US20090118366A1 - Organic compounds - Google Patents

Organic compounds Download PDF

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Publication number
US20090118366A1
US20090118366A1 US11/828,224 US82822407A US2009118366A1 US 20090118366 A1 US20090118366 A1 US 20090118366A1 US 82822407 A US82822407 A US 82822407A US 2009118366 A1 US2009118366 A1 US 2009118366A1
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diastereomer
hydroxy
acetyl
mutilin
cyclohexylsulfanyl
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US11/828,224
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Rosemarie Mang
Werner Heilmayer
Rudolf Badegruber
Dirk Strickmann
Rodger Novak
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Nabriva Therapeutics Forschungs GmbH
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Nabriva Therapeutics Forschungs GmbH
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
    • C07C323/50—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton
    • C07C323/51—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton
    • C07C323/57—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being further substituted by nitrogen atoms, not being part of nitro or nitroso groups
    • C07C323/58—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being further substituted by nitrogen atoms, not being part of nitro or nitroso groups with amino groups bound to the carbon skeleton
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
    • A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
    • A61K31/221—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin with compounds having an amino group, e.g. acetylcholine, acetylcarnitine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/357—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having two or more oxygen atoms in the same ring, e.g. crown ethers, guanadrel
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/10—Anti-acne agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A61P31/06—Antibacterial agents for tuberculosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
    • C07C323/50—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton
    • C07C323/51—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton
    • C07C323/52—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being acyclic and saturated
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
    • C07C323/50—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton
    • C07C323/61—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atom of at least one of the thio groups bound to a carbon atom of a ring other than a six-membered aromatic ring of the carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
    • C07D295/08—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms
    • C07D295/096—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms with the ring nitrogen atoms and the oxygen or sulfur atoms separated by carbocyclic rings or by carbon chains interrupted by carbocyclic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D317/00—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms
    • C07D317/08—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3
    • C07D317/72—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3 spiro-condensed with carbocyclic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/12—Systems containing only non-condensed rings with a six-membered ring
    • C07C2601/14—The ring being saturated
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2603/00—Systems containing at least three condensed rings
    • C07C2603/93—Spiro compounds
    • C07C2603/95—Spiro compounds containing "not free" spiro atoms
    • C07C2603/98—Spiro compounds containing "not free" spiro atoms containing at least one ring with more than six ring members
    • C07C2603/99—Spiro compounds containing "not free" spiro atoms containing at least one ring with more than six ring members containing eight-membered rings

Definitions

  • the present invention relates to organic compounds, namely pleuromutilins.
  • Pleuromutilin a compound of formula A
  • 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.
  • pleuromutilin derivatives according to the invention are compounds of formula (I)
  • n 0 to 4; R is ethyl or vinyl; R 1 is hydrogen or (C 1-6 )alkyl, R 2 is hydrogen or
  • n, R 1 and R 2 are as defined above.
  • 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.
  • the present invention provides a compound of the present invention in the form of a salt and/or solvate.
  • the salts include preferably pharmaceutically acceptable salts, although pharmaceutically unacceptable salts are included, e.g. for preparation/isolation/purification purposes.
  • 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.
  • 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.
  • 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.
  • 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 2 ) n N(R 1 R 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.
  • R 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.
  • the configuration of substituents attached to asymmetric carbon atoms of the mutilin-ring is preferably the same as in natural pleuromutilin.
  • 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.
  • the compounds of the present invention exhibit pharmacological activity and are therefore useful as pharmaceuticals.
  • 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.
  • 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.
  • 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.
  • 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.
  • an antimicrobial such as an antibiotic, e.g. and an anti-anaerobic.
  • the present invention provides a compound of the present invention for use in acne treatment.
  • 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
  • bacteria e.g. selected from Staphylococci, Streptococci, Enterococci;
  • bacteria e.g. selected from Moraxella, Haemophilus, Legionella, Neisseriaceae;
  • 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.
  • 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.
  • Treatment includes treatment and prophylaxis.
  • 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.
  • 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.
  • macrolides such as erythromycins, e.g. clarithromycin or azithromycin.
  • 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 pharmaceutically acceptable salt e.g. an acid addition salt or a base addition salt, e.g. a metal salt
  • 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.
  • 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.
  • 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.
  • the present invention provides a pharmaceutical composition
  • 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.
  • excipient e.g. carrier or diluent
  • the present invention provides a pharmaceutical composition according to the present invention, further comprising another pharmaceutically active agent.
  • 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.
  • 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.
  • 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.
  • the present invention provides a compound of the present invention for use as a veterinary agent.
  • the present invention provides a compound of the present invention for the preparation of a veterinary composition which is useful as a veterinary agent.
  • 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.
  • microbial e.g. bacterial diseases
  • the metabolic stability for compounds of the present invention was determined by using cryopreserved primary human hepatocytes. 1 ⁇ 10 6 cells/mL were incubated in the absence and the presence of 5 and 25 ⁇ g/mL of the test compounds at 37° C., 5% CO 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.
  • 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.
  • 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 1,8 ]tetradecan-9-one.
  • 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.):
  • Pleuromutilin thiol and pleuromutilin tosylate are compounds of formulae:
  • Step A 14-O- ⁇ [(1R, 2R, 4R)-4-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsuffanyl]-acetyl ⁇ -mutilin+(1S, 2S, 4S) diastereomer and 14-O- ⁇ [(1R, 2R, 5S)-5-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 5R) diastereomer and
  • Step B 14-O- ⁇ [(1R, 2R, 4R)4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 4S) diastereomer
  • Step C 14-O- ⁇ [(1R, 2R, 4R)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin hydrochloride+(1S, 2S, 4S) diastereomer hydrochloride
  • Step A 14-O- ⁇ [(1R, 2R, 5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 5R) diastereomer and
  • Example 1 Step A A mixture of 14-O- ⁇ [(1R, 2R, 5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 5R) diastereomer and 14-O- ⁇ [(1R, 2R, 4S)4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 4R) diastereomer (1.12 g, 1.84 mmol) from Example 1 Step A was treated according to the method of Example 1 Step B.
  • Step B 14-O- ⁇ [(1R, 2R, 5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin hydrochloride+(1S, 2S, 5R) diastereomer hydrochloride
  • Example 2 Step A 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.
  • Step A tert-Butyl-dimethyl-(cis-3,4-epoxycyclohexyloxy)-silane
  • 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° C. and stirred for 5 hours at room temperature.
  • the reaction mixture was diluted with dichloromethane and subsequently extracted with 10% NaHSO 3 solution, saturated NaHCO 3 solution and brine. The organic layer was dried over sodium sulfate and filtered.
  • Step B 14-O- ⁇ [(1R, 2R, 5S)-5-(tert-Butyl-dimethyl-silyloxy)-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 5R) diastereomer
  • Step C 14-O- ⁇ [(1R, 2R, 5S)-2,5-Dihydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 5R) diastereomer
  • Step D 14-O- ⁇ [(1R, 2R, 5S)-2-Hydroxy-5-methanesulfonyloxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 5R) diastereomer
  • Step E 14-O- ⁇ [(1R, 2R, 5R)-5-Azido-2-hydroxy-5-methanesulfonyloxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 5S) diastereomer
  • Step F 14-O- ⁇ [(1R, 2R, 5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 5S) diastereomer
  • 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.
  • Step A 14-O- ⁇ [(1R, 2R, 3R)-3-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 3S) diastereomer
  • Step B 14-O- ⁇ [(1R, 2R, 3R)-3-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 3S) diastereomer
  • 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 3 solution and brine. The organic layer was dried over sodium sulfate and filtered.
  • sodium triacetoxyborohydride 750 mg, 3.54 mmol
  • Step A N-Ethyl-N-(cyclohex-3-enyl)-carbamic acid tert-butyl ester
  • Step B N-ethyl-N-(cis-3,4-epoxycyclohexyl)-carbamic acid tert-butyl ester
  • Step C 14-O- ⁇ [(1R, 2R, 5S)-5-(tert-Butoxycarbonyl-ethyl-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 5R) diastereomer
  • 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.
  • Step E 14-O- ⁇ [(1R, 2R, 5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin hydrochloride+(1S, 2S, 5R) diastereomer hydrochloride
  • Step A 14-O- ⁇ [(1R, 2R, 5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilia+(1S, 2S, 5R) diastereomer
  • Step B 14O- ⁇ [(1R, 2R, 5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin hydrochloride+(1S, 2S, 5R) diastereomer hydrochloride
  • Example 2 Step A 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.
  • Example 4 Step F 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.
  • Step A [(1R, 2R, 3R)-2-Hydroxy-3-(2,4,6-trimethyl-benzylsulfanyl)-cyclobexyl]-carbamic acid tert-butyl ester+(1S, 2S, 3S) diastereomer
  • 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
  • Step C [(1R, 2R, 3R)2-(tert-Butyl-dimethyl-silanyloxy)-3-(2,4,6trimethyl-benzylsulfanyl)-cyclohexyl]-ethyl-carbamic acid tert-butyl ester+(1S, 2S, 3S) diastereomer
  • 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
  • Step A 14-O- ⁇ [(1R, 2R, 3R)-3-Diethylamino)-2-(tert-butyl-dimethyl-silanyloxy)-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 35) diastereomer
  • Example 11 14-O- ⁇ [(1R, 2R, 3R)-3-Ethylamino-2-(tert-butyl-dimethyl-silanyloxy)-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 3S) diastereomer (144 mg, 0.222 mmol) from Example 11 Step F was treated with acetaldehyde (25 ⁇ l, 0.444 mmol) according to the method of Example 6.
  • Step B 14-O- ⁇ [(1R, 2R, 3R)-3-Diethylamino-2-hydroxy-cyclohexylsulfanyll-acetyl ⁇ -mutilin+(1S, 2S, 3S) diastereomer
  • Step A 14-O- ⁇ [(7R, 8R)-7-Hydroxy-1,4-dioxa-spiro[4.51dec-8-ylsulfanyl]-acetyl ⁇ -mutilin+(7S, 8S) diastereomer
  • Step B 14-O- ⁇ [(7R, 8R)-7-(tert-Butyl-diphenyl-silanyloxy)-1,4-dioxa-spiro[4.5]dec-8-ylsulfanyl]-acetyl ⁇ -mutilin+(7S, 8S) diastereomer
  • Step C 14O- ⁇ [(1R, 2R)-2-(tert-Butyl-diphenyl-silanyloxy)-4-oxo-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S) diastereomer
  • Step D 14-O- ⁇ [(1R, 2R)-2-(tert-Butyl-diphenyl-silanyloxy)-4-hydroxyimino-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S) diastereomer
  • 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
  • Step F 14-O- ⁇ [(1R, 2R, 4S)-2-(tert-Butyl-diphenyl-silanyloxy)-4-(formyl-hydroxy-amino-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 4R) diastereomer
  • Step G 14-O- ⁇ [(1R, 2R, 4S)-4-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 4R) diastereomer
  • Step B Acetic acid (7R, 8R)-7-(2,4,6-trimethylbenzylsufanyl)-1,4-dioxa-spiro[4.5]dec-8-yl ester+(7S, 8S) diastereomer
  • Step D 14-O- ⁇ [(7R, 8R)-8-Hydroxy-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-acetyl ⁇ -mutilin+ ⁇ 7S, 8S) diastereomer
  • Step F 14-O- ⁇ [(1R, 2R)-2-(tert-Butyl-diphenyl-silanyloxy)-5-oxo-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S) diastereomer
  • Step G 14-O- ⁇ [(1R, 2R)-2-(tert-Butyl-diphenyl-silanyloxy)-5-hydroxyimino-cyclobexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S) diastereomer
  • 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
  • Step I 14-O- ⁇ [(1R, 2R, 5S)-2-(tert-Butyl-diphenyl-silanyloxy)-5-(formyl-hydroxy-amino-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 5R) diastereomer
  • Step A 14-O- ⁇ [(6R, 7R)-6-Hydroxy-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-acetyl ⁇ -mutilin +(6S, 7S) diastereomer
  • Step B 14-O- ⁇ [(1R, 2R)-2-Hydroxy-3-oxo-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 25) diastereomer
  • Step C 14-O-([(1R, 2R)-2-Hydroxy-3-hydroxyimino-cyclohexylsulfanyl]-acetyl ⁇ -mutilin +(1S, 2S) diastereomer
  • Step D 14-O- ⁇ [(1R, 2R, 3R/S)-2-Hydroxy-3-Hydroxyamino-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 3R/S) diastereomer
  • Step E 14-O- ⁇ [(1R, 2R, 3R/S)-3-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl ⁇ -mutilin+(1S, 2S, 3R/S) diastereomer

Abstract

A compound of formula (I)
Figure US20090118366A1-20090507-C00001
wherein
n is 0 to 4;
R is ethyl or vinyl;
R1 is hydrogen or (C1-6)alkyl,
R2 is hydrogen or
    • cycloalkyl including (C3-6)cycloalkyl, or
    • unsubstituted (C1-6)alkyl, or
    • (C1-6)alkyl substituted by one or more of
      • hydroxy; preferably one or two,
      • halogen,
      • (C3-6)cycloalkyl, or
        R1 is hydroxy and R2 is formyl.

Description

    CROSS REFERENCE TO RELATED APPLICATIONS
  • 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.
  • The present invention relates to organic compounds, namely pleuromutilins.
  • Pleuromutilin, a compound of formula A
  • Figure US20090118366A1-20090507-C00002
  • 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.
  • 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.
  • We have now found pleuromutilins with interesting activity combined with an unexpexted remarkable metabolic stability.
  • The pleuromutilin derivatives according to the invention are compounds of formula (I)
  • Figure US20090118366A1-20090507-C00003
  • wherein
    n is 0 to 4;
    R is ethyl or vinyl;
    R1 is hydrogen or (C1-6)alkyl,
    R2 is hydrogen or
      • cycloalkyl including (C3-6)cycloalkyl, or
      • unsubstituted (C1-6)alkyl, or
      • (C1-6)alkyl substituted by one or more of
        • hydroxy; preferably one or two,
        • halogen,
        • (C3-6)cycloalkyl, or
          R1 is hydroxy and R2 is formyl.
  • Preferred compounds of the present invention are
      • a compound of formula (II)
  • Figure US20090118366A1-20090507-C00004
      • a compound of formula (III)
  • Figure US20090118366A1-20090507-C00005
  • and
      • a compound of formula (IV)
  • Figure US20090118366A1-20090507-C00006
  • wherein n, R1 and R2 are as defined above.
  • Particularly preferred is a compound 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-cyclobexylsulfanyl]-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}-mutilin and the (1S, 2S, 4R) diastereomer thereof
  • 14-O-{[(1R, 2R, 5S)-5-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin and the (1S, 2S, 5R) diastereomer thereof
  • and
  • 14-O-{[(1R, 2R, 3R/S)-3-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin and the (1S, 2S, 3R/S) diastereomer thereof.
  • 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.
  • According to another aspect, the present invention provides a compound of the present invention in the form of a salt and/or solvate.
  • The salts include preferably pharmaceutically acceptable salts, although pharmaceutically unacceptable salts are included, e.g. for preparation/isolation/purification purposes.
  • 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.
  • 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.
  • 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.
  • 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 (CH2)nN(R1R2) 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 R2 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.
  • The configuration of substituents attached to asymmetric carbon atoms of the mutilin-ring is preferably the same as in natural pleuromutilin.
  • 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.
  • The compounds of the present invention exhibit pharmacological activity and are therefore useful as pharmaceuticals.
  • 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.
  • 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.
  • 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.
  • 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.
  • In another aspect the present invention provides a compound of the present invention for use in acne treatment.
  • 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
  • diseases mediated by bacteria, e.g. selected from Staphylococci, Streptococci, Enterococci;
  • diseases mediated by bacteria, e.g. selected from Moraxella, Haemophilus, Legionella, Neisseriaceae;
  • diseases mediated by Helicobacter;
  • diseases mediated by Mycobacterium tuberculosis;
  • e.g. diseases mediated by Mycoplasms, Chlamydia and obligatory anaerobes;
  • and for the treatment of acne.
  • 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.
  • 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.
  • Treatment includes treatment and prophylaxis.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • In another aspect the present invention provides a pharmaceutical composition according to the present invention, further comprising another pharmaceutically active agent.
  • 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.
  • 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.
  • In another aspect the present invention provides a compound of the present invention for use as a veterinary agent.
  • 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.
  • 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.
  • Examples 1 to 15 following thereafter exhibit MICs≦2 μg/ml against Staphylococcus aureus ATCC49951 and Streptococcus pneumoniae ATCC49619.
  • The metabolic stability for compounds of the present invention was determined by using cryopreserved primary human hepatocytes. 1×106 cells/mL were incubated in the absence and the presence of 5 and 25 μg/mL of the test compounds at 37° C., 5% CO2 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.
  • 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.
  • For example after 4 h incubation with human hepatocytes at a compound concentration of 5 μg/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
  • 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[5.4.3.01,8]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.):
  • Figure US20090118366A1-20090507-C00007
  • Pleuromutilin thiol and pleuromutilin tosylate are compounds of formulae:
  • Figure US20090118366A1-20090507-C00008
    • Example 1 14-O-{[(1R, 2R, 4R)4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin hydrochloride+(1S, 2S, 4S) diastereomer hydrochloride
  • Step A1. 14-O-{[(1R, 2R, 4R)-4-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsuffanyl]-acetyl}-mutilin+(1S, 2S, 4S) diastereomer and 14-O-{[(1R, 2R, 5S)-5-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer and
  • 14-O-{[(1R, 2R, 4S)-4-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 4R) diastereomer
  • To a solution of 3,4-epoxycyclohexyl-carbamic acid tert-butyl ester (Gómez-Sánchez, E.; Marco-Contelles J. Tetrahedron 2005, 61, 1207-1219.) (4.27 g, 20 mmol) and pleuromutilin thiol (Nagarajan, R. Eli Lilly and Company 1978, U.S. Pat. No. 4,130,709) (7.10 g, 18 mmol) in 200 ml of tetrahydrofuran was added aluminum oxide (40 g, Brockmann activity I, neutral) and the resulting mixture was stirred for 40 hours at room temperature. The suspension was filtered and concentrated under reduced pressure. The residue was subjected to chromatography (silica, cyclohexane/ethyl acetate=1/1) to yield 14-O-{[(1R, 2R, 4R)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 4S) diastereomer (a) (Rf=0.38, 1.34 g, 12%) as well as a mixture of 14-O-{[(1R, 2R, 5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer and 14-O-{[(1R, 2R, 4S)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 4R) diastereomer (b) (Rf=0.26, 2.81 g, 25%) as colorless amorphous foams.
  • (a): 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.35 (s, 9H, tert-butyl), 1.06 (s, 3H, 18-CH3), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.62 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 630 (MNa+), 1237 (2MNa+).
  • (b): 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3, tert-butyl), 1.05 (s, 3H, 18-CH3), 0.82 (d, 3H, 17-CH3, J=7 Hz), 0.62 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 630 (MNa+), 1237 (2MNa+).
  • or Step A2. 14-O-{[(1R, 2R, 4R)-4-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 4S) diastereomer and
  • 14-O-{[(1R, 2R, 5S)-5-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer and
    14-O-{[(1R, 2R, 4S)-4-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 4R) diastereomer
  • 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}-mutilin+(1S, 2S, 4S) diastereomer (Rf=0.40, 3.1 g, 12% yield) as well as a mixture of 14-O-{[(1R, 2R, 5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer and 14-O-{[(1R, 2R, 4S)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 4R) diastereomer (Rf=0.25, 6.35 g, 25%) were obtained as colorless amorphous foams.
  • Step B. 14-O-{[(1R, 2R, 4R)4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 4S) diastereomer
  • To a solution of 14-O-{[(1R, 2R, 4R)-4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 4S) diastereomer (1.34 g, 2.20 mmol) in 75 ml of dichloromethane was added trifluoroacetic acid (4 ml) at 4° C. and stirred for 5 hours at room temperature. The reaction mixture was diluted with dichloromethane and cautiously poured into a saturated NaHCO3 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.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.06 (s, 3H, 18-CH3), 0.82 (d, 3H, 17-CH3, J=7 Hz), 0.62 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 508 (MH+), 530 (MNa+), 1015 (2MH+), 1037 (2MNa+).
  • Step C. 14-O-{[(1R, 2R, 4R)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin hydrochloride+(1S, 2S, 4S) diastereomer hydrochloride
  • 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.
  • 1H NMR (500 MHz, DMSO-d6, δ, ppm, inter alia): 7.6 (bs, 3H, NH3 +), 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-CH3), 1.06 (s, 3H, 18-CH3), 0.82 (d, 3H, 17-CH3, J=7 Hz), 0.62 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 508 (MH+), 530 (MNa+), 1015 (2MH+), 1037 (2MNa+), 542 (MCl−).
    • Example 2 14-O-{[(1R, 2R, 5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin hydrochloride+(1s, 2S, 5R) diastereomer hydrochloride
  • Step A. 14-O-{[(1R, 2R, 5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer and
  • 14-O-{[(1R, 2R, 4S)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 4R) diastereomer
  • A mixture of 14-O-{[(1R, 2R, 5S)-5-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer and 14-O-{[(1R, 2R, 4S)4-tert-butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 4R) diastereomer (1.12 g, 1.84 mmol) from Example 1 Step A was treated according to the method of Example 1 Step B. After work up and chromatography of the reaction mixture (silica, ethyl acetate/methanol/35% ammonia solution=50/50/1) 14-O-{[(1R, 2R, 5S)-5-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer (a) (Rf=0.33, 524 mg, 56% yield) and 14-O-{[(1R, 2R, 4S)-4-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 4R) diastereomer (b) (Rf=0.22, 160 mg, 17%) were obtained as colorless amorphous foams.
  • (a): 1H NMR (400 MHz, DMSO-d6, 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 (νA=3.37, νB=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-CH3), 1.06 (s, 3H, 18-CH3), 0.82 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=7 Hz).
  • (b): 1H NMR (400 MHz, DMSO-d6, δ, 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 (νA=3.33, νB=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-CH3), 1.06 (s, 3H, 18-CH3), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=7 Hz).
  • Step B. 14-O-{[(1R, 2R, 5S)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin hydrochloride+(1S, 2S, 5R) diastereomer hydrochloride
  • 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.
  • 1H NMR (500 MHz, DMSO-d6, δ, ppm, inter alia): 7.7 (bs, 3H, NH3 +), 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-CH3), 1.05 (s, 3H, 18-CH3), 0.82 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 508 (MH+), 530 (MNa+, 1015 (2MH+), 1037 (2MNa+), 542 (MCl−).
    • Example 3 14-O-{[(1R, 2R, 4S)-4-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin hydrochloride+(1S, 2S, 4R) diastereomer hydrochloride
  • 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.
  • 1H NMR (500 MHz, DMSO-d6, δ, ppm, inter alia): 7.8 (bs, 3H, NH3 +), 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-CH3), 1.05 (s, 3H, 18-CH3), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 508 (MH+), 530 (MNa+), 1015 (2MH+), 1037 (2MNa+), 542 (MCl−).
    • Example 4 14-O-{[(1R, 2R, 5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5S) diastereomer
  • Step A. tert-Butyl-dimethyl-(cis-3,4-epoxycyclohexyloxy)-silane
  • 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° C. and stirred for 5 hours at room temperature. The reaction mixture was diluted with dichloromethane and subsequently extracted with 10% NaHSO3 solution, saturated NaHCO3 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 (Rf=0.35, 18.3 g, 79% yield) as colorless oil.
  • 1H NMR (500 MHz, DMSO-d6, δ, 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(CH3)2).
  • Step B. 14-O-{[(1R, 2R, 5S)-5-(tert-Butyl-dimethyl-silyloxy)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer
  • 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.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH2), 2.92 (m, 1H, 1′-H), 2.38 (bs, 1H, 4-H), 1.36 (s, 3H, 15-CH3), 1.05 (s, 3H, 18-CH3), 0.86 (s, 9H, tert-butyl), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=7 Hz), 0.0 (s, 6H, Si(CH3)2).
  • Step C. 14-O-{[(1R, 2R, 5S)-2,5-Dihydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer
  • 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° 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=⅓) to yield the 14-O-{[(1R, 2R, 5S)-2,5-dihydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer (Rf=0.27, 7.07 g, 92% yield) as colorless amorphous foam.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.06 (s, 3H, 18-CH3), 0.82 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 531 (MNa+), 1039 (2MNa+).
  • Step D. 14-O-{[(1R, 2R, 5S)-2-Hydroxy-5-methanesulfonyloxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer
  • 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+(1S, 2S, 5R) diastereomer (Rf=0.15, 2.55 g, 36% yield) as colorless amorphous foam.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.05 (s, 3H, 18-CH3), 0.80 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=7 Hz).
  • Step E. 14-O-{[(1R, 2R, 5R)-5-Azido-2-hydroxy-5-methanesulfonyloxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5S) diastereomer
  • A solution of 14-O-{[(1R, 2R, 5S)-2-hydroxy-5-methanesulfonyloxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer (2.55 g, 4.35 mmol) and sodium azide (0.85 g, 13 mmol) in 30 ml of dimethylformamide was heated for 6 hours at 80° 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}-mutilin+(1S, 2S, 5S) diastereomer (quantitative yield, cyclohexane/ethyl acetate=1/1, Rf=0.35) was obtained as amorphous foam which was directly used for the next step.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.06 (s, 3H, 18-CH3), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=7 Hz).
  • Step F. 14-O-{[(1R, 2R, 5R)-5-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5S) diastereomer
  • 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 (Rf=0.3, 1.74 g, 79% yield) as colorless amorphous foam.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.05 (s, 3H, 18-CH3), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.62 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 508 (MH+), 530 (MNa+), 1037 (2MNa+).
    • Example 5 14-O-{[(1R, 2R, 3R)-3-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin hydrochloride+(1S, 2S, 3S) diastereomer hydrochloride
  • Step A. 14-O-{[(1R, 2R, 3R)-3-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 3S) diastereomer
  • (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}-mutilin+(1S, 2S, 3S) diastereomer (Rf=0.5, 1.32 g, 46%) was obtained as colorless amorphous foam.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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 (νA=3.30, 3.29, νB=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-CH3), 1.05 (s, 3H, 18-CH3), 0.82 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=6 Hz).
  • Step B. 14-O-{[(1R, 2R, 3R)-3-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 3S) diastereomer
  • 14-O-{[(1R, 2R, 3R)-3-tert-Butoxycarbonylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 3S) diastereomer (400 mg, 0.658 mmol) was treated according to the method of Example 1 Step B. After work up and chromatography of the reaction mixture (silica, ethyl acetate/methanol=1/5) 14-O-{[(1R, 2R, 3R)-3-amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 3S) diastereomer (Rf=0.1, 249 mg, 75%) was obtained as colorless amorphous foam.
    • MS-ESI (m/z): 508 (MH+), 530 (MNa+), 1015 (2MH+), 1037 (2MNa+).
      Step C. 14-O-{[(1R, 2R, 3R)-3-Amino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin hydrochloride+(1S, 2S, 4S) diastereomer hydrochloride
  • 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.
  • 1H NMR (400 Mhz, DMSO-d6, δ, ppm, inter alia): 7.8 (bs, 3H, NH3 +), 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 (νA=3.35, νB=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-CH3), 1.05 (s, 3H, 18-CH3), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 508 (MH+), 1015 (2MH+), 542 (MCl−).
    • 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
  • 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 μl, 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 NaHCO3 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.
  • (a): 1H NMR (400 MHz, DMSO-d6, δ, 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 (νA=3.50, 3.42, νB=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, NCH2, 4-H), 1.36 (s, 3H, 15-CH3), 1.05 (s, 3H, 18-CH3), 0.92 (t, 6H, NCH2CH3, J=7 Hz), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 564 (MH+), 586 (MNa+), 562 (M-H)−.
  • (b): 1H NMR (400 MHz, DMSO-d6, δ, 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 (νA=3.48, νB=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-CH3), 1.05 (s, 3H, 18-CH3), 0.95 (t, 3H, NCH2CH3, J=7 Hz), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=6 Hz). MS-ESI (m/z): 536 (MH+), 558 (MNa+), 534 (M-H)−.
    • Example 7 14-O-{[(1R, 2R, 5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin hydrochloride+(1S, 2S, 5R) diastereomer hydrochloride
  • Step A. N-Ethyl-N-(cyclohex-3-enyl)-carbamic acid tert-butyl ester
  • 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 (Rf=0.30, 2.88 g, 58% yield) was obtained as colorless solid.
  • 1H NMR (400 MHz, CDCl6, δ, ppm): 5.61 (m, 2H, double bond), 4.08 (bs, 1H, NCH), 3.15 (m, 2H, NCH2), 2.15, 1.75 (2m, 6H), 1.47 (s, 9H, tert-butyl), 1.13 (t, 3H, NCH2CH3, J=7 Hz).
  • Step B. N-ethyl-N-(cis-3,4-epoxycyclohexyl)-carbamic acid tert-butyl ester
  • 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% NaHSO3 solution, saturated NaHICO3 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 (Rf=0.2, 1.50 g, 49% yield) was obtained.
  • 1H NMR (400 MHz, CDCl6, δ, ppm): 4.0 (bs, 1H, NCH), 3.14 (m, 2H, NCH2), 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, NCH2CH3).
  • Step C. 14-O-{[(1R, 2R, 5S)-5-(tert-Butoxycarbonyl-ethyl-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer
  • 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]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer (Rf=0.4, 2.57 g, 65% yield) was obtained as colorless amorphous foam.
  • MS-ESI (m/z): 536 (MH+), 558 (MNa+), 534 (M-H)−.
    Step D. 14-O-{[(1R, 2R, 5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer
  • 14-O-{[(1R, 2R, 5S)-5-(tert-Butoxycarbonyl-ethyl-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer (2.57 g, 4.04 mmol) was treated according to the method of Example 1 Step B. After work up and chromatography of the reaction mixture (silica, ethyl acetate/methanol/35% ammonia solution=100/100/1) 14-O-{[(1R, 2R, 5S)-5-ethylamino-2-hydroxy-cyclohexylsulfanyl]acetyl}-mutilin+(1 S, 2S, 5R) diastereomer (Rf=0.3, 1.08 g, 50%) was obtained as colorless amorphous foam.
  • 1H NMR (500 MHz, CDCl6, δ, 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, NCH2, J=7 Hz), 2.10 (bs, 1H, 4-H), 1.46 (s, 3H, 15-CH3), 1.18 (s, 3H, 18-CH3), 1.12 (t, 3H, NCH2CH3, J=7 Hz), 0.98 (d, 3H, 17-CH3, J=7 Hz), 0.73 (d, 3H, 16-CH3, J=7 Hz).
  • Step E. 14-O-{[(1R, 2R, 5S)-5-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin hydrochloride+(1S, 2S, 5R) diastereomer hydrochloride
  • 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.
  • 1H NMR (400 MHz, CDCl6, δ, ppm, inter alia): 9.3 (bs, 2H, NH2 +), 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, NCH2), 2.10 (bs, 1H, 4-H), 1.50 (t, 3H, NCH2CH3, J=7 Hz), 1.45 (s, 3H, 15-CH3), 1.18 (s, 3H, 18-CH3), 0.90 (d, 3H, 17-CH3, J=7 Hz), 0.73 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 536 (MH+), 570 (MCl−).
    • Example 8 14-O-{[(1R, 2R, 5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin hydrochloride+(1S, 2S, 5R) diastereomer hydrochloride
  • Step A. 14-O-{[(1R, 2R, 5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilia+(1S, 2S, 5R) diastereomer
  • 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 μl, 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 (Rf=0.2, 230 mg, 87%) was obtained as colorless amorphous foam.
  • 1H NMR (400 MHz, CDCl6, δ, 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, NCH2), 2.11 (bs, 1H, 4-H), 1.46 (s, 3H, 15-CH3), 1.18 (s, 3H, 18-CH3), 0.98 (t, 6H, NCH2CH3, J=7 Hz), 0.88 (d, 3H, 17-CH3, J=7 Hz), 0.73 (d, 3H, 16-CH3, J=7 Hz).
  • Step B. 14O-{[(1R, 2R, 5S)-5-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin hydrochloride+(1S, 2S, 5R) diastereomer hydrochloride
  • 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.
  • 1H NMR (400 MHz, CDCl6, δ, ppm, inter alia): 11.5 (bs, 3H, NH+), 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, NCH2), 2.10 (bs, 1H, 4-H), 1.48 (m, 9H, NCH2CH3, 15-CH3), 1.18 (s, 3H, 18-CH3), 0.89 (d, 3H, 17-CH3, J=7 Hz), 0.73 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 564 (MH+), 586 (MNa+), 1149 (2MNa+), 598 (MCl−).
    • Example 9 14-O-{[(1R, 2R, 4S)-4-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 4R) diastereomer
  • 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 (Rf=0.2, 129 mg, 17%) was obtained as colorless amorphous foam.
  • 1H NMR (500 MHz, DMSO-d6, δ, 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 (νA=3.36, νB=3.22, 22-H, J=15 Hz), 2.72 (m, 2H, 1′-H, 5′-H), 2.47 (m, 2H, NCH2), 2.40 (bs, 1H, 4-H), 1.35 (s, 3H, 15-CH3), 1.04 (s, 3H, 18-CH3), 0.95 (t, 3H, NCH2CH3, J=7 Hz), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.62 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 536 (MH+), 558 (MNa+), 534 (M-H)−.
    • Example 10 14-O-{[(1R, 2R, 5R)-5-Diethylamino-2-hydroxy-cyclohexylsuffanyl]-acetyl}-mutilin+(1S, 2S, 5S) diastereomer
  • 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 (Rf=0.2, 95 mg, 20%) was obtained as colorless amorphous foam.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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, NCH2, 4-H), 1.37 (s, 3H, 15-CH3), 1.06 (s, 3H, 18-CH3), 0.92 (t, 3H, NCH2CH3, J=7 Hz), 0.83 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 564 (MH+), 586 (MNa+), 562 (M-H)−, 598 (MCl−).
    • Example 11 14-O-{[(1R, 2R, 3R)-3-Ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 3S) diastereomer
  • Step A. [(1R, 2R, 3R)-2-Hydroxy-3-(2,4,6-trimethyl-benzylsulfanyl)-cyclobexyl]-carbamic acid tert-butyl ester+(1S, 2S, 3S) diastereomer
  • 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 (Rf=0.25, 5.92g, 23% yield) as colorless amorphous foam.
  • 1H NMR (400 MHz, DMSO-d6, δ, ppm): 6.78 (s, 2H, aromat.-H), 6.15 (bd, OH), 4.95 (bd, NH), 3.75 (d, 1H, SCH2), 3.68 (m, 2H), 3.02 (m, 1H, SCH), 2.30 (s, 9H, CH3), 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
  • 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 ° 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 (Rf=0.25, 3.0 g, 94% yield) was obtained.
  • 1H NMR (500 MHz, DMSO-d6, δ, ppm): 6.80 (s, 2H, aromat-H), 6.20 (bd, NH), 3.90, 3.75, 3.63 (3m, 4H, NCH, OCH, SCH2), 2.98 (m, 1H, SCH), 2.30 (s, 9H, CH3), 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(CH3)2).
  • Step C. [(1R, 2R, 3R)2-(tert-Butyl-dimethyl-silanyloxy)-3-(2,4,6trimethyl-benzylsulfanyl)-cyclohexyl]-ethyl-carbamic acid tert-butyl ester+(1S, 2S, 3S) diastereomer
  • 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+), 1065 (2MNa+).
    Step D. [(1R, 2R, 3R)-2-(tert-Butyl-dimethyl-silanyloxy)-3-mercapto-cyclohexyl]-ethyl-carbamic acid tert-butyl ester+(1S, 2S, 3S) diastereomer
  • 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° C. under an argon atmosphere with sodium (106 mg, 4.60 mmol) and stirred at −78° 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-carbamic acid tert-butyl ester+(1S, 2S, 3S) diastereomer (quantitative yield) which was directly used for the next step.
  • MS-ESI (m/z): 412 (MNa+), 801 (2MNa+).
    Step E. 14-O-{[(1R, 2R, 3R)-3-(tert-Butoxycarbonyl-ethyl-amino)-2-(tert-butyl-dimethyl-silanyloxy)-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 3S) diastereomer
  • Crude [(1R, 2R, 3R)-2-(tert-butyl-dimethyl-silanyloxy)-3-mercapto-cyclohexyl]-ethyl-carbamic 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 NaHCO3 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 (Rf=0.5, 468 mg, 27% yield) was obtained.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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, NCH2), 2.40 (bs, 1H, 4-H), 1.40 (s, 9H, tert-butyl), 1.36, 1.35 (2s, 3H, 15-CH3), 1.05 (s, 3H, 18-CH3), 0.87 (s, 9H, Si-tert-butyl), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.64, 0.62 (2d, 3H, 16-CH3, J=7 Hz) 0.05,-0.05 (2s, 6H, Si(CH3)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
  • 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]-acetyl}-mutilin+(1S, 2S, 3S) diastereomer (a) (Rf=0.6, 144 mg, 36% yield) and 14-O-([(1R, 2R, 3R)-3-ethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl} -mutilin+(1S, 2S, 3S) diastereomer (b) (Rf=0.25, 177 mg, 53% yield) were obtained as colorless solids.
  • (a): MS-ESI (m/z): 672 (MNa+), 1321 (2MNa+).
  • (b): 1H NMR (400 MHz, DMSO-d6, δ, 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 (νA=3.37, νB=3.18, 22-H, J=15 Hz), 3.05 (m, 1H, 3′-H), 2.66 (m, 1H, 1′-H), 2.50 (m, 2H, NCH2), 2.40 (bs, 1H, 4-H), 1.36 (s, 3H, 15-CH3), 1.05 (s, 3H, 18-CH3), 0.98 (t, 3H, NCH2CH3, J=7 Hz), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.62 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 536 (MH+), 558 (MNa+), 1071 (2MH+), 1093 (2MNa+), 534 (M-H)−.
    • Example 12 14-O-{[(1R, 2R, 3R)-3-Diethylamino-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 3S) diastereomer
  • Step A. 14-O-{[(1R, 2R, 3R)-3-Diethylamino)-2-(tert-butyl-dimethyl-silanyloxy)-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 35) diastereomer
  • 14-O-{[(1R, 2R, 3R)-3-Ethylamino-2-(tert-butyl-dimethyl-silanyloxy)-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 3S) diastereomer (144 mg, 0.222 mmol) from Example 11 Step F was treated with acetaldehyde (25 μl, 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 (Rf=0.5, 110 mg, 73%) was obtained as colorless amorphous foam.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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, NCH2), 2.40 (bs, 1H, 4-H), 1.36 (s, 3H, 15-CH3), 1.05 (s, 3H, 18-CH3), 0.87 (m, 15H, NCH2CH3, Si-tert-butyl), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.62, 0.60 (2d, 3H, 16-CH3, J=7 Hz), 0.07 (s, 6H, Si(CH3)2).
  • Step B. 14-O-{[(1R, 2R, 3R)-3-Diethylamino-2-hydroxy-cyclohexylsulfanyll-acetyl}-mutilin+(1S, 2S, 3S) diastereomer
  • 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 NaHCO3 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 (Rf=0.2, 8 mg, 10% yield) as colorless amorphous foam.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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, NCH2), 2.40 (bs, 1n, 4-H), 1.35 (s, 3H, 15-CH3), 1.05 (s, 3H, 18-CH3), 0.90 (m, 6H, NCH2CH3), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.62, 0.61 (2d, 3H, 16-CH3, J=7 Hz). MS-E3SI (m/z): 564 (MH+), 586 (MNa+), 1149 (2MNa+), 562 (M-H)−.
    • Example 13 14-O-{[(1R, 2R, 4S)-4-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 4R) diastereomer
  • Step A. 14-O-{[(7R, 8R)-7-Hydroxy-1,4-dioxa-spiro[4.51dec-8-ylsulfanyl]-acetyl}-mutilin+(7S, 8S) diastereomer
  • 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 (Rf=0.3, 8.40 g, 76% yield) was obtained as colorless amorphous foam.
  • 1H NMR (500 MHz, DMSO-d6, δ, 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, OCH2CH2O), 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-CH3), 1.05 (s, 3H, 18-CH3), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.62 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 573 (MNa+), 549 (M-H)−.
  • Step B. 14-O-{[(7R, 8R)-7-(tert-Butyl-diphenyl-silanyloxy)-1,4-dioxa-spiro[4.5]dec-8-ylsulfanyl]-acetyl}-mutilin+(7S, 8S) diastereomer
  • 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 ° 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-ylsulfanyl]-acetyl}-mutilin+(7S, 8S) diastereomer (Rf=0.7, 8.03 g, 67% yield) was obtained.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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, OCH2CH2O, 1′-H, 11-H, 22-H), 2.40 (bs, 1H, 4-H), 1.39, 1.36 (2s, 3H, 15-CH3), 1.05 (s, 3H, 18-CH3), 1.00 (s, 9H, Si-tert butyl), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.62, 0.60 (2d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 811 (MNa+).
  • Step C. 14O-{[(1R, 2R)-2-(tert-Butyl-diphenyl-silanyloxy)-4-oxo-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S) diastereomer
  • 14-O-{[(7R, 8R)-7-(tert-Butyl-diphenyl-silanyloxy)-1,4-dioxa-spiro[4.5]dec-8-ylsulfanyl]-acetyl}-mutilin+(7S, 8S) diastereomer (8.03 g, 10.2 mmol) was dissolved in 100 ml of dichloromethane and treated with montmorillonite K10(10 g) for 3 days at room temperature. After filtration over celite the reaction mixture was concentrated under reduced pressure and subjected to chromatography (silica, cyclohexane/ethyl acetate 2/1) to yield 14-O-{[(1R, 2R)-2-(tert-butyl-diphenyl-silanyloxy)-4-oxo-cyclohexylsulfanyl]-acetyl)-mutilin+(1S, 2S) diastereomer (Rf=0.38, 5.24 g, 69% yield).
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.06 (s, 3H, 18-CH3), 0.97 (s, 9H, Si-tert-butyl), 0.81 (d, 3H, 17-CH3, J 7 Hz), 0.58 (d, 3H, 16-CH3, J=7 Hz).
  • Step D. 14-O-{[(1R, 2R)-2-(tert-Butyl-diphenyl-silanyloxy)-4-hydroxyimino-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S) diastereomer
  • To a solution of 14-O-{[(1R, 2R)-2-(tert-butyl-diphenyl-silanyloxy)4-oxo-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S) diastereomer (2.50 g, 3.36 mmol) in 10 ml of dimethylformamide was added hydroxylamine hydrochloride (233 mg, 3.36 mmol) and triethylamine (0.47 ml, 3.36 mmol) and stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure, diluted with water and brine and extracted three times with ethyl acetate. The combined organic layers were washed twice with water and dried over sodium sulfate and filtered, The solvent was removed under reduced pressure and crude 14-O-{[(1R, 2R)-2-(tert-butyl-diphenyl-silanyloxy)4-hydroxyimino-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S) diastereomer (quantitative yield, cyclohexane ethyl acetate=2/1, Rf=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
  • 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 NaHCO3 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) (Rf=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) (Rf=0.3, 670 mg, 26% yield).
  • (a): 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.00 (s, 12H, 18-CH3, Si-tert-butyl), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.55 (d, 3H, 16-CH3, J 7 Hz).
  • (b): 1H NMR (400 MHz, DMSO-d6, δ, 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 (νA=3.37, νB=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-CH3), 1.06 (s, 3H, 18-CH3), 1.00 (s, 9H, Si-tert-butyl), 0.83 (d, 3H, 17-CH3, J=7 Hz), 0.64, 0.62 (2d, 3H, 16-CH3, J=7 Hz).
  • Step F. 14-O-{[(1R, 2R, 4S)-2-(tert-Butyl-diphenyl-silanyloxy)-4-(formyl-hydroxy-amino-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 4R) diastereomer
  • 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 μl, 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, Rf=0.5) as colorless solid.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.03 (s, 12H, 18-CH3, Si-tert-butyl), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.55 (d, 3H, 16-CH3, J=6 Hz).
  • Step G. 14-O-{[(1R, 2R, 4S)-4-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 4R) diastereomer
  • 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, Rf=0.4) as colorless solid.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.06 (s, 12H, 18-CH3), 0.81 (d, 3H, 17-CH3, J=7 Hz), 6.63 (d, 3H, 16-CH3, J=6 Hz). MS-ESI (m/z): 574 MNa+), 550 (M-H)−, 1101 (2M-H)−.
    • Example 14 14-O-{[(1R, 2R, 5S)-5-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer
  • Step A. (7R, 8R)-8-(2,4,6-Trimethylbenzylsufanyl)-1,4-dioxa-spiro[4.5]decan-7-ol+(7S, 8S) diastereomer
  • 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 (Rf=0.4, 33 g, 85% yield) was obtained as oil.
  • MS-ESI (m/z): 345 (MNa+), 667 (2MNa+).
  • Step B. Acetic acid (7R, 8R)-7-(2,4,6-trimethylbenzylsufanyl)-1,4-dioxa-spiro[4.5]dec-8-yl ester+(7S, 8S) diastereomer
  • 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° 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 (Rf=0.4, 7.0 g, 22% yield).
  • 1H NMR (500 MHz, CDCl3, δ, ppm): 6.81 (s, 2H, aromat.-H), 4.85 (m, 1H, CHO), 3.96 (m, 4H, OCH2CH2O), AB-system (νA=3.83, νB=3.79, J=11 Hz, SCH2), 2.99 (m, 1H, CHS), 2.36 (s, 6H, CH3), 2.23 (s, 3H, CH3), 2.18 (m, 1H), 2.12 (m, 1H), 2.11 (s, 3H, COCH3), 1.90-1.58 (m, 4H). MS-ESI (m/z): 387 (MNa+).
  • Step C. (7R, 8R)-7-Mercapto-1,4-dioxa-spiro[4.5]decan-8-ol+(7S, 8S) diastereomer
  • 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 (Rf=0.4, 1.36 g, 38%) was obtained.
  • 1H NMR (400 MHz, DMSO-d6, δ, ppm): 4.89 (d, 1H, OH), 3.83 (m, 4H, OCH2CH2O), 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)−.
  • Step D. 14-O-{[(7R, 8R)-8-Hydroxy-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-acetyl}-mutilin+{7S, 8S) diastereomer
  • (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 (Rf=0.25, 1.90 g, 48%) was obtained as colorless amorphous foam.
  • MS-ESI (m/z): 573 (MNa+), 1123 (2MNa+), 549 (M-H)−, 585 (MCl−).
    Step E. 14-O-{[(7R, 8R)-8-(tert-Butyl-diphenyl-silanyloxy)-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-acetyl}-mutilin+(7S, 8S) diastereomer
  • 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 (Rf=0.6, 1.65 g, 61%) was obtained as colorless amorphous foam.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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, OCH2CH2O), 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-CH3), 1.05 (s, 3H, 18-CH3), 1.00 (s, 9H, Si-tert-butyl), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.60, 0.58 (2d, 3H, 16-CH3, J=7 Hz).
  • Step F. 14-O-{[(1R, 2R)-2-(tert-Butyl-diphenyl-silanyloxy)-5-oxo-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S) diastereomer
  • 14-O-{[(7R, 8R)-8-(tert-Butyl-diphenyl-silanyloxy)-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-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}-mutilin+(1S,2S) diastereomer (1.34 g, 86% yield, cyclohexane/ethyl acetate=2/1, Rf=0.3) was obtained as colorless amorphous foam which was directly used for the next step.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.02 (s, 12H, 18-CH3, Si-tert-butyl), 0.81 (d, 3H, 17CH3, J=7 Hz), 0.53 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 767 (MNa+), 779 (MCl−).
  • Step G. 14-O-{[(1R, 2R)-2-(tert-Butyl-diphenyl-silanyloxy)-5-hydroxyimino-cyclobexylsulfanyl]-acetyl}-mutilin+(1S, 2S) diastereomer
  • 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, Rf=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
  • 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) (Rf=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) (Rf=0.25, 560 mg, 41% yield) were obtained.
  • (a): 1H NMR (400 MHz, DMSO-dhd 6, δ, 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 (νA=3.93, νB=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-CH3), 1.00 (s, 12H, 18-CH3, Si-tert-butyl), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.55 (d, 3H, 16-CH3, J=7 Hz).
  • (b) 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.05 (s, 3H, 18-CH3), 0.98 (s, 9H, Si-tert-butyl), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.63, 0.61 (2d, 3H, 16-CH3, J=6 Hz).
  • Step I. 14-O-{[(1R, 2R, 5S)-2-(tert-Butyl-diphenyl-silanyloxy)-5-(formyl-hydroxy-amino-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer
  • 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-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer (135 mg, 61% yield, cyclohexane/ethyl acetate 1/3, Rf=0.65) was obtained as colorless solid.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.03 (s, 12H, 18-CH3, Si-tert-butyl), 0.82, 0.80 (d, 3H, 17-CH3, J=7 Hz), 0.55 (d, 3H, 16-CH3, J=6 Hz).
  • Step J. 14-O-{[(1R, 2R, 5S)-5-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 5R) diastereomer
  • 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}-mutilin+(1S, 2S, 5R) diastereomer (77 mg, 85% yield, dichloromethane methanol=9/1, Rf=0.4) was obtained as colorless solid.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.06 (s, 12H, 18-CH3), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=6 Hz). MS-ESI (m/z): 574 (MNa+), 1125 (MNa+), 550 (M-H)-, 1101 (2M-H)−.
    • Example 15 14-O-{[(1R, 2R, 3R/S)-3-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 3R/S) diastereomer
  • Step A. 14-O-{[(6R, 7R)-6-Hydroxy-1,4-dioxa-spiro[4.5]dec-7-ylsulfanyl]-acetyl}-mutilin +(6S, 7S) diastereomer
  • 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 (Rf=0.5, 15.6 g, 55% yield) was obtained as colorless amorphous foam.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.05 (s, 3H, 18-CH3), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.62 (d, 3H, 16-CH3, J=6 Hz).
  • Step B. 14-O-{[(1R, 2R)-2-Hydroxy-3-oxo-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 25) diastereomer
  • 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 (Rf=0.4, 3.14 g, 22% yield) was obtained as colorless amorphous foam.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.06 (s, 3H, 18-CH3), 0.80 (d, 3H, 17-CH3, J=7 Hz), 0.61 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 529 (MNa+), 1035 (2MNa+.
  • Step C. 14-O-([(1R, 2R)-2-Hydroxy-3-hydroxyimino-cyclohexylsulfanyl]-acetyl}-mutilin +(1S, 2S) diastereomer
  • 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 (Rf=0.2, 1.75 g, 54% yield) was obtained as colorless amorphous foam.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.05 (s, 3H, 18-CH3), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=7 Hz).
  • Step D. 14-O-{[(1R, 2R, 3R/S)-2-Hydroxy-3-Hydroxyamino-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 3R/S) diastereomer
  • 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, 3Rf=2-hydroxy-3-Hydroxyamino-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 3R/S) diastereomer (Rf=0.2, 1.34 g, 65% yield) was obtained as colorless amorphous foam.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.05 (s, 3H, 18-CH3), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=7 Hz);
  • Step E. 14-O-{[(1R, 2R, 3R/S)-3-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin+(1S, 2S, 3R/S) diastereomer
  • 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, Rf=0.5) was obtained as colorless solid.
  • 1H NMR (400 MHz, DMSO-d6, δ, 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-CH3), 1.06 (s, 12H, 18-CH3), 0.81 (d, 3H, 17-CH3, J=7 Hz), 0.63 (d, 3H, 16-CH3, J=7 Hz). MS-ESI (m/z): 574 (MNa+), 1125 (2MNa+), 550 (M-H)−, 1101 (2M-H)−.

Claims (10)

1. A compound of formula (I)
Figure US20090118366A1-20090507-C00009
wherein
n is 0 to 4;
R is ethyl or vinyl;
R1 is hydrogen or (C1-6)alkyl,
R2 is hydrogen or
cycloalkyl including (C3-6)cycloalkyl, or
unsubstituted (C1-6)alkyl, or
(C1-6)alkyl substituted by one or more of
hydroxy; preferably one or two,
halogen,
(C3-6)cycloakyl, or
R1 is hydroxy and R2 is formyl.
2. A compound of formula (II)
Figure US20090118366A1-20090507-C00010
wherein n, R1 and R2 are as defined in claim 1.
3. A compound of formula (III)
Figure US20090118366A1-20090507-C00011
wherein n, R1 and R2 are as defined in claim 1.
4. A compound of formula (IV)
Figure US20090118366A1-20090507-C00012
wherein n, R1 and R2 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}-mutilin and the (1S, 2S, 4R) diastereomer thereof
14-O-{[(1R, 2R, 5S)-5-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin and the (1S, 2S, 5R) diastereomer thereof
and
14-O-{[(1R, 2R, 3R/S)-3-(Formyl-hydroxy-amino)-2-hydroxy-cyclohexylsulfanyl]-acetyl}-mutilin 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.
US11/828,224 2007-03-20 2007-07-25 Organic compounds Abandoned US20090118366A1 (en)

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US13/252,732 Active US8153689B2 (en) 2007-03-20 2011-10-04 Pleuromutilin derivatives for the treatment of diseases mediated by microbes
US13/438,160 Abandoned US20130040954A1 (en) 2007-03-20 2012-04-03 Pleuromutilin derivatives for the treatment of diseases mediated by microbes
US14/283,734 Abandoned US20140256731A1 (en) 2007-03-20 2014-05-21 Pleuromutilin derivatives for the treatment of diseases mediated by microbes
US15/480,756 Abandoned US20170266194A1 (en) 2007-03-20 2017-04-06 Pleuromutilin derivatives for the treatment of diseases mediated by microbes
US16/023,880 Abandoned US20180303842A1 (en) 2007-03-20 2018-06-29 Pleuromutilin derivatives for the treatment of diseases mediated by microbes
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US16/881,117 Abandoned US20200281933A1 (en) 2007-03-20 2020-05-22 Pleuromutilin derivatives for the treatment of diseases mediated by microbes
US17/138,097 Abandoned US20210113571A1 (en) 2007-03-20 2020-12-30 Pleuromutilin derivatives for the treatment of diseases mediated by microbes
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US13/438,160 Abandoned US20130040954A1 (en) 2007-03-20 2012-04-03 Pleuromutilin derivatives for the treatment of diseases mediated by microbes
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US16/023,880 Abandoned US20180303842A1 (en) 2007-03-20 2018-06-29 Pleuromutilin derivatives for the treatment of diseases mediated by microbes
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