present invention provides a compound
of the present invention for use as a veterinary agent.
[0098] In a further aspect the present invention provides a
compound of the present invention for the preparation of a
veterinary composition which is useful as a veterinary agent.
[0099] In another aspect the present invention provides a
veterinary method for the prophylaxis and the treatment of
microbial, e.g. bacterial diseases which comprises administering to
a subject in need of such treatment an effective amount of a
compound of the present invention, e.g. in the form of a veterinary
composition.
[0100] Examples 1 to 37 following thereafter exhibit MICs.ltoreq.2
.mu.g/ml against Staphylococcus aureus ATCC49951 and Streptococcus
pneumoniae ATCC49619.
[0101] The metabolic stability for compounds of the present
invention was determined by using cryopreserved primary human
hepatocytes. 1.times.10.sup.6 cells/mL were incubated in the
absence and the presence of 5 and 25 .mu.g/mL of the test compounds
at 37.degree. C., 5% CO.sub.2 for 4 hours. To evaluate the in vitro
degradation under assay conditions, a sample of each test compound
was incubated also in the absence of hepatocytes. The incubation
was stopped by freezing the reaction mixture. After ultrafiltration
and washing of the filter with acetonitrile, the sample solution
was analyzed for parent compound disappearance or metabolite
appearance using LC/MS (ion trap). The metabolic stability value
corresponds to the detected parent compound in % after
incubation.
[0102] At the compounds of the present invention, the introduction
of the R.sub.3 group, preferably a hydroxy group, in vicinal
position to the sulphur substituent attached to the cyclohexyl ring
reveals unexpected improvements in metabolic stability of the
microbiologically active components. Parent compound or active
metabolite were more stable after incubation with primary human
hepatocytes in comparison to derivatives without the R.sub.3 group,
preferably the hydroxy group, attached to the cyclohexyl moiety of
the pleuromutilin side chain.
[0103] For example after 4 h incubation with human hepatocytes at a
compound concentration of 5 .mu.g/mL, for a mixture of
14-O-{[(1R,2R,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride and the (1S,2S,5R) diastereomer hydrochloride
thereof--Example 2 of the present invention--66% of parent
compounds were found, whereas for mixture of
14-O-{[(1R,3R)-3-Amino-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride and the (1S,3S) diastereomer hydrochloride
thereof--analogous derivative without hydroxyl group--only 24% of
parent compounds could be detected.
EXAMPLES
[0104] The trivial name mutilin refers to the IUPAC systematic name
(1S,2R,3S,4S,6R,7R,8R,14R)-3,6-dihydroxy-2,4,7,14-tetramethyl-4-vinyl-tri-
cyclo[5.4.3.0.sup.1,8]tetradecan-9-one. In the examples,
pleuromutilin derivatives are numbered in analogy to the mutilin
numbering system described by H. Berner (Berner, H.; Schulz, G.;
Schneider H. Tetrahedron 1980, 36, 1807-1811.):
##STR00009##
[0105] Pleuromutilin thiol and pleuromutilin tosylate are compounds
of formulae:
##STR00010##
Example 1
14-O-{[(1R,2R,4R)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride+(1S,2S,4S) diastereomer hydrochloride
Step A1.
14-O-{[(1R,2R,4R)-4-tert-Butoxycarbomylamino-2-hydroxy-cyclohexyl-
sulfanyl]-acetyl}-mutilin+(1S,2S,4S) diastereomer and
14-O-{[(1R,2R,5S)-5-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,5R) diastereomer and
14-O-{[(1R,2R,4S)-4-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,4R) diastereomer
[0106] To a solution of 3,4-epoxycyclohexyl-carbamic acid
tert-butyl ester (Gomez-Sanchez, E.; Marco-Contelles J. Tetrahedron
2005, 61, 1207-1219.) (4.27 g, 20 mmol) and pleuromutilin thiol
(Nagarajan, R. Eli Lilly and Company 1978, U.S. Pat. No. 4,130,709)
(7.10 g, 18 mmol) in 200 ml of tetrahydrofuran was added aluminum
oxide (40 g, Brockmann activity I, neutral) and the resulting
mixture was stirred for 40 hours at room temperature. The
suspension was filtered and concentrated under reduced pressure.
The residue was subjected to chromatography (silica,
cyclohexane/ethyl acetate=1/1) to yield
14-O-{[(1R,2R,4R)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,4S) diastereomer (a) (R.sub.f=0.38, 1.34
g, 12%) as well as a mixture of
14-O-{[(1R,2R,5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,5R) diastereomer and
14-O-{[(1R,2R,4S)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,4R) diastereomer (b) (R.sub.f=0.26, 2.81
g, 25%) as colorless amorphous foams.
[0107] (a): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.74 (d, 1H, NH, J=7 Hz), 6.13 (dd, 1H, 19-H, J=11 Hz and 18
Hz), 5.54 (d, 1H, 14-H, J=8 Hz), 5.05 (m, 2H, 20-H), 4.90 (d, 1H,
2'-OH, J=5 Hz), 4.48 (d, 1H, 11-OH, J=6 Hz), 3.55-3.20 (m, 6H,
1'-H, 2'-H, 4'-H, 11-H, 22-H), 2.40 (bs, 1H, 4-H), 1.36 (s, 3H,
15-CH.sub.3), 1.35 (s, 9H, tert-butyl), 1.06 (s, 3H, 18-CH.sub.3),
0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.62 (d, 3H, 16-CH.sub.3, J=7
Hz). MS-ESI (m/z): 630 (MNa.sup.+), 1237 (2 MNa.sup.+).
[0108] (b): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.70 (d, 1H, N, J=7 Hz), 6.12 (dd, 1H, 19-H, J=11 Hz and 18
Hz), 5.34 (d, 1H, 14-H, J=8 Hz), 5.05 (m, 2H, 20-H), 4.82, 4.78 (d,
1H, 2'-OH, J=4 Hz), 4.48 (d, 1H, 11-OH, J=6 Hz), 3.55-3.20 (m, 5H,
2'-H, 4'/5'-H, 11-H, 22-H), 2.97 (m, 1H, 1'-H), 2.40 (bs, 1H, 4-H),
1.35 (s, 12H, 15-CH.sub.3, tert-butyl), 1.05 (s, 3H, 18-CH.sub.3),
0.82 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.62 (d, 3H, 16-CH.sub.3, J=7
Hz). MS-ESI (m/z): 630 (MNa.sup.+), 1237 (2MNa.sup.+).
or Step A2.
14-O-{[(1R,2R,4R)-4-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,4S) diastereomer and
14-O-{[(1R,2R,5S)-5-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,5R) diastereomer and
14-O-{[(1R,2R,4S)-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]--
acetyl}-mutilin+(1S,2S,4R) diastereomer
[0109] To a solution of 3,4-epoxycyclohexyl-carbamic acid
tert-butyl ester (10 & 47 mmol) and pleuromutilin thiol (16.6
g, 42 mmol) in 200 ml of methanol and 20 ml of dioxane was added 2N
NaOH (21 ml, 42 mmol) and the resulting mixture was stirred for 16
hours at room temperature. After completion of the reaction the pH
was set to 7 with diluted HCl and the reaction mixture was
concentrated under reduced pressure. The residue was diluted with
water and brine and extracted three times with ethyl acetate. The
organic layers were dried over sodium sulfate and filtered. The
filtrate was concentrated under reduced pressure and after
chromatography (silica, cyclohexane/ethyl acetate=1/1)
14-O-{[(1R,2R,4R)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,4S) diastereomer (R.sub.f=0.40, 3.1 g, 12%
yield) as well as a mixture of
14-O-{[(1R,2R,5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,5R) diastereomer and
14-O-{[(1R,2R,4S)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,4R) diastereomer (R.sub.f=0.25, 6.35 g,
25%) were obtained as colorless amorphous foams.
or Step A3.
14-O-{[(1R,2R,4R)-4-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,4S) diastereomer and
14-O-{[(1R,2R,5S)-5-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,5R) diastereomer
[0110] To a solution of Pleuromutilin thiol (9.25 & 23.5 mmol)
in 100 ml of acetonitrile (dried over 4 .ANG. molecular sieve) was
added 1,5-diazabicyclo[4.3.0]non-5-ene (DBN, 2.9 .mu.l, 23.5 mmol)
and after 1 hour of stirring at room temperature under argon
atmosphere the mixture was charged with
syn-3,4-epoxycyclohexyl-carbamic acid tert-butyl ester (4.17 g.
19.5 mmol) and stirred for further 16 hours at room temperature.
The reaction mixture was concentrated under reduced pressure. The
residue was charged with water and brine and extracted three times
with dichloromethane. The organic layers were dried over sodium
sulphate and filtered. The filtrate was concentrated under reduced
pressure and subjected to chromatography (silica, cyclohexane/ethyl
acetate=1/1) to yield
14-O-{([(1R,2R,4R)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexyls-
ulfanyl]-acetyl}-mutilin+(1S,2S,4S) diastereomer (R.sub.f=0.38,
5.07 g, 43%) as well as
14-O-{[(1R,2R,5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,5R) diastereomer (R.sub.f=0.25, 2.95 g,
16.5%) as colorless amorphous foams.
Step B.
14-O-{[(1R,2R,4R)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin+(1S,2S,4S) diastereomer
[0111] To a solution of
14-O-{[(1R,2R,4R)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,4S) diastereomer (1.34 g, 2.20 mmol) in 75
ml of dichloromethane was added trifluoroacetic acid (4 ml) at
4.degree. C. and stirred for 5 hours at room temperature. The
reaction mixture was diluted with dichloromethane and cautiously
poured into a saturated NaHCO.sub.3 solution. The phases were
separated and the aqueous layer was washed two times with
dichloromethane. The combined organic layers are dried over sodium
sulfate and filtered. After chromatography (silica, ethyl
acetate/methanol/35% ammonia solution=50/50/1)
14-O-{[(1R,2R,4R)-4-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(-
1S,2S,4S) diastereomer (745 mg, 67% yield) was obtained as
colorless amorphous foam.
[0112] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.14 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.54 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 2H, 20-H), 4.50 (d, 1H, 11-OH, J=6 Hz), 3.50-3.20
(m, 5H, 2'-H, 4'-H, 11-H, 22-H), 2.55 (m, 1H, 1'-H), 2.40 (bs, 1H,
4-H), 1.35 (s, 3H, 15-CH.sub.3), 1.06 (s, 3H, 18-CH.sub.3), 0.82
(d, 3H, 17-CH.sub.3, J=7 Hz), 0.62 (d, 3H, 16-CH.sub.3, J=7 Hz).
MS-ESI (m/z): 508 (MH.sup.+), 530 (MNa.sup.+), 1015 (2MH.sup.+),
1037 (2MNa.sup.+).
Step C.
14-O-{[(1R,2R,4R)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin hydrochloride+(1S,2S,4S) diastereomer hydrochloride
[0113] A solution of
14-O-{[(1R,2R,4R)-4-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(-
1S,2S,4S) diastereomer (325 mg, 0.64 mmol) in 20 ml of dioxane was
treated with 1N HCl (0.64 ml, 0.64 mmol). After stirring at room
temperature for 30 minutes the solution was lyophilized to obtain
14-O-{[(1R,2R,4R)-4-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride+(1S,2S,4S) diastereomer hydrochloride (quantitative
yield) as colorless amorphous solid.
[0114] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.6 (bs, 3H, NH.sub.3.sup.+), 6.14 (dd, 1H, 19-H, J=11 Hz
and 18 Hz), 5.55 (d, 1H, 14-H, J=8 Hz), 5.05 (m, 2H, 20-H), 4.52
(d, 1H, 11-OH, J=6 Hz), 3.50-3.20 (m, 4H, 2'-H, 11-H, 22-H), 3.03
(m, 1H, 4'-H), 2.53 (m, 1H, 1'-H), 2.40 (bs, 1H, 4-H), 1.37 (s, 3H,
15-CH.sub.3), 1.06 (s, 3H, 18-CH.sub.3), 0.82 (d, 3H, 17-CH.sub.3,
J=7 Hz), 0.62 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 508
(MH.sup.+), 530 (MNa.sup.+), 1015 (2MH.sup.+), 1037 (2MNa.sup.+),
542 (MCl.sup.-).
Example 1A
14-O-{[(1S,2S,4S)-4-Amino-2-hydro-cyclohexylsulfanyl]-acetyl}-mutilin
and
14-O-{[(1R,2R,4R)-4-Amino-2-hydroxy-cyclohexylsulfanyl]acetyl}-mutilin
[0115] The mixture of
14-O-{[(1R,2R,4R)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(-
1S,2S,4S) diastereomer (12 g, 23.6 mmol) from Example 1 Step B was
separated on a chiral column (250.times.20 mm CHIRALCEL OD-H,
n-heptane/ethanol/diethylamine=80/20/0.1) to yield
14-O-{[(1S*,2S*,4S*)-4-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutili-
n (a) (early eluting compound, 4.76 & 37% yield, uncorrected)
and 14-O-{[(1R*,2R*,
4R*)-4-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin (b)
(late eluting compound. 3.63 g, 30% yield, uncorrected) as
colorless amorphous foams.
[0116] (a): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.13 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.54 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 2H, 20-H), 4.50 (d, 1H, 11-OH, J=6 Hz), 3.50-3.20
(m, 5H, 2'-H, 4'-H, 11-H, 22-H), 2.55 (m, 1H, 1'-H), 2.40 (bs, 1H,
4-H), 1.35 (s, 3H, 15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3), 0.82
(d, 3H, 17-CH.sub.3, J=7 Hz), 0.62 (d, 3H, 16-CH.sub.3, J=7 Hz).
MS-ESI (m/z): 508 (MH.sup.+), 530 (MNa.sup.+), 1015 (2MH.sup.+),
1037 (2MNa.sup.+), 506 (M-H).sup.-, 542 (MCl.sup.-).
[0117] (b): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.13 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.54 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 2H, 20-H), 4.50 (d, 1H, 11-OH, J=6 Hz), 3.50-3.20
(m, 5H, 2'-H, 4'-H, 11-H, 22-H), 2.55 (m, 1H, 1'-H), 2.40 (bs, 1H,
4-H), 1.35 (s, 3H, 15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3), 0.82
(d, 3H, 17-CH.sub.3, J=7 Hz), 0.62 (d, 3H, 16-CH.sub.3, J=7 Hz).
MS-ESI (m/z): 508 (MH.sup.+), 530 (MNa.sup.+), 1015 (2MH.sup.+),
1037 (2MNa.sup.+), 506 (M-H).sup.-, 542 (MCl.sup.-).
Example 2
14-O-{[(1R,2R,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride+(1S,2S,5R) diastereomer hydrochloride
Step A.
14-O-{[(1R,2R,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin+(1S,2S,5R) diastereomer and
14-O-{[(1R,2R,4S)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(-
1S,2S,4R) diastereomer
[0118] A mixture of
14-O-{[(1R,2R,5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,5R) diastereomer and
14-O-{[(1R,2R,4S)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,4R) diastereomer (1.12 g, 1.84 mmol) from
Example 1 Step A was treated according to the method of Example 1
Step B. After work up and chromatography of the reaction mixture
(silica, ethyl acetate/methanol/35% ammonia solution=50/50/1)
14-O-{[(1R,2R,5S)-5-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(-
1S,2S,5R) diastereomer (a) (R.sub.f=0.33, 524 mg, 56% yield) and
14-O-{[(1R,2R,4S)-4-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(-
1S,2S,4R) diastereomer (b) (R.sub.f=0.22, 160 mg, 17%) were
obtained as colorless amorphous foams.
[0119] (a): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.13 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.53 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 2H, 20-H), 4.51 (d, 1H, 11-OH, J=6 Hz), 3.48 (m,
1H, 2'-H), 3.42 (m, 1H, 11-H), AB-system (.nu..sub.A=3.37,
.nu..sub.B=3.23, 22-H, J=19 Hz), 2.98 (m, 1H, 1-H), 2.82 (m, 1H,
5'-H), 2.40 (bs, 1H, 4-H), 1.36 (s, 3H, 15-CH.sub.3), 1.06 (s, 3H,
18-CH.sub.3), 0.82 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (d, 3H,
16-CH.sub.3, J=7 Hz).
[0120] (b): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.14 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.54 (d, 1H, 14-H,
J=8 Hz), 5.06 (m, 2H, 20-H), 4.51 (bs, 1H, 11-OH), 3.79 (m, 1H,
2'-H), 3.42 (m, 1H, 11-H), AB-system (.nu..sub.A=3.33,
.sigma..sub.B=3.23, 22-H, J=15 Hz), 3.04 (m, 1H, 4'-H), 2.82 (m,
1H, 1'-H), 2.40 (bs, 1H, 4-H), 1.37 (s, 3H, 15-CH.sub.3), 1.06
(3,3H, 18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (d,
3H, 16-CH.sub.3, J=7 Hz).
Step B.
14-O-{[(1R,2R,5S)-5-Amino-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin hydrochloride+(1S,2S,5R) diastereomer hydrochloride
[0121]
14-O-{[(1R,2R,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mut-
ilin+(1S,2S,5R) diastereomer (516 mg, 1.02 mmol) was treated
according to the method of Example 1 Step C to obtain
14-O-{[(1R,2R,5S)-5-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride+(1S,2S,5R) diastereomer hydrochloride (533 mg, 96%
yield) as colorless amorphous solid.
[0122] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.7 (bs, 3H, NH.sub.3.sup.+), 6.13, 6.12 (2dd, 1H, 19-H,
J=11 Hz and 18 Hz), 5.55 (d, 1H, 14-H, J=8 Hz), 5.05 (m, 2H, 20-H),
4.53 (d, 1H, 11-OH, J=6 Hz), 3.70 (m, 1H, 2'-H), 3.42 (t, 1H, 11-H,
J=6 Hz), 3.35 (m, 2H, 22-H), 3.09 (m, 2H, 1'-H, 5'-H), 2.40 (bs,
1H, 4-H), 1.35 (s, 3H, 15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3),
0.82 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (d, 3H, 16-CH.sub.3, J=7
Hz). MS-ESI (m/z): 508 (MH.sup.+), 530 (MNa.sup.+), 1015
(2MH.sup.+), 1037 (2MNa.sup.+), 542 (MCl.sup.-).
Example 2A
14-O-{[(1R,2R,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
and
14-O-{[(1S,2S,5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutil-
in
[0123] The mixture of
14-O-{[(1R,2R,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(-
1S,2S,5R) diastereomer (4.91 g. 9.67 mmol) from Example 2 Step A
was separated on a. chiral column (250.times.20 mm CHIRALCEL OD-H,
n-heptane/isopropanol/diethylamine=80/20/0.1) to yield
14-O-{[(1R*,2R*,5S)-5-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
(a) (early eluting compound, 2.07 g, 42% yield, uncorrected) and
14-{[(1S*,2S*,5R*)-5-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
(b) (late eluting compound, 2.36 &, 48% yield, uncorrected) as
colorless amorphous foams.
[0124] (a): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.13 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.53 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 2H, 20-H), 4.72 (d, 1H, 2'-OH, J=4 Hz), 4.50 (d,
1H, 11-OH, J=6 Hz), 3.47 (m, 1H, 2'-H), 3.45-3.20 (m, 3H, 11-H,
22-H), 2.98 (m, 1H, 1'-H), 2.80 (m, 1H, 5'-H), 2.40 (bs, 1H, 4-H),
1.36 (s, 3H, 15-CH.sub.3), 1.06 (s, 3H, 18-CH.sub.3), 0.81 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.63 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI
(m/z): 508 (MH.sup.+), 530 (MNa.sup.+), 1015 (2MH.sup.+), 1037
(2MNa.sup.+), 506 (M-H).sup.-, 542 (MCl.sup.-).
[0125] (b): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.13 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.53 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 21, 20-H), 4.72 (d, 1H, 2'-OH, J=4 Hz), 4.50 (d,
1H, 11-OH, J=6 Hz), 3.47 (m, 1H, 2'-H), 3.45-3.20 (m, 3H, 11-H,
22-H), 2.98 (m, 1H, 1'-H), 2.80 (m, 1H, 5'-H), 2.40 (bs, 1H, 4-H),
1.36 (s, 3H, 15-CH.sub.3), 1.06 (s, 3H, 18-CH.sub.3), 0.81 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.63 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI
(m/z): 508 (MH.sup.+), 530 (MNa.sup.+), 1015 (2MH.sup.+), 1037
(2MNa.sup.+), 506 (M-H).sup.-, 542 (MCl.sup.-).
Example 3
14-O-{[(1R,2R,4S)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride+(1S,2S,4R) diastereomer hydrochloride
[0126]
14-O-{[(1R,2R,4S)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mut-
ilin+(1S,2S,4R) diastereomer (152 mg, 0.30 mmol) from Example 2
Step A was treated according to the method of Example 1 Step C to
obtain
14-O-{[(1R,2R,4S)-4-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride+(1S,2S,4R) diastereomer hydrochloride (148 mg, 91%
yield) as colorless amorphous solid.
[0127] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.8 (bs, 3H, NH.sub.3), 6.14, 6.13 (2dd, 1H, 19-H, J=11 Hz
and 18 Hz), 5.55 (d, 1H, 14-H, J=8 Hz), 5.20 (d, 1H, 2'-OH), 5.05
(m, 2H, 20-H), 4.53 (d, 1, 11-OH, J=6 Hz), 3.88 (m, 1H, 2'-H), 3.42
(t, 1H, 11-H, J=6 Hz), 3.32 (m, 2H, 22-H), 3.22 (m, 1H, 4'-H), 2.92
(m, 1H, 1'-H), 2.40 (bs, 1H, 4-H), 1.35 (s, 3H, 15-CH.sub.3), 1.05
(s, 3H, 18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (d,
3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 508 (MH.sup.+), 530
(MNa.sup.+), 1015 (2MH.sup.+), 1037 (2MNa.sup.+), 542
(MCl.sup.-).
Example 4
14-O-{[(1R,2R,5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1-
S,2S,5S) diastereomer
Step A. tert-Butyl-dimethyl-(cis-3,4-epoxycyclohexyloxy)-silane
[0128] To a solution of 3-cyclohexen-1-ol (Amburgey, J. C.; Shuey,
S. W.; Pedersen, I. G.; Hiskey R., Bioorganic Chemistry 1994, 22,
172-197.) (10 & 102 mmol) in 200 ml of dichloromethane was
added vanadyl acetylacetonate (0.5 g, cat.) and tert-butyl
hydroperoxide (20.4 ml 5.5M in decane, 112 mmol) and stirred
overnight at room temperature. The resulting reaction mixture was
treated with tert-butyldimethylsilyl chloride (16.9 g, 112 mmol),
imidazole (9.02 g, 132 mmol) and 4-dimethylaminopyridine (2.49 g,
20 mmol) at 4.degree. C. and stirred for 5 hours at room
temperature. The reaction mixture was diluted with dichloromethane
and subsequently extracted with 10% NaHSO.sub.3 solution, saturated
NaHCO.sub.3 solution and brine. The organic layer was dried over
sodium sulfate and filtered. The filtrate was concentrated under
reduced pressure and subjected to chromatography (silica,
cyclohexane/ethyl acetate 15/1) to yield
tert-butyl-dimethyl-(cis-3,4-epoxycyclohexyloxy)-silane
(R.sub.f=0.35, 18.3 g, 79% yield) as colorless oil.
[0129] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm): 3.55 (m,
1H), 3.00 (m, 2H), 2.15 (m, 1H), 2.00 (m, 1H), 1.80 (m, 1H), 1.50
(m, 1H), 1.35 (m, 1H), 1.35 (m, 1H), 1.25 (m, 1H), 0.83 (s, 9H,
tert-butyl), 0.0 (s, 9H, Si(CH.sub.3).sub.2).
Step B.
14-O-{[(1R,2R,5S)-(tert-Butyl-dimethyl-silyloxy)-2-hydroxy-cyclohe-
xylsulfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer
[0130] tert-Butyl-dimethyl-(cis-3,4-epoxycyclohexyloxy)-silane
(6.41 g, 28 mmol) was treated with pleuromutilin thiol according to
the method of Example 1 Step A2. Crude
14-O-{[(1R,2R,5S)-5-(tert-butyl-dimethyl-silyloxy)-2-hydroxy-cyclohexylsu-
lfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer was obtained as
colorless amorphous foam which was directly used for the next
step.
[0131] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.13 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.52 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 2H, 20-H), 4.78 (dd, 1H, 2'-OH, J=5 Hz and 6 Hz),
4.48 (d, 1H, 11-OH, J=6 Hz), 3.88 (m, 1H, 5'-H), 3.15-3.45 (m, 4H,
2'-H, 11-H, 22-CH.sub.2), 2.92 (m, 1H, 1'-H), 2.38 (bs, 1H, 4-H),
1.36 (s, 3H, 15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3), 0.86 (s, 9H,
tert-butyl), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (d, 3H,
16-CH.sub.3, J=7 Hz), 0.0 (s, 6H, Si(CH.sub.3).sub.2).
Step C.
14-O-{[(1R,2R,5S)-2,5-Dihydroxy-cyclohexylsulfanyl]-acetyl}-mutili-
n+(1S,2S,5R) diastereomer
[0132] To a solution of
14-O-{[(1R,2R,5S)-5-(tert-butyl-dimethyl-silyloxy)-2-hydroxy-cyclohexylsu-
lfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer (9.46 g, 15.1 mmol)
in 25 ml of tetrahydrofuran a mixture of acetic acid and water
(3:1, 100 ml) was added and stirred for 2 days at 40.degree. C. The
reaction mixture was concentrated nearly to dryness under reduced
pressure and the residue was dissolved in ethyl acetate and
submitted to chromatography (silica, cyclohexane/ethyl acetate=1/3)
to yield the
14-O-{[(1R,2R,5S)-2,5-dihydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2-
S,5R) diastereomer (R.sub.f=0.27, 7.07 g, 92% yield) as colorless
amorphous foam.
[0133] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.12 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.53 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 2H, 20-H), 4.72 (dd, 1H, 2'-OH, J=2 Hz and 5 Hz),
4.48 (d, 1H, 11-OH, J=6 Hz), 4.43 (t, 1H, 5'-OH), 3.68 (m, 1H,
5'-H), 3.45-3.20 (m, 4H, 2'-H, 11-H, 22-H), 2.94 (m, 1H, 1'-H),
2.38 (bs, 1H, 4-H), 1.36 (s, 3H, 15-CH.sub.3), 1.06 (s, 3H,
18-CH.sub.3), 0.82 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (d, 3H,
16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 531 (MNa.sup.+), 1039
(2MNa.sup.+).
Step D.
14-O-{[(1R,2R,5S)-2-Hydroxy-5-methanesulfonyloxy-cyclohexylsulfany-
l]-acetyl}-mutilin+(1S,3S,5R) diastereomer
[0134] To a solution of
14-O-{[(1R,2R,5S)-2,5-dihydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2-
S,5R) diastereomer (6.07 g, 11.9 mmol) in 36 ml of pyridine was
added methanesulfonyl chloride (1.1 ml, 14.3 mmol) and the
resulting mixture was stirred overnight at room temperature.
Subsequently the solvent was evaporated under reduced pressure; the
residue was diluted with 1N HCl and extracted three times with
ethyl acetate. The combined organic layers were washed with brine,
dried over sodium sulfate and filtered. The filtrate was
concentrated and purified by column chromatography (silica,
cyclohexane/ethyl acetate=1/1) to yield
14-O-{[(1R,2R,5S)-2-hydroxy-5-methanesulfonyloxy-cyclohexylsulfanyl]-acet-
yl}-mutilin+(1S,2S,5R) diastereomer (R.sub.f=0.15, 2.55 g, 36%
yield) as colorless amorphous foam.
[0135] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.12 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.54 (d, 1H, 14-H,
J=8 Hz), 5.07 (m, 2H, 20-H), 5.00 (t, 1H, 2'-OH, J=5 Hz), 4.78 (m,
1H, 5'-H), 4.50 (d, 1H, 11-OH, J=6 Hz), 3.55-3.25 (m, 4H, 2'-H,
11-H, 22-H), 2.91 (m, 1H, 1'-H), 2.38 (bs, 1H, 4-H). 1.36 (s, 3H,
15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3), 0.80 (d, 3H, 17-CH.sub.3,
J=7 Hz), 0.63 (d, 3H, 16-CH.sub.3, J=7 Hz).
Step E.
14-O-{[(1R,2R,5R)-5-Azido-2-hydroxy-5-methanesulfonyloxy-cyclohexy-
lsulfanyl]-acetyl}-mutilin+(1S,2S,5S) diastereomer
[0136] A solution of
14-O-{[(1R,2R,5S)-2-hydroxy-5-methanesulfonyloxy-cyclohexylsulfanyl]-acet-
yl}-mutilin+(1S,2S,5R) diastereomer (2.55 g, 4.35 mmol) and sodium
azide (0.85 g, 13 mmol) in 30 ml of dimethylformamide was heated
for 6 hours at 80.degree. C. The reaction mixture was diluted with
water and brine and extracted three times with ethyl acetate. The
combined organic layers were washed with water and brine, dried
over sodium sulfate and filtered. The solvent was removed under
reduced pressure and crude
14-O-{[(1R,2R,5R)-5-azido-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(-
1S,2S,5S) diastereomer (quantitative yield, cyclohexane/ethyl
acetate=1/1, R.sub.f=0.35) was obtained as amorphous foam which was
directly used for the next step.
[0137] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.15, 6.13 (2dd, H, 19-H, J=11 Hz and 18 Hz), 5.56, 5.54
(2d, 1H, 14-H, J=8 Hz), 5.05 (m, 2H, 20-H), 4.90 (d, 1H, 2'-OH, J=5
Hz), 4.50, 4.49 (2d, 1H, 11-OH, J=6 Hz), 3.50-3.25 (m, 5H, 2'-H,
5'-H, 11-H, 22-H), 2.64 (m, 1H, 1'-H), 2.40 (bs, 1H, 4-H), 1.36 (s,
3H, 15-CH.sub.3, 1.06 (s, 3H, 18-CH). 0.81 (d, 3H, 17-CH.sub.3, J=7
Hz), 0.63 (d, 3H, 16-CH.sub.3, J=7 Hz).
Step F.
14-O-{[(1R,2R,5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin+(1S,2S,5S) diastereomer
[0138] Triphenylphosphine (1.18 g, 4.50 mmol) was added to a
solution of
14-O-{[(1R,2R,5R)-5-azido-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(-
1S,2S,5S) diastereomer (2.4 & 4.50 mmol) in 30 ml of
tetrahydrofuran and stirred overnight at room temperature.
Subsequently water (approx. 3 ml) was added and the reaction
mixture was heated for 1 hour at reflux. After evaporation of the
solvent the residue was diluted with water and brine and extracted
three times with ethyl acetate. The combined organic layers were
dried over sodium sulfate, filtered and subjected to chromatography
(silica, ethyl acetate/methanol/35% ammonia solution=100/100/1) to
yield
14-O-{[(1R,2R,5R)-5-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(-
1S,2S,5S) diastereomer (R.sub.f=0.3, 1.74 g, 79% yield) as
colorless amorphous foam.
[0139] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.25, 6.65 (2bs, 1H, NH), 6.14 (dd, 1H, 19-H, J=11 Hz and 18
Hz), 5.54 (d, 1H, 14-H, J=8 Hz), 5.04 (m, 2H, 20-H), 4.50 (bs, 1H,
11-OH), 3.55-3.10 (m, 5H, 2'-H, 5'-H, 11-H, 22-H), 2.58 (m, 1H,
1'-H), 2.40 (bs, 1H, 4-H), 1.36 (s, 3H, 15-CH.sub.3), 1.05 (s, 3H,
18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.62 (d, 3H,
16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 508 (MH.sup.+), 530
(MNa.sup.+), 1037 (2MNa.sup.+).
Example 4A
14-O-{[(1R,2R,5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
and
14-O-{[(1S,2S,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutil-
in
[0140] The mixture of
14-O-{[(1R,2R,5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(-
1S,2S,5S) diastereomer (4 g, 7.87 mmol) from Example 4 Step F was
separated on a chiral column (250.times.20 mm CHIRALPAK IC,
n-heptane/tetrahydrofuran/diethylamine=70/30/0.1) to yield
14-O-{[(1S*,2S*,5S*)-5-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutili-
n (late eluting compound, 1.1 g, 28% yield, uncorrected) as
colorless amorphous foam.
[0141] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.13 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.54 (d, 1H, 14-H,
J=8 Hz), 5.06 (m, 2H, 20-H), 5.76 (d, 1H, 2'-OH, J=7 Hz), 4.50 (d,
1H, 11-OH, J=7 Hz), 3.55-3.15 (m, 5H, 2'-H, 11-H, 5'-H, 22-H), 2.48
(m, 1H, 1'-H), 2.40 (bs, 1H, 4-H), 1.36 (s, 3H, 15-CH.sub.3), 1.05
(s, 3H, 18-CH.sub.3). 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.62 (d,
3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 508 (MH.sup.+), 1037
(2MNa.sup.+), 506 (M-H).sup.-, 542 (MCl.sup.-).
Example 5
14-O-{[(1R,2R,3R)-3-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride+(1S,2S,35) diastereomer hydrochloride
Step A.
14-O-{[(1R,2R,3R)-3-tert-Butoxycarbonylamino-2-hydroxy-cyclohexyls-
ulfanyl]-acetyl}-mutilin+(1S,2S,3S) diastereomer
[0142] (cis)-2,3-Epoxycyclohexyl-carbamic acid tert-butyl ester
(O'Brien, P.; Childs, A., C.; Ensor, G. Organic Letters 2003, (26),
4955-4957.) (1 g, 4.69 mmol) was treated with pleuromutilin thiol
according to the method of Example 1 Step A1. After work up and
chromatography of the reaction mixture (silica, cyclohexane/ethyl
acetate=1/1)
14-O-{[(1R,2R,3R)-3-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,3S) diastereomer (R.sub.f=0.5, 1.32 g,
46%) was obtained as colorless amorphous foam.
[0143] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.12 (m, 2H, NH, 19-Hz), 5.54 (d, 1H, 14-H, J=8 Hz), 5.05
(m, 2H, 20-H), 4.96 (d, 1H, 2'-OH, J=4 Hz), 4.50, 4.99 (2d, 1H,
11-OH, J=6 Hz)), 3.65 (m, 1H, 2'-H), 3.57 (m, 1H, 3'-H), 3.42 (t,
1H, 11-H, J=6 Hz), AB-system (.nu..sub.A=3.30, 3.29,
.nu..sub.B=3.23, 3.22, 22-H, J=15 Hz), 3.06 (m, 1H, 1'-H), 2.40
(bs, 1H, 4-H), 1.36 (s, 12H, tert-butyl, 15-CH.sub.3), 1.05 (s, 3H,
18-CH.sub.3), 0.82 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (d, 3H,
16-CH.sub.3, J=6 Hz).
Step B.
14-O-{[(1R,2R,3R)-3-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin+(1S,2S,3S) diastereomer
[0144]
14-O-{[(R,2R,3R)-3-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsuf-
anyl]-acetyl}-mutilin+(1S,2S,3S) diastereomer (400 mg, 0.658 mmol)
was treated according to the method of Example 1 Step B. After work
up and chromatography of the reaction mixture (silica, ethyl
acetate/methanol=1/5)
14-O-{[(1R,2R,3R)-3-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(-
1S,2S,3S) diastereomer (R.sub.f=0.1, 249 mg, 75%) was obtained as
colorless amorphous foam.
[0145] MS-ESI (m/z): 508 (MH.sup.+), 530 (MNa.sup.+), 1015
(2MH.sup.+), 1037 (2MNa.sup.+).
Step C.
14-O-{[(1R,2R,3R)-3-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin hydrochloride+(1S,2S,4S) diastereomer hydrochloride
[0146]
14-O-{[(1R,2R,3R)-3-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mut-
ilin+(1S,2S,3S) diastereomer (249 mg, 0.49 mmol) was treated
according to the method of Example 1 Step C to obtain
14-O-{[(1R,2R,3R)-3-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride+(1S,2S,3S) diastereomer hydrochloride (247 mg, 93%
yield) as colorless amorphous solid.
[0147] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.8 (bs, 3H, NH.sub.3.sup.+), 6.13 (d, 2H, 19-Hz, J=11 Hz
and 18 Hz), 5.80 (d, 1H, 2'-OH, J=4 Hz), 5.55 (d, 1H, 14-H, J=8
Hz), 5.05 (m, 2H, 20-H), 4.55, 4.54 (2d, 1H, 11-OH, J=6 Hz)), 3.87
(m, 1H, 2'-H), 3.42 (t, 1H, 11-H, J=6 Hz), AB-system
(.nu..sub.A=3.35, .nu..sub.B=3.24, 22-H, J=15 Hz), 3.20, 3.13 (2m,
1H, 3'-H, 1'-H), 2.40 (bs, 1H, 4-H), 1.36 (s, 3H, 15-CH.sub.3),
1.05 (s, 3H, 18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63
(d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 508 (MH.sup.+), 1015
(2MH.sup.+), 542 (MCl.sup.-).
Example 6
14-O-{[(1R,2R,4R)-4-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mut-
ilin+(1S,2S,4S) diastereomer and
14-O-{[(1R,2R,4R)-4-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S,2S,4S) diastereomer
[0148] To a solution of
14-O-{[(1R,2R,4R)-4-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(-
1S,2S,4S) diastereomer (900 mg, 1.77 mmol) from Example 1 Step B in
10 ml dichloromethane was added acetaldehyde (2.77 ml, 1M in
dichloromethane) and acetic acid (77 .mu.l, 1.77 mmol) and stirred
for 30 minutes at room temperature. The resulting reaction mixture
was treated with sodium triacetoxyborohydride (750 mg, 3.54 mmol)
and stirred overnight at room temperature, diluted with
dichloromethane and subsequently extracted with NaHCO.sub.3
solution and brine. The organic layer was dried over sodium sulfate
and filtered. The filtrate was subjected to chromatography (silica,
ethyl acetate/methanol/35% ammonia solution=50/50/1) to yield
14-O-{[(1R,2R,4R)-4-diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin+(1S,2S,4S) diastereomer (a) (92 mg, 9% yield) and
14-O-{[(1R,2R,4R)-4-ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S,2S,4S) diastereomer (b) (163 mg, 17% yield) as colorless
amorphous foams.
[0149] (a): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.14 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.55 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 2H, 20-H), 4.83 (d, 1H, 2'-OH, J=4 Hz), 4.47 (d,
1H, 11-OH, J=6 Hz), 3.42 (m, 1H, 11-H), AB-system
(.sigma..sub.A=3.50, 3.42, .nu..sub.B=330, 3.27, 22-H, J=15 Hz),
3.25 (m, 1H, 2'-H), 2.50 (m, 2H, 1'-H, 4'-H), 2.40 (m, 5H,
NCH.sub.2, 4-H), 1.36 (s, 3H, 15-CH.sub.3), 1.05 (s, 3H,
18-CH.sub.3), 0.92 (t, 6H, NCH.sub.2CH.sub.3, J=7 Hz), 0.81 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.63 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI
(m/z): 564 (MH.sup.+), 586 (MNa.sup.+), 562 (M-H).sup.-.
[0150] (b): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.13 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.54 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 2H, 20-H), 4.48 (d, 1H, 11-OH, J=6 Hz), 3.42 (m,
1H, 11-H), AB-system (.nu..sub.A=3.48, .nu..sub.B3.25, 22-H, J=15
Hz), 2.55 (m, 2H, 1'-H, 4'-H), 2.40 (bs, 1H, 4-H), 1.35 (s, 31H,
15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3), 0.95 (t, 3H,
NCH.sub.2CH.sub.3, J=7 Hz), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63
(d, 3H, 16-CH.sub.3, J=6 Hz). MS-ESI (m/z): 536 (MH.sup.+), 558
(MNa.sup.+), 534 (M-H).sup.-.
Example 7
14-O-{[(1R,2R,5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutil-
in hydrochloride+(1S,2S,5R) diastereomer hydrochloride
Step A. N-Ethyl-N-(cyclohex-3-enyl)-carbamic acid tert-butyl
ester
[0151] To a solution of cyclohex-3-enyl-carbamic acid tert-butyl
ester (Kampferer, P.; Vasella, A. Helvetica Chimica Acta 2004, 87,
2764-2789) (4.34 g, 22 mmol) in 20 ml of DMSO was added sodium
hydride (880 mg, 60% dispersion, 22 mmol) and after one hour of
stirring ethyl iodide (1.78 ml, 22 mmol). After further stirring
for 2 hours at room temperature the reaction mixture was
concentrated under reduced pressure. The residue was diluted with
water and brine and extracted three times with ethyl acetate. The
organic layers were dried over sodium sulfate and filtered. The
filtrate was concentrated under reduced pressure and after
chromatography (silica, cyclohexane/ethyl acetate=12/1) the title
compound (R.sub.f=0.30, 2.88 g. 58% yield) was obtained as
colorless solid.
[0152] .sup.1H NMR (400 MHz, CDCl.sub.3, .delta., ppm): 5.61 (m,
2H, double bond), 4.08 (bs, 1H, NCH), 3.15 (m, 2H, NCH.sub.2),
2.15, 1.75 (2m, 6H), 1.47 (s, 9H, tert-butyl), 1.13 (t, 3H,
NCH.sub.2CH.sub.3, J=7 Hz).
Step B. N-ethyl-N-(cis-3,4-epoxycyclohexyl)-carbamic acid
tert-butyl ester
[0153] N-Ethyl-N-(cyclohex-3-enyl)-carbamic acid tert-butyl ester
(2.87 g, 12.7 mmol) was dissolved in 75 ml of dichloromethane and
treated with 3-chloroperbenzoic acid (4.50 g, 70%, 19 mmol). After
stirring at room temperature for 20 hours the reaction mixture was
concentrated under reduced pressure. The residue was diluted with
ethyl acetate and subsequently extracted with 10% NaHSO.sub.3
solution, saturated NaHCO.sub.3 solution and brine. The organic
layer was dried over sodium sulfate and filtered. The filtrate was
concentrated under reduced pressure and after chromatography
(silica, cyclohexane/dioxane=5/1) the title compound (R.sub.f=0.2,
1.50 g, 49% yield) was obtained.
[0154] .sup.1H NMR (400 MHz, CDCl.sub.3, .delta., ppm): 4.0 (bs,
1H, NCH), 3.14 (m, 2H, NCH.sub.2), 3.06 (bs, 2H, epoxide), 2.13,
2.08, 1.88, 1.60, 1.36 (4m, 6H), 1.47 (s, 9H, tert-butyl), 0.08 (t,
3H, NCH.sub.2CH.sub.3).
Step C.
14-O-{[(R,2R,5S)-5-(tert-Butoxycarbonyl-ethyl-amino)-2-hydroxy-cyc-
lohexylsulfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer
[0155] N-Ethyl-N-(cis-3,4-epoxycyclohexyl)-carbamic acid tert-butyl
ester (1.5 g, 6.2 mmol) was treated with pleuromutilin thiol
according to the method of Example 1 Step A1. After work up and
chromatography of the reaction mixture (silica,
cyclohexane/dioxane=3/1)
14-O-{[(1R,2R,5S)-5-(tert-butoxycarbonyl-ethyl-amino)-2-hydroxy-cyclohexy-
lsulfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer (R.sub.f=0.4,
2.57 & 65% yield) was obtained as colorless amorphous foam.
[0156] MS-ESI (m/z): 536 (MH.sup.+), 558 (MNa.sup.+), 534
(M-H).sup.-.
Step D.
14-O-{[(1R,2R,5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-mutilin+(1S,2S,5R) diastereomer
[0157]
14-O-{[(1R,2R,5S)-5-(tert-Butoxycarbonyl)-ethyl-amino)-2-hydroxy-cy-
clohexylsulfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer (2.57 g,
4.04 mmol) was treated according to the method of Example 1 Step B.
After work up and chromatography of the reaction mixture (silica,
ethyl acetate/methanol/35% ammonia solution=100/100/1)
14-O-{[(1R,2R,5S)-5-ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S,2S,5R) diastereomer (R.sub.f=0.3, 1.08 g. 50%) was obtained
as colorless amorphous foam.
[0158] .sup.1H NMR (500 MHz, CDCl.sub.3, .delta., ppm, inter alia):
6.48 (dd, 1H, 19-H, J=100 Hz and 18 Hz), 5.77 (m, 1H, 14-H), 5.36
(m, 1H, 20-H), 5.22 (d, 1H, 20-H, J=17 Hz), 3.45 (d, 1H, 2'-H),
3.37 (d, 1H, 11-H, J=6 Hz), 3.25 (m, 1H, 22-H), 2.97 (m, 1H, 1'-H),
2.91 (m, 1H, 5'-H), 2.63 (q, 2H, NCH.sub.2, J=7 Hz), 2.10 (bs, 1H,
4-Hl), 1.46 (s, 3H, 15-CH.sub.3), 1.18 (s, 3H, 18-CH.sub.3), 1.12
(t, 3H, NCH.sub.2CH.sub.3, J=7 Hz), 0.98 (d, 3H, 17-CH.sub.3, J=7
Hz), 0.73 (d, 3H, 16-CH.sub.3, J=7 Hz).
Step E.
14-O-{[(1R,2R,5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-mutilin hydrochloride+(1S,2S,5R) diastereomer hydrochloride
[0159]
14-O-{[(1R,2R,5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl-
}-mutilin+(1S,2S,5R) diastereomer (86 mg, 0.16 mmol) was treated
according to the method of Example 1 Step C to obtain
14-O-{[(1R,2R,5S)-5-ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin hydrochloride+(1S,2S,5R) diastereomer hydrochloride (83 mg, 90%
yield) as colorless amorphous solid.
[0160] .sup.1H NMR (400 MHz, CDCl.sub.3, .delta., ppm, inter alia):
9.3 (bs, 2H, NH.sub.2.sup.+), 6.45 (m, 1H, 19-H), 5.73 (d, 1H,
14-H, J=10 Hz), 5.35 (m, 1H, 20-H), 5.22 (d, 1H, 22-H, J=18H), 3.85
(m, 1H, 2'-H), 3.33 (m, 3H, 11-H, 22-H), 3.07 (m, 2H, NCH.sub.2),
2.10 (bs, 1H, 4-H), 1.50 (t, 3H, NCH.sub.2CH.sub.3, J=7 Hz), 1.45
(s, 3H, 15-CH.sub.3), 1.18 (s, 3H, 18-CH.sub.3), 0.90 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.73 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI
(m/z): 536 (MH.sup.+), 570 (MCl.sup.-).
Example 8
14-O-{[(1R,2R,5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mut-
ilin hydrochloride+(1S,2S,5R) diastereomer hydrochloride
Step A.
14-O-{[(1R,2R,5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-ace-
tyl}-mutilin+(1S,2S,5R) diastereomer
[0161]
14-O-{[(1R,2R,5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl-
}-mutilin+(1S,2S,5R) diastereomer (250 mg, 0.47 mmol) from Example
7 Step D was treated with acetaldehyde (53 .mu.l, 0.93 mmol)
according to the method of Example 6. After work up and
chromatography of the reaction mixture (silica, ethyl
acetate/methanol=1/1)
14-O-{[(1R,2R,5S)-5-diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin+(1S,2S,5R) diastereomer (R.sub.f=0.2, 230 mg, 87%) was
obtained as colorless amorphous foam.
[0162] .sup.1H NMR (400 MHz, CDCl.sub.3, .delta., ppm, inter alia):
6.48 (dd, 1H, 19-H, J=11 Hz and 17 Hz), 5.77 (d, 1H, 14-H, J=8 Hz),
5.36 (m, 1H, 20-H), 5.22 (d, 1H, 20-H, J=17 Hz), 3.57, 3.36, 3.21,
3.03, 2.72 (5m, 6H, 1'-H, 2'-H, 5'-H, 11-H, 22-H), 2.59 (m, 4H,
NCH.sub.2), 2.11 (bs, 1H, 4-H), 1.46 (s, 3H, 15-CH.sub.3), 1.18 (s,
3H, 18-CH.sub.3), 0.98 (t, 6H, NCH.sub.2CH.sub.3, J=7 Hz), 0.88 (d,
3H, 17-CH.sub.3, J=7 Hz), 0.73 (d, 3H, 16-CH.sub.3, J=7 Hz).
Step B.
14-O-{[(1R,2R,5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-ace-
tyl}-mutilin hydrochloride+(1S,2S,5R) diastereomer
hydrochloride
[0163]
14-O-{[(1R,2R,5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acet-
yl}-mutilin+(1S,2S,5R) diastereomer (230 mg, 0.41 mmol) was treated
according to the method of Example 1 Step C to obtain
14-O-{[(1R,2R,5S)-5-diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin hydrochloride+(1S,2S,5R) diastereomer hydrochloride (223 mg,
91% yield) as colorless amorphous solid.
[0164] .sup.1H NMR (400 MHz, CDCl.sub.3, .delta., ppm, inter alia):
11.5 (bs, 3H, NH.sup.+), 6.46 (dd, 1H, 19-H, J=11 Hz and 17 Hz),
5.74 (d, 1H, 14-H, J=8 Hz), 5.34 (m, 1H, 20-H), 5.22 (d, 1H, 22-H,
J=17H), 3.98 (m, 1H, 2'-H), 3.60-2.90 (m, 9H, 1'-H, 5'-H, 11-H,
22-H, NCH.sub.2), 2.10 (bs, 1H, 4-H), 1.48 (m, 91,
NCH.sub.2CH.sub.3, 15-CH.sub.3), 1.18 (s, 3H, 18-CH.sub.3), 0.89
(d, 3H, 17-CH.sub.3, J=7 Hz), 0.73 (d, 3H, 16-CH.sub.3, J=7 Hz).
MS-ESI (m/z): 564 (MH.sup.+), 586 (MNa.sup.+), 1149 (2MNa.sup.+),
598 (MCl.sup.-).
Example 9
14-O-{[(1R,2R,4S)-4-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mut-
ilin+(1S,2S,4R) diastereomer
[0165]
14-O-{[(1R,2R,4S)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mut-
ilin+(1S,2S,4R) diastereomer (680 mg, 1.34 mmol) from Example 2
Step A was treated according to the method of Example 6. After work
up and chromatography of the reaction mixture (silica, ethyl
acetate/methanol=1/1)
14-O-{[(1R,2R,4S)-4-diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin+(1S,2S,4R) diastereomer (R.sub.f=0.2, 129 mg, 17%) was
obtained as colorless amorphous foam.
[0166] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia) 6.13 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.54 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 2H, 20-H), 4.70 (d, 1H, 2'-OH), 4.49 (d, 1H,
11-OH, J=6 Hz), 3.70 (m, 1H, 2'-H), 3.42 (t, 1H, 11-H, J=6 Hz),
AB-system (.nu..sub.A=3.36, .nu..sub.B=3.22, 22-H, J=15 Hz), 2.72
(m, 2H, 1'-H, 4'-H), 2.47 (m, 2H, NCH.sub.2), 2.40 (bs, 1H, 4-H),
1.35 (s, 3H, 15-CH.sub.3), 1.04 (s, 3H, 18-CH.sub.3), 0.95 (t, 3H,
NCH.sub.2CH.sub.3, J=7 Hz), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.62
(d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 536 (MH.sup.+), 558
(MNa.sup.+), 534 (M-H).sup.-.
Example 10
14-O-{[(1R,2R,5R)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mut-
ilin+(1S,2S,5S) diastereomer
[0167]
14-O-{[(1R,2R,5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mut-
ilin+(1S,2S,5S) diastereomer (420 mg, 0.827 mmol) from Example 4
Step F was treated according to the method of Example 6. After work
up and chromatography of the reaction mixture (silica, ethyl
acetate/methanol/35% ammonia solution=50/50/1)
14-O-{[(1R,2R,5R)-5-diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin+(1S,2S,5S) diastereomer (R.sub.f=0.2, 95 mg, 20%) was
obtained as colorless amorphous foam.
[0168] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.13 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.55 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 2H, 20-H), 4.79 (m, 1H, 2'-OH), 3.55-3.15 (m,
2'-H, 5'-H, 11-H, 22-H), 2.58 (m, 1H, 1'-H), 2.40 (m, 5H,
NCH.sub.2, 4-H), 1.37 (s, 3H, 15-CH.sub.3), 1.06 (s, 3H,
18-CH.sub.3), 0.92 (t, 3H, NCH.sub.2CH.sub.3, J=7 Hz), 0.83 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.63 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI
(m/z): 564 (MH.sup.+), 586 (MNa.sup.+), 562 (M-H).sup.-, 598
(MCl.sup.-).
Example 11
14-O-{[(1R,2R,3R)-3-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutil-
in+(1S,2S,3S) diastereomer
Step A.
[(1R,2R,3R)-2-Hydroxy-3-(2,4,6-trimethyl-benzylsulfanyl)-cyclohexy-
l]-carbamic acid tert-butyl ester+(1S,2S,3S) diastereomer
[0169] To a solution of (cis)-2,3-epoxycyclohexyl-carbamic acid
tert-butyl ether (O'Brien, P.; Childs, A., C.; Ensor, G. Organic
Letters 2003, 5(26), 4955-4957.) (14.9 g, 68.9 mmol) and
2,4,6-trimethylbenzyl mercaptan (11.5 g, 68.9 mmol) in 50 ml of
methanol was added 10N NaOH (5 ml, 50 mmol) and the resulting
mixture was stirred for 16 hours at room temperature. The reaction
mixture was diluted with water and brine and extracted with ethyl
acetate three times. The organic layers were dried over sodium
sulfate and filtered. The filtrate was subjected to chromatography
(silica, cyclohexane/ethyl acetate=5/1) to yield
[(1R,2R,3R)-2-hydroxy-3-(2,4,6-trimethyl-benzylsulfanyl)-cyclohexyl]-carb-
amic acid tert-butyl ester+(1S,2S,3S) diastereomer (R.sub.f=0.25;
5.92 g, 23% yield) as colorless amorphous foam.
[0170] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm): 6.78 (s,
2H, aromat.-H), 6.15 (bd, OH), 4.95 (bd, NH), 3.75 (d, 1H,
SCH.sub.2), 3.68 (m, 2H), 3.02 (m, 1H, SCH), 2.30 (s, 9H,
CH.sub.3), 2.30, 1.90, 1.40 (3m, 6H), 1.35 (s, 9H, tert-butyl).
Step B.
[(1R,2R,3R)-2-(tert-Butyl-dimethyl-silanyloxy)-3-(2,4,6-trimethyl--
benzylsulfanyl)-cyclohexyl]-carbamic acid tert-butyl
ester+(1S,2S,3S) diastereomer
[0171] A solution of
[(1R,2R,3R)-2-hydroxy-3-(2,4,6-trimethyl-benzylsulfanyl)-cyclohexyl]-carb-
amic acid tert-butyl ester+(1S,2S,3S) diastereomer (2.46 & 6.49
mmol) in 50 ml of dimethylformamide was treated with
tert-butyldimethylsilyl chloride (978 mg, 6.49 mmol) and imidazole
(552 mg, 8.11 mmol) and stirred at 80.degree. C. for 5 days. The
reaction mixture was concentrated under reduced pressure. The
residue was diluted with 0.1 N HCl and extracted three times with
ethyl acetate. The combined organic layers were washed with brine,
dried over sodium sulfate and filtered. After chromatography
(silica, cyclohexane/ethyl acetate=10/1)
[(1R,2R,3R)-2-(tert-butyl-dimethyl-silanyloxy)-3(2,4,6-trimethyl-benzylsu-
lfanyl)-cyclo-hexyl]-carbamic acid tert-butyl ester+(1S,2S,3S)
diastereomer (R.sub.f=0.25, 3.0 g 94% yield) was obtained.
[0172] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm): 6.80 (s,
2H, aromat-H), 6.20 (bd, NH), 3.90, 3.75, 3.63 (3m, 4H, NCH, OCH,
SCH.sub.2), 2.98 (m, 1H, SCH), 2.30 (s, 9H, CH.sub.3), 1.90, 1.50,
1.33 (3m, 6H), 1.35 (s, 9H, C-tert-butyl), 0.85 (s, 9H,
Si-tert-butyl), 0.0 (s, 6H, Si(CH.sub.3).sub.2).
Step C.
[(1R,2R,3R)-2-(tert-Butyl-dimethyl-silanyloxy)-3-(2,4,6-trimethyl--
benzylsulfanyl)-cyclohexyl]-ethyl-carbamic acid tert-butyl
ester+(1S,2S,3S) diastereomer
[0173] A solution of
[(1R,2R,3R)-2-(tert-butyl-dimethyl-silanyloxy)-3-(2,4,6-trimethyl-benzyls-
ulfanyl)-cyclohexyl]-carbamic acid tert-butyl ester+(1S,2S,3S)
diastereomer (3.0 &g, 6.08 mmol) was treated with ethyl iodide
according to the method of Example 7 Step A. After work up and
chromatography of the reaction mixture (silica, cyclohexane/ethyl
acetate=3/1)
[(1R,2R,3R)-2-(tert-butyl-dimethyl-silanyloxy)-3-(2,4,6-trimethyl-benzyls-
ulfanyl)-cyclohexyl]-ethyl-carbamic acid tert-butyl
ester+(1S,2S,3S) diastereomer (1.20 g, 38%) was obtained.
[0174] MS-ESI (m/z): 544 (MNa.sup.+), 1065 (2MNa.sup.+).
Step D.
[(1R,2R,3R)-2-(tert-Butyl-dimethyl-silanyloxy)-3-mercapto-cyclohex-
yl]-ethyl-carbamic acid tert-butyl ester+(1S,2S,3S)
diastereomer
[0175] A solution of
[(1R,2R,3R)-2-(tert-butyl-dimethyl-silanyloxy)-3-(2,4,6-trimethyl-benzyls-
ulfanyl)-cyclohexyl]-ethyl-carbamic acid tert-butyl
ester+(1S,2S,3S) diastereomer (1.20 g. 2.30 mmol) in 10 ml of
tetrahydrofuran and 20 ml of liquid ammonia was treated at
-78.degree. C. under an argon atmosphere with sodium (106 mg, 4.60
mmol) and stirred at -78.degree. C. for one hour. Then solid
ammonium chloride was added and the reaction mixture was warmed to
room temperature, diluted with tetrahydrofuran and flushed with
nitrogen. The residual mixture was filtered and concentrated under
reduced pressure to yield crude
[(R,2R,3R)-2-(tert-butyl-dimethyl-silanyloxy)-3-mercapto-cyclohexyl]-ethy-
l-carbamic acid tert-butyl ester+(1S,2S,3S) diastereomer
(quantitative yield) which was directly used for the next step.
MS-ESI (m/z): 412 (MNa.sup.+), 801 (2MNa.sup.+).
Step E
14-O-{[(1R,2R,3R)-3-(tert-Butoxycarbonyl-ethyl-amino)-2-(tert-butyl-
-dimethyl-silanyloxy)-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,3S)
diastereomer
[0176] Crude
[(1R,2R,3R)-2-(tert-butyl-dimethyl-silanyloxy)-3-mercapto-cyclohexyl]-eth-
yl-carbamic acid tert-butyl ester+(18S,2S,3S) diastereomer (895 mg,
2.30 mmol) was dissolved in 30 ml of tetrahydrofuran and treated
subsequently with pleuromutilin tosylate (979 mg, 1.84 mmol) and
potassium tert-butoxide (206 mg, 1.84 mmol) and the resulting
mixture was stirred for 16 hours at room temperature. After
evaporation of the solvent the residue was diluted with 1N HCl and
extracted three times with ethyl acetate. The combined organic
layers were washed with NaHCO.sub.3 solution and brine, dried over
sodium sulfate and filtered. After chromatography (silica,
cyclohexane/ethyl acetate=10/1)
14-O-{[(1R,2R,3R)-3-(tert-butoxycarbonyl-ethyl-amino-2-(tert-butyl-dimeth-
yl-silanyloxy)-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,3S)
diastereomer (R.sub.f=0.5, 468 mg, 27% yield) was obtained.
[0177] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.14 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.55 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 2H, 20-H), 4.49 (m, 1H, 11-OH), 3.94 (m, 1H,
2'-H), 3.43 (t, 1H, 11-H, J=6 Hz), 3.28, 3.04 (2m, 5H, 1'-H, 22-H,
NCH.sub.2), 2.40 (bs, 1H, 4-H), 1.40 (s, 9H, tert-butyl), 1.36,
1.35 (2s, 3H, 15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3), 0.87 (s, 9H,
Si-tert-butyl), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.64, 0.62 (2d,
3H, 16-CH.sub.3, J=7 Hz) 0.05, -0.05 (2s, 6H,
Si(CH.sub.3).sub.2).
Step F.
14-O-{[(1R,2R,3R)-3-Ethylamino-2-(tert-butyl-dimethyl-silanyloxy)--
cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,3S) diastereomer and
14-O-{[(1R,2R,3R)-3-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S,2S,3S) diastereomer
[0178]
14-O-{[(1R,2R,3R)-3-(tert-Butoxycarbonyl-ethyl-amino)-2-(tert-butyl-
-dimethyl-silanyloxy)-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,3S)
diastereomer (468 mg, 0.624 mmol) was treated with trifluoroacetic
acid overnight according to the method of Example 1 Step B. After
work up and chromatography of the reaction mixture (silica, ethyl
acetate/methanol=1/2)
14-O-{[(1R,2R,3R)-3-ethylamino-2-(tert-butyl-dimethyl-silanyloxy)-cyclohe-
xylsulfanyl]-acetyl}-mutilin+(1S,2S,3S) diastereomer (a)
(R.sub.f=0.6, 144 mg, 36% yield) and
14-O-{[(1R,2R,3R)-3-ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S,2S,3S) diastereomer (b) (R.sub.f=0.25, 177 mg, 53% yield)
were obtained as colorless solids.
[0179] (a): MS-ESI (m/z): 672 (MNa.sup.+), 1321 (2MNa.sup.+).
[0180] (b): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.14 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.54 (d, 1H, 14-H,
3-8 Hz), 5.05 (m, 2H, 20-H), 4.76 (m, 1H, 2'-OH), 4.49, 4.48 (2d,
1H, 11-OH, J=6 Hz), 3.55 (m, 1H, 2'-H), 3.42 (t, 1H, 11-H, 3=6 Hz),
AB-system (.nu..sub.A=3.37, .nu..sub.B=3.18, 22-H, J=15 Hz), 3.05
(m, 1H, 3'-H), 2.66 (m, 1H, 1'-H), 2.50 (m, 2H, NCH.sub.2), 2.40
(bs. 1H, 4-H), 1.36 (s, 3H, 15-CH.sub.3), 1.05 (s, 3H,
18-CH.sub.3), 0.98 (t, 3H, NCH.sub.2CH.sub.3, J=7 Hz), 0.81 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.62 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI
(m/z): 536 (MH.sup.+), 558 (MNa.sup.+), 1071 (2MH.sup.+), 1093
(2MNa.sup.+), 534 (M-H).sup.-.
Example 12
14-O-{[(1R,2R,3R)-3-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mut-
ilin+(18, 28, 35) diastereomer
Step A.
14-O-{[(1R,2R,3R)-3-Diethylamino)-(tert-butyl-dimethyl-silanyloxy)-
-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,3S) diastereomer
[0181]
14-O-{[(1R,2R,3R)-3-Ethylamino-2-(tert-butyl-dimethyl-silanyloxy)-c-
yclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,3S) diastereomer (144 mg,
0.222 mmol) from Example 11 Step F was treated with acetaldehyde
(25 .mu.l, 0.444 mmol) according to the method of Example 6. After
work up and chromatography of the reaction mixture (silica, ethyl
acetate/methanol=2/1)
14-O-{[(1R,2R,3R)-3-diethylamino)-2-(tert-butyldimethyl-silanyloxy)-cyclo-
hexylsulfanyl]-acetyl}-mutilin+(1S,2S,3S) diastereomer
(R.sub.f=0.5, 110 mg, 73%) was obtained as colorless amorphous
foam.
[0182] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.14 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.53 (d, 1H, 14-H,
J=7 Hz), 5.05 (m, 2H, 20-1H), 4.98 (m, 1H, 11-OH), 3.97 (m, 1H,
2'-H), 3.42 (t, 1H, 11-H, J=6 Hz), 3.24 (m, 2H, H-22), 3.00 (m, 1H,
1'-H), 2.70 (m, 1H, 3'-H), 2.55 (m, 4H, NCH.sub.2), 2.40 (be, 1H,
4-H), 1.36 (s, 3H, 15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3), 0.87
(m, 15H, NCH.sub.2CH.sub.3, Si-tert-butyl), 0.81 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.62, 0.60 (2d, 3H, 16-CH.sub.3, J=7 Hz),
0.07 (s, 6H, Si(CH.sub.3).sub.2).
Step B.
14-O-({[(1R,2R,3R)-3-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-ac-
etyl}-mutilin+(1S,2S,3S) diastereomer
[0183] To a solution of
14-O-{[(1R,2R,3R)-3-diethylamino-2-(tert-butyl-dimethyl-silanyloxy)-cyclo-
hexylsulfanyl]-acetyl}-mutilin+(1S,2S,3S) diastereomer (101 mg,
0.149 mmol) in 5 ml of tetrahydrofuran was added tetrabutylammonium
fluoride (0.44 ml, 1M in tetrahydrofuran, 0.447 mmol). After
stirring at room temperature for 2 days the reaction mixture was
concentrated under reduced pressure, diluted with NaHCO.sub.3
solution and extracted two times with ethyl acetate. The combined
organic layers were dried over sodium sulfate and filtered. The
filtrate was subjected to chromatography (silica, ethyl
acetate/methanol=1/2) to obtain
14-O-{[(1R,2R,3R)-3-diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin+(1S,2S,3S) diastereomer (R.sub.f=0.2, 8 mg, 10% yield) as
colorless amorphous foam.
[0184] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.15, 6.14 (2dd, 1H, 19-H, J=1 Hz and 18 Hz), 5.54 (d, 1H,
14-H, J=8 Hz), 5.05 (m, 2H, 20-H), 4.99, 4.42 (2m, 2H, 11-OH,
2'-OH), 3.87 (m, 1H, 2'-H), 3.42 (t, 1H, 11-H, J=6 Hz), 3.25 (m,
2H, H-22), 3.05 (m, 1H, 1'-H), 2.60 (m, 3H, 3'-H, NCH.sub.2), 2.40
(bs, 1H, 4-H), 1.35 (s, 3H, 15-CH.sub.3), 1.05 (s, 3H,
18-CH.sub.3). 0.90 (m, 6H, NCH.sub.2CH.sub.3), 0.81 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.62, 0.61 (2d, 3H, 16-CH.sub.3, J=7 Hz).
MS-ESI (m/z): 564 (MH.sup.+), 586 (MNa.sup.+), 1149 (2MNa.sup.+),
562 (M-H).sup.-.
Example 13
14-O-{[(1R,2R,4S)-4-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-a-
cetyl}-mutilin+(1S,2S,4R) diastereomer
Step A1.
14-O-{[(7R,8R)-7-Hydroxy-1,4-dioxa-spiro[4.5]dec-8-ylsulfanyl]-ac-
etyl}-mutilin+(7S,5S) diastereomer
[0185] 7,8-Epoxy-1,4-dioxa-spiro[4.5]decane (Zhang, L.; Koreeda, M.
Organic Letters 2002, 4(21), 3755-3758.) (6.25 g, 40 mmol) and
pleuromutilin thiol (8 g, 20 mmol) were treated according to the
method of Example 1 Step A2. After work up and chromatography of
the reaction mixture (silica, cyclohexane/ethyl acetate=1/1)
14-O-{[(7R,8R)-7-hydroxy-1,4-dioxa-spiro[4.5]dec-8-ylsulfanyl]-acetyl}-mu-
tilin+(7S,8S) diastereomer (R.sub.f=0.3, 8.40 g, 76% yield) was
obtained as colorless amorphous foam.
[0186] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.14 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.55 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 2H, 20-H), 4.95 (d, 1H, 2'-OH, J=6 Hz), 4.50 (d,
1H, 11-OH, J=6 Hz), 3.82 (m, 4H, OCH.sub.2CH.sub.2O), 3.55-3.25 (m,
4H, 2'-H, 11-H, H-22), 2.58, (m, 1H, 1'-H), 2.40 (bs, 1H, 4-H),
1.35, 1.34 (2s, 3H, 15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3), 0.81
(d, 3H, 17-CH.sub.3, J=7 Hz), 0.62 (d, 3H, 16-CH.sub.3, J=7 Hz).
MS-ESI (m/z): 573 (MNa.sup.+), 549 (M-H).sup.-.
or Step A2.
14-O-{[(7R,8R)-7-Hydroxy-1,4-dioxa-spiro[4.5]dec-8-ylsulfanyl]-acetyl}-mu-
tilin+(7S,8S) diastereomer and
14-O-{[(7R,8R)-8-Hydroxy-14-dioxa-spiro[4.5]dec-7-ylsulfanyl]-acetyl}-mut-
ilin+(7S,8S) diastereomer
[0187] 7,8-Epoxy-1,4-dioxa-spiro[4.5]decane (6.24 g, 39.95 mmol)
and pleuromutilin thiol (16.4 g, 41.7 mmol) were treated according
to the method of Example 1 Step A3. After work up and
chromatography of the reaction (silica, ethyl acetate/toluene=1/1)
a mixture of
14-O-{([(7R,8R)-7-hydroxy-1,4-dioxa-spiro[4.5]dec-8-ylsulfanyl]-acetyl}-m-
utilin+(7S,8S) diastereomer as well as
14-O-{[(7R,8R)-8-hydroxy-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-acetyl}-mu-
tilin+(7S,8S) diastereomer (R.sub.f=0.24, 4.40 g, 20% yield) was
obtained as colorless solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6,
.delta., ppm, inter alia): 6.14, 6.12 (2dd, 1H, 19-H, J=1 Hz and 18
Hz), 5.55, 5.53 (2d, 1H, 14-H, J=8 Hz), 5.05 (m, 2H, 20-H), 4.95,
4.87 (d+dd, 1H, 2'-OH, J=6 Hz), 4.50, 4.49 (2d, 1H, 11-OH, J=6 Hz),
3.83 (m, 4H, OCH.sub.2CH.sub.2O), 3.55-3.25 (m, 4H, 2'-H, 11-H,
H-22), 2.77, 2.57 (2m, 1H, 1'-H), 2.40 (bs, 1H, 4-H), 1.36, 1.36
(2s, 3H, 15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3), 0.81 (d, 31,
17-CH.sub.3, J=7 Hz), 0.62 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI
(m/z): 573 (MNa.sup.+), 549 (M-H).sup.-.
Step B.
14-O-{[(7R,8R)-7-(tert-Butyl-diphenyl-silanyloxy)-1,4-dioxa-spiro[-
4.5]dec-8-ylsulfanyl]-acetyl}-mutilin+(7S,8S) diastereomer
[0188] A solution of
14-O-{[(7R,8R)-7-hydroxy-1,4-dioxa-spiro[4.5]dec-8-ylsulfanyl]-acetyl}-mu-
tilin+(7S,8S) diastereomer (8.4 g, 15.3 mmol) from Step A1 in 50 ml
of dimethylformamide was treated with tert-butyldiphenylsilyl
chloride (5.16 ml, 19.8 mmol) and imidazole (1.66 g, 24.4 mmol) and
stirred overnight at 80.degree. C. The reaction mixture was
concentrated under reduced pressure. The residue was diluted with
water and brine and extracted three times with ethyl acetate. The
combined organic layers were washed with water and brine, dried
over sodium sulfate and filtered. After chromatography (silica,
cyclohexane/ethyl acetate=1/1)
14-O-{[(7R,8R)-7-(tert-butyl-diphenyl-silanyloxy)-1,4-dioxa-spiro[4.5]dec-
-8-ylsulfanyl]-acetyl}-mutilin+(7S,8S) diastereomer (R.sub.f=0.7,
8.03 g, 67% yield) was obtained.
[0189] .sup.1H NMR 0.400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.7-7.35 (m, 10H, aromat.-H), 6.15, 6.13 (2dd, 1H, 19-H,
J=11 Hz and 18 Hz), 5.57, 5.53 (2d, 1H, 14-H, J=7 Hz), 5.05 (m, 2H,
20-H), 4.50 (m, 1H, 11-OH), 3.30 (m, H, 2'-H), 3.70-2.80 (m, 8H,
OCH.sub.2CH.sub.2O, 1'-H, 11-H, 22-H), 2.40 (bs, 1H, 4-H), 1.39,
1.36 (2s, 3H, 15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3), 1.00 (s, 9H,
Si-tert-butyl), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.62, 0.60 (2d,
3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 811 (MNa.sup.+).
Step C.
14-O-{[(1R,2R)-2-(tert-Butyl-diphenyl-silanyloxy)-4-oxo-cyclohexyl-
sulfanyl]-acetyl}-mutilin+(1S,2S) diastereomer
[0190]
14-O-{[(7R,8R)-7-(tert-Butyl-diphenyl-silanyloxy)-1,4-dioxa-spiro[4-
.5]dec-8-ylsulfanyl]-acetyl}-mutilin+(7S,8S) diastereomer (8.03 g,
10.2 mmol) was dissolved in 100 ml of dichloromethane and treated
with montmorillonite K10 (10 g) for 3 days at room temperature.
After filtration over celite the reaction mixture was concentrated
under reduced pressure and subjected to chromatography (silica,
cyclohexane/ethyl acetate=2/1) to yield
14-O-{[(1R,2R)-2-(tert-butyl-diphenyl-silanyloxy)-4-oxo-cyclohexylsulfany-
l]-acetyl}-mutilin+(1S,2S) diastereomer (R.sub.f=0.38, 5.24 g, 69%
yield).
[0191] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.65-7.40 (m, 10H, aromat.-H), 6.15, 6.13 (2dd, 1H, 19-H,
J=11 Hz and 18 Hz), 5.53 (d, 1H, 14-H, J=7 Hz), 5.00 (m, 2H, 20-H),
4.47 (m, 1H, 11-OH), 4.24 (m, 1H, 2'-H), 3.41 (t, 1H, 11-H, J=6
Hz), 3.20-3.00 (m, 21, 22-H), 2.60 (m, 1H, 1'-H), 2.40 (bs, 1H,
4-H), 1.35, 1.33 (2s, 3H, 15-CH.sub.3), 1.06 (s, 3H, 18-CH.sub.3),
0.97 (s, 9H, Si-tert-butyl), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz),
0.58 (d, 3H, 16-CH.sub.3, J=7 Hz).
Step D.
14-O-{[(1R,2R)-2-(t-Butyl-diphenyl-silanyloxy)-4-hydroxyimino-cycl-
ohexylsulfanyl]-acetyl}-mutilin+(1S,2S) diastereomer
[0192] To a solution of
14-O-{[(1R,2R)-2-(t-butyl-diphenyl-silanyloxy)-4-oxo-cyclohexylsulfanyl]--
acetyl}-mutilin+(1S,2S) diastereomer (2.50 g, 3.36 mmol) in 10 ml
of dimethylformamide was added hydroxylamine hydrochloride (233 mg,
3.36 mmol) and triethylamine (0.47 ml, 3.36 mmol) and stirred at
room temperature overnight. The reaction mixture was concentrated
under reduced pressure, diluted with water and brine and extracted
three times with ethyl acetate. The combined organic layers were
washed twice with water and dried over sodium sulfate and filtered.
The solvent was removed under reduced pressure and crude
14-O-{[(1R,2R)-2-(tert-butyl-diphenyl-silanyloxy)-4-hydroxyimino-cyclohex-
ylsulfanyl]-acetyl}-mutilin+(1S,2S) diastereomer (quantitative
yield, cyclohexane/ethyl acetate=2/1, R.sub.f=0.25, 0.35) is
obtained which was used for the next step without further
purification.
Step E.
14-O-{[(1R,2R,4S)-2-(tert-Butyl-diphenyl-silanyloxy)-4-hydroxyamin-
o-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,4R) diastereomer and
14-O-{[(1R,2R,4R)-2-(tert-Butyl-diphenyl-silanyloxy)-4-hydroxyamino-cyclo-
hexylsulfanyl]-acetyl}-mutilin+(1S,2S,4S) diastereomer
[0193]
14-O-{[(R,2R)-2-(tert-Butyl-diphenyl-silanyloxy)-4-hydroxyimino-cyc-
lohexylsulfanyl]-acetyl}-mutilin+(1S,2S) diastereomer (2.55 g, 3.36
mmol) was dissolved in 10 ml of acetic acid and treated with sodium
cyanoborohydride (210 mg, 3.36 mmol) for 90 minutes at room
temperature. The reaction mixture was concentrated under reduced
pressure. The residue was diluted with NaHCO.sub.3 solution and
extracted three times with ethyl acetate. The combined organic
layers were dried over sodium sulfate and filtered. The filtrate
was submitted to chromatography (silica, cyclohexane/ethyl
acetate=2/3) to yield
14-O-{[(1R,2R,4S)-2-(tert-butyl-diphenyl-silanyloxy)-4-hydroxyamino-cyclo-
hexylsulfanyl]-acetyl}-mutilin+(1S,2S,4R) diastereomer (a)
(R.sub.f=0.5, 590 mg, 23% yield) and
14-O-{[(1R,2R,4R)-2-(tert-butyldiphenyl-silanyloxy)-4-hydroxyamino-cycloh-
exylsulfanyl]-acetyl}-mutilin+(1S,2S,4S) diastereomer (b)
(R.sub.f=0.3, 670 mg, 26% yield).
[0194] (a): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.6-7.35 (m, 10H, aromat.-H), 6.93 (bs, 1H, NH/OH), 6.12,
6.08 (2dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.50 (m, 2H, 14-H, NH/OH),
5.00 (m, 2H, 20-H), 4.47 (m, 1H, 11-OH), 3.95 (m, 1H, 2'-H), 3.40
(t, 1H, 11-H, J=6 Hz), 3.10-2.60 (m, 4H, 1'-H, 4'-H, 22-H), 2.40
(bs, 1H, 4-H), 1.31, 1.30 (2s, 3H, 15-CH.sub.3), 1.00 (s, 12H,
18-CH.sub.3, Si-tert-butyl), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz),
0.55 (d, 3H, 16-CH.sub.3, J=7 Hz).
[0195] (b): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.7-7.35 (m, 10H, aromat.-H), 6.85 (s, 1H, NH/OH) 6.16, 6.04
(2dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.55 (m, 2H, 14-H, NH/OH), 5.05
(m, 2H, 20-H), 4.49 (d, 1H, 11-OH, J=6 Hz), 3.55 (n, 1H, 2'-H),
3.42 (t, 1H, 11-H, J=6 Hz), AB-system (.nu..sub.A=3.37,
.nu..sub.B=3.18, 22-H, J=14 Hz), 2.88 (m, 1H, 1'-H), 2.54 (m, 1H,
4'-H), 2.40 (bs, 1, 4-H), 1.39, 1.37 (2s, 3H, 15-CH.sub.3), 1.06
(s, 3H, 18-CH.sub.3), 1.00 (s, 9H, Si-tert-butyl), 0.83 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.64, 0.62 (2d, 3H, 16-CH.sub.3, J=7 Hz).
Step F.
14-O-{[(1R,2R,4S)-2-(tert-Butyl-diphenyl-silanyloxy)-4-(formyl-hyd-
roxy-amino-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,4R)
diastereomer
[0196] To a solution of
14-O-{[(1R,2R,4S)-2-(tert-butyl-diphenyl-silanyloxy)-4-hydroxyamino-cyclo-
hexylsulfanyl]-acetyl}-mutilin+(1S,2S,4R) diastereomer (474 mg,
0.622 mmol) in 15 ml of tert-butyl methyl ether was added
2,2,2-trifluoroethyl formate (594 .mu.l, 6.22 mmol) and heated to
reflux for 5 hours. The reaction mixture was cooled to room
temperature and added dropwise to 150 ml of heptane. The resulting
precipitate was isolated by filtration to give
14-O-{[(1R,2R,4S)-2-(tert-butyl-diphenyl-silanyloxy)-4-(formyl-hydro-
xy-amino-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,4R)
diastereomer (307 mg, 62% yield, cyclohexane/ethyl acetate 1/3,
R.sub.f=0.5) as colorless solid.
[0197] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 9.6, 9.2 (2bs, 1H, NOH), 8.2, 7.9 (2s, 1H, CHO), 7.65-7.35
(m, 10H, aromat.-H), 6.12, 6.08 (2dd, 1H, 19-H, J=11 Hz and 18 Hz),
5.50 (d, 1H, 14-H, J=7 Hz), 5.05 (m, 21, 20-H), 4.47 (m, 1H,
11-OH), 3.40 (t, 1H, 11-H, J=6 Hz), 2.37 (bs, 1H, 4-H), 1.31, 1.30
(2s, 3H, 15-CH.sub.3), 1.03 (s, 12H, 18-CH.sub.3, Si-tert-butyl),
0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.55 (d, 3H, 16-CH.sub.3, J=6
Hz).
Step G.
14-O-{[(1R,2R,4S)-4-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsul-
fanyl]-acetyl}-mutilin+(1S,2S,4R) diastereomer
[0198]
14-O-{[(1R,2R,4S)-2-(tert-Butyl-diphenyl-silanyloxy)-4-(formyl-hydr-
oxy-amino-cyclohexyl-sulfanyl]-acetyl}-mutilin+(1S,2S,4R)
diastereomer (215 mg, 0.272 mmol) in 10 ml of tetrahydrofuran was
treated with tetrabutylammonium fluoride (1.36 ml, 1M in THF, 1.36
mmol) and stirred for 24 hours at room temperature. The reaction
was diluted with a mixture of water, NaHCO.sub.3 solution and brine
(1:1:1) and extracted three times with ethyl acetate. The combined
organic layers were dried over sodium sulfate, filtered and
concentrated under reduced pressure. The residue was added dropwise
to 250 ml heptane. The resulting precipitate was isolated by
filtration to yield
14-O-{[(1R,2R,4S)-4-(formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]--
acetyl}-mutilin+(1S,2S,4R) diastereomer (97 mg, 65% yield,
dichloromethane/methanol=9/1, R.sub.f=0.4) as colorless solid.
[0199] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 9.65, 9.25 (2bs, 1H, NOH), 8.2, 7.9 (2s, 1H, CHO), 6.13 (m,
1H, 19-H), 5.54 (d, 1H, 14-H, J=8 Hz), 5.05 (m, 2H, 20-H), 4.49 (d,
1H, 11-OH, J=6 Hz), 3.42 (t, 1H, 11-H, J=6 Hz), 2.40 (bs, 1H, 4-H),
1.36 (s, 3H, 15-CH.sub.3), 1.06 (a, 12H, 18-CH.sub.3), 0.81 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.63 (d, 3H, 16-CH.sub.3, J=6 Hz). MS-ESI
(m/z): 574 (MNa.sup.+), 550 (M-H).sup.-, 1101 (2M-H).sup.-.
Example 14
14-O-{[(1R,2R,5S)-5-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-a-
cetyl}-mutilin+(1S,2S,5R) diastereomer
Step A.
(7R,8R)-8-(2,4,6-Trimethylbenzylsulfanyl)-1,4-dioxa-spiro[4.5]deca-
n-7-ol+(7S,8S) diastereomer
[0200] 7,8-Epoxy-1,4-dioxa-spiro[4.5]decane (Zhang, L; Koreeda, M.
Organic Letters 2002, 4(21), 3755-3758.) (22 g, 120 mmol) was
treated with 2,4,6-trimethylbenzyl mercaptan (20 g, 120 mmol)
according to the method of Example 1 Step A2. After work up and
chromatography of the reaction mixture (silica, cyclohexane/ethyl
acetate=2/1)
(7R,8R)-8-(2,4,6-trimethylbenzylsulfanyl)-1,4-dioxa-spiro[4.5]decan-7-ol+-
(7S,8S) diastereomer (R.sub.f=0.4, 33 g, 85% yield) was obtained as
oil.
[0201] MS-ESI (m/z): 345 (MNa.sup.+), 667 (2MNa.sup.+).
Step B. Acetic acid
(7R,8R)-7-(2,4,6-trimethylbenzylsulfanyl)-1,4-dioxa-spiro[4.5]dec-8-yl
ester+(7S,8S) diastereomer
[0202] To a solution of triphenylphosphine (26.5 g, 101 mmol) in
500 ml of tetrahydrofuran under argon atmosphere was added
isopropyl azodicarboxylate (19.6 ml, 101 mmol) and stirred for 30
minutes. Then
(7R,8R)-8-(2,4,6-trimethylbenzylsulfanyl)-1,4-dioxa-spiro[4.5]decan-7-ol+-
(7S,8S) diastereomer (27.7 g. 86 mmol) in 150 ml of tetrahydrofuran
and acetic acid (7.7 ml, 135 mmol) were added and the reaction
mixture was heated to 80.degree. C. for 24 hours. The resulting
reaction mixture was concentrated under reduced pressure and
subjected to chromatography (silica, cyclohexane/ethyl
acetate/methanol=3/1) to yield acetic acid
(7R,8R)-7(2,4,6-trimethylbenzylsulfanyl)-1,4-dioxa-spiro[4.5]dec-8-yl
ester+(7S,8S) diastereomer (R.sub.f=0.4, 7.0 g, 22% yield).
[0203] .sup.1H NMR (500 MHz, CDCl.sub.3, .delta., ppm): 6.81 (s,
2H, aromat.-H), 4.85 (m, 1H, CHO), 3.96 (m, 4H,
OCH.sub.2CH.sub.2O), AB-system (.nu..sub.A=3.83, .nu..sub.B=3.79,
J=11 Hz, SCH.sub.2), 2.99 (m, 1H, CHS), 2.36 (s, 6H, CH.sub.3),
2.23 (s, 3H, CH.sub.3), 2.18 (m, 1H), 2.12 (m, 1H), 2.11 (s, 3H,
COCH.sub.3), 1.90-1.58 (m, 4H). MS-ESI (m/z): 387 (MNa.sup.+).
Step C. (7R,8R)-7-Mercapto-1,4-dioxa-spiro[4.5]decan-8-ol+(7S,8S)
diastereomer
[0204] Acetic acid
(7R,8R)-7-(2,4,6-trimethylbenzylsulfanyl)-1,4-dioxa-spiro[4.5]dec-8-yl
ester+(7S,8S) diastereomer (6.33 g, 17.4 mol) was treated with
sodium (1.6 g 69.5 mmol) according to the method of Example 11 Step
D. After work up and chromatography of the reaction mixture
(silica, cyclohexane/ethyl acetate=1/1)
(7R,8R)-7-mercapto-1,4-dioxa-spiro[4.5]decan-8-ol+(7S,8S)
diastereomer (R.sub.f=0.4, 1.36 g, 38%) was obtained.
[0205] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm): 4.89 (d,
1H, OH), 3.83 (m, 41H, OCH.sub.2CH.sub.2O), 3.17 (m, 1H, CHO), 2.76
(m, 1H, CHS), 2.43 (s, 1H, SH), 1.90-1.30, 6H). MS-ESI (m/z): 189
(M-H).sup.-.
Step D.
14-O-{[(7R,8R)-8-Hydroxy-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-ace-
tyl}-mutilin+(7S,5S) diastereomer
[0206] (7R,8R)-7-Mercapto-1,4-dioxa-spiro[4.5]decan-8-ol+(7S,8S)
diastereomer (1.36 g, 7.15 mmol) was treated with pleuromutilin
tosylate (3.8 g, 7.15 mmol) according to the method of Example 11
Step E. After work up and chromatography of the reaction mixture
(silica, cyclohexane/ethyl acetate=1/1)
14-O-{[(7R,8R)-8-hydroxy-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-acetyl}-mu-
tilin+(7S,8S) diastereomer (R.sub.f=0.25, 1.90 g, 48%) was obtained
as colorless amorphous foam.
[0207] MS-ESI (m/z): 573 (MNa.sup.+), 1123 (2MNa.sup.+), 549
(M-H).sup.-. 585 (MCl.sup.-).
Step E.
14-O-{[(7R,8R)-8-(tert-Butyl-diphenyl-silanyloxy)-1,4-dioxa-spiro[-
4.5]dec-7-ylsulfanyl]-acetyl}-mutilin+(7S,8S) diastereomer
[0208]
14-O-{[(7R,8R)-8-Hydroxy-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-acet-
yl}-mutilin+(7S,8S) diastereomer (1.90 g, 3.45 mmol) was treated
according to the method of Example 13 Step B. After work up and
chromatography of the reaction mixture (silica, cyclohexane/ethyl
acetate=3/2)
14-O-{[(7R,8R)-8-(tert-butyl-diphenyl-silanyloxy)-1,4-dioxa-spiro[4.5]dec-
-7-ylsulfanyl]-acetyl}-mutilin+(7S,8S) diastereomer (R.sub.f=0.6,
1.65 g, 61%) was obtained as colorless amorphous foam.
[0209] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.7-7.35 (m, 10H, aromat.-H), 6.13, 6.12 (2dd, 1H, 19-H,
J=11 Hz and 18 Hz), 5.53 (d, 1H, 14-H, J=7 Hz), 5.05 (m, 2H, 20-H),
4.50 (m, 1H, 11-OH), 3.78 (m, 4H, OCH.sub.2CH.sub.2O), 3.70 (m, 1H,
1'-H), 3.42 (m, 1H, 11-H), 3.05 (m, 3H, 2'-H, 22-H), 2.40 (bs, 1H,
4-H), 1.36, 1.34 (2s; 3H, 15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3),
1.00 (s, 9H, Si-tert-butyl), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz),
0.60, 0.58 (2d, 3H, 16-CH.sub.3, J=7 Hz).
Step F.
14-O-{[(1R,2R)-2-(tert-Butyl-diphenyl-silanyloxy)-5-oxo-cyclohexyl-
sulfanyl]-acetyl}-mutilin+(1S,2S) diastereomer
[0210]
14-O-{[(7R,8R)-8-(tert-Butyl-diphenyl-silanyloxy)-1,4-dioxa-spiro[4-
.5]dec-7-ylsulfanyl]-acetyl}-mutilin+(7S,8S) diastereomer (1.65 g,
2.09 mmol) was treated according to the method of Example 13 Step
C. Crude
14-O-{[(1R,2R)-2-(tert-butyl-diphenyl-silanyloxy)-5-oxo-cyclohexylsulfany-
l]-acetyl}-mutilin+(1S,2S) diastereomer (1.34 g. 86% yield,
cyclohexane/ethyl acetate=2/1, R.sub.f=0.3) was obtained as
colorless amorphous foam which was directly used for the next
step.
[0211] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.7-7.35 (m, 10H, aromat.-H), 6.11, 6.09 (2dd. 1H, 19-H,
J=11 Hz and 18 Hz), 5.48 (d, 1H, 14-H, J=7 Hz), 4.98 (m, 2H, 20-H),
4.47 (m, 1H, 11-OH), 4.03 (m, 1H, 1'-H), 3.45-2.95 (m, 4H, 11-H,
2'-H, 22-H), 2.37 (bs, 1H, 4-H), 1.31, 1.29 (2s, 3H, 15-CH.sub.3),
1.02 (s, 12H, 18-CH.sub.3, Si-tert-butyl), 0.81 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.53 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI
(m/z): 767 (MNa.sup.+), 779 (MCl.sup.-).
Step G.
14-O-{[(1R,2R)-2-(tert-Butyl-diphenyl-silanyloxy)-5-hydroxyimino-c-
yclohexylsulfanyl]-acetyl}-mutilin+(1S,2S) diastereomer
[0212]
14-O-{[(1R,2R)-2-(tert-Butyl-diphenyl-silanyloxy)-5-oxo-cyclohexyls-
ulfanyl]-acetyl}-mutilin+(1S,2S) diastereomer (1.34 g, 1.80 mmol)
was treated according to the method of Example 13 Step D. Crude
14-O-{[(1R,2R)-2-(tert-butyl-diphenyl-silanyloxy)-5-hydroxyimino-cyclohex-
ylsulfanyl]-acetyl}-mutilin+(1S,2S) diastereomer (quantitative
yield, cyclohexane/ethyl acetate=1/1, R.sub.f=0.6) was obtained as
colorless amorphous foam which was directly used for the next
step.
Step H.
14-O-{[(1R,2R,5S)-2-(tert-Butyl-diphenyl-silanyloxy)-5-hydroxyamin-
o-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer and
14-O-{[(1R,2R,5R)-2-(tert-butyl-diphenyl-silanyloxy)-5-hydroxyamino-cyclo-
hexylsulfanyl]-acetyl}-mutilin+(1S,2S,5S) diastereomer
[0213]
14-O-{[(1R,2R)-2-(tert-butyl-diphenyl-silanyloxy)-5-hydroxyimino-cy-
clohexylsulfanyl]-acetyl}-mutilin+(1S,2S) diastereomer (2.55 g,
3.36 mmol) was treated according to the method of Example 13 Step
E. After work up and chromatography of the reaction mixture
(silica, cyclohexane/ethyl acetate=1/3)
14-O-{[(1R,2R,5S)-2-(tert-butyl-diphenyl-silanyloxy)-5-hydroxyamino-cyclo-
hexylsulfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer (a)
(R.sub.f=0.4, 220 mg, 16% yield) and
14-O-{[(1R,2R,5R)-2-(tert-butyl-diphenyl-silanyloxy)-5-hydroxyamino-cyclo-
hexylsulfanyl]-acetyl}-mutilin+(1S,2S,5S) diastereomer (b)
(R.sub.f=0.25, 560 mg, 41% yield) were obtained.
[0214] (a): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.65-7.35 (m, 10H, aromat.-H), 7.00 (be, 1H, NH/OH), 6.11,
6.09 (2dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.50 (d, 1H, 14-H, J=8
Hz), 5.00 (m, 2H, 20-H), 4.47 (m, 1H, 11-OH), 3.80 (m, 1H, 2'-H),
3.40 (t, 1H, 11-H, J=6 Hz), 3.00 (m, 1H, 1'-H), AB-system
(.nu..sub.A=3.93, .nu..sub.B=3.80, 22-H, J=15 Hz), 2.68 (m, 1H,
5'-H), 2.40 (bs, 1H, 4-H), 1.31, 1.29 (2s, 3H, 15-CH.sub.3), 1.00
(s, 12H, 18-CH.sub.3, Si-tert-butyl), 0.81 (d, 3H, 17-CH.sub.3, J=7
Hz), 0.55 (d, 3H, 16-CH.sub.3, J=7 Hz).
[0215] (b): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.7-7.33 (m, 10H, aromat.-H), 6.97 (s, 1H, NH/OH), 6.16,
6.14 (2dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.56 (m, 2H, 14-H), 5.40
(bs, 1H, NH/OH), 5.07 (m, 2H, 20-H), 4.49, 4.48 (2d, 1H, 11-H, J=6
Hz), 3.48 (m, 1H, 2'-H), 3.43 (t, 11-H, 11-H, J=6 Hz), 3.24 (m, 2H,
22-H), 2.79 (m, 1H, 1'-H), 2.40 (bs, 1H, 4-H), 2.33 (m, 1H, 5'-H),
1.38, 1.35 (2s, 3H, 15-CH.sub.3), 1.05 (a, 3H, 18-CH.sub.3), 0.98
(s, 9H, Si-tert-butyl), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63,
0.61 (2d, 3H, 16-CH.sub.3, J=6 Hz).
Step I.
14-O-{[(1R,2R,5S)-2-tert-Butyl-diphenyl-silanyloxy)-5-(formyl-hydr-
oxy-amino-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,5R)
diastereomer
[0216]
14-O-{[(1R,2R,5S)-2-(tert-butyl-diphenyl-silanyloxy)-5-hydroxyamino-
-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer (215
mg, 0.282 mmol) was treated according to the method of Example 13
Step F. Isolation of the precipitate by filtration resulted in
14-O-{[(1R,2R,5S)-2-(tert-butyl-diphenyl-silanyloxy)-5-(formyl-hydroxy-am-
ino-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer
(135 mg, 61% yield, cyclohexane/ethyl acetate=1/3, R.sub.f=0.65) as
colorless solid.
[0217] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 9.8, 9.3 (2bs, 1H, NOH), 8.2, 7.9 (2bs, 1H, CHO), 7.60-7.35
(m, 10H, aromat.-H), 6.11, 6.09 (2dd, 1H, 19-H, J=11 Hz and 18 Hz),
5.50 (d, 1H, 14-H, J=8 Hz), 5.00 (m, 2H, 20-H), 4.47 (d, 1H, 11-OH,
J=6 Hz), 3.40 (t, 1H, 11-H, J=6 Hz), 2.37 (bs, 1H, 4-H), 1.32, 1.30
(2s, 3H, 15-CH.sub.3), 1.03 (s, 12H, 18-CH.sub.3, Si-tert-butyl),
0.82, 0.80 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.55 (d, 3H, 16-CH.sub.1,
J=6 Hz).
Step J.
14-O-{[(1R,2R,5S)-5-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsul-
fanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer
[0218]
14-O-{[(1R,2R,5S)-2-(tert-Butyl-diphenyl-silanyloxy)-5-(formyl-hydr-
oxy-amino-cyclohexyl-sulfanyl]-acetyl}-mutilin+(1S,2S,5R)
diastereomer (130 mg, 0.164 mmol) was treated according to the
method of Example 13 Step G. Isolation of the precipitate by
filtration resulted in
14-O-{[(1R,2R,5S)-5-(formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]--
acetyl}-mutilin+(1S,2S,5R) diastereomer (77 mg, 85% yield,
dichloromethane/methanol=9/1, R.sub.f=0.4) as colorless solid.
[0219] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 9.7, 9.3 (2bs, 1H, NOH), 8.2, 7.9 (2s, 1H, CHO), 6.13 (dd,
1H, 19-H, J=11 Hz and 18 Hz), 5.53 (d, 1H, 14-H, J=8 Hz), 5.06 (m,
2H, 20-H), 4.91 (d, 1H, 2'-OH), 4.49 (d, 1H, 11-OH, J=6 Hz), 4.2,
3.7 (2m, 2H, 2'-H, 5'-H), 3.41 (t, 1H, 11-H, J=6 Hz), 3.28 (m, 2H,
22-H), 3.13 (m, 1H, 1'-H), 2.40 (bs, 1H, 4-1), 1.35 (s, 3H,
15-CH.sub.3), 1.06 (s, 12H, 18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3,
J=7 Hz), 0.63 (d, 3H, 16-CH.sub.3, J=6 Hz). MS-ESI (m/z): 574
(MNa.sup.+), 1125 (MNa.sup.+), 550 (M-H).sup.-, 1101
(2M-H).sup.-.
Example 15
14-O-{[(1R,2R,3R/S)-3-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-
-acetyl}-mutilin+(1S,2S,3R/S) diastereomer
Step A.
14-O-{[(6R,7R)-6-Hydroxy-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-ace-
tyl}-mutilin+(6S,7S) diastereomer
[0220] 6,7-Epoxy-1,4-dioxa-spiro[4.5]decane (Vankar, Y. D.; Reddy
M. V.; Chaudhuri, N. C. Tetrahedron 1994, 50(37), 11057-11078.)
(16.24 g, 104 mmol) and pleuromutilin thiol (20.5 g, 52 mmol) were
treated according to the method of Example 1 Step A1. After work up
and chromatography of the reaction mixture (silica,
cyclohexane/dioxane=2/1)
14-O-{[(6R,7R)-6-hydroxy-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-acetyl}-mu-
tilin+(6S,7S) diastereomer (R.sub.f=0.5, 15.6 g, 55% yield) was
obtained as colorless amorphous foam.
[0221] .sup.1H NMR (400 MHz, DMSO-d.delta., ppm, inter alia): 6.13
(dd, H, 19-H, J=11 Hz and 18 Hz), 5.53 (d, 1H, 14-H, J=7 Hz), 5.05
(m, 2H, 20-H), 4.90 (m, 1H, 2'-OH), 4.47 (m, 1H, 11-OH), 3.97 (m,
1H, 2'-H), 3.32 (m, 1H, 11-H), 3.50-3.20 (m, 2H, 22-H), 2.80 (m,
1H, 1'-H), 2.40 (be, 1H, 4-H), 1.35, 1.34 (2s, 3H, 15-CH.sub.3),
1.05 (s, 3H, 18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.62
(d, 3H, 16-CH.sub.3, J=6 Hz).
Step B.
14-O-{[(1R,2R)-2-Hydroxy-3-oxo-cyclohexylsulfanyl]-acetyl}-mutilin-
+(1S,2S) diastereomer
[0222]
14-O-{[(6R,7R)-6-Hydroxy-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-acet-
yl}-mutilin+(6S,7S) diastereomer (15.6 g, 28.4 mmol) was treated
according to the method of Example 13 Step C. After work up and
chromatography of the reaction mixture (silica, cyclohexane/ethyl
acetate=1/1)
14-O-{[(1R,2R)-2-hydroxy-3-oxo-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S-
) diastereomer (R.sub.f=0.4, 3.14 g, 22% yield) was obtained as
colorless amorphous foam.
[0223] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.13 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.55 (d, 1H, 14-H,
J=8 Hz), 5.23 (m, 1H, 2'-OH), 5.05 (m, 2H, 20-H), 4.49 (d, 1H,
11-OH, J=6 Hz), 4.00 (m, 1H, 2'-H), 3.50-3.30 (m, 3H, 11-H, 22-H),
2.86 (m, 1H, 1'-H), 2.40 (bs, 1H, 4-H), 1.35 (s, 3H, 15-CH.sub.3),
1.06 (s, 3H, 18-CH.sub.3), 0.80 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.61
(d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 529 (MNa.sup.+), 1035
(2MNa.sup.+).
Step C.
14-O-{[(1R,2R)-2-Hydroxy-3-hydroxyimino-cyclohexylsulfanyl]-acetyl-
}-mutilin+(1S,2S) diastereomer
[0224]
14-O-{[(1R,2R)-2-Hydroxy-3-oxo-cyclohexylsulfanyl]-acetyl}-mutilin+-
(1S,2S) diastereomer (3.14 g, 6.19 mmol) was treated according to
the method of Example 13 Step D. After work up and chromatography
of the reaction mixture (silica, cyclohexane/ethyl acetate=1/1)
14-O-{[(1R,2R)-2-hydroxy-3-hydroxyimino-cyclohexylsulfanyl]-acetyl}-mutil-
in+(1S,2S) diastereomer (R.sub.f=0.2, 1.75 g, 54% yield) was
obtained as colorless amorphous foam.
[0225] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 10.5 (s, 1H, NOH), 6.13, 6.12 (2dd, 1H, 19-H, J=1 Hz and 18
Hz), 5.54 (d, 1H, 14-H, J=8 Hz), 5.33 (d, 1H, 2'-OH, J=4 Hz), 5.05
(m, 2H, 20-H), 4.50 (m, 1H, 11-OH), 3.96 (m, 1H, 2'-H), 3.42 (t,
1H, 11-H, J=6 Hz), 3.25 (m, 2H, 22-H), 3.14 (m, 1H, 1'-H), 2.40
(bs, 1H, 4-H), 1.35 (s, 3H, 15-CH.sub.3), 1.05 (s, 3H,
18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (d 3H, 3H,
16-CH.sub.3, J=7 Hz).
Step D.
14-O-{[(1R,2R,3R/S)-2-Hydroxy-3-hydroxyamino-cyclohexylsulfanyl]-a-
cetyl}-mutilin+(1S,2S,3R/S) diastereomer
[0226]
14-O-{[(1R,2R)-2-Hydroxy-3-hydroxyimino-cyclohexylsulfanyl]-acetyl}-
-mutilin+(1S,2S) diastereomer (1.75 g, 3.35 mmol) was treated
according to the method of Example 13 Step E. After work up and
chromatography of the reaction mixture (silica, ethyl
acetate/methanol=10/1)
14-O-{[(1R,2R,3R/S)-2-hydroxy-3-hydroxyamino-cyclohexylsulfanyl]-acetyl}--
mutilin+(1S,2S,3R/S) diastereomer (R.sub.f=0.2, 1.34 g, 65% yield)
was obtained as colorless amorphous foam.
[0227] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.1 (bs, 1H, NH/OH), 6.12, 6.11 (2dd, 1H, 19-H, J=11 Hz and
18 Hz), 5.53 (d, 1H, 14-H, J=8 Hz), 5.05 (m, 2H, 20-H), 4.90 (m,
1H, 2'-OH), 4.5 (m, 1H, 11-OH), 3.41 (t, 1H, 11-H, J=6 Hz), 3.73,
3.53, 3.30, 3.14, 3.01, 2.87 (6m, 5H, 1'-H, 2'-H, 3'-H, 22-H), 2.40
(bs, 1H, 4-H), 1.35 (2s, 3H, 15-CH.sub.3), 1.05 (s, 3H,
18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (d, 3H,
16-CH.sub.3, J=7 Hz).
Step E.
14-O-{[(1R,2R,3R/S)-3-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexyls-
ulfanyl]-acetyl}-mutilin+(1S,2S,3R/S) diastereomer
[0228]
14-O-{[(1R,2R,3R/S)-2-hydroxy-3-hydroxyamino-cyclohexylsulfanyl]-ac-
etyl}-mutilin+(1S,2S,3R/S) diastereomer (899 mg, 1.72 mmol) was
treated according to the method of Example 13 Step F. After
isolation of the precipitate by filtration
14-O-{[(1R,2R,3R/S)-3-(formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,3R/S) diastereomer (724 mg, 76% yield,
dichloromethane/methanol=9/1, R.sub.f=0.5) was obtained as
colorless solid.
[0229] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 9.6, 9.9.4, 9.1 (3bs, 1H, NOH), 8.2, 7.9 (2s, 1H, CHO),
6.13, 6.11 (2dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.54 (d, 1H, 14-H,
J=8 Hz), 5.24 (m, 1H, 2'-OH), 5.05 (m, 2H, 20-H), 4.49 (m, 1H,
11-OH), 3.86, 3.60 (2m, 1H, 2'-H), 3.39 (t, 1H, 11-H, J=6 Hz),
3.28, 3.13, 2.64 (3m, 4H, 1'-H, 3'-H, 22-H), 2.38 (bs, 1H, 4-H),
1.36 (s, 3H, 15-CH.sub.3), 1.06 (s, 12H, 18-CH.sub.3), 0.81 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.63 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI
(m/z): 574 (MNa.sup.+), 1125 (2MNa.sup.+), 550 (M-H).sup.-, 1101
(2M-H).sup.-.
Example 16
14-O-{[(1R,2R,5S)-2-Hydroxy-5-methylamino-cyclohexylsulfanyl]-acetyl}-muti-
lin hydrochloride+(1S,2S,5R) diastereomer hydrochloride
Step A. N-Methyl-N-(cyclohex-3-enyl)-carbamic acid tert-butyl
ester
[0230] Cyclohex-3-enyl-carbamic acid tert-butyl ester (Kampferer,
P.; Vasella, A. Helvetica Chimica Acta 2004, 87, 2764-2789) (3 g,
15.2 mmol) and methyl iodide (0.95 ml, 15.2 mmol) were treated for
1 hour according to the method of Example 7 Step A. After work up
and chromatography of the reaction mixture (silica,
cyclohexane/ethyl acetate=10/1) the title compound (R.sub.f=0.22,
2.04 g, 64% yield) was obtained as colorless solid.
[0231] .sup.1H NMR (200 MHz, CDCl.sub.3, .delta., ppm): 5.64 (bs,
2H, double bond), 4.17 (bs, 1H, NCH), 2.74 (s, 3H, NCH.sub.3),
2.13, 1.70 (2m, 6H), 1.47 (s, 9H, tert-butyl).
Step B. N-Methyl-N-(cis-3,4-epoxycyclohexyl)-carbamic acid
tert-butyl ester
[0232] N-Methyl-N-(cyclohex-3-enyl)-carbamic acid tert-butyl ester
(2 g, 9.5 mmol) and 3-chloroperbenzoic acid (2.2 g, 70%, 8.9 mmol)
were treated for 1 hour according to the method of Example 7 Step
B. After work up the crude title compound (silica,
cyclohexane/ethyl acetate=3/1, R.sub.f=0.25, 1.70 g, 79% yield) was
obtained.
[0233] .sup.1H NMR (200 MHz, CDCl.sub.3, .delta., ppm): 4.0 (bs,
1H, NCH), 3.15 (bs, 2H, epoxide), 2.67 (s, 3H, NCH.sub.3),
2.30-1.10 (m, 6H), 1.45 (s, 9H, tert-butyl).
Step C.
14-O-{[(1R,2R,5S)-(tert-Butoxycarbonyl-methyl-amino)-hydroxy-cyclo-
hexylsulfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer
[0234] N-Methyl-N-(cis-3,4-epoxycyclohexyl)-carbamic acid
tert-butyl ester (1.7 g, 7.5 mmol) was treated with pleuromutilin
thiol (2.95 g, 7.5 mmol) according to the method of Example 1 Step
A3. After work up and chromatography of the reaction mixture
(silica, cyclohexane/ethyl acetate=2/1)
14-O-{[(1R,2R,5S)-5-(tert-Butoxycarbonyl-methyl-amino)-2-hydroxy-cyclohex-
ylsulfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer (R.sub.f=0.23,
1.3 g. 28% yield) was obtained as colorless solid.
[0235] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.12 (2dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.54 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 2H, 20-H), 4.9 (d, 1H, 2'-OH, J=4 Hz), 4.47 (d,
1H, 11-OH, J=6 Hz), 3.97 (m, 1H, 5'-H), 3.70 (bs, 1H, 2'-H), 3.42
(m, 1H, 11-H), 3.28 (m, 2H, 22-H), 3.11 (m, 1H, 1'-H), 2.62 (s, 3H,
NCH.sub.3), 2.40 (bs, 1H, 4-H), 1.37 (s, 9H, tert-butyl), 1.35 (s,
3H, 15-CH.sub.3), 1.04 (s, 3H, 18-CH.sub.3), 0.81 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.63 (d, 3H, 16-CH.sub.3, J=7 Hz).
Step D.
14-O-{[(1R,2R,5S)-2-Hydroxy-5-methylamino-cyclohexylsulfanyl]-acet-
yl}-mutilin+(1S,2S,5R) diastereomer
[0236]
14-O-{[(1R,2R,5S)-5-(tert-Butoxycarbonyl-methyl-amino)-2-hydroxy-cy-
clohexylsulfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer (1.3 g,
2.1 mmol) was treated according to the method of Example 1 Step B.
After work up and chromatography of the reaction mixture (silica,
dichloromethane/methanol/i-propanol/water/acetic acid=80/20/6/3/2)
with subsequent basic extraction
14-O-{[(1R,2R,5S)-2-hydroxy-5-methylamino-cyclohexylsulfanyl]-acetyl}-mut-
ilin+(1S,2S,5R) diastereomer (R.sub.f=0.4, 690 mg, 63%) was
obtained as colorless solid.
[0237] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.14 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.54 (d, 1H, 14-H,
J=8 Hz), 5.06 (m, 2H, 20-H), 4.81 (bs, 1H, 2'-OH), 4.52 (d, 1H,
11-OH, J=6 Hz), 3.51 (m, 1H, 2'-H), 3.43 (m, 1H, 11-H), 3.30 (m,
2H, 22-H), 3.00 (m, 1, 1'-H), 2.63 (m, 1H, 5'-H), 2.41 (bs, 1H,
4-H), 2.29 (s, 3H, NCH.sub.3), 1.37 (s, 3H, 15-CH.sub.3), 1.06 (s,
3H, 18-CH.sub.3), 0.82 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.64 (d, 3H,
16-CH.sub.3, J=7 Hz).
Step E.
14-O-{[(1R,2R,5S)-2-Hydroxy-5-methylamino-cyclohexylsulfanyl]-acet-
yl}-mutilin hydrochloride+(1S,2S,5R) diastereomer hydrochloride
[0238]
14-O-{[(1R,2R,5S)-2-Hydroxy-5-methylamino-cyclohexylsulfanyl]-acety-
l}-mutilin+(1S,2S,5R) diastereomer (690 mg, 1.32 mmol) was treated
according to the method of Example 1 Step C to obtain
14-O-{[(1R,2R,5S)-2-hydroxy-5-methylamino-cyclohexylsulfanyl]-acetyl}-mut-
ilin hydrochloride+(1S,2S,5R) diastereomer hydrochloride (731 mg,
quantitative yield) as colorless solid.
[0239] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 8.65 (bs, 2H, NH.sub.2.sup.+), 6.14 (2dd, 1H, 19-H, J=11 Hz
and 18 Hz), 5.54 (d, 1H, 14-H, J=8 Hz), 5.06 (m, 2H, 20-H), 4.51
(d, 1H, 11-OH, J=6 Hz), 3.72 (m, 1H, 2'-H), 3.43 (m, 1H, 11-H),
3.37-3.00 (m, 4H, 22-H, 1'-H, 5'-H), 2.50 (s, 3H, NCH.sub.3), 2.40
(bs, 1H, 4-H), 1.36 (s, 3H, 15-CH.sub.3), 1.06 (s, 3H,
18-CH.sub.3), 0.82 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (d, 3H,
16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 522 (MH.sup.+), 556
(MCl.sup.-).
Example 17
14-O-{[(1R,2R,5S)-5-Allylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutil-
in hydrochloride+(1S,2S,5R) diastereomer hydrochloride
Step A. N-Allyl-N-(cyclohex-3-enyl)-carbamic acid tert-butyl
ester
[0240] Cyclohex-3-enyl-carbamic acid tert-butyl ester (Kampferer,
P.; Vasella, A. Helvetica Chimica Acta 2004, 87, 2764-2789) (3 g,
15.2 mmol) and allyl iodide (1.4 ml, 15.2 mmol) were treated
overnight according to the method of Example 7 Step A. After work
up and chromatography of the reaction mixture (silica,
cyclohexane/ethyl acetate=10/1) the title compound (R.sub.f=0.55,
2.0 g, 55% yield) was obtained as colorless solid.
[0241] .sup.1H NMR (200 MHz, CDCl.sub.3, .delta., ppm): 5.60 (m,
2H, double bond), 5.80, 5.10, 3:64 (3m, 5H, allyl), 4.18 (bs, 1H,
NCH), 2.14, 1.74 (2m, 6H), 1.45 (s, 9H, tert-butyl).
Step B. N-Allyl-N-(cis-3,4-epoxycyclohexyl)-carbamic acid
tert-butyl ester
[0242] N-Allyl-N-(cyclohex-3-enyl)-carbamic acid tert-butyl ester
(2 g, 8.4 mmol) and 3-chloroperbenzoic acid (2.2 g. 70%, 8.9 mmol)
were treated overnight according to the method of Example 7 Step B.
After work up the crude title compound (silica, cyclohexane/ethyl
acetate=3/1, R.sub.f=0.31, 1.90 g, 89% yield) was obtained.
[0243] .sup.1H NMR (200 MHz, CDCl.sub.3, .delta., ppm): 5.76, 5.10,
3.66 (3m, 5H, allyl), 4.04 (bs, 1H, NCH), 3.12 (bs, 2H, epoxide),
2.30-1.20 (m, 6H), 1.47 (s, 9H, tert-butyl).
Step C.
14-O-{[(R,2R,5S)-5-(tert-Butoxycarbonyl-allyl-amino)-2-hydroxy-cyc-
lohexylsulfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer
[0244] N-Allyl-N-(cis-3,4-epoxycyclohexyl)-carbamic acid tert-butyl
ester (1.9 g, 7.5 mmol) was treated with Pleuromutilin thiol (2.95
g, 7.5 mmol) according to the method of Example 1 Step A2. After
work up and chromatography of the reaction (silica,
cyclohexane/ethyl acetate=3/1->1/1) a mixture of
14-O-{[(1R,2R,5S)-5-(tert-Butoxycarbonyl-allyl-amino)-2-hydroxy-cyclohexy-
lsulfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer and
Pleuromutilin disulfide (cyclohexane/ethyl acetate=1/1,
R.sub.f=0.21, 2.49 g) was obtained.
[0245] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.12 (2dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.74 (m, 1H,
NCH.sub.2CHCH.sub.2), 5.54 (d, 1H, 14-H, J=8 Hz), 5.05 (m, 4H,
20-H, NCH.sub.2CHCH.sub.2), 4.87 (d, 1H, 2'-OH, J=3 Hz), 4.49 (d,
1H, 11-OH, J=6 Hz), 3.95 (m, 1H, 5'-H), 3.68 (bs, 3H, 2'-H,
NCH.sub.2CHCH.sub.2), 3.42 (t, 1H, 11-H, J=6 Hz), 3.26 (m, 2H,
22-H), 3.09 (m, 1H, 1'-H), 2.39 (bs, 1H, 4-H), 1.3.7 (s, 9H,
tert-butyl), 1.35 (s, 3H, 15-CH.sub.3), 1.05 (a, 31, 18-CH.sub.3),
0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (d, 3H, 16-CH.sub.3, J=7
Hz). MS-ESI (m/z): 670 (MNa.sup.+), 1317 (2MNa.sup.+), 646
(M-H).sup.-.
Step D.
14-O-{[(1R,2R,5S)-5-Allylamino-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-mutilin+(1S,2S,5R) diastereomer
[0246] The mixture of
14-O-{[(1R,2R,5S)-5-(tert-Butoxycarbonyl-allyl-amino)-2-hydroxy-cyclohexy-
lsulfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer and
Pleuromutilin disulfide (2.4 g) was treated according to the method
of Example 1 Step B. After work up and chromatography of the
reaction mixture (silica,
dichloromethane/methanol/i-propanol/water/acetic acid=80/20/6/3/2)
with subsequent basic extraction
14-O-{[(1R,2R,5S)-5-allylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S,2S,5R) diastereomer (R.sub.f=0.5, 250 mg) was obtained as
colorless solid.
[0247] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.14 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.82 (m, 1H,
NCH.sub.2CHCH.sub.2), 5.55 (d, 1H, 14-H, J=8 Hz), 5.10 (m, 4H,
20-H, NCH.sub.2CHCH.sub.2), 4.77 (m, 1H, 2'-OH), 4.51 (d, 1H, 1-OH,
J=6 Hz), 3.50-3.10 (m, 6H, 2'-H, 11-H, 22-H, NCH.sub.2CHCH.sub.2),
2.99 (m, 1H, 1'-H), 2.68 (m, 1H, 5'-H), 2.40 (bs, 1H, 4-H), 1.37
(s, 3H, 15-CH.sub.3), 1.06 (s, 3H, 18-CH.sub.3), 0.82 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.63 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI
(m/z): 548 (MH.sup.+), 546 (M-H).sup.-, 582 (MCl.sup.-).
Step E.
14-O-{[(1R,2R,5S)-5-Allylamino-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-mutilin hydrochloride+(1S,2S,5R) diastereomer hydrochloride
[0248]
14-O-{[(1R,2R,5S)-5-Allylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl-
}-mutilin+(1S,2S,5R) diastereomer (250 mg, 0.46 mmol) was treated
according to the method of Example 1 Step C to obtain
14-O-{[(1R,2R,5S)-2-hydroxy-5-ethylamino-cyclohexylsulfanyl]-acetyl}-muti-
lin hydrochloride+(1S,2S,5R) diastereomer hydrochloride (273 mg,
quant.yield uncorrected) as colorless solid.
[0249] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 8.85 (bs, 2H, NH.sub.2*), 6.13 (2dd, 1H, 19-H, J=11 Hz and
18 Hz), 5.87 (m, 1H, NCH.sub.2CHCH.sub.2), 5.55 (d, 1H, 14-H, J=8
Hz), 5.47, 5.37 (2d, 2H, NCH.sub.2CHCH.sub.2, J=17 Hz and 10 Hz),
5.06 (m, 2H, 20-H), 4.51 (d, 1H, 11-OH, J=6 Hz), 3.72 (m, 1H,
2'-H), 3.56 (d, 2H, NCH.sub.2CHCH.sub.2, J=6 Hz), 3.43 (t, H, 11-H,
J=6 Hz), 3.34 (m, 2H, 22-H), 3.13 (m, 2H, 1'-H, 5'-H), 2.40 (bs,
1H, 4-H), 1.36 (s, 31H, 15-CH.sub.3), 1.06 (s, 3H, 18-CH.sub.3),
0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (d, 3H, 16-CH.sub.3, J=7
Hz). MS-ESI (m/z): 548 (MH.sup.+), 582 (MCl.sup.-).
Example 18
14-O-{[(1R,2R,5S)-2-Hydroxy-5-(2-methoxy-ethylamino)-cyclohexylsulfanyl]-a-
cetyl}-mutilin hydrochloride+(1S,2S,5R) diastereomer
hydrochloride
Step A. N-(2-methoxy-ethyl)-N-(cyclohex-3-enyl)-carbamic acid
tert-butyl ester
[0250] Cyclohex-3-enyl-carbamic acid tert-butyl ester (Kampferer,
P.; Vasella, A. Helvetica Chimica Acta 2004, 87, 2764-2789) (3 g,
15.2 mmol) and 2-bromoethyl methyl ether (1.43 ml, 15.2 mmol) were
treated overnight to the method of Example 7 Step A. After work up
and chromatography of the reaction mixture (silica,
cyclohexane/ethyl acetate=7/1) the title compound (R.sub.f=0.33,
1.2 & 31% yield) was obtained as colorless solid.
[0251] .sup.1H NMR (200 MHz, CDCl.sub.3, .delta., ppm): 5.61 (m,
2H, double bond), 4.10 (bs, 1H, NCH), 3.50-3.15 (m, 7H,
NCH.sub.2CH.sub.2OCH.sub.3), 2.15, 1.72 (2m, 6H), 1.47 (s, 9H,
tert-butyl).
Step B. N-2-Methoxy-ethyl)-N-(cis-3,4-epoxycyclohexyl)-carbamic
acid tert-butyl ester
[0252] N-(2-Methoxy-ethyl)-N-(cyclohex-3-enyl)-carbamic acid
tert-butyl ester (1.2, 4.7 mmol) and 3-chloroperbenzoic acid (1.2
g, 70%, 4.87 mmol) were treated over the weekend according to the
method of Example 7 Step B. After work up the crude title compound
(silica, cyclohexane/ethyl acetate=3/1, R.sub.f=0.33, 1.08 &
85% yield) was obtained.
[0253] .sup.1H NMR (200 MHz, CDCl.sub.3, .delta., ppm): 3.94 (bs,
1H, NCH), 3.50-3.05 (m, 9H, NCH.sub.2CH.sub.2OCH.sub.3, epoxide),
2.30-1.20 (m, 6H), 1.45 (s, 9H, tert-butyl).
Step C.
14-{[(1R,2R,5S)-5-(tert-Butoxycarbonyl-(2-methoxy-ethyl)-amino)-2--
hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,5R)
diastereomer
[0254] N-(2-Methoxy-ethyl)-N-(cis-3,4-epoxycyclohexyl)-carbamic
acid tert-butyl ester (1.08 g, 4.0 mmol) was treated with
pleuromutilin thiol (1.57 g, 4.0 mmol) according to the method of
Example 1 Step A2. After work up and chromatography of the reaction
mixture (silica, cyclohexane/ethyl acetate=1/2)
14-O-{[(1R,2R,5S)-5-(tert-Butoxycarbonyl-(2-methoxy-ethyl)-amino)-2-hydro-
xy-cyclohexylsulfanyl]-acetyl})-mutilin+(1S,2S,5R) diastereomer
(R.sub.f=0.5, 500 mg, 19% yield) was obtained as colorless
solid.
[0255] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.13 (2dd, 1H, 19-H, J=111 Hz and 18 Hz), 5.54 (d, 1H, 14-H,
J=8 Hz), 5.05 (m, 2H, 20-H), 4.88 (d, 1H, 2'-OH, J=4 Hz), 4.48 (d,
1H, 11-OH, J=6 Hz), 3.85 (m, 1H, 5'-H), 3.68 (be, 1H, 2'-H), 3.42
(t, 1H, 1-H, J=6 Hz), 3.35-3.05 (m, 10H, 22-H,
NCH.sub.2CH.sub.2OCH.sub.3, 1'-H), 2.40 (bs, 1H, 4-H), 1.38 (s, 9H,
tert-butyl), 1.35 (s, 3H, 15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3),
0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (d, 3H, 16-CH.sub.3, J=7
Hz).
Step D.
14-O-{[(1R,2R,5S)-2-Hydroxy-5-(2-methoxy-ethylamino)-cyclohexylsul-
fanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer
[0256]
14-{[(1R,2R,5S)-5-(tert-Butoxycarbonyl-(2-methoxyethyl)-amino)-2-hy-
droxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer
(500 mg, 0.75 mmol) was treated according to the method of Example
1 Step B. After work up and chromatography of the reaction mixture
(silica, dichloromethane/methanol/i-propanol/water/acetic
acid=80/20/6/3/2) with subsequent basic extraction
14-O-{[(1R,2R,5S)-2-hydroxy-5-(2-methoxy-ethylamino)-cyclohexylsulfanyl]--
acetyl}-mutilin+(1S,2S,5R) diastereomer (R.sub.f=0.6, 330 mg, 78%)
was obtained as colorless solid.
[0257] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.14 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.54 (d, 1H, 14-H,
J=8 Hz), 5.06 (m, 2H, 20-H), 4.74 (m, H, 2'-OH), 4.51 (2d, 1H,
11-OH, J=6 Hz), 3.50-3.20 (m, 9H, 11-H, 2'-H, 22-H,
NCH.sub.2CH.sub.2OCH.sub.3), 2.97 (m, 1H, 1'-H), 2.63 (m, 3H, 5'-H,
NCH.sub.2CH.sub.2OCH.sub.3), 2.40 (bs, 1H, 4-H), 1.37 (s, 3H,
15-CH.sub.3), 1.06 (s, 3H, 18-CH.sub.3), 0.82 (d, 3H, 17-CH.sub.3,
J=7 Hz), 0.63 (d, 31, 16-CH.sub.3, J=7 Hz).
Step E.
14-O-{[(1R,2R,5S)-2-Hydroxy-5-(2-methoxy-ethylamino)-cyclohexylsul-
fanyl]-acetyl}-mutilin hydrochloride+(1S,2S,5R) diastereomer
hydrochloride
[0258]
14-O-{[(1R,2R,5S)-2-Hydroxy-5-(2-methoxy-ethylamino)-cyclohexylsulf-
anyl]-acetyl}-mutilin+(1S,2S,5R) diastereomer (330 mg, 0.58 mmol)
was treated according to the method of Example 1 Step C to obtain
14-O-{[(1R.
2R,5S)-2-hydroxy-5-(2-methoxy-ethylamino)-cyclohexylsulfanyl]-acetyl}-mut-
ilin hydrochloride+(1S,2S,5R) diastereomer hydrochloride (355 mg,
quant. yield, uncorrected) as colorless solid.
[0259] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 8.65 (bs, 2H, NH.sub.2.sup.+), 6.13 (2dd, 1H, 19-H, J=11 Hz
and 18 Hz), 5.55 (d, 1, 14-H, J=8 Hz), 5.06 (m, 2H, 20-H), 4.51 (d,
1H, 11-OH, J=6 Hz), 3.71 (m, 1H, 2'-H), 3.57 (m, 5H,
NCH.sub.2CH.sub.2OCH.sub.3), 3.42 (t, 1H, 11-H, J=6 Hz), 3.33 (m,
2H, 22-H), 3.20-3.00 (m, 4H, 1'-H, 5'-H, NCH.sub.2CH.sub.2OCH),
2.40 (bs, 1H, 4-H), 1.36 (s, 3H, 15-CH.sub.3), 1.05 (s, 3-,
18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (2d, 3H,
16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 566 (MH.sup.+), 600
(MCl.sup.-).
Example 19
14-O-{[(1R,2R,4R*)-2-Hydroxy-4-(2-hydroxy-ethylamino)-cyclohexylsulfanyl]--
acetyl}-mutilin hydrochloride+(1S,2S,4S) diastereomer
hydrochloride
Step A.
14-O-{[(1R,2R,4R/S)-2-(tert-Butyl-diphenyl-silanyloxy)-4-(2-hydrox-
y-ethylamino)-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,4R/S)
diastereomer
[0260] To a solution of
14-O-{[(1R,2R)-2-(tert-butyl-diphenyl-silanyloxy)-4-oxo-cyclohexylsulfany-
l]-acetyl})-mutilin+(1S,2S) diastereomer (1.50 g, 2.01 mmol) from
Example 13 Step C in 20 ml of dichloromethane was added
ethanolamine (0.12 ml, 2.01 mmol) and titanium(IV)isopropoxide (0.7
ml, 2.52 mmol) and stirred for 2 hours at room temperature. The
resulting reaction mixture was treated with sodium cyanoborohydride
(126 mg, 2 mmol) overnight at room temperature, diluted with
further dichloromethane and extracted with NaHCO.sub.3 solution.
The organic layer was dried over sodium sulphate and filtered. The
filtrate was subjected to chromatography (silica,
dichloromethane/methanol=30/1) to yield
14-O-{[(1R,2R,4R/S)-2-(tert-butyl-diphenyl-silanyloxy)-4-(2-hydroxy-ethyl-
amino)-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,4R/S)
diastereomer (R.sub.f=0.3, 230 mg, 14% yield).
[0261] .sup.1H NMR (500 MHz, CDCl.sub.3, .delta., ppm, inter alia):
7.8-7.30 (m, 10H, aromat.-H), 6.47 (m, 1H, 19-H), 5.75, 5.69 (2d,
1H, 14-H, J=8 Hz), 5.40-5.15 (m, 1H, 20-H), 4.04, 3.65 (2m, 1H,
2'-H), 3.64, 3.51 (2m, 2H, NCH.sub.2CH.sub.2OH), 3.36 (m, 1H,
11-H), 2.74, 2.54 (2m, 2H, NCH.sub.2CH.sub.2OH), 2.11 (bs, 1H, 4-H,
1.44, 1.45 (2s, 3H, 15-CH.sub.3), 1.17, 1.16 (s, 3H, 18-CH.sub.3),
1.08 (s, 9H, Si-tert-butyl), 0.88 (2d, 3H, 17-CH.sub.3), 0.75-0.65
(m, 3H, 16-CH.sub.3). MS-ESI (m/z): 790 (MH.sup.+), 824
(MCl.sup.-).
Step B.
14-O-{[(1R,2R,4R*)-2-Hydroxy-4-(2-hydroxy-ethylamino)-cyclohexylsu-
lfanyl]-acetyl}-mutilin+(1S,2S,4S*) diastereomer
[0262] To a solution of
14-O-{[(1R,2R,4R/S)-2-(tert-butyl-diphenyl-silanyloxy)-4-(2-hydroxy-ethyl-
amino)-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,4R/S)
diastereomer (230 mg, 0.29 mmol) in 15 ml of acetonitrile was
treated with HF (40% aqueous, 2 drops) and stirred overnight at
room temperature. The reaction was charged with NaHCO.sub.3
solution and extracted with dichloromethane. The combined organic
layers were dried over sodium sulfate, filtered and concentrated
under reduced pressure. After chromatography (silica.
dichloromethane/methanol=6/1) the title compound (R.sub.f=0.4, 50
mg, 31% yield) was obtained.
[0263] .sup.1H NMR (400 MHz, CDCl.sub.3, .delta., ppm, inter alia):
6.48 (dd, 1H, 19-H, J=11 Hz and 17 Hz), 5.76, 5.68 (2d, 1H, 14-H,
J=8 Hz), 5.40-5.15 (m, 1H, 20-H), 3.66 (t, 2H, NCH.sub.2CH.sub.2OH,
J=5 Hz), 3.50-3.15 (m, 4H, 2'-H, 11-H, 22H), 2.80 (m, 2H,
NCH.sub.2CH.sub.2OH), 2.63 (m, 2H, 1'-H, 4'-H), 2.11 (bs, 1H, 4-H),
1.46 (s, 3H, 15-CH.sub.3), 1.18 (s, 3H, 18-CH.sub.3), 0.89 (d, 3H,
17-CH.sub.2), 0.73 (2d, 3H, 16-CH.sub.3). MS-ESI (m/z): 552
(MH.sup.+).
Step C.
14-O-{[(1R,2R,4R*)-2-Hydroxy-4-(2-hydroxy-ethylamino)-cyclohexylsu-
lfanyl]-acetyl}-mutilin hydrochloride+(1S,2S,4S*) diastereomer
hydrochloride
[0264]
14-O-{[(1R,2R,4R*)-2-Hydroxy-4-(2-hydroxy-ethylamino)-cyclohexylsul-
fanyl]-acetyl}-mutilin+(1S,2S,4S*) diastereomer (50 mg, 0.091 mmol)
was treated according to the method of Example 1 Step C to obtain
14-O-{[(1R,2R,4R)-2-Hydroxy-4-(2-hydroxy-ethylamino-cyclohexylsulfanyl]ac-
etyl}-mutilin hydrochloride+(1S,2S,4S*) diastereomer hydrochloride
(43 mg, 80% yield) as colorless solid.
[0265] .sup.1H NMR (500 MHz, CDCl.sub.3, .delta., ppm, inter alia):
6.46 (dd, 1H, 19-H, J=11 Hz and 17 Hz), 5.76 (d, 1H, 14-H, J=8 Hz),
5.37 (d, 1H, 20-H, J=11 Hz), 5.23 (d, 1H, 20-H, J=17 Hz), 3.95 (m,
2H, NCH.sub.2CH.sub.2OH, J=5 Hz), 3.58 (m, 1H, 2'-H), 3.40-3.10 (m,
5H, 11-H, 22H, NCH.sub.2CH.sub.2OH), 2.72 (m, 1H, 1'-H), 2.11 (bs,
1H, 4-H), 1.46 (s, 3H, 15-CH.sub.3), 1.18 (s. 3H, 18-CH.sub.3),
0.89 (d, 3H, 17-CH.sub.3, 0.1-7 Hz), 0.73 (d, 3H, 16-CH.sub.3, J=7
Hz). MS-ESI (m/z): 552 (MH.sup.+), 586 (MCl.sup.-).
Example 20
14-O-{[(1R,2R,4R*)-4-Cyclohexylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-
-mutilin hydrochloride+(1S,2S,4S*) diastereomer hydrochloride
Step A.
14-O-{[(1R,2R,4R*)-2-(tert-Butyl-diphenyl-silanyloxy)-4-(2-hydroxy-
-ethylamino)-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,4S*)
diastereomer
[0266]
14-O-{[(1R,2R)-2-(tert-butyl-diphenyl-silanyloxy)-4-oxo-cyclohexyls-
ulfanyl]-acetyl}-mutilin+(1S,2S) diastereomer (1.50 g, 2.01 mmol)
from Example 13 Step C was reacted with cyclohexylamine (0.23 ml,
1.01 mmol) according to the method of Example 19 Step A. After work
up and chromatography of the reaction mixture (silica,
dichloromethane/methanol=30/1)
14-O-{[(1R,2R,4R*)-2-(tert-butyl-diphenyl-silanyloxy)-4-cyclohexylamino-c-
yclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,4S*) diastereomer
(R.sub.f=0.13, 150 mg, 9% yield) was obtained.
[0267] .sup.1H NMR (500 MHz, CDCl.sub.3, .delta., ppm, inter alia):
7.8-7.3 (m, 10H, aromat.-H), 6.48 (dd, 1H, 19-H, J=11 Hz and 17
Hz), 5.76 (d, 1H, 14-H, J=8 Hz), 5.40-5.15 (m, 1H, 20-H), 3.62 (m,
1H, 2'-H), 3.40-3.10 (m, 3H, 11-H, 22-H), 2.77 (m, 1H, 1'-H), 2.11
(bs. 1H, 4-H), 1.46 (s, 3H, 15-CH.sub.3), 1.16 (s, 3H,
18-CH.sub.3), 1.07 (s, 9H, Si-tert-butyl), 0.88 (2d, 3H,
17-CH.sub.3), 0.74, 0.73 (2d, 3H, 16-CH.sub.3). MS-ESI (m/z): 828
(MH.sup.+), 862 (MCl.sup.-).
Step B.
14-O-{[(1R,2R,4R*)-4-Cyclohexyl-2-hydroxy-cyclohexylsulfanyl]-acet-
yl}-mutilin+(1S,2S,4S*) diastereomer
[0268]
14-O-{[(1R,2R,4R*)-2-(tert-butyldiphenyl-silanyloxy)-4-cyclohexylam-
ino-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,4S*) diastereomer
(100 mg, 0.121 mmol) was treated with HF (40% aqueous, 30 drops)
for 5 hours according to the method of Example 19 Step B. After
work up and chromatography of the reaction mixture (silica,
dichloromethane/methanol=10/1) the title compound (R.sub.f=0.13, 23
mg, 32% yield) was obtained.
[0269] .sup.1H NMR (500 MHz, CDCl.sub.3, .delta., ppm, inter alia):
6.46 (dd, 1H, 19-H, J=11 Hz and 17 Hz), 5.75 (m, 1H, 14-H),
5.40-5.15 (m, 1H, 20-H), 3.49 (m, 1H, 2'-H), 3.35 (m, 1H, 11-H),
AB-system (.nu..sub.A=3.30, .nu..sub.B=3.20, 22-H, J=15 Hz), 3.01
(m, 1H, 4'-H), 2.67 (m, 1H, 1'-H) 2.09 (bs, 1H, 4-H), 1.45 (s, 3H,
15-CH.sub.3) 1.16 (s, 3H, 18-CH.sub.3), 0.87 (d, 3H, 17-CH.sub.3,
J=7 Hz), 0.72 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 591
(MH.sup.+).
Step C.
14-O-{[(1R,2R,4R*)-4-Cyclohexyl-2-hydroxy-cyclohexylsulfanyl]-acet-
yl}-mutilin hydrochloride+(1S,2S,4S*) diastereomer
hydrochloride
[0270]
14-O-{[(1R,2R,4R*)-4-Cyclohexyl-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-mutilin+(1S,2S,4S*) diastereomer (23 mg, 0.039 mmol) was treated
according to the method of Example 1 Step C to obtain
14-O-{[(1R,2R,4R*)-4-Cyclohexyl-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mut-
ilin hydrochloride+(1S,2S,4S*) diastereomer hydrochloride (26 mg,
quantitative yield, uncorrected) as colorless solid.
[0271] .sup.1H NMR (500 MHz, CDCl.sub.3, .delta., ppm, inter alia):
9.3 (bs, 2H, NH.sub.2.sup.+), 6.46 (dd, 1H, 19-H, J=11 Hz and 17
Hz), 5.75 (d, 1H, 14-H, J=8 Hz), 5.38 (d, 1H, 20-H, J=12 Hz), 5.22
(d, 1H, 20-H, J=17 Hz), 3.50-3.00 (m, 6H, 2'-H, 11-H, 22H, NcHex),
2.65 (m, 2H, 1'-H, 3a'-H), 2.10 (bs, 1H, 4-H), 1.45 (s, 3H,
15-CH.sub.3), 1.18 (s, 3H, 18-CH.sub.3), 0.88 (d, 3H, 17-CH.sub.3,
J=7 Hz), 0.73 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 591
(MH.sup.+), 624 (MCl.sup.-).
Example 21
14-O-{[(1R,2R,4R*)-4-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl-
}-mutilin+(1S,2S,4S*) diastereomer
Step A.
14-O-{[(1R,2R,4R*)-2-(ret-Butyl-diphenyl-silanyloxy)-4-cyclopropyl-
amino-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,4S*)
diastereomer
[0272]
14-O-{[(1R,2R)-2-(tert-butyl-diphenyl-silanyloxy)-4-oxo-cyclohexyls-
ulfanyl]-acetyl}-mutilin+(1S,2S) diastereomer (750 mg, 1.01 mmol)
from Example 13 Step C was reacted with cyclopropylamine (0.07 ml,
1.01 mmol) in 40 ml of dichloromethane according to the method of
Example 19 Step A. After the treatment of sodium cyanoborohydride
ethanol was added (0.7 ml) and the mixture was stirred overnight at
room temperature. After work up and chromatography of the reaction
mixture (silica, dichloromethane/methanol=30/1)
14-O-{[(1R,2R,4R*)-2-(tert-butyl-diphenyl-silanyloxy-4-cyclopropylamino-c-
yclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,4S*) diastereomer
(R.sub.f=0.35, 283 mg, 36% yield) was obtained.
[0273] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.65-7.35 (m, 10H, aromat.-H), 6.12 (m, 1H, 19-H), 5.5.2,
5.51 (2d, 1H, 14-H, J=8 Hz), 5.00 (m, 1H, 20-H), 4.50 (t, 1H,
11-OH, J=5.5 Hz), 3.94 (m, 1H, 2'-H), 3.41 (m, 1H, 11-H), 3.05-2.80
(m, 4H, 22-H, 1'-H, 4'-H), 2.39 (bs, 1H, 4-H), 1.86 (m, 1H, cPr),
1.33 (2s, 3H, 15-CH.sub.3), 1.12 (s. 12H, 18-CH.sub.3,
Si-tert-butyl), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.57, 0.56 (2d,
3H, 16-CH.sub.3, J=7 Hz), 0.25, 0.06 (2m, 4H, cPr). MS-ESI (m/z):
786 (M1H), 784 (M-H).sup.-.
Step B. 14-O-{[(1R,
R,4R*)-4-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]acetyl}-mutilin+(1-
S,2S,4S*) diastereomer
[0274]
14-O-{[(1R,2R,4R*)-2-(tert-butyl-diphenyl-silanyloxy)-4-cyclopropyl-
-amino-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S,4S*) diastereomer
(223 mg, 0.284 mmol) was treated overnight with tetrabutylammonium
fluoride according to the method of Example 13 Step G. After work
up and chromatography of the reaction mixture (silica,
dichloromethane/methanol=10/1) the title compound (R.sub.f=0.2, 10
mg, 6% yield) was obtained.
[0275] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.13 (dd, 1H, 19-H, J=11 Hz and 17 Hz), 5.50 (d, 1H, 14-H,
J=7 Hz), 5.05 (m, 1H, 20-H), 4.87 (m, 1H, 2'-OH), 4.50 (d, 1H,
11-OH, J=6 Hz), 3.55-3.20 (m, 4H, 22-H, 2'-H, 11-H), 2.50 (m, 2H,
1'-H, 4'-H), 2.40 (bs, 1H, 4-H), 2.01 (m, 1H, cPr), 1.35 (s, 3H,
15-CH.sub.3), 1.04 (s, 3H, 18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3,
J=7 Hz), 0.62 (d, 3H, 16-CH.sub.3, J=6 Hz), 0.32, 0.15 (2m, 4H,
cPr). MS-ESI (m/z): 548 (MH.sup.+), 1095 (2MH.sup.+), 1117
(2MNa.sup.+), 582 (MCl.sup.-).
Example 22
14-O-{[(1R,2R,5S*)-4-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl-
}-mutilin hydrochloride+(1S,2S,5R*) diastereomer hydrochloride
Step A.
14-O-{[(1R,2R)-2-Hydroxy-4-oxo-cyclohexylsulfanyl]-acetyl}-mutilin-
+(1S,2S) diastereomer and
14-O-{[(1R,2R)-2-Hydroxy-5-oxo-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S-
) diastereomer
[0276]
14-O-{[(7R,8R)-7-hydroxy-1,4-dioxa-spiro[4.5]dec-8-ylsulfanyl]-acet-
yl}-mutilin+(7S,8S) diastereomer as well as
14-O-{[(7R,8R)-8-hydroxy-1,4-dioxa-spiro[4.5]de-7-ylsulfanyl]-acetyl}-mut-
ilin+(7S,8S) diastereomer (3.96 g, 7.19 mmol) was dissolved in 50
ml of dioxane and treated with 4N HCl (5 ml, 20 mmol) for 6 hours
at room temperature. The reaction mixture was concentrated under
reduced pressure, charged with NaHCO.sub.3 solution and extracted
tree times with ethyl acetate. The organic layers were dried over
sodium sulphate and filtered. The filtrate was concentrated under
reduced pressure and subjected to chromatography (silica,
cyclohexane/dioxane=2/1) to yield a mixture of the title compounds
(R.sub.f=0.30, 860 mg, 24% yield).
[0277] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.13 (m, 1H, 19-H), 5.56, 5.54 (2d, 1H, 14-H, J=8 Hz), 5.38,
5.32 (2m, 1H, 2'-OH), 5.05 (m, 2H, 20-H), 4.50 (d, 1H, 11-OH, J=5
Hz), 3.95, 3.83 (2m, 1H, 2'-H), 3.50-3.20 (m, 3H, 11-H, 22-H),
3.17, 3.07 (2m, 1H, 1-H), 2.40 (bs, 1H, 4-H), 1.35, 1.33 (2s, 3H,
15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3,
J=7 Hz), 0.63, 0.62 (2d, 3H, 16-CH.sub.3). 529 (MNa.sup.+), 505
(M-H).sup.-.
Step B.
14-O-{[(1R,2R,5S*)-5-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,4R*) diastereomer and
14-O-{[(1R,2R,4S*)-4-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-mutilin+(1S,2S,4R*) diastereomer
[0278]
14-O-{[(1R,2R)-2-Hydroxy-4-oxo-cyclohexylsulfanyl]-acetyl}-mutilin+-
(1S,2S) diastereomer and
14-O-{[(1R,2R)-2-hydroxy-5-oxo-cyclohexylsulfanyl]acetyl}-mutilin+(1S,2S)
diastereomer (250 mug, 0.493 mmol) was reacted with
cyclopropylamine (0.03 ml, 0.493 mmol) in 15 ml of dichloromethane
according to the method of Example 19 Step A. After the treatment
of sodium cyanoborohydride ethanol was added (0.7 ml) and the
mixture was stirred overnight at room temperature. After work up
and chromatography of the reaction mixture (silica,
dichloromethane/methanol=20/1)
14-O-{[(1R,2R,5S*)-4-cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-mutilin+(1S,2S,5R*) diastereomer (a)
(dichloromethane/methanol=10/1, R.sub.f=0.22, 34 mg, 13% yield) and
14-O-{[(1R,2R,4S*)-4-cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-mutilin+(1S,2S,4R*) diastereomer (b)
(dichloromethane/methanol=10/1, R.sub.f=0.13, 26 mg, 4% yield) were
obtained.
[0279] (a): .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, infer
alia): 6.13 (m, 1H, 19-H, J=11 and 18 Hz). 5.53 (d, 1H, 14-H, J=8
Hz), 5.06 (m, 8H, 20-H). 4.67 (t, 1H, 2'-OH), 4.47 (d, 1H, 11-OH,
J=6 Hz), 3.50-3.20 (m, 4H, 11-H, 2'-H, 22-H) 2.92 (m, 1H, 1'-H),
2.71 (m, 1H, 5'-H), 2.39 (bs, 1H, 4-H), 1.96 (m, 1H, cPr), 1.36 (s,
3H, 15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3), 0.81 (d, 3H, 17-C, J=7
Hz), 0.62 (di, 3H, 16-CH.sub.3, J=7 Hz), 0.32, 0.16 (2m, 4H, cPr).
MS-ESI (m/z): 548 (MH.sup.+), 546 (M-H).sup.-.
[0280] (b): .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.13 (m, 1H, 19-H, J=11 and 18 Hz), 5.54 (d, 1H, 14-H, J=8
Hz), 5.05 (m, 1H, 20-H), 4.70 (d, 1H, 2'-OH; J=5 Hz), 4.47 (d, 1H,
11-OH, J=6 Hz), 3.68 (m, 1H, 2'-H), 3.45-3.15 (m, 3H, 11-H, 22-H)
2.86 (m, 1H, 4'-H), 2.71 (m, 1H, 1'-H), 2.39 (bs, 1H, 4-H), 1.96
(m, 1H, cPr), 1.35 (s, 3H, 15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3),
0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.62 (d, 3H, 16-CH.sub.3, J=7
Hz), 0.32, 0.15 (2m, 4H, cPr). MS-ESI (m/z): 548 (MH.sup.+), 54.6
(M-H).sup.-.
Step C.
14-O-{[(1R,2R,5S*)-5-Cyclopropylamino-cyclohexylsulfanyl]acetyl}-m-
utilin hydrochloride+(1S,2S,5R*) diastereomer hydrochloride
[0281]
14-O-{[(1R,2R,5S*)-4-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-
-acetyl}-mutilin+(1S,2S,5R*) diastereomer (34 mg, 0.062 mmol) was
treated according to the method of Example 1 Step C to obtain
14-O-{[(1R,2R,5R*)-4-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-mutilin hydrochloride+(1S,2S,5S*) diastereomer hydrochloride (24
mg, 66% yield) as colorless solid.
[0282] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.13 (m, H, 19-H, J=11 and 18 Hz), 5.55 (d, 1H, 14-H, J=8
Hz), 5.05 (m, 1H, 20-H), 5.00 (m, 1H, 2-OH), 4.54 (d, 1H, 11-OH,
J=6 Hz), 3.66 (m, 1H, 2'-H), 3.45-3.05 (m, 5H, 11-H, 22-H, 1'-H,
5'-H), 2.41 (bs, 1H, 4-H), 1.36 (s, 3H, 15-CH.sub.3), 1.06 (s, 3H,
18-CH.sub.3), 0.82 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (m, 7H,
16-CH.sub.3, cPr). MS-ESI (m/z): 548 (MH.sup.+), 582
(MCl.sup.-).
Example 23
14-O-{[(1R,2R,4S)-4-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-
-mutilin hydrochloride+(1S,2S,4R*) diastereomer hydrochloride
[0283]
14-O-{[(1R,2R,4S*)-4-Cyclopropylamino-2-hydroxy-cyclohexylsulfanyl]-
-acetyl}-mutilin+(1S,2S,4R*) diastereomer (10 mg, 0.018 mmol) from
Example 21 Step B was treated according to the method of Example 1
Step C to obtain the title compound (20 mg, quantitative yield,
uncorrected) as colorless solid.
[0284] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 8.6 (bs, 2H, NH.sub.2.sup.+), 6.13 (m, 1H, 19-H, J=11 and 18
Hz), 5.55 (d, 1H, 14-H, J=8 Hz), 5.26 (m, 1H, 2'-OH), 5.05 (m, 1H,
20-H), 4.52 (d, 1H, 11-OH, J=6 Hz), 3.93 (m, 1H, 2'-H), 3.45-3.20
(m, 4H, 11-H, 22-H, 4'-H), 2.95 (m, 1H, 1'-H), 2.64 (m, 1H, cPr),
2.40 (be, 1H, 4-H), 1.36 (s, 3H, 15-CH.sub.3), 1.05 (s, 3H,
18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.74 (m, 4H, cPr),
0.62 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 548 (MH.sup.+),
582 (MCl.sup.-).
Example 24
14-O-{[(1R,2R,5R*)-2-Hydroxy-5-morpholin-4-yl-cyclohexylsulfanyl]-acetyl}--
mutilin hydrochloride+(1S,2S,5S*) diastereomer hydrochloride
Step A,
14-O-{[(1R,2R,5R*)-2-Hydroxy-5-morpholin-4yl-cyclohexylsulfanyl]ac-
etyl}-mutilin+(1S,2S,5S*) diastereomer and
14-O-{[(1R,2R,5S*)-2-Hydroxy-5-morpholin-4-yl-cyclohexylsulfanyl]-acetyl}-
-mutilin+(1S,2S,5R*) diastereomer
[0285]
14-{[(1R,2R)-2-Hydroxy-4-oxo-cyclohexylsulfanyl]-acetyl}-mutilin+(1-
S,2S) diastereomer and
14-O-{[(1R,2R)-2-hydroxy-5-oxo-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S-
) diastereomer (270 mg, 0.533 mmol) from Example 22 Step A was
reacted with morpholine (0.05 ml, 0.533 mmol) in 10 ml of
dichloromethane according to the method of Example 19 Step A. After
addition of sodium cyanoborohydride ethanol was added (0.6 ml) and
the mixture was stirred overnight at room temperature. After work
up and chromatography of the reaction mixture (silica,
dichloromethane/methanol=20/1)
14-O-{[(1R,2R,5R*)-2-Hydroxy-5-morpholin-4-yl-cyclohexylsulfanyl]-acetyl}-
-mutilin+(1S,2S,5S*) diastereomer (a)
(dichloromethane/methanol=10/1, R.sub.f=0.32, 23 mg, 7% yield) and
14-O-{[(1R,2R,5S*)-2-Hydroxy-5-morpholin-4-yl-cyclohexylsulfanyl]-acetyl}-
-mutilin+(1S,2S,5R*) diastereomer (b)
(dichloromethane/methanol=10/1, R.sub.f=0.27, 40 mg, 13% yield)
were obtained.
[0286] (a): .sup.1H NMR (500 MHz, CDCl.sub.3, .delta., ppm, inter
alia): 6.47 (m, 1H, 19-H), 5.77, 5.75 (2d, 1H, 14-H, J=8 Hz), 5.35
(dd, 1H, 20-H, J=3 and 11 Hz), 5.21 (d, 1H, 20-H, J=17 Hz), 3.70
(s, 4H, morpholine), 3.40-3.15 (m, 4H, 2'-H, 11-H, 22H) 2.53 (m,
5H, 1'-H, morpholine), 2.10 (bs, 1H, 4-H), 1.45 (s, 3H,
15-CH.sub.3), 1.17 (s, 3H, 18-CH.sub.3), 0.87 (d, 3H, 17-CH.sub.3,
J=7 Hz), 0.73, 0.72 (2d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z):
578 (MH.sup.+), 600 (MNa.sup.+), 576 (M-H).sup.-, 612
(MCl.sup.-).
[0287] (b): .sup.1H NMR (500 MHz, CDCl.sub.3, .delta., ppm, inter
alia): 6.47 (m, 1H, 19-H), 5.77, 5.75 (2d, 1H, 14-H, J=8 Hz),
5.40-5.15 (m, 2H, 20-H), 3.70 (s, 4H, morpholine), 3.47 (m, 1H,
2'-H), 3.35 (m, 1H, 11-H), 3.22 (m, 2H, 22-H), 2.98 (m, 1H, 11'-H),
2.54, 2.45 (2m, 4H, morpholine), 2.10 (bs, 1H, 4-H), 1.45 (s, 3H,
15-CH.sub.3), 1.17 (s, 3H, 18-CH.sub.3), 0.88 (d, 3H, 17-CH.sub.3,
J=6 Hz), 0.72 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 578
(MH.sup.+), 600 (MNa.sup.+), 576 (M-H).sup.-, 612 (MCl.sup.-).
Step B.
14-O-{[(1R,2R,5R*)-2-Hydroxy-5-morpholin-4-yl-cyclohexylsulfanyl]--
acetyl}-mutilin hydrochloride+(1S,2S,5S*) diastereomer
hydrochloride
[0288]
14-O-{[(1R,2R,5R*)-2-Hydroxy-5-morpholin-4-yl-cyclohexylsulfanyl]-a-
cetyl}-mutilin+(1S,2S,5S*) diastereomer (10 ms, 0.017 mmol) was
treated according to the method of Example 1 Step C to obtain
14-O-{[(1R,2R,5R*)-2-Hydroxy-5-morpholin-4-yl-cyclohexylsulfanyl]-acetyl}-
-mutilin hydrochloride+(1S,2S,5S*) diastereomer hydrochloride (20
mg, quantitative yield, uncorrected) as colorless solid.
[0289] .sup.1H NMR (500 MHz, CDCl.sub.3, .delta., ppm, inter alia):
13 (bs, 1H, NH.sup.+), 6.47 (m, 1H, 19-H); 5.78, 5.76 (2d, 1H,
14-H, J=9 Hz), 5.36 (dd, 1H, 20-H, J=4 and 11 Hz), 5.23 (d, 1H,
20-H; J=17 Hz), 4.40, 3.98 (2bs, 4H, morpholine), 3.45-3.20 (m, 4H,
2'-H, 11-H, 22-H), 2.91, 2.56 (2m, 5H, morpholine, 1'-H), 2.11 (bs,
1H, 4-H), 1.46 (s, 3H, 15-CH.sub.3), 1.20, 1.19 (2s, 3H,
18-CH.sub.3), 0.89 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.74, 0.73 (2d,
3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 578 (MH.sup.+), 600
(MNa.sup.+), 612 (MCl.sup.-).
Example 25
14-O-{[(1R,2R,5S*)-2-Hydroxy-5-morpholin-4-yl-cyclohexylsulfanyl]-acetyl}--
mutilin hydrochloride+(1S,2S,5R*) diastereomer hydrochloride
[0290]
14-O-{[(1R,2R,5S)-2-Hydroxy-5-morpholin-4-yl-cyclohexylsulfanyl]-ac-
etyl}-mutilin+(1S,2S,5R*) diastereomer (26 mg, 0.045 mmol) from
Example 24 Step A was treated according to the method of Example 1
Step C to obtain the title compound (15 mg, 54% yield) as colorless
solid.
[0291] .sup.1H NMR (500 MHz, CDCl.sub.3, .delta., ppm, inter alia):
12.8 (bs, 1H, NH.sup.+), 6.47 (m, 1H, 19-H), 5.75 (m, 1H, 14-H),
5.40-5.15 (m, 2H, 20-H), 4.40, 3.98 (2bs, 4H, morpholine, 2'-H),
3.50-3.15 (m, 6H, 5'-H, 11-H, 22-H, morpholine), 2.98 (m, 2H,
morpholine), 2.11 (bs, 1H, 4-H), 1.46 (s, 3H, 15-CH.sub.3), 1.19
(s, 3H, 18-CH.sub.3), 0.89 (d, 3H, 17-CH.sub.3; J=7 Hz), 0.74 (d,
3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 578 (MH.sup.+), 600
(MNa.sup.+), 612 (MCl.sup.-).
Example 26
14-O-{[(1R,2R,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-9,20-dihyd-
ro-mutilin hydrochloride+(1S,2S,5R) diastereomer hydrochloride
Step A.
14-O-{[(1R,2R,5S)-5-tert-Butoxycarbonylamino-2-hydroxy-cyclohexyls-
ulfanyl]-acetyl}-19,20-dihydro-mutilin+(1S,2S,5R) diastereomer
[0292] To a solution of
14-O-{[(1R,2R,5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,5R) diastereomer (1 g, 1.65 mmol) from
Example 1 Step A in 20 ml of ethanol was added palladium on
charcoal (10%, 515 mg, 0.48 mmol) and hydrogenated overnight at
room temperature. The reaction mixture was treated with
dichloromethane, filtered and the filtrate was concentrated to
dryness under reduced pressure to obtain
14-O-{[(1R,2R,5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-19,20-dihydro-mutilin+(1S,2S,5R) diastereomer (1 g,
quantitative yield) as colorless solid.
[0293] MS-ESI (m/z): 632 (MNa.sup.+).
Step B.
14-O-{[(1R,2R,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19-
,20-dihydro-mutilin+(1S,2S,5R) diastereomer
[0294]
14-O-{[(1R,2R,5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsu-
lfanyl]-acetyl}-19,20-dihydro-mutilin+(1S,2S,5R) diastereomer (1 g,
1.64 mmol) was treated according to the method of Example 1 Step B.
After work up and chromatography of the reaction mixture (silica,
ethyl acetate/methanol/35% ammonia solution=33/66/1)
14-O-{[(1R,2R,5S)-5-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19,20-dih-
ydro-mutilin+(1S,2S,5R) diastereomer (R.sub.f=0.35, 590 mg, 71%
yield) was obtained as colorless solid.
[0295] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 5.51 (d, 1H, 14-H, J=8 Hz), 4.74 (bs, 1H, 2'-OH), 4.37 (m,
1H, 11-OH), 3.49 (m, 1H, 2'-H), 3.45-3.15 (m, 3H, 11-H, 22-H), 3.00
(m, 1H, 1'-H), 2.82 (m, 1H, 5'-H), 2.35 (bs, 1H, 4-H), 1.34 (s, 3H,
15-CH.sub.3), 0.85 (s, 3H, 18-CH.sub.3), 0.80 (d, 3H, 17-CH.sub.3,
J=7 Hz), 0.63 (m, 6H, 16-CH.sub.3, 20-H).
Step C.
14-O-{[(1R,2R,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19-
,20-dihydro-mutilin hydrochloride+(1S,2S,5R) diastereomer
hydrochloride
[0296]
14-O-{[(1R,2R,5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19,-
20-dihydro-mutilin+(1S,2S,5R) diastereomer (590 mg, 1.16 mmol) was
treated according to the method of Example 1 Step C to obtain
14-O-{[(1R,2R,5S)-5-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19,20-dih-
ydro-mutilin hydrochloride+(1S,2S,5R) diastereomer hydrochloride
(566 mg, 89% yield) as colorless solid.
[0297] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.9 (bs, 3H, NH.sub.3.sup.+), 5.52 (d, 1H, 14-H, J=8 Hz),
3.80-3.00 (m, 6H, 2'-H, 11-H, 22-H, 1'-H, 5'-H), 2.35 (bs, 1H,
4-H), 1.35 (s, 3H, 15-CH.sub.3) 0.85 (s, 3H, 18-CH.sub.3), 0.80 (d,
3H, 17-CH.sub.3, J=7 Hz), 0.63 (m, 6H, 16-CH.sub.3, 20-H). MS-ESI
(m/z): 510 (MH.sup.+), 544 (MCl.sup.-).
Example 27
14-O-{[(1R,2R,5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19,20-
-dihydro-mutilin hydrochloride+(1S,2S,5R) diastereomer
hydrochloride
Step A. 19,20-Dihydro-pleuromutilin thiol
[0298] A solution of 19,20-dihydro-pleuromutilin tosylate (Egger,
H.; Reinshagen, H. Journal of Antibiotics 1976, 29, 915-927.) (11.5
g, 22.2 mmol) in 50 ml of acetone was treated with thiourea (1.69
g, 22.2 mmol) under reflux for 1.5 hours. The reaction mixture was
evaporated to dryness under reduced pressure and dissolved in
ethanol. The solution was charged with sodium metabisulfite (4.57
g, 24.0 mmol) dissolved in 20 ml of water, and 100 ml of ethyl
acetate. The biphasic mixture was refluxed for 1.5 hours under
vigorous stirring. After cooling to room temperature the phases
were separated and the aqueous phase was extracted three times with
ethyl acetate. The combined organic layers wen dried over sodium
sulfate, filtered and the solvent evaporated under reduced
pressure. After chromatography (silica, cyclohexane/ethyl
acetate=2/1) 19,20-dihydro-pleuromutilin thiol (cyclohexane/ethyl
acetate=4/3, R.sub.f=0.24, 3 g, 34% yield) were obtained.
[0299] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 5.53 (d, 1H, 14-H, J=8 Hz), 4.40 (d, 1H, 11-OH, J=6 Hz),
3.36 (t, 1H, 11-H, J=6 Hz), 3.25 (m, 2H, 22-H), 2.85 (t, 1H, SH,
J=8 Hz), 2.38 (bs, 1H, 4-H), 1.37 (s, 3H, 15-CH.sub.3), 0.87 (s,
3H, 18-CH.sub.3), 0.83 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.65 (m, 6H,
16-CH.sub.3, 20-H).
Step B.
14-O-{[(1R,2R,5S)-5-(tert-Butoxycarbonyl-ethyl-amino)-2-hydroxy-cy-
clohexylsulfanyl]-acetyl}-19,20-dihydro-mutilin+(1S,2S,5R)
diastereomer
[0300] N-Ethyl-N-(cis-3,4-epoxycyclohexyl)-carbamic acid tert-butyl
ester (2.8 g, 11.6 mmol) from Example 7 Step B was treated with
19,20-dihydro-pleuromutilin thiol (4.60 g, 11.6 mmol) according to
the method of Example 1 Step A3 over the weekend at room
temperature. After work up and chromatography of the reaction
mixture (silica, cyclohexane/ethyl acetate=1/1)
14-O-{[(1R,2R,5S)-(tert-Butoxycarbonyl-ethyl-amino)-2-hydroxy-cyclohexyls-
ulfanyl]-acetyl}-19,20-dihydro-mutilin+(1S,2S,5R) diastereomer
(R.sub.f=0.35, 1.98 g, 27% yield) was obtained.
Step C.
14-O-{[(1R,2R,5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-19,20-dihydro-mutilin+(1S,2S,5R) diastereomer
[0301]
14-O-{[(1R,2R,5S)-5-(tert-Butoxycarbonyl-ethyl-amino)-2-hydroxy-cyc-
lohexylsulfanyl]-acetyl}-19,20-dihydro-mutilin+(1S,2S,5R)
diastereomer (1.98 g, 3.10 mmol) was treated according to the
method of Example 1 Step B. After work up and chromatography of the
reaction mixture (silica, ethyl acetate/methanol/35% ammonia
solution=100/10/1)
14-O-{[(1R,2R,5S)-5-ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19,2-
0-dihydro-mutilin+(1S,2S,5R) diastereomer (ethyl
acetate/methanol/35% ammonia solution=100/100/1, R.sub.f=0.7, 150
mg, 9% yield) was obtained as colorless solid.
[0302] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 5.52 (d, 1H, 14-H, J=7 Hz), 4.72 (m, 1H, 2'-OH), 4.36 (d,
1H, 11-OH, J=6 Hz), 3.50-3.15 (m, 4H, 2'-H, 11-H, 22-H), 2.97 (m,
1H, 1'-H), 2.62 (m, 1H, 5'-H), 2.47 (m, 2H, NCH.sub.2), 2.35 (bs,
1H, 4-H), 1.35 (a, 3H, 15-CH.sub.3), 0.98 (m, 3H,
NCH.sub.2CH.sub.3), 0.85 (s, 3H, 18-CH.sub.3), 0.80 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.63 (m, 6H, 16-CH.sub.3, 20-H). MS-ESI
(m/z): 538 (MH.sup.+), 560 (MNa.sup.+), 536 (M-H).sup.-, 572
(MCl.sup.-).
Step D.
14-O-{[(1R,2R,5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acety-
l}-19,20-dihydro-mutilin hydrochloride+(1S,2S,5R) diastereomer
hydrochloride
[0303]
14-O-{[(1R,2R,5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl-
}-19,20-dihydro-mutilin+(1S,2S,5R) diastereomer (38 mg, 0.071 mmol)
was treated according to the method of Example 1 Step C to obtain
the title compounds (40 mg, quantitative yield) as colorless
solid.
[0304] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 5.53 (d, 1H, 14-H, J=8 Hz), 4.97 (m, 1H, 2'-OH), 4.40 (d,
1H, 1-OH, J=6 Hz), 3.64 (m, 1H, 2'-H), 3.45-3.20 (m, 3H, 11-H,
22-H), 2.98 (m, 1H, 1'-H), 2.94 (m, 1H, 5'-H), 2.88 (m, 2H,
NCH.sub.2), 2.37 (bs, 1H, 4-H), 1.34 (s, 3H, 15-CH.sub.3), 0.98 (t,
3H, NCH.sub.2CH.sub.3, J=7 Hz), 0.85 (s, 3H, 18-CH.sub.3), 0.80 (d,
3H, 17-CH.sub.3, J=7 Hz), 0.63 (m, 6H, 16-CH.sub.3, 20-H). MS-ESI
(m/z): 538 (MH.sup.+).
Example 28
14-O-{[(1R,2R,5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19,20-dihy-
dro-mutilin+(1S,2S,5S) diastereomer
Step A.
14-O-{[(1R,2R,5S)-(tert-Butyl-dimethyl-silyloxy)-2-hydroxy-cyclohe-
xylsulfanyl]-acetyl}-19,20-dihydro-mutilin+(1S,2S,5R)
diastereomer
[0305] tert-Butyl-dimethyl-(cis-3,4-epoxycyclohexyloxy)-silane (864
mg, 3.78 mmol) from Example 4 Step A was treated with
19,20-dihydro-pleuromutilin thiol (1.5 g, 3.78 mmol) from Example
27 Step A according to the method of Example 1 Step A3. After work
up and chromatography of the reaction mixture (silica,
cyclohexane/ethyl acetate=3:1)
14-O-{[(1R,2R,5S)-5-(tert-butyl-dimethyl-silyloxy)-2-hydroxy-cyclohexylsu-
lfanyl]-acetyl}-19,20-dihydro-mutilin+(1S,2S,5R) diastereomer
(cyclohexane/ethyl acetate=1/1, R.sub.f=0.45, 1.2 g, 51% yield) was
obtained as colorless solid.
[0306] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 5.50 (d, 1H, 14-H, J=8 Hz), 4.81 (m, 1H, 2'-OH), 4.38 (d,
1H, 11-OH, J=6 Hz), 3.88 (m, 1H, 5'-H), 3.50-3.20 (m, 4H, 2'-H,
11-H, 22-H), 2.95 (m, 1H, 1'-H), 2.34 (bs, 1H, 4-H), 1.34 (s, 3H,
15-CH.sub.3), 0.84 (m, 12H, 18-CH.sub.3, tert-butyl), 0.80 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.63 (m, 6H, 16-CH.sub.3, 20-H), 0.02 (s, 6H,
Si(CH.sub.3).sub.2).
Step B.
14-O-{[(1R,2R,5S)-2,5-Dihydroxy-cyclohexylsulfanyl]-acetyl}-19,20--
dihydro-mutilin+(1S,2S,5R) diastereomer
[0307]
14-O-{[(1R,2R,5S)-5-(tert-butyl-dimethyl-silyloxy)-2-hydroxy-cycloh-
exylsulfanyl]-acetyl}-19,20-dihydro-mutilin+(1S,2S,5R) diastereomer
(1.2 g, 1.92 mmol) was treated according to the method of Example 4
Step C. After work up and chromatography of the reaction mixture
(silica, cyclohexane/ethyl acetate=1/4)
14-O-{[(1R,2R,5S)-2,5-dihydroxy-cyclohexylsulfanyl]-acetyl}-19,20-dihydro-
-mutilin+(1S,2S,5R) diastereomer (cyclohexane/ethyl acetate=1:1,
R.sub.f=0.2, 720 mg, 73% yield) was obtained as colorless
solid.
[0308] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 5.50 (d, 1H, 14-H, J=8 Hz), 4.74 (d, 1H, 2'-OH, J=3 Hz),
4.42 (m, 1H, 5'-OH), 4.38 (d, 1H, 11-OH, J=6 Hz), 3.67 (m, 1H,
5'-H), 3.50-320 (m, 4H, 2'-H, 11-H, 22-H), 2.96 (m, 1H, 1'-H), 2.34
(bs, 1H, 4-H), 1.34 (s, 3H, 15-CH.sub.3), 0.84 (s, 3H,
18-CH.sub.3), 0.80 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63 (m, 6H,
16-CH.sub.3, 20-H).
Step C.
14-O-{[(1R,2R,5S)-2-Hydroxy-5-methanesulfonyloxy-cyclohexylsulfany-
l]-acetyl}-19,20-dihydro-mutilin+(1S,2S,5R) diastereomer
[0309]
14-O-{[(1R,2R,5S)-2,5-dihydroxy-cyclohexylsulfanyl]-acetyl}-19,20-d-
ihydro-mutilin+(1S,2S,5R) diastereomer (720 mg, 1.41 mmol) was
treated according to the method of Example 4 Step D. After work up
and chromatography of the reaction mixture (silica,
cyclohexane/ethyl acetate=1/1)
14-O-{[(1R,2R,5S)-2-hydroxy-5-methanesulfonyloxy-cyclohexylsulfanyl]-acet-
yl}-19,20-dihydro-mutilin+(1S,2S,5R) diastereomer
(cyclohexane/ethyl acetate=1/2, R.sub.f=0.4, 640 g, 77% yield) was
obtained as colorless solid.
[0310] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 5.51 (d, 1H, 14-H, J=8 Hz), 4.79 (m, 1H, 5'-H), 4.38 (bs,
1H, 11-OH), 3.60-3.20 (m, 7H, 2'-H, 11-H, 22-H, SO.sub.2CH.sub.3),
2.93 (m, 1H, 1'-H), 2.35 (bs, 1H, 4-H), 1.35 (s, 3H, 5-CH.sub.3),
0.84 (s, 3H, 18-CH.sub.3), 0.80 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.63
(m, 6H, 16-CH.sub.3, 20-H).
Step D.
14-O-{[(11R,2R,5R)-5-Azido-2-hydroxy-cyclohexylsulfanyl]-acetyl}-1-
9,20-dihydro-mutilin+(1S,2S,5S) diastereomer
[0311]
14-O-{[(1R,2R,5S)-2-hydroxy-5-methanesulfonyloxy-cyclohexylsulfanyl-
]-acetyl}-19,20-dihydro-mutilin+(1S,2S,5R) diastereomer (640 mg,
1.09 mmol) was treated with sodium aside according to the method of
Example 4 Step E. After work up crude
14-O-{[(1R,2R,5R)-5-azido-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19,20-dih-
ydro-mutilin+(1S,2S,5S) diastereomer (quantitative yield,
cyclohexane/ethyl acetate=1/2, R.sub.f=0.7) was obtained which was
directly used for the next step.
Step E.
14-O-{[(1R,2R,5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19-
,20-dihydro-mutilin+(1S,2S,5S) diastereomer
[0312]
14-O-{[(1R,2R,5R)-5-Azido-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19,-
20-dihydro-mutilin+(1S,2S,5S) diastereomer (584 mg, 1.09 mmol) was
treated with triphenylphosphine (342 mg, 1.30 mmol) according to
the method of Example 4 Step F. After work up and chromatography
(silica, dichloromethane/methanol/i-propanol/water/acetic
acid=80/20/6/3/2) with subsequent basic extraction
14-O-{[(1R,2R,5R)-5-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-19,20-dih-
ydro-mutilin+(1S,2S,5S) diastereomer (silica,
dichloromethane/methanol/35% ammonia solution=100/10/1,
R.sub.f=0.3, 50.5 mg, 9% yield) was obtained as colorless foam.
[0313] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 5.51 (d, 1H, 14-H, J=8 Hz), 4.77 (m, 1H, 2'-OH), 4.38 (d,
1H, 11-OH, J=8 Hz), 3.60-3.15 (m, 4H, 2'-H, 11-H, 22-H), 2.60 (m,
1H, 1'-H), 2.50 (m, 1H, 5'-H), 2.35 (bs, 1H, 4-H), 1.34 (s, 3H,
15-CH.sub.3), 0.84 (s, 3H, 18-CH.sub.3), 0.80 (d, 3H, 17-CH.sub.3,
J=7 Hz), 0.63 (m, 6H, 16-CH.sub.3, 20-H). MS-ESI (m/z): 510
(MH.sup.+).
Example 29
14-O-{[(1R,2R)-4-Aminomethyl-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
diasteromers+(1S,2S) diastereomers
Step A. Cyclohex-3-emylmethyl-carbamic acid tert-butyl ester
[0314] To a solution of C-Cyclohex-3-enyl-methylamine (3.28 g, 29.5
mmol) and N-methyl-morpholine (2.98 g, 29.5 mmol) in 70 ml of
anhydrous dichloromethane was added di-tert-butyldicarbonate (6.44
g, 29.5 mmol) under cooling. The resulting mixture was stirred for
20 hours at room temperature and the solvent was removed under
reduced pressure. The residue was diluted with ethyl acetate and
washed with 1N HCl. The aqueous phase was extracted three times
with ethyl acetate and the combined organic phases were washed with
water and brine. The resulting organic phase was dried over
magnesium sulfate, filtered and the solvent was removed under
reduced pressure to yield 6.57 g of a brown oil. Column
chromatography (silica, petrol ether/ethyl acetate=12/1 to 8/1)
resulted in cyclohex-3-enylmethyl-carbamic acid tert-butyl ester
(petrol ether/ethyl acetate=10/1, R.sub.f=0.62, 3.06 g, 49% yield)
as colorless solid.
[0315] .sup.1H NMR (200 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.90-6.83 (m, 1H, NH), 5.66-5.63 (m, 2H, olef. H), 2.86 (t,
2H, CH.sub.2N, J=6 Hz), 221-1.50 (m, 6H, 2.times.CH.sub.2),
2.21-0.95 (m, 16H, tert-butyl, 3.times.CH.sub.2 and CH).
Step B. (7-Oxa-bicyclo[4.1.0]hept-3-ylmethyl)-carbamic acid
tert-butyl ester
[0316] To a solution of cyclohex-3-enylmethyl-carbamic acid
tert-butyl ester (1.5 g, 7.10 mmol) in 20 ml of anhydrous
dichloromethane was added 3-chloroperoxybenzoic acid (2.45 g, 14.2
mmol) under cooling. The resulting mixture was stirred for 19 hours
at room temperature and washed with saturated sodium bicarbonate
and 0.5M aqueous solution of sodium thiosulphate. The aqueous phase
was extracted three times with dichloromethane and the combined
organic phases were washed with brine. The resulting organic phase
was dried over magnesium sulfate, filtered and the solvent was
removed under reduced pressure to yield 1.48 g of crude product.
Column chromatography of 3.17 g of crude product (silica, petrol
ether/ethyl acetate=3/1) resulted in
(7-oxa-bicyclo[4.1.0]hept-3-ylmethyl)-carbamic acid tart-butyl
ester (R.sub.f=0.19, 2.68 g, 81% yield) as colorless solid.
[0317] .sup.1H NMR (200 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.84-6.78 (m, 1H, NH), 3.09-3.03 (m, 2H, CHO), 2.71-2.70 (m,
2H, CH.sub.2N), 1.37 (s, 9H, tert-butyl). MS-ESI (m/z): 250
(MNa.sup.+), 477 (2MNa.sup.+).
Step C. 14-O{[(1R,2R)
4-(tert-Butoxycarbonylamino-methyl)-2-hydroxy-cyclohexylsulfanyl])-acetyl-
}-mutilin diasteromers+(1S,2S) diastereomers
[0318] To a solution of
(7-oxa-bicyclo[4.1.0]hept-3-ylmethyl)-carbamic acid tert-butyl
ester (1.34 g, 5.90 mmol) and Pleuromutilin thiol (2.32 g, 5.90
mmol) in 25 ml of methanol was added 2M NaOH (2.95 ml, 5.90 mmol)
drop wise under cooling. The resulting mixture was stirred at room
temperature overnight and the solvent was removed under reduced
pressure. The residue was diluted with ethyl acetate and washed
with brine. The aqueous phase was extracted three times with ethyl
acetate and the combined organic phases were washed with brine. The
resulting organic phase was dried over magnesium sulfate, filtered
and the solvent was removed under reduced pressure to yield 3.86 g
of a crude product Column chromatography (silica, petrol
ether/ethyl acetate=1/1) resulted in
14-O-{[(1R,2R)-4-(tert-butoxycarbonylamino-methyl)-2-hydroxy-cyclohexylsu-
lfanyl]-acetyl}-mutilin diasteromers+(1S,2S) diastereomers
(R.sub.f=0.24, 2.11 g 58% yield) as colorless solid.
[0319] .sup.1H NMR (500 MHz, CDCl.sub.3, .delta., ppm, inter alia):
6.32-6.40 (m, 1H, 19-H), 5.80-5.71 (m, 1H, 14-H), 5.40-5.15 (m, 2H,
20-H), 4.64, 4.55 (2bs, 1H, NH), 3.79-3.67, 3.58-3.45 (2m, 1H,
2'-H), 3.40-3.31 (m, 1H, 11-H), 3.29-3.11 (m, 2H, 22-H), 3.10-2.92
(m, 2H, CH.sub.2N), 2.89-2.77, 2.74-2.64 (2m, 1H, 1'-H), 1.45 (s,
3H, 15-CH.sub.3), 1.43 (s, 9H, tert-butyl), 1.17 (s, 3H,
18-CH.sub.3), 0.78-0.66 (m, 3H, 16-CH.sub.3). MS-ESI (m/z): 644
(MNa.sup.+).
Step D.
14-O-{[(1R,2R)-4-Aminomethyl-2-hydroxy-cyclohexylsulfanyl]-acetyl}-
-mutilin diasteromers+(1S,2S) diastereomers
[0320] To a solution of
14-O-{[(R,2R)-4-(tert-butoxycarbonylamino-methyl)-2-hydroxy-cyclohexylsul-
fanyl]-acetyl}-mutilin diasteromers+(1S,2S) diastereomers (1.14 g,
1.83 mmol) in 20 ml of anhydrous dichloromethane was added 20 ml of
1M HCl in diethyl ether drop wise under cooling. The resulting
mixture was stirred at room temperature for two days and the
solvent was removed under reduced pressure. The residue was diluted
with dichloromethane and washed with a saturated solution of sodium
bicarbonate. The aqueous phase was extracted three times with
dichloromethane. The resulting organic phases were combined and
dried over magnesium sulfate, filtered and the solvent was removed
under reduced pressure. Column chromatography (silica,
dichloromethane/methanol=5/1 to 1/1) resulted in
14-O-{[(1R,2R)-4-aminomethyl-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutili-
n diasteromers+(1S,2S) diastereomers (39 mg, 8% yield) as colorless
solid. .sup.1H NMR (400 MHz, CDCl.sub.3, .delta., ppm, inter alia):
6.58-6.38 (m, 1H, 19-H), 5.87-5.68 (m, 1H, 14-H); 5.44-5.12 (m, 2H,
20-H), 4.52-4.34 (m, 1H, 1'-H), 2.68-2.52 (m, 2H, CH.sub.2N), 1.46
(s, 3H, 15-CH.sub.3), 1.19 (s, 3H, 18-CH.sub.3), 0.89 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.74 (d, 3H, 16-CH, J=7 Hz). MS-ESI (m/z):
544 (MNa.sup.+).
Example 30
14-O-{[(5-Amino-2-chloro-cyclohexylsulfanyl]-acetyl}-mutilin
acetate and
14-O-{[4-Amino-2-chloro-cyclohexylsulfanyl]-acetyl}-mutilin
acetate
Step A. (1R,3S,6S)(7-thia-bicyclo[4.1.0]hept-3-yl)carbamic acid
tert-butyl ester+(1S,3R,6R) diastereomer
[0321] To a solution of syn-3,4-epoxycyclohexyl-carbamic acid
tert-butyl ester (3.55 g, 16.6 mmol) and tetrabutylammonium
chloride (500 mg, 1.80 mmol) in 50 ml of tert-butyl methyl ether
was added a solution of potassium thiocyanate (8.07 g. 83.0 mmol)
in 50 ml of water. The resulting mixture was stirred for 7 days at
room temperature and the phases were separated. The aqueous phase
was extracted twice with ethyl acetate and the combined organic
phases were washed with brine. The resulting organic phase was
dried over magnesium sulfate, filtered and the solvent was removed
under reduced pressure to yield 4.33 g of a crude product as a
colorless solid. Column chromatography (silica, petrol ether/ethyl
acetate=7/1 to 3/1) resulted in
(1R,3S,6S)-(7-thia-bicyclo[4.1.0]hept-3-yl)-carbamic acid
tert-butyl ester+(1S,3R,6R) diastereomer (petrol ether/ethyl
acetate=5/1, R.sub.f=0.54, 1.88 g, 49% yield based on recovered
starting material) as colorless crystals. Fp=105-108.degree. C.
[0322] .sup.1H NMR (500 MHz, CDCl.sub.3, .delta., ppm, inter alia):
4.40 (bs, 1H, NH), 3.78 (bs, 1H, 1-H), 3.28-3.20 (m, 1H, 3-H),
3.18-3.09 (m, 1H, 4-H), 2.54 (dd, 1H, 2a-H, J=5 Hz and J=15 Hz),
2.40-2.18 (m, 2H, 5-H), 1.98-1.87 (m, 1H, 2b-H), 1.86-1.72 (m, 1H,
6a-H), 1.58 (s, 9H, tert-butyl), 1.34-1.20 (m, 1H, 6b-H). MS-ESI
(m/z): 252 (MNa.sup.+).
Step B.
14-O-{[(1R,5R,5R)-3-oxo-2-oxa-4-aza-bicyclo[3.3.1]non-8-ylsulfanyl-
]-acetyl}-mutilin+(1S,5S,8S) diastereomer
[0323] To a solution of
(1R,3S,6S)-(7-thia-bicyclo[4.1.0]hept-3-yl)-carbamic acid
tert-butyl ester+(1S,3R,6R) diastereomer (2.95 g, 12.9 mmol) in 160
ml of dichloromethane was added p-toluene sulfonic acid (1.21 g,
6.50 mmol) under cooling. The resulting mixture was stirred at room
temperature over night and the solvent was removed under reduced
pressure to yield 3.26 g of a colorless solid. The crude product
was subsequently dissolved in 150 ml of anhydrous tetrahydrofuran,
and pleuromutilin tosylate (13.1 g, 24.6 mmol) followed by
1,8-diazabicyclo[5.4.0]undec-7-ene (DBU, 4.2 ml, 28.2 mmol) was
added under cooling. The resulting mixture was stirred at room
temperature overnight and water was added. The mixture was
extracted four times with ethyl acetate and the combined organic
phases were washed with water and brine, dried over magnesium
sulfate and the solvent was removed under reduced pressure to yield
9.32 g of a colorless solid. Chromatography (silica,
dichloromethane/methanol=19/1) resulted in
14-O-{[(1R,5R,8R)-3-oxo-2-oxa-4-aza-bicyclo[3.3.1]non-8-ylsulfanyl]-acety-
l})-mutilin+(1S,5S,8S) diastereomer (dichloromethane/methanol=20/1,
R.sub.f=0.45, 4.36 g, 63% yield) as colorless solid. .sup.1H NMR
(500 MHz, CDCl.sub.3, .delta., ppm, inter alia): 6.46 (dd, 1H,
19-H, J=7 Hz and J=11 Hz), 5.77 (d, 1H, 14-H, J=8 Hz), 5.52 (bs,
1H, NH), 5.40-5.17 (m, 2H, 20-H), 4.68-4.55 (m, 1H, 2'-H), 3.63
(bs, 1H, 4'-H), 3.41-3.30 (m, 2, 1'-H, 1-H), 3.27-3.12 (m, 2H,
22-H), 1.45 (s, 3H, 15-CH.sub.3), 1.18 (s, 3H, 18-CH.sub.3), 0.89
(d, 3H, 17-CH.sub.3, J=7 Hz), 0.73 (dd, 3H, 16-CH.sub.3, J=2 Hz and
J=7 Hz). MS-ESI (m/z): 556 (MNa.sup.+).
Step C. 14-O-{[5-Amino-2-chloro-cyclohexylsulfanyl]-acetyl}-mutilin
acetate and
14-O-{[4-Amino-2-chloro-cyclohexylsulfanyl]-acetyl}-mutilin
acetate
[0324] To a solution of
14-O-{[(1R,5R,8R)-3-oxo-2-oxa-4-aza-bicyclo[3.3.1]non-8-ylsulfanyl]-acety-
l}-mutilin+(1S,5S,8S) diastereomer (500 mg, 0.94 mmol) in 2.5 ml of
dioxane was added 6M HCl (7 ml) under cooling. The resulting
mixture was stirred for 23 hours and added to a saturated solution
of sodium bicarbonate. The resulting solution was extracted twice
with ethyl acetate and the combined organic phases were washed with
brine. The resulting organic phase was dried over magnesium
sulfate, filtered and the solvent was removed under reduced
pressure to yield 473 mg of crude product. Column chromatography
(silica, dichloromethane/methanol=10/1 containing 1% of acetic acid
and dichloromethane/methanol/diisopropylether/water/acetic
acid=80/20/6/3/2) resulted in
14-O-{[5-Amino-2-chloro-cyclohexylsulfanyl]acetyl}-mutilin acetate
(a) (dichloromethane/methanol/diisopropylether/water/acetic
acid=80/20/6/3/2, R.sub.f=0.5, 178 mg, 36% yield) and
14-O-{[4-Amino-2-chloro-cyclohexylsulfanyl]acetyl}-mutilin acetate
(b) (dichloromethane/methanol/diisopropylether/water/acetic
acid=80/20/613/2, R.sub.f=0.43, 91 mg, 18% yield) as a colorless
solids.
[0325] (a): .sup.1H NMR (500 MHz, CDCl.sub.3, .delta., ppm, inter
alia): 6.46 (dd, 1H, 19-H, J=11 Hz and J=17 Hz), 5.78 (d, 1H, 14-H,
J=8 Hz), 5.45-5.15 (m, 2H, 20-H), 4.32 (m, 1H, 2'-H), 3.41-3.30 (m,
2H, 11-H, 1'-H), 3.28-3.14 (m, 2H, 22-H), 3.13-3.0 (m, 1H, 5'-H),
2.02 (s, 3H, CH.sub.3 of acetate), 1.46 (s, 3H, 15-CH.sub.3), 1.19
(s, 3H, 18-CH.sub.3), 0.89 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.73 (d,
3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 526 (MH.sup.+).
[0326] (b): .sup.1H NMR (500 MHz, CDCl.sub.3, .delta., ppm, inter
alia): 6.49 (dd, 1H, 19-H, J=1 Hz and J=17 Hz), 5.77 (d, 1H, 14-H,
J=8 Hz), 5.40-5.15 (m, 2H, 20-H), 3.88-3.73 (m, 1H, 2'-H),
3.50-3.15 (m, 3H, 11-H, 22-H), 3.00-2.70 (m, 2H, 1'-H, 4'-H), 2.03
(s, 3H, CH, of acetate), 1.46 (a, 3H, 15-CH.sub.3), 1.18 (s, 3H,
18-CH.sub.3), 0.88 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.74 (d, 3H,
16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 526 (MH.sup.+).
Example 31
14-O-[(4-Amino-1-hydroxy-cyclohexylmethylsulfanyl)-acetyl]-mutilin
hydrochloride
Step A. (1-Oxa-spiro[2.5]oct-6-yl)-carbamic add tert-butyl
ester
[0327] (4-Methylene-cyclohexyl)-carbamic acid tert-butyl ester
(Raju, B. et al, Bioorganic and Medicinal Chemistry Letters 2004,
14(12), 3103-3107) (2.3 g, 10.9 mmol) was treated with
3-chloroperbenzoic acid (70% purity, 3.76 g, 21.8 mmol,
uncorrected) according to the method of Example 7 Step B. After
work up the title compound was obtained (2.3 g, 93% yield) as a
yellow solid.
[0328] .sup.1H NMR (DMSO-d.sub.6, 400 MHz, .delta., ppm): 6.77 (bd,
1H, NH, J=7 Hz), 3.35 (m, 1H), 2.55, 2.51 (2s, 2H), 1.86-1.75 (m,
2H), 1.73-1.67 (m, 2H), 1.47-1.28 (m, 2H), 1.36 (s, 9H,
tert-butyl), 1.22-1.13 (m, 2H).
Step B.
14-O-[(4-tert-Butoxycarbonylamino-1-hydroxy-cyclohexylmethylsulfan-
yl)-acetyl]-mutilin
[0329] Pleuromutilin thiol (2.6 g, 6.59 mmol) was treated with
(1-Oxa-spiro[2.5]oct-6-yl)-carbamic acid tert-butyl ester (1 g 4.40
mmol) according to the method of Example 1 Step A3. After workup
and chromatography of the mixture (silica, toluene/ethyl
acetate=5/1->3/1) the title compound (toluene/ethyl acetate=1/1,
R.sub.f=0.44, 0.41 g, 15% yield, uncorrected) was obtained as a
colorless foam.
[0330] .sup.1H NMR (DMSO-d.sub.6, 500 MHz, .delta., ppm, inter
alia): 6.66 (d, 1H, NH, J=8 Hz), 6.14 (dd, 1H, 19-H, J=18 Hz and 11
Hz), 5.54 (d, 1H, 14-H, J=8 Hz), 5.10-5.02 (m, 2H, 20-H), 4.49 (bs,
1H, 11-OH), 4.23 (bs, 1H), 3.46-3.28 (m, 3H), 3.08 (bs, 3H, 4'-H),
2.60 (s, 2H, COHCH.sub.2S), 2.40 (bs, 1H, 4-H), 2.18 (m, 1H),
2.12-2.02 (m, 3H), 1.69-1.54 (m, 31), 1.52-1.43 (m, 3H), 1.35 (s,
3H, 15-CH.sub.3), 1.35 (s, 9H, tert-butyl), 1.32-1.21 (m, 4H), 1.08
(s, 3H, 18-CH.sub.3), 1.14-0.98 (m, 1H), 0.81 (d, 3H, 17-CH.sub.3,
J=7 Hz), 0.62 (d, 3H, 16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 644
(MNa.sup.+).
Step C.
14-O-[(4-Amino-1-hydroxy-cyclohexylmethylsulfanyl)-acetyl]-mutilin
[0331]
14-O-[(4-tert-Butoxycarbonylamino-hydroxy-cyclohexylmethylsulfanyl)-
-acetyl]-mutilin (0.36 g, 0.58 mmol) was treated with
trifluoroacetic acid (0.72 ml) according to the method of Example 1
Step B. After workup and chromatography of the mixture (silica,
ethyl acetate/methanol/NH.sub.4OH (25%)=50/50/1) the title compound
(R.sub.f=0.04, 0.13 g, 43% yield) was obtained as colorless
foam.
[0332] .sup.1H NMR (DMSO-d.sub.6, 400 MHz, .delta., ppm, inter
alia): 6.13 (dd, 1H, 19-H, J=18 Hz and 11 Hz), 5.53 (d, 1H, 14-H,
J=8 Hz), 5.10 (d, 1H, 20-H, J=2 Hz), 5.02 (d, 1H, 20-H, J=2 Hz),
4.49 (bs, 1H), 4.21 (bs, 1H), 3.42 (m, 1H), 3.23 (q, 2H, H-22, J=14
Hz), 2.60 (s, 2H, COHCH.sub.2S), 2.47-2.36 (m, 2H), 2.23-2.14 (m,
1H), 2.12-2.04 (m, 3H). 1.68-0.54 (m, 3H), 1.41-1.20 (m, 5H), 1.35
(s, 3H, 15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3), 1.03 (m, 1H), 0.81
(d, 3H, 17-CH.sub.3, J=7 Hz), 0.62 (d, 3H, 16-CH.sub.3, J=7 Hz).
MS-ESI (m/z)=522 (MH.sup.+), 544 (MNa.sup.+).
Step D.
14-O-[(4-Amino-1-hydroxy-cyclohexylmethylsulfanyl)-acetyl]-mutilin
hydrochloride
[0333] A solution of
14-O-[(4-amino-1-hydroxy-cyclohexylmethylsulfanyl)-acetyl]-mutilin
(0.1 g, 0.19 mmol) in 1 ml dioxane was treated with aqueous
hydrochloric acid (0.05 M, 11.6 ml, 0.58 mmol) under stirring
according to the method of Example 1 Step C. After 1 hour the
mixture was lyophilized overnight to give the title compound (107
mg, 99% yield) as white solid.
[0334] .sup.1H NMR (DMSO-d.sub.6, 400 MHz, .delta., ppm, inter
alia): 7.88 (s, 3H, NH.sub.3.sup.+), 6.13 (dd, 1H, 19-H, J=18 Hz
and 11 Hz), 5.52 (d, 1H, J=8 Hz), 5.08 (dd, 1H, 20-H, J=5 Hz and 18
Hz), 5.03 (dd, 1H, 20-H, J=5 Hz and 11 Hz), 3.25 (q, 2H, 22-H, J=14
Hz), 2.85 (m, 1H, 4'-H), 2.62 (s, 2H, COHCH.sub.2S), 2.40 (s, 1H,
4-H), 2.23-2.03 (m, 4H), 1.71-1.56 (m, 6H), 1.47 (m, 1H), 1.36 (s,
3H, 15-CH.sub.3), 1.41-1.19 (m, 4H), 1.05 (s, 3H, 18-CH.sub.3),
0.00 (m, 1H), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.62 (d, 3H,
16-CH.sub.3, J=7 Hz).
Example 32
14-O-{[(R,2R)-2-Hydroxy-5-(3-methylamino-propyl)-cyclohexylsulfanyl]-acety-
l}-mutilin+(1S,2S) diastereomer and
14-O-{[(R,2R)-2-Hydroxy-4-(3-methylamino-propyl)-cyclohexylsulfanyl]-acet-
yl}-mutilin+(1S,2S) diastereomer
Step A. 3-Cyclohex-3-enyl-N-methyl-propionamide
[0335] Methyl amine (8 M in EtOH, 75 ml, 600 mmol) was added to a
mixture of 3-cyclohex-3-enyl-propionic acid methyl ester (German
patent DE 4023848 A1 19920130) (20.0 g, 119 mmol) in 75 mL of
methanol. The mixture was stirred at room temperature for one day.
Additional Methyl amine (8 M in EtOH, 40 ml, 320 mmol) was added to
the mixture and stirring continued for one more day. The mixture
was concentrated and the residue was taken up in ethyl acetate,
washed with 0.5 M aqueous HCl, dried and stripped of the solvent to
give the title compound (19.50 g, 98% yield, uncorrected) as a pale
orange solid.
[0336] .sup.1H NMR (CDCl.sub.3, 200 MHz, .delta., ppm): 5.58 (s,
2H, olefinic H), 5.57 (m, 1H, NH), 2.74 (d, 3H, NCH.sub.3, J=5 Hz),
2.20-1.90 (m, 5H), 1.75-1.40 (m, 5H), 1.15 (m, 1H).
Step B. (3-Cyclohex-3-enyl-propyl)-methyl-amine
[0337] A solution of 3-cyclohex-3-enyl-N-methyl-propionamide (15.5
& 92.7 mmol) in 55 ml of tetrahydrofuran was added dropwise
over a period of 25 min to a suspension of lithium aluminium
hydride (95% purity, 5.3 & 139 mmol, corrected) in 120 ml of
tetrahydrofuran at 0.degree. C. under stirring. The mixture was
refluxed for 4 hours, stirred overnight at room temperature and
quenched with 2 M aqueous NaOH, diluted with tetrahydrofuran,
stirred and filtered. The filtrate was concentrated and the residue
was acidified with 1 M aqueous HCl and washed with dichloromethane.
The aqueous phase was basified with 1 M aqueous NaOH, and extracted
with ethyl acetate. The organic extract was dried and concentrated
to obtain the title compound (9.38 & 66% yield) as pale yellow
oil.
[0338] .sup.1H NMR (CDCl.sub.3, 200 MHz, .delta., ppm): 5.58 (d,
2H, olefinic K, J=2 Hz), 2.50 (t, 2H, J=7 Hz), 2.36 (s, 3H,
NCH.sub.3), 2.15-1.90 (m, 3H). 1.75-1.05 (m, 9H).
Step C. (3-Cyclohex-3-enyl-propyl)-methyl-carbamic add tert-butyl
ester
[0339] Ethyl-diisopropyl-amine (11.3 ml, 66.0 mmol) and
di-tert-butyl-dicarbonate (14.4 g, 66.0 mmol) were added to a
solution of (3-cyclohex-3-enyl-propyl)-methyl-amine (7.50 &
48.9 mmol) in 75 ml of dioxane. The mixture was stirred at room
temperature for 3 days, diluted with ethyl acetate and washed with
cold 0.1 M aqueous HCl, and saturated aqueous sodium bicarbonate,
dried over sodium sulfate and concentrated in vacuo to give a
mixture. After chromatography of the mixture (silica,
dichloromethane) the title compound (14.24 & quantitative
yield, uncorrected) was obtained as pale yellow oil.
[0340] .sup.1H NMR (CDCl.sub.3, 200 MHz, .delta., ppm): 5.65 (d,
2H, olefinic H, J=2 Hz), 3.19 (t, 2H, CH.sub.2N, J=7 Hz), 2.84 (s,
3H, NCH.sub.3), 2.15-1.90 (m, 3H), 1.75-130 (m, 5H), 1.45 (s, 9H,
tert-butyl), 130-1.05 (m, 3H).
Step D. Methyl-[3-(7-oxa-bicyclo[4.1.0]hept-3-yl-propyl]-carbamic
acid tert-butyl ester
[0341] (3-Cyclohex-3-enyl-propyl)-methyl-carbamic acid tert-butyl
ester (14.24 g, 56.2 mmol) was treated with 3-chloroperbenzoic acid
(70% purity, 14.8 g, 60 mmol, corrected) according to the method of
Example 7 Step B and stirred for 3 hours at room temperature. After
work up the title compound (14.6 g, 96% yield, uncorrected) was
obtained as pale yellow oil.
[0342] .sup.1H NMR (CDCl.sub.3, 200 MHz, .delta., ppm): 3.15-3.00
(m, 4H), 2.75 (s, 3H, NCH.sub.3), 2.15-0.70 (m, 11H), 1.38 (s, 9H,
tert-butyl).
Step E.
14-O-{{(1R,2R)-5-[3-(tert-Butoxycarbonyl-methyl-amino)-propyl]-2-h-
ydroxy-cyclohexylsulfanyl}-acetyl}-mutilin+(1S,2S) diastereomer and
14-O-{{(1R,2R)-4-[3-(tert-Butoxycarbonyl-methyl-amino)-propyl]-2-hydroxy--
cyclohexylsulfanyl}-acetyl}-mutilin+(1S,2S) diastereomer
[0343] Methyl-[3-(7-oxa-bicyclo[4.1.0]hept-3-yl)-propyl]-carbamic
acid tert-butyl ester (3.00 g, 11.1 mmol) was treated with
pleuromutilin thiol (6.57 g, 16.7 mmol) according to the method of
Example 1 Step A3 and stirred for 3 days at room temperature. After
workup and chromatography of the mixture (silica, cyclohexane/ethyl
acetate 1:1) a mixture of the title compounds (R.sub.f=0.29, 3.20
g, 43% yield, uncorrected) was obtained as white foam. The mixture
was taken to the next step.
Step F.
14-O-{[(1R,2R)-2-Hydroxy-5-(3-methylamino-propyl)-cyclohexylsulfan-
yl]-acetyl}-mutilin+(1S,2S) diastereomer and
14-O-{[(1R,2R)-2-Hydroxy-4-(3-methylamino-propyl)-cyclohexylsulfanyl]-ace-
tyl}-mutilin+(1S,2S) diastereomer
[0344] HCl (4 M in dioxane, 5.30 ml, 21.2 mmol) was added to the
mixture of compounds from Example 32 Step E (5.63 g, 8.48 mmol,
uncorrected) in 50 ml of dioxane. The mixture was stirred for 4
hours and stripped of the solvent. The residue was pinioned between
dichloromethane and saturated aqueous bicarbonate and the organic
layer was separated, dried and stripped of the solvent to give a
mixture. After chromatography (silica, dichloromethane/methanol/28%
aq. NH.sub.3=91/6/3)
14-O-{[(1R,2R)-2-Hydroxy-5-(3-methylamino-propyl)-cyclohexylsulfanyl]-ace-
tyl}-mutilin+(1S,2S) diastereomer (a)
(dichloromethane/methanol/28%-aq. NH.sub.3=88/8/4, R.sub.f=0.18,
117 mg, 3% yield, uncorrected) and
14-O-{[(1R,2R)-2-Hydroxy-4-(3-methylamino-propyl)cyclohexylsulfanyl]-acet-
yl}-mutilin (b) (dichloromethane/methanol/28%-aq. NH.sub.3=88/8/4,
R.sub.f=0.10, 172 mg, 4% yield, uncorrected) were obtained as white
foams.
[0345] (a): .sup.1H NMR: (400 MHz, DMSO-d.sub.6, .delta., ppm,
inter alia): 6.12 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.53 (d, 1H,
14-H, J=8 Hz), 5.1-5.0 (m, 2H, 20-H), 4.74 (bs, 1H, 2'-OH), 4.49
(bs, 1H, 11-OH), 3.59 (m, 1H), 3.42 (m, 1H, 11-H), 3.20 (m), 2.90
(m, 1H), 2.42-2.36 (m, 3H), 2.24 (s, 3H, CH.sub.3--N), 2.21-2.01
(m), 1.72-1.56 (m), 1.55-1.10 (m), 1.36 (s, 3H, 15-CH.sub.3), 1.05
(s, 3H, 18-CH.sub.3), 1.00 (m, 1H), 0.81 (d, 3H, 17-CH.sub.3, J=7
Hz), 0.62 (d, 3H, 16-CH.sub.3, J=7 Hz).
[0346] (b): .sup.1H NMR: (500 MHz, DMSO-d.sub.6, .delta., ppm,
inter alia): 6.13 (m, 1H, 19-H), 5.54 (d, 1H, H-14, J=9 Hz),
5.10-5.00 (m, 2H, 20-H), 4.74 (bs, 1H, 2'-OH), 4.48 (bs, 1H,
11-OH), 3.70 (bs, 1H, 2'-H), 3.41 (m, 1H, 11-H), 3.20 (m, 2H,
H-22), 2.85 (m, 1H), 2.43-2.37 (m, 3H), 2.24 (s, 3H, CH.sub.3--N),
2.22-1.89 (m), 1.70-0.96 (m), 1.35 (s, 3H, 15-CH.sub.3), 1.05 (s,
3H, 18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.62 (d, 3H,
16-CH.sub.3, J=7 Hz).
Example 33
14-O-{[(1R,2R)-5-(3-Amino-propyl)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin+(1S,2S) diastereomer and
14-O-{[(1R,2R)-4-(3-Amino-propyl)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-m-
utilin+(1S,2S) diastereomer
Step A. N-tert-Butoxycarbonyl-(3-cyclohex-3-enyl-propyl)-carbamic
acid tert-butyl ester
[0347] Sodium hydride (60% in mineral oil, 2.5 g, 62.5 mmol,
corrected) was added in several portions over a period of 15 min to
an ice-cold mixture of di-tert-butyl-iminodicarboxylate (22.0 g,
100 mmol) in 60 ml of dimethyl formamide and 180 ml of
tetrahydrofuran under stirring. A solution of toluene-4-sulfonic
acid 3-cyclohex-3-enyl-propyl ester (Marvell, E.; Stunner, D.;
Kunston, R. Journal of Organic Chemistry 1968, 33, 2991-2993) (14.8
g, 50.0 mmol) in a mixture of 15 ml of dimethyl formamide and 45 ml
of tetrahydrofuran was charged to it dropwise over 30 minutes. The
mixture was stirred for 7 hours at 70.degree. C. and 16 hours at
room temperature, diluted with water and extracted with tert-butyl
methyl ether. The organic extract was washed with water, brine and
stripped of the solvent to give a mixture. After chromatography of
the mixture (silica, toluene/cyclohexane=75/25) the title compound
(ethyl acetate/toluene=15/85, R.sub.f=0.19, 14.7 g 86% yield,
uncorrected) was obtained as clear yellow oil.
[0348] .sup.1H NMR (CDCl.sub.3, 200 MHz, .delta., ppm): 5.57 (d,
2H, olefinic H, J=2 Hz), 3.46 (t, 21, CH.sub.2N, J=8 Hz), 2.15-1.90
(m, 3H), 1.75-130 (m, 5H), 1.43 (s, 18H, tert-butyl), 1.30-1.00 (m,
3H).
Step B.
N-tert-Butoxycarbonyl-[3(7-oxa-bicyclo[4.1.0]hept-3-yl)-propyl]-ca-
rbamic acid tert-butyl ester
[0349] N-tert-Butoxycarbonyl-(3-cyclohex-3-enyl-propyl)-carbamic
acid tert-butyl ester (12.2 g, 29.4 mmol, corrected) was treated
with 3-chloroperbenzoic acid (70% purity, 8.7 g, 35.3 mmol,
corrected) according to the method of Example 7 Step B and stirred
at room temperature for 4.5 hours. After work up the tide compound
(9.38 g, 90% yield) was obtained as light yellow oil.
[0350] .sup.1H NMR (CDCl.sub.3, 200 MHz, .delta., ppm): 3.43 (t,
2H, CH.sub.2N, J=8 Hz), 3.06 (bs, 2H), 2.15-0.70 (m, 11H), 1.43 (s,
18H, tert-butyl).
Step C.
14-O-{[(1R,2R)-(3-N,N-Bis-(tert-butoxycarbonyl-amino-propyl)-2-hyd-
roxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S) diastereomer and
14-O-{[(1R,2R)-4-(3-N,N-Bis-(tert-butoxycarbonyl)-amino-propyl)-2-hydroxy-
-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,2S) diastereomer
[0351]
N-tert-Butoxycarbonyl-[3-(7-oxa-bicyclo[4.1.0]hept-3-yl)-propyl]-ca-
rbamic acid tert-butyl ester (3.00 g, 844 mmol) was treated with
pleuromutilin thiol (5.00 g. 12.7 mmol) according to the method of
Example 1 Step A3 and stirred for 24 h. After chromatography of the
mixture (silica, petroleum benzene/ethyl acetate=7/3.fwdarw.1/1) a
mixture of the title compounds (petroleum benzene/ethyl
acetate=3/2, R.sub.f=0.30, 3.68 &g 58% yield, uncorrected) was
obtained as white foam.
[0352] .sup.1H NMR (DMSO-d.sub.6, 500 MHz, .delta., ppm, inter
alia): 6.12 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.53 (d, 1H, 14-H,
J=8 Hz), 5.10-5.00 (m, 2H, 20-H), 4.75 (bs, 1H), 4.48 (bs, 1H,
11-OH), 3.69 (m, 0.5H), 3.60 (m, 0.5H), 3.45-3.38 (m, 3H), 3.26 (d,
1H, 22-H, J=12 Hz), 3.21 (d, 1H, 22-H, J=13 Hz), 2.91 (m, 0.5H),
2.85 (m, 0.5H), 2.39 (bs, 1H, 4-H), 2.18 (dd, 1H, 2-H, J=11 Hz and
19 Hz), 2.12-2.01 (m, 3H), 1.92 (m, 1H), 1.70-0.96 (m), 1.41 (s,
18H, tert-butyl), 1.35 (s, 3H, 15-CH.sub.3), 1.05 (s, 3H,
18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.62 (d, 3H,
16-CH.sub.3, J=7 Hz).
Step D.
14-O-{[(1R,2R)-5-(3-Amino-propyl)-2-hydroxy-cyclohexylsulfanyl]-ac-
etyl}-mutilin+(1S,2S) diastereomer and
14-O-{[(1R,2R)-4-(3-Amino-propyl)-2-hydroxy-cyclohexylsufanyl]-acetyl}-mu-
tilin+(1S,2S) diastereomer
[0353] A mixture of the mixture from Example 33 Step C (250 mg,
0.33 mmol) was treated with trifluoroacetic acid (6 ml) according
to the method of Example 1 Step B, and stirred at room temperature
for 5 days. After work up and chromatography (silica,
dichloromethane/methanol/28% aq NH.sub.4OH=86:10:4) a mixture of
the title compounds (R.sub.f=0.08 and 0.12, 50 mg, 27% yield,
uncorrected) was obtained as white foam.
[0354] .sup.1H NMR (DMSO-d.sub.6, 500 MHz, .delta., ppm, inter
alia): 6.14 (dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.55 (d, 1H, 14-H,
J=8 Hz), 5.13-5.02 (m, 2H, 20-H), 4.75 (bs, 1H), 4.50 (bs, 1H,
11-OH), 3.72 (m, 0.5H), 3.61 (m, 0.5H), 3.50-3.05 (m), 2.92 (m,
0.5H), 2.86 (m, 0.5H), 2.47 (m, 2H, CH.sub.2N), 2.41 (bs, 1H, 4-H),
2.19 (dd, 1H, 2-H), 2.13-2.03 (m, 3H), 1.94 (m, 1H), 1.73-0.82 (m),
1.37 (s, 3H, 15-CH.sub.3), 1.06 (s, 3H, 18-CH.sub.3), 0.82 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.63 (d, J=7 Hz, 3H, 16-CH.sub.3). MS-ESI
(m/z): 550 (MH.sup.+), 572 (MNa.sup.+).
Example 34
14-O-[(4-Amino-2-oxo-cyclohexylsulfanyl)-acetyl]-mutilin
Step A. Thiobenzoic acid
S-((1R,2R,4R)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexyl)
ester+(1S,2S,4S) diastereomer and Thiobenzoic acid
S-((1R,2R,5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclo hexyl)
ester+(1S,2S,5R) diastereomer
[0355] To a solution of syn-3,4-epoxycyclohexyl-carbamic acid
tert-butyl ester (63.3 g, 0.29 mol) in 630 ml of toluene was added
thiobenzoic acid (105.13 ml, 0.90 mmol) followed by tetrabutyl
ammonium chloride monohydrate (2.66 g, 9.00 mmol). The mixture was
stirred under argon for 3.5 hours and was charged with saturated
aqueous sodium bicarbonate, stirred for 10 min and the organic
phase was separated. The organic phase was washed with saturated
aqueous sodium bicarbonate, brine, dried and the solvent was
removed under vacuum to obtain the crude mixture of the title
compounds. The mixture was crystallized from a mixture of
toluene/heptane (1/1) to give thiobenzoic acid
S-((1R,2R,4R)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexyl)
ester+(1S,2S,4S) diastereomer (a) (22.1 g, 21% yield) as a solid.
The mother liquor was chromatographed (silica, toluol/ethyl
acetate=8/1->7/1) to obtain thiobenzoic acid
S-((1R,2R,4R)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexyl)
ester+(1S,2S,4S) diastereomer (a) (toluene/ethyl acetate=3/1,
R.sub.f=0.35, 4.09 g, 4% yield) as a solid and thiobenzoic acid
S-((1R,2R,5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclohexyl)
ester+(1S,2S,5R) diastereomer (b) (toluene/ethyl acetate=3/1,
R.sub.f=0.25, 16.57 g, 16% yield) as oil.
[0356] (a): .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm): 7.88
(d, 2H, aromatic H, J=7 Hz), 7.66 (t, 1H, aromatic H, J=7 Hz), 7.53
(t, 2H, aromatic H, J=8 Hz), 6.82 (d, 1H, NH, J=8 Hz), 5.08 (d, 1H,
OH, J=6 Hz), 3.41 (m, 1H), 3.33-3.28 (m, 2H), 2.08 (bd, 1H), 1.98
(m, 1H), 1.75 (m, 1H), 1.36 (s, 9H, tert-butyl), 1.49-1.18 (m,
3H).
[0357] (b): .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm): 7.91
(d, 2H, aromatic H, J=8 Hz), 7.67 (t, 1H, aromatic H, J=7 Hz), 7.53
(t, 2H, aromatic H, J=8 Hz), 6.85 (d, 1H, NH, J=7 Hz), 5.12 (d, 1H,
OH, J=3 Hz), 3.89 (d, 1H, J=4 Hz), 3.62 (bs, 1H), 3.41 (bs, 1H),
2.10 (m, 1H), 1.69-1.48 (m, 5H), 1.35 (s, 9H, tert-butyl).
Step B. Thiobenzoic acid
S-(4-tert-butoxycarbonylamino-2-oxo-cyclohexyl) ester
[0358] A mixture of thiobenzoic acid
S-((1R,2R,4R)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexyl)
ester+(1S,2S,4S) diastereomer (5 g, 14.2 mmol), 4 .ANG.-molecular
sieve (3 g) 1,1-dihydro-1,1,1-triacetoxy-1,2-benziodoxol-3(1H)-one
(6.33 g, 15 mmol) in 100 ml of dichloromethane was stirred under
argon at 5.degree. C. for 1 hour and 1 hour at room temperature.
The mixture was filtered over celite, dried and concentrated under
vacuum to give the crude product. After chromatography of the
mixture (silica, toluene/ethyl acetate=7/1) the title compound
(toluene/ethyl acetate 3/1, R.sub.f=0.48, 4.18 g, 84% yield) was
obtained as a white solid.
[0359] .sup.1H NMR (DMSO-d 200 MHz, .delta., ppm): 7.83 (d, 2H,
aromatic H, J=7 Hz,), 7.71 (t, 1H, aromatic H, J=7 Hz), 7.57 (t,
2H, aromatic H, J=7 Hz), 7.16 (bd, 1H, NH, J=8 Hz), 4.48 (m, 1H),
3.98, 3.7 (2m, 1H), 2.58 (m, 2H), 2.31-1.68 (m, 4H), 1.39 (s, 9H,
tert-butyl).
Step C.
14-O-[(2-Benzoyloxy-4-tert-butoxycarbonylamino-cyclohex-1-en-ylsul-
fanyl) acetyl]-mutilin
[0360] A mixture of thiobenzoic acid
S-(4-tert-butoxycarbonylamino-2-oxo-cyclohexyl) ester (2 g, 5.72
mmol), pleuromutilin tosylate (3.96 g, 7.44 mmol), potassium
carbonate (1.58 g, 11.44 mmol) and tetrabutyl ammonium chloride
monohydrate (0.2 g, 0.68 mmol) in 20 ml of dimethyl formamide and 2
ml of water was stirred for 24 hours. The mixture was taken up in
ethyl acetate, washed with aqueous saturated sodium bicarbonate and
brine, dried and concentrated under vacuum. After chromatography of
the mixture (silica, toluene/ethyl acetate=5/1) the title compound
(toluene/ethylacetate=3/1, R.sub.f=0.24, 2.36 g. 58% yield) was
obtained as white foam.
[0361] .sup.1H NMR (DMSO-d.sub.6, 400 MHz, .delta., ppm, inter
alia): 7.99 (d, 2H, aromatic H, J=7 Hz), 7.70 (t, 1H, aromatic H,
J=7 Hz), 7.54 (t, 2H, aromatic H, J=8 Hz), 6.91 (bd 1H, NH, J=5
Hz), 6.13 (dd, 1H, 19-H, J=18 Hz and 1Hz), 5.53 (dd, 1H, 14-H, J=3
Hz and 8 Hz), 5.10 (dd, 1H, 20-H, J=2 Hz and 17 Hz), 5.04 (dd, 1H,
20-H, J=2 Hz and 11 Hz), 4.49 (d, 1H, 11-OH, J=6 Hz), 3.64 (bs,
4'-H, 1H), 3.49-3.35 (m, 3H), 2.46-2.34 (m, 3H), 2.28-2.02 (m, 4H),
1.87 (m, 1H), 1.67-1.42 (m, 5H), 1.41-1.36 (m, 1H), 1.37 (s, 9H,
tert-butyl), 1.34 (a, 3H, 15-CH.sub.3), 1.23 (m, 1H), 1.06 (s, 3H,
18-CH.sub.3), 1.00 (m, 1H), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.58
(d, 3H, 16-CH.sub.3, J=7 Hz, 3H). MS-ESI (m/z): 732
(MNa.sup.+).
Step D.
14-O-[(4-tert-Butoxycarbonylamino-2-oxo-cyclohexylsulfanyl)-acetyl-
]-mutilin
[0362] A solution of
14-O-[(2-benzoyloxy-4-tert-butoxycarbonylamino-cyclohex-1-en-ylsulfanyl)--
acetyl]-mutilin (0.8 g. 1.13 mmol) in 8 ml of methanol and 1.25 ml
of aqueous 1M sodium hydroxide (1.25 mmol) was stirred for 0.5
hours. The mixture was diluted with water and extracted with
dichloromethane. The organic extract was dried over magnesium
sulfate and concentrated under vacuum to give a mixture. After
chromatography of the mixture (silica, toluene/ethyl acetate=5/1)
the title compound (toluene/ethyl acetate=3/1, R.sub.f=0.13, 0.57
g, 83% yield) was obtained as white foam.
[0363] .sup.1H NMR (DMSO-d.sub.6, 400 MHz, .delta., ppm, inter
alia): 7.04 (m, 1H, NH), 6.12 (m, 1H, 19-H), 5.53 (m, 1H, 14-H),
5.09-5.02 (m, 2H, 20-H), 4.49 (m, 1H, 11-OH), 3.65 (m, 1H, 4'-H),
3.40 (m, 1H), 3.32-3.14 (m, 2H), 2.39 (s, 1M, 4-H), 2.40-0.95 (m),
1.35 (s, 9H, tert-butyl), 1.34 (s, 3H, 15-CH.sub.3), 1.05 (s, 3H,
18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.60 (d; 3H,
16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 628 (MNa.sup.+).
Step E.
14-O-[(4-Amino-2-oxo-cyclohexylsulfanyl)-acetyl]-mutilin
[0364]
14-O-[(4-tert-Butoxycarbonylamino-2-oxo-cyclohexylsulfanyl)-acetyl]-
-mutilin (0.55 g, 0.91 mmol) was treated with 2.3 ml of 4M HCl in
dioxane (9.2 mmol) according to the method of Example 32 Step F and
stirred for 2 hours at room temperature. After work up the title
compound (0.42 g, 91% yield) was obtained as white foam.
[0365] .sup.1H NMR (DMSO-d.sub.6, 400 MHz, .delta., ppm, inter
alia): 6.13 (m, 1H, 19-H), 5.53 (d, 1H, 14-H, J=4 Hz), 5.05 (m, 2H,
20-H), 4.48 (bs, 1H, 1-OH), 3.55-3.10 (m), 2.41-2.30 (m), 2.22-2.13
(m, 2H), 2.12-1.99 (m, 3H), 1.69-1.55 (m, 3H), 1.55-1.20 (m), 1.35
(s, 3H, 15-CH.sub.3), 1.05 (s, 3H, 18-CH.sub.3), 0.99 (m, 1H), 0.81
(d, 3H, 17-CH.sub.3, J=7 Hz), 0.61 (d, 3H, 16-CH.sub.3).
Example 35
14-O-[(5-Amino-2-oxo-cyclohexylsulfanyl)-acetyl]-mutilin succinic
acid salt
Step A. Thiobenzoic acid
S-(5-tert-butoxycarbonylamino-2-oxo-cyclohexyl) ester
[0366] Thiobenzoic acid
S-((1R,2R,5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclohexyl)
ester+(1S,2S,5R) diastereomer (8.7 g, 24.8 mmol) from Example 34
Step A was treated with
1,1-dihydro-1,1,1-triacetoxy-1,2-benziodoxol-3(1H)-one (11.02 g, 26
mmol) according to the method of Example 34 Step B. After work up
and chromatography (silica, toluene/ethyl acetate=6/1) of the
mixture the title compound (toluene/ethyl acetate=3/1,
R.sub.f=0.43, 7.15 g, 83% yield) was obtained as a white solid.
[0367] .sup.1H NMR (DMSO-d.sub.6, 200 MHz, .delta., ppm): 7.91 (d,
2H, aromatic H, J=7 Hz), 7.71 (t, 1H, aromatic H, J=7 Hz), 7.55 (t,
2H, aromatic H, J=7 Hz), 6.97 (d, 1H, NH, J=7 Hz), 4.70 (m, 1H),
4.02 (m, 1H), 2.79 (m, 1H), 2.46-2.2 (m, 2H). 2.18-1.93 (m, 2H),
1.75 (m, 1H), 1.39 (s, 9H, tert-butyl).
Step B.
14-O-[(2-Benzoyloxy-5-tert-butoxycarbonylamino-cyclohex-1-en-ylsul-
fanyl)-acetyl]-mutilin
[0368] Thiobenzoic acid
S-(5-tert-butoxycarbonylamino-2-oxo-cyclohexyl) ester (1 g, 2.86
mmol) was treated with pleuromutilin tosylate (1.98 g, 3.72 mmol)
according to the method of Example 34 Step C. After work up and
chromatography (silica, toluene/ethyl acetate=5/1) of the mixture
the title compound (toluene/ethyl acetate=3/1, R.sub.f=0.23, 1.33
g, 65% yield) was obtained as white foam.
[0369] .sup.1H NMR (DMSO-d.sub.6, 500 MHz, .delta., ppm, inter
alia): 8.00 (d, 2H, aromatic H, J=8 Hz), 7.70 (t, 1H, aromatic H,
J=7 Hz), 7.54 (t, 2H, aromatic H, J=8 Hz), 6.96 (d, 0.5H, J=8 Hz),
6.92 (d, 0.5H, J=8 Hz), 6.12 (dd, 1H, 19-H, J=18 Hz and 1 Hz), 5.52
(dd, 1H, 14-H, J=3 Hz and 8 Hz), 5.08 (dd, 1H, H-20, J=2 Hz and 18
Hz), 5.05 (dd, 1H, H-20, J=2 Hz and 11 Hz), 4.51 (t or 2d, 1H, J=6
Hz), 3.63 (bs, 1H), 3.47-3.33 (m, 3H), 2.55 (m, 1H), 2.44-2.32 (m,
2H), 2.29-2.14 (m, 3H), 2.10-2.01 (m, 3H), 1.84 (m, 1H), 1.68-1.57
(m, 3H), 1.46 (m, 1H), 1.39 (s, 9H, tert-butyl), 1.34, 1.32 (2s,
3H, 15-CH.sub.3), 1.29-1.18 (m, 3H), 1.05 (s, 3H, 18-CH.sub.3),
0.98 (m, 1H), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.58 (d, 3H,
16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 732 (MNa.sup.+).
Step C.
14-O-[(5-tert-Butoxycarbonylamino-2-oxo-cyclohexylsulfanyl)-acetyl-
]-mutilin
[0370]
14-O-[(2-Benzoyloxy-5-tert-butoxycarbonylamino-cyclohex-1-en-ylsulf-
anyl)-acetyl]-mutilin (1 g, 1.41 mmol) was treated with 1.55 ml of
aqueous 1M sodium hydroxide (1.55 mmol) according to Example 34
Step D. After work up and chromatography of the mixture (silica,
toluene/ethyl acetate=5/1) the title compound (toluene/ethyl
acetate=3/1, R.sub.f=0.22, 0.6 g, 70% yield) was obtained as white
foam.
[0371] .sup.1H NMR (DMSO-d.sub.6, 400 MHz, .delta., ppm, inter
alia): 6.89 (d, 0.5H, NH, J=6 Hz), 6.86 (d, 0.5 H, J=8 Hz),
6.17-6.05 (m, 1H, 19-H), 5.51 (d, 1H, 14-H, J=8 Hz), 5.09-4.99 (m,
2-, 20-H), 4.47 (m, 1H, 11-OH), 3.92, 3.55 (2m, 1H, 1'-H), 3.79 (m,
1H, 5-H), 3.40 (t, 1H, 11-H, J=6 Hz), 3.35-3.12 (m, 2H), 2.77, 2.50
(2m, 1H), 2.41-1.90 (m), 1.71-1.18 (m), 1.37 (s, 9H, tert-butyl),
1.35 (s, 3H, 15-CH.sub.3), 1.04 (s, 3H, 18-CH.sub.3), 0.99 (m, 1H),
0.80 (d, 3H, 17-CH.sub.3, J 7 Hz), 0.59 (d, 3H, 16-CH.sub.3, J=6
Hz).
Step D.
14-O-[(5-Amino-2-oxo-cyclohexylsulfanyl)-acetyl]-mutilin
[0372]
14-O-[(5-tert-Butoxycarbonylamino-2-oxo-cyclohexylsulfanyl)-acetyl]-
-mutilin (0.3 g. 0.5 mmol) was treated with 1.25 ml of 4M HCl in
dioxane (5 mmol) according to the method of Example 32 Step F and
stirred for 2 hours. After work up the title compound (0.26 g,
quantitative yield, uncorrected) was obtained as white foam and was
directly taken to the next step.
[0373] MS-ESI (m/z): 506.0 (MH.sup.+).
Step E. 14-O-[(5-Amino-2-oxo-cyclohexylsulfanyl)-acetyl]-mutilin
succinic acid salt
[0374] A solution of succinic acid (59 mg, 0.5 mmol) in 5 ml of
iso-propanol was added under stirring over a period of 5 min to a
solution of
14-O-[(5-Amino-2-oxo-cyclohexylsulfanyl)-acetyl]-mutilin (0.26 g,
0.5 mmol) in 10 ml of methyl tert-butyl ether and 5 ml of
isopropanol. The mixture was stirred for 4 h and stripped of the
solvent. The residue was dissolved in 2 ml of isopropanol and 20 ml
of methyl tert-butyl ether were added to it and stirred for 1 hour.
The precipitate was filtered, washed with 5 ml of methyl tert-butyl
ether and dried under vacuum to obtain the title compound (0.18 g,
58% yield) as a white solid.
[0375] .sup.1H NMR (DMSO-d.sub.6, 400 MHz, .delta., ppm, inter
alia): 6.18-6.05 (m, 1H, H-19), 5.52 (d, 1H, 14-H, J=7 Hz),
5.12-4.98 (m, 21, 20-H), 4.50 (bs, 1H, 1-OH), 3.89, 3.53 (2m, 1H,
1'-H), 3.42-3.14 (m), 2.39 (s, 1H), 2.27 (s, 2H), 2.23-1.96 (m),
1.69-1.17 (m), 1.34 (s, 3H, 15-CH.sub.3), 1.08 (as, 3H,
18-CH.sub.3), 1.13-0.93 (m), 0.80 (d, 3H, 17-CH.sub.3, J=7 Hz),
0.59 (d, 3H, 16-CH.sub.3, J=6 Hz).
Example 36
14-O-{[(6R,8R)-8-Amino-1,4-dioxa-spiro[4.5]dec-6-ylsulfanyl]-acetyl}-mutil-
in succinic acid salt+(6S,8S) diastereomer succinic acid salt
Step A. Thiobenzoic acid
S-((6R,8R)-8-tert-butoxycarbonylamino-1,4-dioxa-spiro[4.5]dec-6-yl)
ester+(6S,5S) diastereomer
[0376] A mixture of the compound of Example 35 Step A (2.00 g, 5.72
mmol), 1,2-ethandiol (2.0 ml, 35.8 mmol), 25 ml of dichloromethane
and BF.sub.3 (48% purity, 0.60 ml, 4.25 mmol) was stirred for 2
days. The mixture was diluted with dichloromethane and washed with
saturated aqueous sodium bicarbonate and brine, dried and stripped
of the solvent. After chromatography (silica, ethyl
acetate/cyclohexane=1/9->15/85) the title compound
(toluene/ethyl acetate=9/1, R.sub.f=0.28, 632 mg, 28% yield) was
obtained as foam.
[0377] .sup.1H NMR (DMSO-d.sub.6, 400 MHz, .delta., ppm): 7.89 (m,
2H, aromatic H), 7.66 (t, 1H, aromatic H, J=8 Hz), 7.53 (t, 2H,
aromatic H, J=8 Hz), 6.86 (d, 1H, NH, J=8 Hz), 4.04-3.83 (m, 5H),
3.48 (m, 1H), 2.09-1.40 (m, 6H), 1.35 (s, 9H, tert-butyl).
Step B. ((6R,8R)-6-Mercapto-1,4-dioxa-spiro[4.5]dec-8-yl)-carbamic
acid tert-butyl ester+(6S,8S) diastereomer
[0378] A mixture of thiobenzoic acid
S-((6R,8R)-8-tert-butoxycarbonylamino-1,4-dioxa-spiro[4.5]dec-6-yl)
ester+(6S,8S) diastereomer (695 mg, 1.77 mmol) and hydrazine (80%
aq. solution, 0.10 ml, 2.65 mmol) in 7 ml of dichloromethane was
stirred for 24 hours. The mixture was diluted with dichloromethane
and washed with 1M aqueous HCl, dried and stripped of the solvent
to obtain the title compound (490 mg, 95% yield) as yellow-gray
oil.
[0379] .sup.1H NMR (CDCl.sub.3, 400 MHz, .delta., ppm, inter alia):
4.34 (bs, 1H, NH), 4.13-4.05 (m, 2H), 3.95-3.85 (m, 2H), 3.49 (bs,
1H), 2.98 (m, 1H), 2.25 (m, 1H), 1.90-1.75 (m, 2H), 1.69-1.41 (m),
1.37 (s, 9H, tert-butyl).
Step C.
14-O-{[(6R,8R)-8-tert-Butoxycarbonylamino-1,4-dioxa-spiro[4.5]dec--
6-ylsulfanyl]-acetyl}-mutilin+(6S,8S) diastereomer
[0380] ((6R,8R)-6-Mercapto-1,4-dioxa-spiro[4.5]dec-8-yl)-carbamic
acid tert-butyl ester+(6S,8S) diastereomer (410 mg, 1.42 mmol) was
treated with pleuromutilin tosylate (910 mg, 1.71 mmol) according
to the method of Example 1 Step A3 and stirred for 2.5 hours. The
mixture was concentrated in vacuo and the residue was taken up in
ethyl acetate, washed with water, 0.1M aqueous HCl, aqueous sodium
bicarbonate and brine, dried and stripped of the solvent. After
chromatography (silica, ethyl acetate/toluene=1/4) of the mixture
the title compound (R.sub.f=0.18, 613 mg, 66% yield) was obtained
as pale yellow foam.
[0381] .sup.1H NMR (DMSO-d.sub.6, 400 MHz, .delta., ppm, inter
alia): 6.77 (bm, 1H, NH), 6.13 (dd, 1H, 19-H, J=11 Hz and 18 Hz),
5.54 (d, 1H, 14-H, J=8 Hz), 5.11-5.02 (m, 2H, H-20), 4.47 (m, 1H,
11-OH), 4.04-3.81 (m, 4H), 3.39 (m, 1H, 11-H), 3.35-3.17 (m, 3H),
3.02 (m, 1H), 239 (s, 1H, 4-H), 2.25-1.93 (m), 1.75-1.18 (m), 1.36
(2s, 12H, 15-CH.sub.3, tert-butyl), 1.05-0.95 (m), 1.05 (s, 3H,
18-CH.sub.3), 0.81 (d, 3H, 17-CH.sub.3, J=7 Hz), 0.62 (d, 3H,
16-CH.sub.3, J=7 Hz). MS-ESI (m/z): 672 (MNa.sup.+).
Step D.
14-O-{[(6R,8R)-8-Amino-1,4-dioxa-spiro[4.5]dec-6-ylsulfanyl]-acety-
l}-mutilin+(6S,8S) diastereomer
[0382]
14-O-{[(6R,8R)-8-tert-Butoxycarbonylamino-1,4-dioxa-spiro[4.5]dec-6-
-ylsulfanyl]-acetyl}-mutilin+(6S,8S) diastereomer (250 mg, 0.39
mmol) was treated with 1.5 ml trifluoroacetic acid according to the
method of Example 1 Step B and stirred for 6 hours at mom
temperature. After work up and chromatography (silica,
dichloromethane/methanol=19/1) the title compound
(dichloromethane/methanol=9/1, R.sub.f=0.15-0.54, 160 mg 76% yield)
was obtained as white foam, which was directly taken to the next
step.
Step E.
14-O-{[(6R,8R)-8-Amino-1,4-dioxa-spiro[4.5]dec-6-ylsulfanyl]-acety-
l}-mutilin succinic acid salt+(6S,8S) diastereomer succinic acid
salt
[0383]
14-O-{[(6R,8R)-8-Amino-1,4-dioxa-spiro[4.5]dec-6-ylsulfanyl]-acetyl-
}-mutilin+(6S,8S) diastereomer (120 mg, 0.22 mmol) was treated with
succinic acid (25.7 mg, 0.22 mmol) according to the method of
Example 35 Step E to obtain the title compound (100 mg, 69% yield)
as a pale yellow solid.
[0384] .sup.1H NMR (DMSO-d.sub.6, 400 MHz, .delta., ppm, inter
alia): 6.16, 6.15 (2dd, 1H, 19-H, J=11 Hz and 18 Hz), 5.56 (d, 1H,
14-H, J=8 Hz), 5.14-5.02 (m, 2H, 20-H), 4.53 (bs), 4.08-3.85 (m),
3.48-3.21 (m), 3.12-3.01 (m), 2.42 (bs, 1H, 4-H), 2.30 (s, 4H,
succinic acid), 2.26-2.02 (m), 1.84-1.21 (m), 1.38 (s, 3H,
15-CH.sub.3), 1.08 (s, 3H, 18-CH.sub.3, 0.83 (d, 3H, 17-CH.sub.3,
J=7 Hz), 0.64 (d, 3H, 16-CH.sub.3, J=7 Hz).
Example 37
14-O-{[5-Amino-2-methoxy-cyclohexylsulfanyl]-acetyl}-mutilin and
14-O-{[4-Amino-2-methoxy-cyclohexylsulfanyl]-acetyl}-mutilin
Step A.
14-O-{[5-tert-Butoxycarbonylamino-2-methoxy-cyclohexylsulfanyl]-ac-
etyl}-mutilin and
14-O-{([4-Butoxycarbonylamino-2-methoxy-cyclohexylsulfanyl]-acetyl}-mutil-
in
[0385] An ice-cold solution of
14-O-{[(1R,2R,5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S,2S,5R) diastereomer (4.00 & 6.58 mmol)
from Example 1 Step A in 120 ml of dichloromethane was treated with
BF.sub.3 (48% purity, 0.20 ml, 1.65 mmol) followed by
trimethylsilyl diazomethane (2M in hexane, 0.82 ml, 1.64 mmol)
under stirring. After 20 minutes 3 additional amounts of BF.sub.3
(48% purity, 0.20 ml, 1.65 mmol) followed by 3 additional amounts
of trimethylsilyl diazomethane (2M in hexane, 0.82 ml, 1.64 mmol)
were added each time in time interval of 20 min. The mixture was
stirred at room temperature for 30 min, charged with 200 ml of
saturated aqueous sodium bicarbonate and stirred. The organic phase
was separated and washed with saturated aqueous sodium bicarbonate,
dried and stripped of the solvent to give a mixture. After
chromatography (silica, ethyl acetate/cyclohexane=1/4->1/1) a
mixture of the title compounds (0.49 g. 12% yield) was obtained as
a mixture as white foam.
[0386] .sup.1H NMR (DMSO-d.sub.6, 500 MHz, .delta., ppm, inter
alia): 6.72 (bm, 1H, NH), 6.17-6.09 (2dd, 1H, 19-H, J=11 Hz and 18
Hz), 5.54 (2d, 1H, 14-H, J=8 Hz), 5.10-5.00 (m, 2H, 20-H), 4.49 (d,
1 l-OH, J=6 Hz), 3.50-3.17 (m), 3.21 (2s, 3H, OMe), 2.40 (bs, 1H,
4-H), 2.26-2.01 (m), 1.73-1.56 (m), 1.53-0.96 (m), 1.35 (2s, 12H,
15-CH.sub.3, tert-butyl), 1.05 (s, 3H, 18-CH.sub.3), 0.81 (d, 3H,
17-CH.sub.3, J=7 Hz), 0.62 (d, 3H, 16-CH.sub.3, J=7 Hz).
Step B.
14-O-{[5-tert-Butoxycarbonylamino-2-methoxy-cyclohexylsulfanyl]-ac-
etyl}-mutilin and
14-O-{[4-tert-Butoxycarbonylamino-2-methoxy-cyclohexylsulfanyl]-acetyl}-m-
utilin
[0387]
14-O-{[5-tert-Butoxycarbonylamino-2-methoxy-cyclohexylsulfanyl]-ace-
tyl}-mutilin and 14
O-{[4-tert-Butoxycarbonylamino-2-methoxy-cyclohexylsulfanyl]-acetyl}-muti-
lin (80 mg, 0.13 mmol) was treated with 0.32 ml of 4M HCl in
dioxane (1.28 mmol) according to the method of Example 32 Step F
and stirred for 2 hours. After work up and chromatography (silica,
ethyl acetate/methanol/28% aq NH.sub.3=500/100/1) a mixture of the
title compounds (12 mg, 18% yield, uncorrected) was obtained as
white foam.
[0388] .sup.1H NMR (DMSO-d.sub.6, 500 MHz, .delta., ppm, inter
alia): 6.18-6.05 (m, 1H, 19-H), 5.55 (m, 1H, 14-H), 5.10-5.00 (m,
2H, 20-H), 4.58, 4.50 (2m, 1H, 11-OH), 3.55-3.20 (m), 2.86-2.72
(m), 2.44, 2.40 (2s, 1H, 4-H), 2.22-1.99 (m), 1.78-1.20 (m), 1.06,
1.05 (2s, 3H, 18-CH.sub.3), 1.03 (m, 1H), 0.81 (m, 3H,
17-CH.sub.3), 0.62 (m, 3H, 18-CH.sub.3). MS-ESI (m/z): 522
(MH.sup.+).
* * * * *