U.S. patent application number 11/828224 was filed with the patent office on 2009-05-07 for organic compounds.
This patent application is currently assigned to Nabriva Therapeutics Forschungs GmbH. Invention is credited to Rudolf Badegruber, Werner Heilmayer, Rosemarie Mang, Rodger Novak, Dirk Strickmann.
Application Number | 20090118366 11/828224 |
Document ID | / |
Family ID | 38668870 |
Filed Date | 2009-05-07 |
United States Patent
Application |
20090118366 |
Kind Code |
A1 |
Mang; Rosemarie ; et
al. |
May 7, 2009 |
ORGANIC COMPOUNDS
Abstract
A compound of formula (I) ##STR00001## wherein n is 0 to 4; R is
ethyl or vinyl; R.sub.1 is hydrogen or (C.sub.1-6)alkyl, R.sub.2 is
hydrogen or cycloalkyl including (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, halogen,
(C.sub.3-6)cycloalkyl, or R.sub.1 is hydroxy and R.sub.2 is
formyl.
Inventors: |
Mang; Rosemarie; (Wien,
AT) ; Heilmayer; Werner; (Zillingtal, AT) ;
Badegruber; Rudolf; (Wien, AT) ; Strickmann;
Dirk; (Wien, AT) ; Novak; Rodger; (Wien,
AT) |
Correspondence
Address: |
KNOBBE MARTENS OLSON & BEAR LLP
2040 MAIN STREET, FOURTEENTH FLOOR
IRVINE
CA
92614
US
|
Assignee: |
Nabriva Therapeutics Forschungs
GmbH
Wien
AT
|
Family ID: |
38668870 |
Appl. No.: |
11/828224 |
Filed: |
July 25, 2007 |
Current U.S.
Class: |
514/529 ;
560/125 |
Current CPC
Class: |
A61K 31/215 20130101;
C07C 323/61 20130101; A61P 31/06 20180101; C07C 2601/14 20170501;
C07C 323/52 20130101; A61P 31/04 20180101; C07D 317/72 20130101;
A61P 17/10 20180101; A61K 31/357 20130101; A61P 31/00 20180101;
A61K 31/5375 20130101; C07D 295/096 20130101; C07C 2603/99
20170501 |
Class at
Publication: |
514/529 ;
560/125 |
International
Class: |
A61K 31/215 20060101
A61K031/215; C07C 229/22 20060101 C07C229/22 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 20, 2007 |
EP |
07450053.9 |
Claims
1. A compound of formula (I) ##STR00009## wherein n is 0 to 4; R is
ethyl or vinyl; R.sub.1 is hydrogen or (C.sub.1-6)alkyl, R.sub.2 is
hydrogen or cycloalkyl including (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, halogen,
(C.sub.3-6)cycloakyl, or R.sub.1 is hydroxy and R.sub.2 is
formyl.
2. A compound of formula (II) ##STR00010## wherein n, R.sub.1 and
R.sub.2 are as defined in claim 1.
3. A compound of formula (III) ##STR00011## wherein n, R.sub.1 and
R.sub.2 are as defined in claim 1.
4. A compound of formula (IV) ##STR00012## wherein n, R.sub.1 and
R.sub.2 are as defined in claim 1.
5. A compound according to claim 1, selected from the group
consisting of 14-O-{[(1R, 2R,
4R)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl }-mutilin and the
(1S, 2S 4S) diastereomer thereof 14-O-{[(1R, 2R,
5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin and the
(1S, 2S, 5R) diastereomer thereof 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 and the
(1S, 2S, 5S) diastereomer thereof 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 }-mutilin
and the (1S, 2S, 4S) diastereomer thereof 14-O-{[(1R, 2R,
4R)-4-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin and
the (1S, 2S, 4S) diastereomer thereof 14-O-{[(1R, 2R,
5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl }-mutilin and
the (1S, 2S, 5R) diastereomer thereof 14-O-{[(1R, 2R,
5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl }-mutilin
and the (1S, 2S, 5R) diastereomer thereof 14-O-{[(1R, 2R,
4S)-4-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl }-mutilin
and the (1S, 2S, 4R) diastereomer thereof 14-O-{[(1R, 2R,
5R)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl }-mutilin
and the (1S, 2S, 5S) diastereomer thereof 14-O-{[(1R, 2R,
3R)-3-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl }-mutilin and
the (1S, 2S, 3S) diastereomer thereof 14-O-{[(1R, 2R,
3R)-3-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
and the (1S, 2S, 3S) diastereomer thereof 14-O-{[(1R, 2R,
4S)-4-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutili-
n and the (1S, 2S, 4R) diastereomer thereof 14-O-{[(1R, 2R,
5S)-5-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutili-
n and the (1S, 2S, 5R) diastereomer thereof and 14-O-{[(1R, 2R,
3R/S)-3-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin and the (1S, 2S, 3R/S) diastereomer thereof.
6. The compound according to claim 1 the form of a salt and/or
solvate.
7. A compound according to claim 1 for use as a pharmaceutical drug
substance.
8. 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 claim 1.
9. A pharmaceutical drug composition comprising a compound of claim
1, in association with at least one pharmaceutical excipient.
10. A pharmaceutical drug composition according to claim 9, further
comprising another pharmaceutically active agent.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This applications claims to benefit and priority to European
patent application 07450053.9, filed on Mar. 20, 2007, which is
hereby incorporated by reference in its entirety.
[0002] The present invention relates to organic compounds, namely
pleuromutilins.
[0003] 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.
[0004] 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.
[0005] We have now found pleuromutilins with interesting activity
combined with an unexpexted remarkable metabolic stability.
[0006] The pleuromutilin derivatives according to the invention are
compounds of formula (I)
##STR00003##
wherein n is 0 to 4; R is ethyl or vinyl; R.sub.1 is hydrogen or
(C.sub.1-6)alkyl, R.sub.2 is hydrogen or [0007] cycloalkyl
including (C.sub.3-6)cycloalkyl, or [0008] unsubstituted
(C.sub.1-6)alkyl, or [0009] (C.sub.1-6)alkyl substituted by one or
more of [0010] hydroxy; preferably one or two, [0011] halogen,
[0012] (C.sub.3-6)cycloalkyl, or R.sub.1 is hydroxy and R.sub.2 is
formyl.
[0013] Preferred compounds of the present invention are [0014] a
compound of formula (II)
[0014] ##STR00004## [0015] a compound of formula (III)
##STR00005##
[0015] and [0016] a compound of formula (IV)
##STR00006##
[0016] wherein n, R.sub.1 and R.sub.2 are as defined above.
[0017] Particularly preferred is a compound selected from the group
consisting of
[0018] 14-O-{[(1R, 2R,
4R)4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin and the
(1S, 2S, 4S) diastereomer thereof
[0019] 14-O-{[(1R, 2R,
5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl }-mutilin and the
(1S, 2S, 5R) diastereomer thereof
[0020] 14-O-{[(1R, 2R,
4S)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin and the
(1S, 2S, 4R) diastereomer thereof
[0021] 14-O-{[(1R, 2R,
5R)-5-Amino-2-hydroxy-cyclobexylsulfanyl]-acetyl}-mutilin and the
(1S, 2S, 5S) diastereomer thereof
[0022] 14-O-{[(1R, 2R,
3R)-3-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin and the
(1S, 2S, 3S) diastereomer thereof
[0023] 14-O-{[(1R, 2R,
4R)-4-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
and the (1S, 2S, 4S) diastereomer thereof
[0024] 14-O-{[(1R, 2R,
4R)4-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin and
the (1S, 2S, 4S) diastereomer thereof
[0025] 14-O-{[(1R, 2R,
5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin and
the (1S, 2S, 5R) diastereomer thereof
[0026] 14-O-{[(1R, 2R,
5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
and the (1S, 2S, 5R) diastereomer thereof
[0027] 14-O-{[(1R, 2R,
4S)-4-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
and the (1S, 2S, 4R) diastereomer thereof
[0028] 14-O-{[(1R, 2R,
5R)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
and the (1S, 2S, 5S) diastereomer thereof
[0029] 14-O-{[(1R, 2R,
3R)-3-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin and
the (1S, 2S, 3S) diastereomer thereof
[0030] 14-O-{[(1R, 2R,
3R)-3-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
and the (1S, 2S, 3S) diastereomer thereof
[0031] 14-O-{[(1R, 2R,
4S)4-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
and the (1S, 2S, 4R) diastereomer thereof
[0032] 14-O-{[(1R, 2R,
5S)-5-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutili-
n and the (1S, 2S, 5R) diastereomer thereof
[0033] and
[0034] 14-O-{[(1R, 2R,
3R/S)-3-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin and the (1S, 2S, 3R/S) diastereomer thereof.
[0035] 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.
[0036] According to another aspect, the present invention provides
a compound of the present invention in the form of a salt and/or
solvate.
[0037] The salts include preferably pharmaceutically acceptable
salts, although pharmaceutically unacceptable salts are included,
e.g. for preparation/isolation/purification purposes.
[0038] 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, suceinic acid, salicylic acid,
azelaic acid, 2-[(2,6-dichlorophenyl)amino]bezene acetic acid,
hydrochloric acid, deuterochloric acid, preferably hydrochloric
acid.
[0039] 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.
[0040] 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 enatiomers 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.
[0041] For example, in a compound of formula I the carbon atom of
the cycloalkyl ring which is attached to the sulphur atom, the
carbon atom of the cycloalkyl ring which is attached to the hydroxy
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.
[0042] The configuration of substituents attached to asymmetric
carbon atoms of the mutilin-ring is preferably the same as in
natural pleuromutilin.
[0043] 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.
[0044] The compounds of the present invention exhibit
pharmacological activity and are therefore useful as
pharmaceuticals.
[0045] 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 epidermidis, Staphylococcus haemolyticus, and
Streptococci, e.g. Streptococcus pyogenes, Streptococcus
pneumoniae, Enterococci, e.g. Enterococcus faecium 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.
[0046] 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-A , 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.
[0047] 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.
[0048] 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.
[0049] In another aspect the present invention provides a compound
of the present invention for use in acne treatment.
[0050] 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
[0051] diseases mediated by bacteria, e.g. selected from
Staphylococci, Streptococci, Enterococci;
[0052] diseases mediated by bacteria, e.g. selected from Moraxella,
Haemophilus, Legionella, Neisseriaceae;
[0053] diseases mediated by Helicobacter;
[0054] diseases mediated by Mycobacterium tuberculosis;
[0055] e.g. diseases mediated by Mycoplasms, Chlamydia and
obligatory anaerobes;
and for the treatment of acne.
[0056] 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.
[0057] 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.
[0058] Treatment includes treatment and prophylaxis.
[0059] For antimicrobial and acne treatment, the appropriate dosage
will, of course, vary depending upon, for example, the chemical
nature and the pharmakokinetic 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.
[0060] 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.
[0061] 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.
[0062] 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.
[0063] 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.
[0064] 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,
stabilizers, wetting agents and/or emulsifiers, solubilizers, salts
for regulating osmotic pressure and/or buffers.
[0065] In another aspect the present invention provides a
pharmaceutical composition according to the present invention,
further comprising another pharmaceutically active agent.
[0066] 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.
[0067] 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.
[0068] In another aspect the present invention provides a compound
of the present invention for use as a veterinary agent.
[0069] 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.
[0070] 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.
[0071] Examples 1 to 15 following thereafter exhibit MICs.ltoreq.2
.mu.g/ml against Staphylococcus aureus ATCC49951 and Streptococcus
pneumoniae ATCC49619.
[0072] 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.
[0073] At the compounds of the present invention, the introduction
of a hydroxy group in ortho position to the sulphur substituent in
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
a hydroxy group in the cyclohexyl moiety of the pleuromutilin side
chain.
[0074] 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,
3S)-3-Amino-cyclohexylsulfanyl]-acetyl}-mutilin hydrochloride and
the (1S, S3R) diastereomer hydrochloride thereof--analogous
derivative without hydroxyl group--only 24% of parent compounds
could be detected.
EXAMPLES
[0075] 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-tricyclo[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.):
##STR00007##
[0076] Pleuromutilin thiol and pleuromutilin tosylate are compounds
of formulae:
##STR00008##
Example 1
14-O-{[(1R, 2R,
4R)4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride+(1S, 2S, 4S) diastereomer hydrochloride
[0077] Step A1. 14-O-{[(1R, 2R,
4R)-4-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsuffanyl]-acetyl}-muti-
lin+(1S, 2S, 4S) diastereomer and 14-O-{[(1R, 2R,
5S)-5-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S, 2S, 5R) diastereomer and
14-O-{[(1R, 2R,
4S)-4-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S, 2S, 4R) diastereomer
[0078] 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}-muti-
lin+(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}-muti-
lin+(1S, 2S, 5R) diastereomer and 14-O-{[(1R, 2R,
4S)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S, 2S, 4R) diastereomer (b) (R.sub.f=0.26, 2.81 g, 25%) as
colorless amorphous foams.
[0079] (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 (2MNa.sup.+).
[0080] (b): .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.70 (d, 1H, NH, 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,
1'-H, 2'-H, 4'-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.+).
[0081] or Step A2. 14-O-{[(1R, 2R,
4R)-4-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S, 2S, 4S) diastereomer and
14-O-{[(1R, 2R,
5S)-5-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S, 2S, 5R) diastereomer and 14-O-{[(1R, 2R,
4S)-4-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S, 2S, 4R) diastereomer
[0082] To a solution of 3,4-epoxycyclohexyl-carbamic acid
tert-butyl ester (10 g, 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}-muti-
lin+(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}-muti-
lin+(1S, 2S, 5R) diastereomer and 14-O-{[(1R, 2R,
4S)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S, 2S, 4R) diastereomer (R.sub.f=0.25, 6.35 g, 25%) were
obtained as colorless amorphous foams.
[0083] Step B. 14-O-{[(1R, 2R,
4R)4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S,
4S) diastereomer
[0084] To a solution of 14-O-{[(1R, 2R,
4R)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(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.
[0085] .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.+).
[0086] Step C. 14-O-{[(1R, 2R,
4R)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride+(1S, 2S, 4S) diastereomer hydrochloride
[0087] 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.
[0088] .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 2
14-O-{[(1R, 2R,
5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride+(1s, 2S, 5R) diastereomer hydrochloride
[0089] Step A. 14-O-{[(1R, 2R,
5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S,
5R) diastereomer and
14-O-{[(1R, 2R,
4S)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S,
4R) diastereomer
[0090] A mixture of 14-O-{[(1R, 2R,
5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S, 2S, 5R) diastereomer and 14-O-{[(1R, 2R,
4S)4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutil-
in+(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.
[0091] (a): .sup.1H NMR (400 MHz, DMSO-d.sub.6, 67, 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).
[0092] (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,
.nu..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 (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).
[0093] Step B. 14-O-{[(1R, 2R,
5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride+(1S, 2S, 5R) diastereomer hydrochloride
[0094] 14-O-{[(1R, 2R,
5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(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.
[0095] .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, 11H, J=6
Hz), 3.35 (m, 2H, 22-H), 3.09 (m, 2H, 1'-H, 5'-H), 2.40 (bs, 11H,
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 3
14-O-{[(1R, 2R,
4S)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride+(1S, 2S, 4R) diastereomer hydrochloride
[0096] 14-O-{[(1R, 2R,
4S)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(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.
[0097] .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.8 (bs, 3H, NH.sub.3.sup.+), 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, 1H, 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, 4-4H), 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+(1S, 2S,
5S) diastereomer
[0098] Step A.
tert-Butyl-dimethyl-(cis-3,4-epoxycyclohexyloxy)-silane
[0099] To a solution of 3-cyclohexen-1-ol (Amburgey, J. C.; Shuey,
S. W.; Pedersen, L. G.; Hiskey R., Bioorganic Chemistry 1994, 22,
172-197.) (10 g, 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 pure protected
tert-butyl-dimethyl-(cis-3,4-epoxycyclohexyloxy)-silane
(R.sub.f=0.35, 18.3 g, 79% yield) as colorless oil.
[0100] .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)-5-(tert-Butyl-dimethyl-silyloxy)-2-hydroxy-cyclohexylsulfanyl]-acetyl-
}-mutilin+(1S, 2S, 5R) diastereomer
[0101] 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-cyclohexylsulfanyl]-acetyl-
}-mutilin+(1S, 2S, 5R) diastereomer was obtained as colorless
amorphous foam which was directly used for the next step.
[0102] .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}-mutilin+(1S, 2S, 5R)
diastereomer
[0103] To a solution of 14-O-{[(1R, 2R,
5S)-5-(tert-butyl-dimethyl-silyloxy)-2-hydroxy-cyclohexylsulfanyl]-acetyl-
}-mutilin+(1S, 2S, 5R) diastereomer (9.46 g, 15.1 mmol) in 25 ml of
THF 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, 2S, 5R)
diastereomer (R.sub.f=0.27, 7.07 g, 92% yield) as colorless
amorphous foam.
[0104] .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-cyclohexylsulfanyl]-acetyl}-mutilin+(1-
S, 2S, 5R) diastereomer
[0105] To a solution of 14-O-{[(1R, 2R,
5S)-2,5-dihydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 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]-acetyl}-mutilin+(1-
S, 2S, 5R) diastereomer (R.sub.f=0.15, 2.55 g, 36% yield) as
colorless amorphous foam.
[0106] .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-cyclohexylsulfanyl]-acetyl}-mu-
tilin+(1S, 2S, 5S) diastereomer
[0107] A solution of 14-O-{[(1R, 2R,
5S)-2-hydroxy-5-methanesulfonyloxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1-
S, 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-5-methanesulfonyloxy-cyclohexylsulfanyl]-acetyl}-mu-
tilin+(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.
[0108] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.15, 6.13 (2dd, 1H, 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.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 F. 14-O-{[(1R, 2R,
5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S,
5S) diastereomer
[0109] Triphenylphosphine (1.18 g, 4.50 mmol) was added to a
solution of 14-O-{[(1R,
2R,5R)-5-azido-2-hydroxy-5-methanesulfonyloxy-cyclohexylsulfanyl]-acetyl}-
-mutilin+(1S, 2S, 5S) diastereomer (2.4 g, 4.50 mmol) in 30 ml of
THF 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.
[0110] .sup.1H NMR (400 MHz, DMSO-d6, .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 5
14-O-{[(1R, 2R,
3R)-3-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride+(1S, 2S, 3S) diastereomer hydrochloride
[0111] Step A. 14-O-{[(1R, 2R,
3R)-3-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S, 2S, 3S) diastereomer
[0112] (cis)-2,3-Epoxycyclohexyl-carbamic acid tert-butyl ester
(O'Brien, P.; Childs, A., C.; Ensor, G. Organic Letters 2003,
5(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}-muti-
lin+(1S, 2S, 3S) diastereomer (R.sub.f=0.5, 1.32 g, 46%) was
obtained as colorless amorphous foam.
[0113] .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, lH,
1-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'-1), 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}-mutilin+(1S, 2S,
3S) diastereomer
[0114] 14-O-{[(1R, 2R,
3R)-3-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(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. [0115] 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}-mutilin
hydrochloride+(1S, 2S, 4S) diastereomer hydrochloride
[0116] 14-O-{[(1R, 2R,
3R)-3-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(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.
[0117] .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-cyclobexylsulfanyl]-acetyl}-mutilin+(1S,
2S, 4S) diastereomer and 14-O-{[(1R, 2R,
4R)-4-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,
2S, 4S) diastereomer
[0118] 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}-mutilin+(1S,
2S, 4S) diastereomer (a) (92 mg, 9% yield) and 14-O-{[(1R, 2R,
4R)-4-ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,
2S, 4S) diastereomer (b) (163 mg, 17% yield) as colorless amorphous
foams.
[0119] (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 (.nu..sub.A=3.50,
3.42, .nu..sub.B=3.30 ,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.-.
[0120] (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.B=3.25,22-H, J=15
Hz), 2.55 (m, 2H, 1'-H, 4'-H), 2.40 (bs, 1H, 4-H), 1.35 (s, 3H,
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}-mutilin
hydrochloride+(1S, 2S, 5R) diastereomer hydrochloride
[0121] Step A. N-Ethyl-N-(cyclohex-3-enyl)-carbamic acid tert-butyl
ester
[0122] 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.
[0123] .sup.1H NMR (400 MHz, CDCl.sub.6, .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
[0124] 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 NaHICO.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.
[0125] .sup.1H NMR (400 MHz, CDCl.sub.6, .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-{[(1R, 2R,
5S)-5-(tert-Butoxycarbonyl-ethyl-amino)-2-hydroxy-cyclohexylsulfanyl]-ace-
tyl}-mutilin+(1S, 2S, 5R) diastereomer
[0126] 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-cyclohexylsulfanyl]-ace-
tyl}-mutilin+(1S, 2S, 5R) diastereomer (R.sub.f=0.4, 2.57 g, 65%
yield) was obtained as colorless amorphous foam.
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]-acetyl}-mutilin+(1S,
2S, 5R) diastereomer
[0127] 14-O-{[(1R, 2R,
5S)-5-(tert-Butoxycarbonyl-ethyl-amino)-2-hydroxy-cyclohexylsulfanyl]-ace-
tyl}-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}-mutilin+(1 S,
2S, 5R) diastereomer (R.sub.f=0.3, 1.08 g, 50%) was obtained as
colorless amorphous foam.
[0128] .sup.1H NMR (500 MHz, CDCl.sub.6, .delta., ppm, inter alia):
6.48 (dd, 1H, 19-H, J=10 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-H), 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]-acetyl}-mutilin
hydrochloride+(1S, 2S, 5R) diastereomer hydrochloride
[0129] 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}-mutilin
hydrochloride+(1S, 2S, 5R) diastereomer hydrochloride (83 mg, 90%
yield) as colorless amorphous solid.
[0130] .sup.1H NMR (400 MHz, CDCl.sub.6, .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}-mutilin
hydrochloride+(1S, 2S, 5R) diastereomer hydrochloride
[0131] Step A. 14-O-{[(1R, 2R,
5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilia+(1S,
2S, 5R) diastereomer
[0132] 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}-mutilin+(1S,
2S, 5R) diastereomer (R.sub.f=0.2, 230 mg, 87%) was obtained as
colorless amorphous foam.
[0133] .sup.1H NMR (400 MHz, CDCl.sub.6, .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. 14O-{[(1R, 2R,
5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin
hydrochloride+(1S, 2S, 5R) diastereomer hydrochloride
[0134] 14-O-{[(1R, 2R,
5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-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}-mutilin
hydrochloride+(1S, 2S, 5R) diastereorner hydrochloride (223 mg, 91%
yield) as colorless amorphous solid.
[0135] .sup.1H NMR (400 MHz, CDCl.sub.6, .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, 9H,
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}-mutilin+(1S,
2S, 4R) diastereomer
[0136] 14-O-{[(1R, 2R,
4S)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl} -mutilin+(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}-mutilin+(1S,
2S, 4R) diastereomer (R.sub.f=0.2, 129 mg, 17%) was obtained as
colorless amorphous foam.
[0137] .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, 5'-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-cyclohexylsuffanyl]-acetyl}-mutilin+(1S,
2S, 5S) diastereomer
[0138] 14-O-{[(1R, 2R,
5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(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}-mutilin+(1S,
2S, 5S) diastereomer (R.sub.f=0.2, 95 mg, 20%) was obtained as
colorless amorphous foam.
[0139] .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}-mutilin+(1S,
2S, 3S) diastereomer
[0140] Step A. [(1R, 2R,
3R)-2-Hydroxy-3-(2,4,6-trimethyl-benzylsulfanyl)-cyclobexyl]-carbamic
acid tert-butyl ester+(1S, 2S, 3S) diastereomer
[0141] 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]-carbamic
acid tert-butyl ester+(1S, 2S, 3S) diastereomer (R.sub.f=0.25,
5.92g, 23% yield) as colorless amorphous foam.
[0142] .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, 38)
diastereomer
[0143] A solution of [(1R, 2R,
3R)-2-hydroxy-3-(2,4,6-trimethyl-benzylsulfanyl)-cyclohexyl]-carbamic
acid tert-butyl ester+(1S, 2S, 3S) diastereomer (2.46 g, 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-benzylsulfanyl--
cyclo-hexyl]-carbamic acid tert-butyl ester+(1S, 2S, 3S)
diastereomer (R.sub.f=0.25, 3.0 g, 94% yield) was obtained.
[0144] .sup.1H NMR (500 MHz, DMSO-d6, .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-(2,4,6trimethyl-benzylsulfanyl)-c-
yclohexyl]-ethyl-carbamic acid tert-butyl ester+(1S, 2S, 3S)
diastereomer
[0145] To a solution of [(1R, 2R,
3R)-2-(tert-butyl-dimethyl-silanyloxy)-3-(2,4,6-trimethyl-benzylsulfanyl)-
-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-benzylsulfanyl)-
-cyclohexyl]-ethyl-carbamic acid tert-butyl ester+(1S, 2S, 3S)
diastereomer (1.20 g, 38%) was obtained.
MS-ESI (m/z): 544 (MNa.sup.+), 1065 (2MNa.sup.+). Step D. [(1R, 2R,
3R)-2-(tert-Butyl-dimethyl-silanyloxy)-3-mercapto-cyclohexyl]-ethyl-carba-
mic acid tert-butyl ester+(1S, 2S, 3S) diastereomer
[0146] A solution of [(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 (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 [(1R, 2R,
3R)-2-(tert-butyl-dimethyl-silanyloxy)-3-mercapto-cyclohexyl]-ethyl-carba-
mic 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
[0147] Crude [(1R, 2R,
3R)-2-(tert-butyl-dimethyl-silanyloxy)-3-mercapto-cyclohexyl]-ethyl-carba-
mic acid tert-butyl ester+(1S, 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-dimethyl-silanyloxy-
)-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 3S) diastereomer
(R.sub.f=0.5, 468 mg, 27% yield) was obtained.
[0148] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.14 (dd, 1H, 19-H, J=1 Hz and 18 Hz), 5.55 (d, 1 H, 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-OH, J=6 Hz), 3.28, 3.04 (2m, 7H, 1'-H, 3'-H,
11-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-Etlhylamino-2-(tert-butyl-dimethyl-silanyloxy)-cyclohexylsulfanyl]--
acetyl}-mutilin+(1S, 2S, 3S) diastereomer and 14-O-([(1R, 2R,
3R)-3-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,
2S, 3S) diastereomer
[0149] 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)-cyclohexylsulfanyl]-a-
cetyl}-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}
-mutilin+(1S, 2S, 3S) diastereomer (b) (R.sub.f=0.25, 177 mg, 53%
yield) were obtained as colorless solids.
[0150] (a): MS-ESI (m/z): 672 (MNa.sup.+), 1321 (2MNa.sup.+).
[0151] (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), 5.05 (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, J=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}-mutilin+(1S,
2S, 3S) diastereomer
[0152] Step A. 14-O-{[(1R, 2R,
3R)-3-Diethylamino)-2-(tert-butyl-dimethyl-silanyloxy)-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S, 2S, 35) diastereomer
[0153] 14-O-{[(1R, 2R,
3R)-3-Ethylamino-2-(tert-butyl-dimethyl-silanyloxy)-cyclohexylsulfanyl]-a-
cetyl}-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-butyl-dimethyl-silanyloxy)-cyclohexylsulfanyl-
]-acetyl}-mutilin+(1S, 2S, 3S) diastereomer (R.sub.f=0.5, 110 mg,
73%) was obtained as colorless amorphous foam.
[0154] .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-H), 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 (bs, 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-cyclohexylsulfanyll-acetyl}-mutilin+(1S,
2S, 3S) diastereomer
[0155] To a solution of 14-O-{[(1R, 2R,
3R)-3-diethylamino-2-(tert-butyl-dimethyl-silanyloxy)-cyclohexylsulfanyl]-
-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 THF, 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}-mutilin+(1S,
2S, 3S) diastereomer (R.sub.f=0.2, 8 mg, 10% yield) as colorless
amorphous foam.
[0156] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 6.15, 6.14 (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), 4.99, 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, 1n, 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-E3SI (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]-acetyl}-mutili-
n+(1S, 2S, 4R) diastereomer
[0157] Step A. 14-O-{[(7R,
8R)-7-Hydroxy-1,4-dioxa-spiro[4.51dec-8-ylsulfanyl]-acetyl}-mutilin+(7S,
8S) diastereomer
[0158] 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}-mutilin+(7S,
8S) diastereomer (R.sub.f=0.3, 8.40 g, 76% yield) was obtained as
colorless amorphous foam.
[0159] .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,
S=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 (mn, 1H, 1'-H), 2.40 (bs, 1H, 4-H),
1.35, 1.34 (2s, 3H1, 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.-.
Step B. 14-O-{[(7R,
8R)-7-(tert-Butyl-diphenyl-silanyloxy)-1,4-dioxa-spiro[4.5]dec-8-ylsulfan-
yl]-acetyl}-mutilin+(7S, 8S) diastereomer
[0160] A solution of 14-O-{[(7R,
8R)-7-hydroxy-1,4-dioxa-spiro[4.5]dec-8-ylsulfanyl]-acetyl}-mutilin+(7S,
8S) diastereomer (8.4 g, 15.3 mmol) 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 with ethyl acetate three times. 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-ylsulfan-
yl]-acetyl}-mutilin+(7S, 8S) diastereomer (R.sub.f=0.7, 8.03 g, 67%
yield) was obtained.
[0161] .sup.1H NMR (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, 1H, 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. 14O-{[(1R,
2R)-2-(tert-Butyl-diphenyl-silanyloxy)-4-oxo-cyclohexylsulfanyl]-acetyl}--
mutilin+(1S, 2S) diastereomer
[0162] 14-O-{[(7R,
8R)-7-(tert-Butyl-diphenyl-silanyloxy)-1,4-dioxa-spiro[4.5]dec-8-ylsulfan-
yl]-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-cyclohexylsulfanyl]-acetyl)--
mutilin+(1S, 2S) diastereomer (R.sub.f=0.38, 5.24 g, 69%
yield).
[0163] .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, 2H, 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-(tert-Butyl-diphenyl-silanyloxy)-4-hydroxyimino-cyclohexylsulfanyl]-
-acetyl}-mutilin+(1S, 2S) diastereomer
[0164] To a solution of 14-O-{[(1R,
2R)-2-(tert-butyl-diphenyl-silanyloxy)4-oxo-cyclohexylsulfanyl]-acetyl}-m-
utilin+(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-cyclohexylsulfanyl]--
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-hydroxyamino-cyclohexylsulfanyl]-
-acetyl}-mutilin+(1S, 2S, 4R) diastereomer and 14-O-{[(1R, 2R,
4R)-2-(tert-Butyl-diphenyl-silanyloxy)-4-hydroxyamino-cyclohexylsulfanyl]-
-acetyl}-mutilin+(1S, 2S, 4S) diastereomer
[0165] 14-O-{[(1R,
2R)-2-(tert-Butyl-diphenyl-silanyloxy)-4-hydroxyimino-cyclohexylsulfanyl]-
-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-cyclohexylsulfanyl]-
-acetyl} -mutilin+(1S, 2S, 4R) diastereomer (a) (R.sub.f=0.5, 590
mg, 23% yield) and 14-O-{[(1R, 2R,
4R)-2-(tert-butyl-diphenyl-silanyloxy)-4-hydroxyamino-cyclohexylsulfanyl]-
-acetyl}-mutilin+(1S, 2S, 4S) diastereomer (b) (R.sub.f=0.3, 670
mg, 26% yield).
[0166] (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, 11H, 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).
[0167] (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, 3-11 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 (m, 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,
5'-H), 2.40 (bs, 1H, 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-hydroxy-amino-cyclohexyl-
sulfanyl]-acetyl}-mutilin+(1S, 2S, 4R) diastereomer
[0168] To a solution of 14-O-{[(1R, 2R,
4S)-2-(tert-butyl-diphenyl-silanyloxy)-4-hydroxyamino-cyclohexylsulfanyl]-
-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-diphenylsilanyloxy)-4-(formyl-hydroxy-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.
[0169] .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=11Hz and 18 Hz),
5.50 (d, 1H, 14-H, J=7 Hz), 5.05 (m, 2H, 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-cyclohexylsulfanyl]-acetyl}-mutili-
n+(1S, 2S, 4R) diastereomer
[0170] 14-O-{[(1R, 2R,
4S)-2-(tert-Butyl-diphenyl-silanyloxy)-4-(formyl-hydroxy-amino-cyclohexyl-
-sulfanyl]-acetyl}-mutilin+(1S, 2S, 4R) diastereomer (215 mg, 0.272
mmol) in 10 ml of tetrahydrofuran was treated with
tetrabutylammoniun 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, NaHCO3 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+(1 S,
28, 4R) diastereomer (97 mg, 65% yield,
dichloromethane/methanol=9/1, R.sub.f=0.4) as colorless solid.
[0171] .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 (s, 12H, 18-CH.sub.3), 0.81 (d, 3H,
17-CH.sub.3, J=7 Hz), 6.63 (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]-acetyl}-mutili-
n+(1S, 2S, 5R) diastereomer
[0172] Step A. (7R,
8R)-8-(2,4,6-Trimethylbenzylsufanyl)-1,4-dioxa-spiro[4.5]decan-7-ol+(7S,
8S) diastereomer
[0173] 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-trirethylbenzyl 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-trimethylbenzylsufanyl)-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.
[0174] MS-ESI (m/z): 345 (MNa.sup.+), 667 (2MNa.sup.+).
Step B. Acetic acid (7R,
8R)-7-(2,4,6-trimethylbenzylsufanyl)-1,4-dioxa-spiro[4.5]dec-8-yl
ester+(7S, 8S) diastereomer
[0175] 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-trimethylbenzylsufanyl)-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-trimethylbenzylsufanyl)-1,4-dioxa-spiro[4.5]dec-8-yl
ester+(7S, 8S) diastereomer (R.sub.f=0.4, 7.0 g, 22% yield).
[0176] .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
[0177] Acetic acid (7R,
8R)-7-(2,4,6-trimethylbenzylsufanyl)-1,4-dioxa-spiro[4.5]dec-8-yl
ester+(7S, 8S) diastereomer.(6.33 g, 17.4 mmol) 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.
[0178] .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm): 4.89 (d,
1H, OH), 3.83 (m, 4H, 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]-acetyl}-mutilin+{7S,
8S) diastereomer
[0179] (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}-mutilin+(7S,
8S) diastereomer (R.sub.f=0.25, 1.90 g, 48%) was obtained as
colorless amorphous foam.
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-ylsulfan-
yl]-acetyl}-mutilin+(7S, 8S) diastereomer
[0180] 14-O-{[(7R&
8R)-8-Hydroxy-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-acetyl}-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 (silia, cyclohexane/ethyl acetate=3/2)
14-O-{[(7R,
8R)-8-(tert-butyl-diphenyl-silanyloxy)-1,4dioxa-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.
[0181] .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,
2'-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-cyclohexylsulfanyl]-acetyl}--
mutilin+(1S, 2S) diastereomer
[0182] 14-O-{[(7R,
8R)-8-(tert-Butyl-diphenyl-silanyloxy)-1,4-dioxa-spiro[4.5]dec-7-ylsulfan-
yl]-acetyl}-mutilin+(1S, 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-&phenyl-silanyloxy)-5-oxo-cyclohexylsulfanyl]-acetyl}-m-
utilin+(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.
[0183] .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, 2'-H), 3.40 (m, 1H, 11-H), 3.04
(m, 3H, 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, 17CH.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-cyclobexylsulfanyl]-
-acetyl}-mutilin+(1S, 2S) diastereomer
[0184] 14-O-{[(1R,
2R)-2-(tert-Butyl-diphenyl-silanyloxy)-5-oxo-cyclohexylsulfanyl]-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-cyclohexylsulfanyl]-
-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-hydroxyamino-cyclohexylsulfanyl]-
-acetyl}-mutilin+(1S, 2S, 5R) diastertomer and 14-O-{[(1R, 2R,
5R)-2-(Iert-Butyl-diphenyl-silanyloxy)-5-hydroxyamino-cyclohexylsulfanyl]-
-acetyl}-mutilin+(1S, 2S, 5S) diastereomer
[0185] 14-O-{[(1R,
2R)-2-(tert-butyl-diphenyl-silanyloxy)-5-hydroxyimino-cyclohexylsulfanyl]-
-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-cyclohexylsulfanyl]-
-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-cyclohexylsulfanyl]-
-acetyl}-mutilin+(1S, 2S, 5S) diastereomer (b) (R.sub.f=0.25, 560
mg, 41% yield) were obtained.
[0186] (a): .sup.1H NMR (400 MHz, DMSO-dhd 6, .delta., ppm, inter
alia): 7.65-7.35 (m, 19H, aromat.-H), 7.00 (bs, 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).
[0187] (b) .sup.1H NMR (400 MHz, DMSO-d.sub.6, .delta., ppm, inter
alia): 7.7-7.35 (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-OH, J=6
Hz), 3.48 (m, 1H, 2'-H), 3.43 (t, 1H, 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, 4'-H),
1.38, 1.35 (2s, 3H, 15-CH.sub.3), 1.05 (s, 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-hydroxy-amino-cyclohexyl-
sulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer
[0188] 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. After
isolation of the precipitate by filtration 14-O-{[(1R, 2R,
5S)-2-(tert-butyl-diphenyl-silanyloxy)-5-(formyl-hydroxy-amino-cyclohexyl-
sulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer (135 mg, 61%
yield, cyclohexane/ethyl acetate 1/3, R.sub.f=0.65) was obtained as
colorless solid.
[0189] .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.3,
J=6 Hz).
Step J. 14-O-{[(1R, 2R,
5S)-5-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutili-
n+(1S, 2S, 5R) diastereomer
[0190] 14-O-{[(1R, 2R,
5S)-2-(tert-Butyl-diphenyl-silanyloxy)-5-(formyl-hydroxy-amino-cyclohexyl-
-sulfanyl]-acetyl}-mutilin+(1 S, 2S, 5R) diastereomer (130 mg,
0.164 mmol) was treated according to the method of Example 13 Step
G. After isolation of the precipitate by filtration 14-O-{[(1R, 2R,
5S)-5-(formyl-hydroxy-a&rino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutil-
in+(1S, 2S, 5R) diastereomer (77 mg, 85% yield, dichloromethane
methanol=9/1, R.sub.f=0.4) was obtained as colorless solid.
[0191] .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, iH, 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, 1'-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-H), 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)-, 1101 (2M-H).sup.-.
Example 15
14-O-{[(1R, 2R,
3R/S)-3-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S, 2S, 3R/S) diastereomer
[0192] Step A. 14-O-{[(6R,
7R)-6-Hydroxy-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-acetyl}-mutilin
+(6S, 7S) diastereomer
[0193] 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}-mutilin+(6S,
7S) diastereomer (R.sub.f=0.5, 15.6 g, 55% yield) was obtained as
colorless amorphous foam.
[0194] .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=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 (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=6 Hz).
Step B. 14-O-{[(1R,
2R)-2-Hydroxy-3-oxo-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 25)
diastereomer
[0195] 14-O-{[(6R,
7R)-6-Hydroxy-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-acetyl}-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.
[0196] .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
[0197] 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}-mutilin+(1S,
2S) diastereomer (R.sub.f=0.2, 1.75 g, 54% yield) was obtained as
colorless amorphous foam.
[0198] .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=11 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,
16-CH.sub.3, J=7 Hz).
Step D. 14-O-{[(1R, 2R,
3R/S)-2-Hydroxy-3-Hydroxyamino-cyclohexylsulfanyl]-acetyl}-mutilin+(1S,
2S, 3R/S) diastereomer
[0199] 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.sub.f=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.
[0200] .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-cyclohexylsulfanyl]-acetyl}-muti-
lin+(1S, 2S, 3R/S) diastereomer
[0201] 14-O-{[(1R, 2R,
3R/S)-2-hydroxy-3-Hydroxyamino-cyclohexylsulfanyl]-acetyl}-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.
[0202] .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=11Hz 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.-.
* * * * *