Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
  • C07D498/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains three hetero rings
  • C07D498/20—Spiro-condensed systems
• • 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
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
  • C07D513/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains three hetero rings
  • C07D513/16—Peri-condensed systems
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
  • C07D513/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains three hetero rings
  • C07D513/20—Spiro-condensed systems

Definitions

• the present invention relates to macrocyclic indole compounds, to pharmaceutical compositions containing them, to their use in the prevention and treatment of hepatitis C infections and to methods of preparation of such compounds and compositions.
• HCV Hepatitis C
• Molecolare P. Angeletti SpA specifically discloses compounds of the formula (A) wherein R 1 is carboxy and R 2 is hydrogen, A is cyclohexyl, Z is oxygen, W and Y are both CH 2 and X is a spiro-azetidine ring optionally substituted on the ring nitrogen atom.
• R 1 is selected from hydrogen, hydroxyl, fluoro or a Ci_4 alkoxy group or R 1 is linked to R 3 to form a 4, 5 or 6-membered aliphatic ring optionally containing one N or O atom;
• R 2 is selected from cyclopentyl or cyclohexyl optionally substituted by a fluoro;
• R 3 is hydrogen or is linked to R 1 as hereinbeforedefmed
• R 4 is hydrogen, a Ci_4 alkyl group optionally substituted by a group NR 8 R 9 , in which R 8 and R 9 are each hydrogen or a Ci_4alkyl group or R 4 is linked to R 6 to form a (CH 2 )I n NR 10 SO 2 N group wherein m is an integer selected from 2 or 3 and R 10 is a Ci_4 alkyl group; R 5 is hydrogen or a Ci_4 alkyl group;
• R 6 is hydroxyl or is linked to R 4 as hereinbefore defined;
• R 7 is hydrogen or an aliphatic ring of 4-7 ring atoms containing one or two heteroatoms selected from O, N or S or a group S(O), S(O) 2 ,,NH or NCi_ 4 alkyl and optionally substituted by fluoro or R 7 is a 5-10-membered heteroaromatic ring system containing 1 to 4 heteroatoms independently selected from N, O and S, which ring is optionally substituted by one or two halo atoms or Ci_4 alkyl or Ci_4 alkoxy groups;
• Y is a single bond or a CH 2 group; when R 4 is linked to R 6 and m is 3 then Y is a single bond; when R 4 is linked to R 6 and m is 2 then Y is a CH 2 group; n is an integer from zero to four; with the proviso that when R 1 is hydrogen, n is not zero; and pharmaceutically acceptable salts thereof.
• R 1 is selected from hydrogen, hydroxyl, fluoro or a methoxy group or R 1 is linked to R to form a 5 or 6-memebered aliphatic ring optionally conaining one N or O atom such as a fur an ring.
• R 2 is selected from cyclohexyl or cyclohexyl optionally substituted by a fluoro and preferably R 2 is cyclohexyl.
• R 3 is hydrogen or linked to R 1 to form a 5 or 6-memebered aliphatic ring optionally conaining one N or O atom such as a furan ring.
• R 3 is hydrogen when R 4 is linked to R 6 .
• R 4 is a Ci_4 alkyl group optionally substituted by a group NR 8 R 9 or is linked to R 6 as hereinbefore defined and R 10 is a methyl group.
• R 7 is a five- or six-membered aliphatic ring optionally containing 1 or 2 heteroatoms independently selected from N and O and S, and which ring is optionally substituted by one or two fluoro.
• R 7 is 1-piperidino, 1- morpholino or 1-pyrrolidino optionally substituted by one or two fluoro.
• R 7 is a 5-10-membered heteroaromatic ring system containing 1 , 2 or 3 heteroatoms independently selected from N, O and S and which ring is optionally substituted by one or two fluoro atoms or a methyl, ethyl or methoxy group.
• R 7 is hydrogen.
• n is zero or one or three and preferably n is one to three.
• the present invention provides the compound of the formula
• n, R 1 , R 2 and R 10 are as hereinbefore defined
• XY 1 is a group CH 2 NR 5 or NR 5 CH 2 wherein R 5 is as hereinbefore defined
• Het is an aliphatic ring of 4-7 ring atoms containing one or two heteroatoms selected from O, N or S or a group S(O), S(O) 2 , NH or NC 1-4 alkyl, Het being optionally substituted by 1-3 fluoro or Het is a 5-10-membered heteroaromatic ring system containing 1 to 4 heteroatoms independently selected from N, O and S, which ring is optionally substituted by one or two halo atoms or Ci_4 alkyl or Ci_4 alkoxy groups, and pharmaceutically acceptable salts thereof.
• R 1 is hydrogen, hydroxyl or fluoro, and preferably hydrogen.
• Het is a 5- or 6-membered aliphatic ring system containing 1 or 2 heteroatoms independently selected from N, and O which ring is optionally substituted by one or two halo atoms, suitably fluoro, or a Ci_4 alkyl or Ci_4 alkoxy group,, and pharmaceutically acceptable salts thereof.
• Het is 1-piperidino, 1-morpholino or 1-pyrrolidino optionally substituted by one or two fluoro.
• the present invention provides the compound of the formula (III):
• R 2 to R 6 are as hereinbefore defined and Y 2 is a single bond or a CH 2 group and R lb is linked to R 3 to form a 4, 5 or 6-memebered aliphatic ring optionally conaining one N or O atom, and pharmaceutically acceptable salts thereof.
• R lb is linked to R 3 to form a 5 or 6- memebered aliphatic ring optionally containing one N or O atom such as a furan ring.
• the present invention provides the compound of the formula (IV):
• alkyl as a group or part of a group means that the group is straight or branched.
• suitable alkyl groups include methyl, ethyl, n-propyl, i- propyl, n-butyl, s-butyl and t-butyl.
• suitable alkenyl groups include vinyl and allyl.
• aliphatic ring as a group or part of a group means a ring system which contains 4 to 7 ring atoms and single or double bonds which do not impart aromaticity to the ring.
• the ring may optionally contain 1 or 2 heteroatoms independently selected from N and O and S or a group S(O), S(O) 2 ,,NH or NCi_4alkyl.
• Particular examples of such groups include pyrrolino, morpholino, and piperidino.
• heteroaryl as a group or part of a group means a 5-10 membered heteroaromatic ring system containing 1 to 4 heteroatoms selected from N, O and S.
• Such groups include pyrrolyl, furanyl, thienyl, pyridyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazolyl, oxadiazolyl, thiadiazolyl, triazinyl, tetrazolyl, indolyl, benzothienyl, benzimidazolyl, benzofuryl, quinolinyl and isoquinolinyl.
• substituents may be present.
• substituents may be attached to the compounds or groups which they substitute in a variety of ways, either directly or through a connecting group of which the following are examples: amine, amide, ester, ether, thioether, sulfonamide, sulfamide, sulfoxide, urea, thiourea and urethane.
• an optional substituent may itself be substituted by another substituent, the latter being connected directly to the former or through a connecting group such as those exemplified above.
• Specific compounds within the scope of this invention include: (7R)- 14-cyclohexyl- 18,21 -dimethyl-4-(piperidin- 1 -ylmethyl)-7,8-dihydro-6/f-7, 11 - (propanoiminoethanoiminothioiminomethano)indolo[l ,2-e] [1 ,5]benzoxazocin- 15-one 17,17- dioxide,
• the salts of the compounds of formula (I) will be non-toxic pharmaceutically acceptable salts.
• Other salts may, however, be useful in the preparation of the compounds according to the invention or of their non-toxic pharmaceutically acceptable salts.
• Suitable pharmaceutically acceptable salts of the compounds of this invention include acid addition salts which may, for example, be formed by mixing a solution of the compound according to the invention with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, fumaric acid, /?-toluenesulfonic acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, carbonic acid, phosphoric acid or sulfuric acid.
• a pharmaceutically acceptable acid such as hydrochloric acid, fumaric acid, /?-toluenesulfonic acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, carbonic acid, phosphoric acid or sulfuric acid.
• Salts of amine groups may also comprise quaternary ammonium salts in which the amino nitrogen atom carries a suitable organic group such as an alkyl, alkenyl, alkynyl or aralkyl moiety.
• suitable pharmaceutically acceptable salts thereof may include metal salts such as alkali metal salts, e.g. sodium or potassium salts; and alkaline earth metal salts, e.g. calcium or magnesium salts.
• the salts may be formed by conventional means, such as by reacting the free base form of the product with one or more equivalents of the appropriate acid in a solvent or medium in which the salt is insoluble, or in a solvent such as water which is removed in vacuo or by freeze drying or by exchanging the anions of an existing salt for another anion on a suitable ion exchange resin.
• the present invention includes within its scope prodrugs of the compounds of formula (I) above.
• prodrugs will be functional derivatives of the compounds of formula (I) which are readily convertible in vivo into the required compound of formula (I).
• Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs", ed. H. Bundgaard, Elsevier, 1985.
• a prodrug may be a pharmacologically inactive derivative of a biologically active substance (the "parent drug” or “parent molecule”) that requires transformation within the body in order to release the active drug, and that has improved delivery properties over the parent drug molecule.
• the transformation in vivo may be, for example, as the result of some metabolic process, such as chemical or enzymatic hydrolysis of a carboxylic, phosphoric or sulfate ester, or reduction or oxidation of a susceptible functionality.
• the present invention includes within its scope solvates of the compounds of formula (I) and salts thereof, for example, hydrates.
• the present invention also includes within its scope any enantiomers, diastereomers, geometric isomers and tautomers of the compounds of formula (I). It is to be understood that all such isomers and mixtures thereof are encompassed within the scope of the invention.
• the present invention further provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in therapy.
• the present invention also provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for treatment or prevention of infection by hepatitis C virus in a human or animal.
• the invention provides the use of a compound of formula (I) as defined above, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treatment or prevention of infection by hepatitis C virus in a human or animal.
• a further aspect of the invention provides a pharmaceutical composition
• a pharmaceutical composition comprising a compound of formula (I) as defined above, or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable carrier.
• the composition may be in any suitable form, depending on the intended method of administration. It may for example be in the form of a tablet, capsule or liquid for oral administration, or of a solution or suspension for administration parenterally.
• compositions optionally also include one or more other agents for the treatment of viral infections such as an antiviral agent, or an immunomodulatory agent such as ⁇ -, ⁇ - or ⁇ -interferon.
• agents for the treatment of viral infections such as an antiviral agent, or an immunomodulatory agent such as ⁇ -, ⁇ - or ⁇ -interferon.
• the invention provides a method of inhibiting hepatitis C virus polymerase and/or of treating or preventing an illness due to hepatitis C virus, the method involving administering to a human or animal (preferably mammalian) subject suffering from the condition a therapeutically or prophylactically effective amount of the pharmaceutical composition described above or of a compound of formula (I) as defined above, or a pharmaceutically acceptable salt thereof.
• Effective amount means an amount sufficient to cause a benefit to the subject or at least to cause a change in the subject's condition.
• the dosage rate at which the compound is administered will depend on a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age of the patient, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition and the host undergoing therapy. Suitable dosage levels may be of the order of 0.02 to 5 or 10 g per day, with oral dosages two to five times higher. For instance, administration of from 1 to 50 mg of the compound per kg of body weight from one to three times per day may be in order. Appropriate values are selectable by routine testing.
• An additional aspect of the invention provides a method of preparation of a pharmaceutical composition, involving admixing at least one compound of formula (I) as defined above, or a pharmaceutically acceptable salt thereof, with one or more pharmaceutically acceptable adjuvants, diluents or carriers and/or with one or more other therapeutically or prophylactically active agents.
• the present invention also provides a process for the preparation of compounds of formula (I).
• the compounds of the present invention can be administered by any means that produces contact of the active agent with the agent's site of action. They can be administered by one or more conventional means available for use in conjunction with pharmaceuticals, either as individual therapeutic agents or in a combination of therapeutic agents. They can be administered alone, but typically are administered with a pharmaceutical carrier selected on the basis of the chosen route of administration and standard pharmaceutical practice.
• the compounds of the invention can, for example, be administered by one or more of the following: orally, parenterally (including subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques), by inhalation (such as in a spray form), or rectally, in the form of a unit dosage of a pharmaceutical composition containing an effective amount of the compound and conventional non-toxic pharmaceutically-acceptable carriers, adjuvants and vehicles.
• Liquid preparations suitable for oral administration e.g., suspensions, syrups, elixirs and the like
• Solid preparations suitable for oral administration can be prepared according to techniques known in the art and can employ such solid excipients as starches, sugars, kaolin, lubricants, binders, disintegrating agents and the like.
• Parenteral compositions can be prepared according to techniques known in the art and typically employ sterile water as a carrier and optionally other ingredients, such as solubility aids.
• injectable solutions can be prepared according to methods known in the art wherein the carrier comprises a saline solution, a glucose solution or a solution containing a mixture of saline and glucose. Further description of methods suitable for use in preparing pharmaceutical compositions of the present invention and of ingredients suitable for use in said compositions is provided in Remington's Pharmaceutical Sciences, 18 th edition (ed. A. R. Gennaro, Mack Publishing Co., 1990).
• the present invention also relates to a method of inhibiting HCV NS5B activity, inhibiting HCV viral replication and/or HCV viral production, treating HCV infection and/or reducing the likelihood or severity of symptoms of HCV infection with a compound of the present invention in combination with one or more therapeutic agents and a pharmaceutical composition comprising a compound of the present invention and one or more therapeutic agents selected from the group consisting of a HCV antiviral agent, an immunomodulator, and an anti-infective agent.
• Such therapeutic agents active against HCV include, but are not limited to, ribavirin, levovirin, viramidine, thymosin alpha- 1, R7025 (an enhanced interferon (Roche)), interferon- ⁇ , interferon- ⁇ , pegylated interferon- ⁇ (peginterferon- ⁇ ), a combination of interferon- ⁇ and ribavirin, a combination of peginterferon- ⁇ and ribavirin, a combination of interferon- ⁇ and levovirin, and a combination of peginterferon- ⁇ and levovirin.
• the combination of pegylated- interferon and ribaviron represents the current Standard of Care for HCV treatment.
• Interferon- ⁇ includes, but is not limited to, recombinant interferon- ⁇ 2a (such as ROFERON interferon available from Hoffmann- LaRoche, Nutley, NJ), pegylated interferon- ⁇ 2a (PEGASYS), interferon- ⁇ 2b (such as INTRON-A interferon available from Schering Corp., Kenilworth, NJ), pegylated interferon- ⁇ 2b
• interferon- ⁇ 2a such as ROFERON interferon available from Hoffmann- LaRoche, Nutley, NJ
• PEGASYS pegylated interferon- ⁇ 2a
• interferon- ⁇ 2b such as INTRON-A interferon available from Schering Corp., Kenilworth, NJ
• the compounds of the invention may also be administered in combination with the antiviral agent NS5B polymerase inhibitor R7128 (Roche).
• the compounds of the present invention also may be combined for the treatment of HCV infection with antiviral 2'-C-branched ribonucleosides disclosed in Rogers E. Harry-O'Kuru et al., A Short, Flexible Route toward 2 '-C-Br -cinched Ribonucleosides, 62 J. ORG. CHEM. 1754-59 (1997); Michael S. Wolfe & Rogers E. Harry-O'Kuru, A Concise Synthesis of 2'-C- Methylribonucleosides, 36(42) TETRAHEDRON LETTERS 7611-14 (1995); U.S. Patent
• Such 2'-C-branched ribonucleosides include, but are not limited to, 2'-C-methyl-cytidine, 2'-C-methyl-uridine, 2'-C-methyl-adenosine, 2'-C-methyl- guanosine, and 9-(2-C-methyl- ⁇ -D-ribofuranosyl)-2,6-diaminopurine, and the corresponding amino acid ester of the ribose C-2', C-3', and C-5' hydroxyls and the corresponding optionally substituted cyclic 1,3 -propanediol esters of the 5 '-phosphate derivatives.
• the compounds of the present invention may also be administered in combination with an agent that is an inhibitor of HCV NS3 serine protease.
• HCV NS3 serine protease is an essential viral enzyme and has been described to be an excellent target for inhibition of HCV replication.
• Exemplary substrate and non- substrate based inhibitors of HCV NS3 protease inhibitors are disclosed in International Patent Application Publications WO 98/22496, WO 98/46630, WO 99/07733, WO 99/07734, WO 99/38888, WO 99/50230, WO 99/64442, WO 00/09543, WO 00/59929, WO 02/48116, WO 02/48172, WO 2008/057208 and WO 2008/057209, in British Patent No. GB 2 337 262, and in U.S. Patent Nos. 6,323,180 and 7,470,664.
• the compounds of the present invention may also be combined for the treatment of HCV infection with nucleosides having anti-HCV properties, such as those disclosed in International Patent Application Publications WO 02/51425, WO 01/79246, WO 02/32920, WO 02/48165 and WO 2005/003147 (including R1656, (2'R)-2'-deoxy-2'-fluoro-2'-C- methylcytidine, shown 77); WO 01/68663; WO 99/43691; WO 02/18404 and WO 2006/021341, and U.S. Patent Application Publication US 2005/0038240, including 4'-azido nucleosides such as R1626, 4'-azidocytidine; U.S.
• the compounds of the present invention may also be administered in combination with an agent that is an inhibitor of HCV NS5B polymerase.
• HCV NS5B polymerase inhibitors that may be used as combination therapy include, but are not limited to, those disclosed in International Patent Application Publications WO 02/057287, WO 02/057425, WO 03/068244, WO 2004/000858, WO 04/003138 and
• HCV polymerase inhibitors include, but are not limited to, valopicitabine (NM-283; Idenix) and 2'-F-2'-beta-methylcytidine (see also WO 2005/003147).
• additional nucleoside HCV NS5B polymerase inhibitors that are used in combination with the present HCV NS5B inhibitors are selected from the following compounds: 4-amino-7-(2-C-methyl- ⁇ -D-arabinofuranosyl)-7H-pyrrolo[2,3-(i]pyrimidine; 4- amino-7-(2-C-methyl- ⁇ -D-ribofuranosyl)-7H-pyrrolo[2,3-(i]pyrimidine; 4-methylamino-7-(2-C- methyl- ⁇ -D-ribofuranosyl)-7H-pyrrolo[2,3-(i]pyrimidine; 4-dimethylamino-7-(2-C-methyl- ⁇ -D- ribofuranosyl)-7/f-pyrrolo[2,3-J]pyrimidine; 4-cyclopropylamino-7-(2-C-methyl- ⁇ -D- ribofuranosyl)-7H-pyrrolo[2,3-(i]pyrimidine; 4-a
• the compounds of the present invention may also be combined for the treatment of ⁇ CV infection with non-nucleoside inhibitors of ⁇ CV polymerase such as those disclosed in U.S. Patent Applciation Publications US 2006/0100262 and US 2009/0048239; International Patent Application Publications WO 01/77091, WO 01/47883, WO 02/04425, WO 02/06246, WO 02/20497, WO 2005/016927 (in particular JTK003), WO 2004/041201, WO 2006/066079, WO 2006/066080, WO 2008/075103, WO 2009/010783 and WO 2009/010785; the content of each is incorporated herein by reference in its entirety.
• additional non-nucleoside ⁇ CV NS5B polymerase inhibitors that are used in combination with the present ⁇ CV NS5B inhibitors are selected from the following compounds: 14-cyclohexyl-6-[2-(dimethylamino)ethyl]-7-oxo-5,6,7,8- tetrahydroindolo[2,l- ⁇ ][2,5]benzodiazocine-l 1-carboxylic acid; 14-cyclohexyl-6-(2-morpholin- 4-ylethyl)-5,6,7,8-tetrahydroindolo[2,l- ⁇ ][2,5]benzodiazocine-l 1-carboxylic acid; 14- cyclohexyl-6-[2-(dimethylamino)ethyl]-3-methoxy-5,6,7,8-tetrahydroindolo[2,l- ⁇ ] [2,5]benzodiazocine-l 1-carboxylic acid;
• the present ⁇ CV NS5B polymerase inhibitors are used in combination with non-nucleoside ⁇ CV NS5A inhibitors and pharmaceutically acceptable salts thereof.
• Compounds of formula (I) wherein R 4 is linked to R 6 may be prepared by: a) internal ring closure of the compound of formula (V): where R 1 , R 2 , R 3 , R 6 and R 7 are as defined in relation to formula (I), and M' and M" have suitable precursor functionality to form group X- Y 1 as defined in relation to formula (II).
• M' can be CHO and M" can be HN(CH 3 )-CH 2 -, where the reaction is carried out in the presence of a mild reducing agent, such as sodium cyanoborohydride, in a suitable solvent mixture, such as aqueous methanol, at pH 5-6.
• R 1 , R 2 , R 3 , R 6 and R 7 are as defined in relation to formula (I), and X 1 and Y 2 are either X and Y 1 as defined in formula (II) or suitable precursors.
• the reaction is conveniently carried out in the presence of a coupling reagent, such as EDC, and an additive, such as DMAP, in a suitable solvent such as DMF, DCM and mixtures thereof.
• the compound of formula (I) where NR 5 and/or NR 10 comprises an N-H group may be converted into the compound of formula (I) where NR 5 and/or NR 10 comprises an N-CH 2 CH 3 or an N- CH(CH 3 ) 2 group by using acetaldehyde or acetone, respectively.
• alkylation of the N-H group may be carried out using a suitable alkyl halide and a base, such as potassium carbonate, in a suitable solvent, such as acetone, MeCN or DMF.
• Methods A, B and C Three general strategies were employed for assembly of compounds from the macrocyclic class (Methods A, B and C); Method C can be regarded as an extension of Methods A, and B.
• Functional group manipulation and macrocyclization sets up the macrocycle.
• Functional groups on the macrocycle can then be manipulated post-closure, eg, via reductive amination, alkylation, amide reduction, amide formation etc.
• the precursor assembled in Method A was modified at R 1 if appropriate, and then the M" functionality was converted into the desired linker terminating in the protected sulfamide.
• modification of the pre-cursor functionality W can be performed at this juncture, or can also be performed prior to manipulation of R 1 or M". Unmasking of the acid and sulfamide was followed by cyclization and optionally by further manipulation of the functional groups in the macrocycle.
• Method D Two general strategies may be employed for assembly of spirocyclic compounds (Methods D, and E). Method E can be considered an extension of D.
• Method D a suitably functionalized keto-tether is assembled first (as described in WO 2006/046030). Allylation sets up the quaternary centre. Activation of the double bond with subsequent cyclization generates the spiro-system with a suitable handle for further functionalization. Nucleophilic displacement and functional group manipulation introduces the side chain. Following ester cleavage, preparative HPLC allows for separation of the relative cis- and trans- diastereoisomers on the dihydro-spirofuran ring, as their racemates. Separation of the enantiomers could be realized via chiral SFC.
• Method E functional group manipulation of the spirocyclic intermediate from method D afforded the primary amine. Installation of the macrocyclic linker and R 1 on the nitrogen (in analogous fashion to what is described in WO 2009/010783 and WO 2009/010785), was followed by sequential ester deprotection and unmasking of the sulfamide. Macrocyclization under peptide coupling conditions afforded the desired targets.
• any of the above synthetic sequences it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules concerned. This may be achieved by means of conventional protecting groups, such as those described in Protective Groups in Organic Chemistry, ed. J.F.W. McOmie, Plenum Press, 1973; and T.W. Greene & P. G. M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 4th edition, 2007.
• the protecting groups may be removed at a convenient subsequent stage using methods known from the art.
• the present invention is illustrated further by the following non-limiting examples.
• the compounds of the invention were tested for inhibitory activity against the HCV RNA dependent RNA polymerase (NS5B) in an enzyme inhibition assay as described in example i).
• the compounds generally have ICso's below 100 nM in this assay.
• Incorporation of radioactive UMP was measured as follows. The standard reaction (50 ⁇ l) was carried out in a buffer containing 20 mM tris/HCl pH 7.5, 5 mM MgCl 2 , 1 mM DTT, 50 mM NaCl, 0.03% N-octylglucoside, 1 ⁇ Ci [ 3 H]-UTP (40 Ci/mmol, NEN), 10 ⁇ M UTP and 10 ⁇ g/ml poly(A) or 5 ⁇ M NTPs and 5 ⁇ g/ml heteropolymeric template. Oligo(U)i2 (1 ⁇ g/ml, Genset) was added as a primer in the assay working on PoIy(A) template.
• the final NS5B enzyme concentration was 5 nM.
• the order of assembly was: 1) compound, 2) enzyme, 3) template/primer, 4) NTP.
• Cell clones that stably maintain subgenomic HCV replicon were obtained by transfecting Huh-7 cells with an RNA replicon identical to l3 77 neo/NS3-37wt described by Lohmann et al. (1999) (EMBL-genbank No. AJ242652), followed by selection with neomycin sulfate (G418). Viral replication was monitored by measuring the expression of the NS3 protein by an ELISA assay performed directly on cells grown in 96 wells microtiter plates (CeIl-ELISA) using the anti-NS3 monoclonal antibody 10E5/24 (as described in published International application WO 02/59321).
• Ai absorbance value of HBIlO cells supplemented with the indicated inhibitor concentration.
• Ao absorbance value of HBIlO cells incubated without inhibitor.
• b absorbance value of Huh-7 cells plated at the same density in the same microtiter plates and incubated without inhibitor.
• n Hill coefficient.
• a standard MTT assay as described by T. Mosmann (Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays; J Immunol Methods. 1983, 65, 55-63) was used.
• the assay is based on the capacity of mitochondrial dehydrogenase enzymes in living cells to convert the yellow water- soluble substrate 3-(4,5-dimethilthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) into a dark blue formazan product, which is insoluble in water.
• the amount of formazan produced is directly proportional to the cell number in a range of cell line.
• Reagents were usually obtained directly from commercial suppliers (and used as supplied) but a limited number of compounds from in-house corporate collections were utilised. In the latter case the reagents are readily accessible using routine synthetic steps that are either reported in the scientific literature or are known to those skilled in the art.
• Mass spectral (MS) data were obtained on a Perkin Elmer API 100, or Waters MicroMass ZQ, operating in negative (ES " ) or positive (ES + ) ionization mode and results are reported as the ratio of mass over charge (m/z) for the parent ion only.
• Preparative scale HPLC separations were carried out on a Waters Delta Prep 4000 separation module, equipped with a Waters 486 absorption detector or on an automated Waters Fraction Lynx or Gilson preparative system. In all cases compounds were eluted with linear gradients of water and MeCN both containing 0.1% TFA using flow rates between 15 and 40 mL/min.
• aq. saturated solution
• TBAF tetrabutyl ammonium fluoride
• TBTU O-benzotriazol-l-yl- ⁇ /, ⁇ /, ⁇ f', ⁇ f'-tetramethyluronium tetrafluoroborate
• TFA trifluoroacetic acid
• THF tetrahydrofuran
• THP tetrahydropyranyl
• TMS trimethylsilyl
• Ts /? ⁇ ra-toluene sulfonyl.
• Step 1 methyl 4-azidobutanoate To a solution of methyl-4-bromobutyrate in DMSO (0.6 M) was added with stirring NaN 3 (1.5 eq). The suspension was heated (45-50 0 C, oil bath) with stirring for 5 h. After cooling, H 2 O was added and the mixture extracted with Et 2 O. The organic extracts were washed with brine and dried over Na 2 SO 4 . The solvent was removed in vacuo and the crude oil used without further purification (99%).
• Step 3 (4S)-3-(4-azidobutanoyl)-4-benzyl- 1 ,3-oxazolidin-2-one Et 3 N was added to a stirred mixture of 4-azidobutanoic acid in THF (0.3 M) cooled to -78 0 C and stirring was continued at this temperature for 10 min. Pivaloyl chloride (1.3 eq) was added dropwise. The resulting mixture was warmed to 0 0 C and stirred for 1 h before re-cooling to - 78°C.
• Step 4 (4S)-3-[(2R)-4-azido-2-(hvdroxymethyl)butanoyl]-4-benzyl-l,3-oxazolidin-2-one
• Step 6 (2S)-4-azido-2-( ⁇ [tert-butyl(dimethyl)silyl]oxy ⁇ methyl)butan-l-ol
• Et 2 O 0.117 M
• MeOH 1.1 eq
• LiBH 4 1.1 eq
• the mixture was stirred for 30 min at 0 0 C.
• the mixture was warmed to RT and stirred for 1 h.
• the mixture was then cooled to 0 0 C and sat. aq. NaHCO 3 was carefully added over 30 min.
• the biphasic mixture was stirred vigorously for 1 h at 0 0 C and then poured into H 2 O.
• Step 2 (2R)-5-azido-2-( ⁇ [tert-butyl(dimethyl)silyl] oxyimethvDpentyl methanesulfonate (25)-5-Azido-2-( ⁇ [tert-butyl(dimethyl)silyl]oxy ⁇ methyl)pentan-l-ol was treated in analogous fashion to the synthesis of intermediate A, step 7, to afford the title compound.
• Step 1 l-(methoxymethoxy)-2-methylbenzene
• a solution of NaH (60% dispersion in mineral oil; 1.2 eq) in Et 2 OZDMF (5:1, 0.2M) a solution of o-cresol in Et 2 O (1 eq, 2M) was added dropwise over 15 min.
• a solution of MOMCl in Et 2 O (1.1 eq, 2M) was added and the mixture was stirred for 30 min then poured into H 2 O and extracted with Et 2 O. The combined organic extracts were washed with IN NaOH, H 2 O and brine, dried over Na 2 SO 4 , filtered and evaporated in vacuo to give the title compound. The product was used in the next step without further purification.
• Step 1 methyl 3-cvclohexyl-2-(3-formyl-2-methoxyphenyl)-lH-indole-6-carboxylate
• Methyl 2-bromo-3-cyclohexyl-lH-indole-6-carboxylate prepared as in International patent application WO 2004/087714
• (3-formyl-2-methoxyphenyl) boronic acid 1.5 eq
• Pd(PPlIs) 2 Cl 2 0.1 eq
• dioxane 0.08M
• the solution was degassed and flushed with Ar.
• An aq. solution of Na 2 CO 3 solution (2 M, 1 eq) was added and the mixture was heated to 110 0 C for 2 h. All volatiles were evaporated in vacuo and the residual material was dissolved in EtOAc, filtered over Celite and evaporated in vacuo.
• Step 2 methyl 3-cvclohexyl-2-(3-formyl-2-hvdroxyphenyl)-lH-indole-6-carboxylate
• BBr 3 1 M in DCM, 2 eq
• MeOH an excess of MeOH was added dropwise and stirring continued for 20 min.
• the volatiles were evaporated and the mixture partitioned between EtOAc and IN HCl.
• the organic layer was washed with brine, dried over Na 2 SO 4 , filtered and concentrated in vacuo.
• Step 4 methyl 2-(2- ⁇ f(2R)-5-azido-2-(hvdroxymethyl)pentylJoxy ⁇ -3-formylphenyl)-3-cvclohexyl- lH-indole-6-carboxylate
• Step 5 methyl 2-(2- ⁇ [(2R)-5-azido-2- ⁇ [(methylsulfonyl)oxy]methyl ⁇ pentyl] oxy ⁇ -3- formylphenvD-S-cvclohexyl-lH-indole- ⁇ -carboxylate
• Step 6 methyl (7R)-7-(3-azidopropyl)- 14-cvclohexyl-4-formyl-7 ' ,8-dihvdro-6H-indolo [ 1 ,2- e] [1 ,5]benzoxazocine-l 1-carboxylate
• DMA 0.5 M
• CS 2 CO 3 2.0 eq
• DMA final concentration 0.050 M
• Step 8 methyl (7R)- 7-/3- / (tert-butoxycarbonyl) amino) ] propyl ⁇ - 14-cvclohexyl-4-(piperidin- 1- ylmethyl)-7 ,8-dihvdro-6H-indolo [ 1 ,2-e] [ 1 , 5 Jbenzoxazocine-11-carboxylate
• EtOAc/MeOH 3/1, 0.025 M
• BoC 2 O 1.5 eq
• 10 wt % palladium on carbon 0.1 eq
• Step 10 fert-butyl (3-IY7i?V14-cvclohexyl-l l-(IY2.2- dimethoxyethyl)(methyl)sulfamoyl "
• Step 11 (7R)- 14-cvclohexyl- 18,21 -dimethyl-4-(piperidin- 1 -ylmethvD-7 ⁇ 8-dihvdro-6H-7 ', 11 ' -
• Example 2 (7R)- 14-cyclohexyl- 18,2 l-dimethyl-4- [3-(pyrrolidin- l-yl)propyl] -7,8-dihydro- 6H-7,ll-(propanoiminoethanoiminothioiminomethano)indolo[l,2-e] [l,5]benzoxazocin-15- one 17,17-dioxide
• Step 1 methyl (7R)-7-(3-azidopropyl)-14-cyclohexyl-4-f(lE)-3-methoxy-3-oxoprop-l-en-l-ylJ- 7,8-dihvdro-6H-indolo[l,2-e] [1 ,5]benzoxazocine-l 1-carboxylate
• methyl methyl (7R)-7-(3- azidopropyl)-14-cyclohexyl-4-formyl-7,8-dihydro-6 ⁇ -indolo[l,2-e][l,5]benzoxazocine-l l- carboxylate Example 1, Step 6
• Step 2 methyl (7R)- 7- ⁇ -aminopropyl)- 14-cvclohexyl-4-(3-methoxy-3-oxopropyl)- 7, 8-dihvdro- 6H-indolofl,2-eJfl,5Jbenzoxazocine-l 1-carboxylate
• Step 3 methyl (7R)-l4-cvclohexyl-4-(3-hvdroxypropyl)- 7- [3-(methylamino) propyl]- 7, 8-dihvdro- 6H-indolof 1 ,2-eJ [1 ,5 Jbenzoxazocine-11 -carboxylate
• THF tetrahydrofuran
• 2,2,2 trifluoroethyl acetate 2.0 eq
• Step 4 methyl (7R)- 7-/3- f(tert-butoxycarbonyl) (methyl) amino) ] propyl ⁇ - 14-cvclohexyl-4-(3- hydroxypropyl)-7,8-dihydro-6H-indolofl,2-eJfl,5Jbenzoxazocine-ll-carboxylate
• DCM 0.05M
• Et 3 N 2.9 eq
• DMAP 0.1 eq
• BoC 2 O BoC 2 O
• Step 6 tert-butyl (3- ⁇ (7R)-14-cyclohexyl-ll- ⁇ [(2.2- dimethoxyethyl)(methyl)sulfamoylJcarbamoyl ⁇ -4-f3-(pyrrolidin-l-yl)propylJ-7,8-dihydro-6H- indolo 11 ,2-e] '/ 1 ,5]benzoxazocin-7-yl ⁇ propyl)methylcarbamate
• Step 7 (7S)-14-cvclohexyl-18.21-dimethyl-4-[3-(pyrrolidin-l-yl)propyl]-7.8-dihvdro-6H-7.11- (propanoiminoethanoiminothioiminomethano)indolo[l,2-e] [1 ,5Jbenzoxazocin-l 5-one 17,17- dioxide
• Step 1 methyl 3-cvclohexyl-2-[2-(methoxymethoxy)-3-methylphenyl]-lH-indole-6-carboxylate Methyl 2-bromo-3-cyclohexyl-lH-indole-6-carboxylate (prepared as described in International patent application WO 2004/087714), [2-(methoxymethoxy)-3-methylphenyl]boronic acid (Intermediate D; 1 eq) and bis(triphenylphosphinepalladium(II) chloride (0.1 eq) were dissolved in dioxane (0.1M). The solution was degassed and flushed with argon.
• Step 2 methyl 3-cvclohexyl-2-(2-hvdroxy-3-methylphenyl)-lH-indole-6-carboxylate
• MeOH aqueous 3N HCl
• the mixture was warmed to 80 0 C for 1.5 h.
• the mixture was cooled and all volatiles were evaporated in vacuo.
• the residual material was dissolved in EtOAc and the solution washed with sat. aq. NaHCO 3 and brine. After drying over Na 2 SO 4 and filtering, all volatiles were evaporated in vacuo leaving a beige amorphous solid (quant).
• (ES + ) m/z 364 (M+H) + .
• Step 3 (S)-methyl 3-cvclohexyl-2-[3-methyl-2-(oxiran-2-ylmethoxy)phenyl]-lH-indole-6- carboxylate
• CsF 3 eq
• 10 mins (iS)-(+)-glycidyl tosylate (1.2 eq) was added and the mixture stirred at RT overnight.
• the mixture was diluted with EtOAc and washed with IN aqueous HCl, sat. aq. NaHCO 3 and brine.
• Step 4 methyl (7S)-14-cvclohexyl-7-hvdroxy-4-methyl-7,8-dihvdro-6H-indolo[l,2- e] (1 ,5] benzoxazocine- 11-carboxylate
• Step 6 methyl 14-cyclohexyl-7-hydroxy-4-methyl-7-(prop-2-en-l-yl)-7,8-dihydro-6H-indolofl,2- e] [1 ,5] benzoxazocine- 11-carboxylate
• Step 7 methyl 14-cyclohexyl- 7 -hydroxy-4-methyl- 7-(2-oxoethyl)- 7, 8-dihydro-6H-indolofl ,2- e] '/ 1 ,5] benzoxazocine- 11-carboxylate
• osmium tetroxide 0.02 eq
• sodium periodate 3.0 eq
• the reaction was stirred at RT for 2 h, diluted with EtOAc and washed with H 2 O (x 2) and brine.
• the combined organic extracts were dried over Na 2 SO 4 , filtered and concentrated in vacuo to give a residue that was used directly in the next step.
• (ES + ) m/z 462 (M+H) + .
• Step 8 methyl 7- ⁇ 2-[ ⁇ 3-[(tert-butoxycarbonyl) (methyl) amino] propyl ⁇ (methyl) amino] ethyl ⁇ - 14- cyclohexyl-7-hydroxy-4-methyl-7,8-dihydro-6H-indolofl,2-e]fl,5]benzoxazocine-ll-carboxylate
• MeOH 0.1 M
• tert-butyi methyl[3- (methylamino)propyl]carbamate (Intermediate E) (1.5 eq) and acetic acid (1.0 eq).
• Step 10 methyl 7- ⁇ 2-l(3-U(tert- butoxycarbonvDsulfamoyl] (methyl) aminolpropyl) (methyl) amino] ' ethyl ⁇ - 14-cvclohexyl-7- hydroxy-4-methyl-7,8-dihydro-6H-indolo[l,2-e][l,5]benzoxazocine-ll-carboxylate
• THF 0.05 M
• (tert- butoxycarbonyl) ⁇ [4-(dimethyliminio)pyridin-l(4H)-yl]sulfonyl ⁇ azanide prepared as described in Org. Lett.
• Step 11 7- ⁇ 2-[(3- ⁇ ( (tert-butoxycarbonvDsulfamoyl) ' (methyl) amino lpropy I) (methyl) amino] 'ethyl ⁇ -
• Step 12 14-cvclohexyl-7-hvdroxy-4-methyl-7- [2-(methyl ⁇ 3- fmethyl(sulfamoyl)amino]propyl ⁇ amino)ethyl]-7,8-dihvdro-6H-indolofl,2- e] [1 ,5] benzoxazocine- 11-carboxylic acid
• Step 13 (7R, 7S)-14-cvclohexyl-7-hvdroxy-4J8,22-trimethyl-7,8-dihvdro-6H-7,ll- (ethanoiminopropanoiminothioiminomethano)indolo[l,2-e][l,5]benzoxazocin-15-one 17,17- dioxide
• Step 1 methyl 14-cvclohexyl-7-oxo-7,8-dihvdro-6H-indolofl,2-eJfl,5Jbenzoxazocine-ll- carboxylate
• Step 2 methyl 7 -alhl-14-cvclohexyl- 7 -hydroxy- 7, 8-dihvdro-6H-indolo[l ,2-eJ [1 ,5J benzoxazocine- 11 -carboxylate
• Methyl 14-cyclohexyl-7-oxo-7,8-dihydro-6H-indolo[l,2-e][l,5]benzoxazocine-l 1-carboxylate was dissolved in dry DCM (0.065M) and treated dropwise with TiCl 4 (1.5 eq). After stirring the dark solution at room temperature for 5 minutes neat allyltrimethylsilane (1.1 eq) was added at 0 0 C and stirring was continued at this temperature for 1 h. The reaction was quenched with cold water and the aqueous phase extracted with EtOAc.
• Step 3 methyl 14-cvclohexyl-7-hvdroxy-7-(3-hvdroxypropyl)-7 ,8-dihvdro-6H-indolo [ 1 ,2- e] [l,5]benzoxazocine-ll-carboxylate
• Step 4 methyl 14-cvclohexyl-7-hvdroxy-7-(3-oxopropyl)-7 ,8-dihvdro-6H-indolo [ 1 ,2- ejfl,5jbenzoxazocine-ll-carboxylate
• Step 5 (-) and (+) methyl 7-(3-[(2-[(tert- butoxycarbonyl) (methyl) amino] ethyl ⁇ (methyl) amino) ] propyl ⁇ - 14-cvclohexyl- 7 -hydroxy- 7, 8- dihvdro-6H-indolo [ 1 ,2-e] fl,5Jbenzoxazocine-l 1-carboxylate
• Step 6 methyl (7R or 7S)-14-cvclohexyl-7-hvdroxy-7-(3- ⁇ methyl[2-
• Step 7 methyl (7 R or 7S)-7-i3-fi2-fU(tert- butoxycarbonyl) amino) ] sulfonyl ⁇ (methyl) amino) l ethyl ⁇ (methyl) amino) ] propyl ⁇ - 14-cvclohexyl-7- hvdroxy-7 ,8-dihvdro-6H-indolo[l ,2-e] [1 ,5]benzoxazocine-l 1-carboxylate
• Step 8 (7R or 7 S) -7- ⁇ 3- [ ⁇ 2- [ ⁇ [(tert-butoxycarbonyl) amino] sulfonyl ⁇ (methyl) amino] ethyl ⁇ (methyl)amino] propyl ⁇ - 14-cvclohexyl- 7 -hydroxy- 7, 8-dihvdro-6H-indolo[l,2- e] [1 ,5]benzoxazocine-l 1-carboxylic acid
• Step 9 (7R or 7S)-7- ⁇ 3-[ ⁇ 2-[(aminosulfonyl) (methyl)amino] ethyl ⁇ (methyl)amino]propyl ⁇ - 14- cvclohexyl-7-hvdroxy-7 ,8-dihvdro-6H-indolo [ 1 ,2-e] [1 ,5]benzoxazocine-l 1-carboxylic acid
• DCM/TFA (4/1) 0.05 M
• Volatiles were removed in vacuo and the residue dissolved in EtOAc and washed with sat. aq. NaHCO 3 , brine, dried over Na 2 SO 4 filtered and evaporated in vacuo to give the title compound as a yellow glass (60%).
• (ES + ) m/z 599 (M+H) + .
• Step 10 (7R or 7S)-14-cvclohexyl-7-hvdroxy-18.21-dimethyl-7.8-dihvdro-6H-7.11- (propanoiminoethanoiminothioiminomethano)indolo [ 1 ,2-e] [ 1 ,5] benzoxazocin- 15-one 17,17- dioxide
• Step 1 (7S or 7R)-14-cvclohexyl-7 -hydroxy- 18.21 -dimethyl-7 ,8-dihvdro-6H-7.11-
• Example 6 14-cyclohexyl-7-fluoro- 18,2 l-dimethyl-7,8-dihydro-6H-7, 11- (propanoiminoethanoiminothioiminomethano)indolo[l,2-e] [l,5]benzoxazocin-15-one 17,17- dioxide
• Step 1 methyl 7 7 -allyl-14-cvclohexyl- ' / 7 - ⁇ uoro-7 ' ,8-dihvdro-6H-indolo 17 ,2-e) '/ 7 ,5]benzoxazocine- 11-carboxylate
• Step 2 methyl 14-cvclohexyl-7-fluoro-7-(3-hvdroxypropyl)-7 ,8-dihydro-6H-indolo [ 1 ,2- e] [1 ,5]benzoxazocine-l 1-carboxylate
• Step 3 methyl 14-cvclohexyl-7-fluoro-7-(3-oxopropyl)-7 ,8-dihvdro-6H-indolo [ 1 ,2- e] [1 ,5] benzoxazocine-11-carboxylate
• Step 4 methyl 7- ⁇ 3-[ ⁇ 2-[(tert-butoxycarbonyl) (methyl) amino] 'ethyl ⁇ (methyl) amino) ] propyl ⁇ - 14- cvclohexyl-7 7 -fluoro-7 ' ,8-dihvdro-6H-indolo >/ 7 ,2-e) '/ 7 ,5] benzoxazocine-11-carboxylate
• MeOH (0.1M) tert-butyl methyl[2- (methylamino)ethyl] carbamate prepared as described in Europoean patent application EPO296811) (1.15 eq) and acetic acid (3 eq) followed
• Step 5 methyl 14-cvclohexyl-7-fluoro-7-(3- ⁇ methyl[2-(methylamino)ethyl]amino ⁇ propyl)-7,8- dihvdro-6H-indolo [ 1 ,2-e] fl,5]benzoxazocine-l 1-carboxylate
• Step 6 methyl 7-(3-[(2-[([(tert- butoxycarbonyl) amino] 'sulfonyl) (methyl) amino] 'ethyl ⁇ (methyl) amino) ] propyl ⁇ - 14-cvclohexyl-7 - fiuoro-7,8-dihvdro-6H-indolo[l,2-e][l,5]benzoxazocine-l 1-carboxylate To a solution of methyl 14-cyclohexyl-7-fluoro-7-(3- ⁇ methyl[2-
• Step 7 7- ⁇ 3-[ ⁇ 2-[ ⁇ [(tert- butoxycarbonyl) amino) 'sulfonyl ⁇ (methyl) amino) 'ethyl ⁇ (methyl) amino) ] propyl ⁇ - 14-cvclohexyl-7 - fiuoro-7,8-dihvdro-6H-indolo[l,2-e][l,5]benzoxazocine-ll-carboxylic acid
• Step 8 7- ⁇ 3-[ ⁇ 2-[(aminosulfonyl) (methyl) amino] ethyl ⁇ (methyl) amino] propyl ⁇ - 14-cyclohexyl- 7- fluoro-7,8-dihvdro-6H-indolo[l,2-e][l,5]benzoxazocine-ll-carboxylic acid
• Step 9 14-cvclohexyl-7-fluoro-18.21-dimethyl-7 7 .8-dihvdro-6H-7 7 Jl- (propanoiminoethanoiminothioiminomethano)indolo [ 1 ,2-e] [ 1 ,5] benzoxazocin- 15-one 17,17- dioxide
• Step 1 methyl 14-cvclohexyl- 7- (2, 3-dibromopropyl)- 7 -hydroxy- 7, 8-dihvdro-6H-indolo[l , 2- el ' 'I r l ,5]benzoxazocine-l 1-carboxylate
• Step 2 methyl 4-bromo-14'-cvclohexyl-4,5-dihvdro-3H-spiro[furan-2, 7'-indolo[l,2- e] [1 ,5] benzoxazocine] - 1 l '-carboxylate
• Step 3 methyl 4-azido-14'-cvclohexyl-4,5-dihvdro-3H-spiroffuran-2, 7'-indolofl,2- e] [1 ,5] benzoxazocine] - 1 l '-carboxylate
• Step 4 methyl 4-amino-14'-cvclohexyl-4,5-dihvdro-3H-spiroffuran-2, 7'-indolofl,2- e] [1,5] benzoxazocine] -1 l '-carboxylate
• Step 5 methyl 14'-cvclohexyl-4-(dimethylamino)-4 ,5-dihvdro-3H-spiro [furan-2 , 7'-indolofl,2- e]f 1,5] benzoxazocine] -11 '-carboxylate
• Aqueous formaldehyde solution (37 wt %; 6 eq) was added to a solution in methanol (0.07 M) of the preceding compound.
• the pH of the reaction medium was adjusted to circa pH 5 - 6 by the drop wise addition of acetic acid.
• Sodium cyanoborohydride (6 eq) was introduced and the reaction mixture left to stir at RT overnight.
• the reaction mixture was partitioned between EtOAc and sat. aq. NaHCO 3 .
• the organics were washed with brine, dried (Na 2 SO 4 ), filtered and concentrated in vacuo to afford the product as a clear glass that was taken on without further purification.
• (ES + ) m/z 489 (M+H) + .
• Step 6 14 '-cvclohexyl-4- (dimethylamino)-4, 5-dihvdro-3H-spiro [furan-2, 7 '-indolofl , 2- el f 1,5] benzoxazocine] -11 '-carboxylic acid 2M NaOH (aq) (20 eq) was added to a solution of the foregoing compound (0.01 M) in a 2/1 mixture of THF/methanol. The reaction was stirred with heating at 60 0 C for 2 hours before being allowed to cool to RT and partitioned between EtOAc and IN HCl (aq), ensuring that the aqueous phase was acidic.
• Example 7a (2S,4R) or (2R,4S)-14'-cyclohexyl-4-(dimethylamino)-4,5-dihy(iro-3H- spirolfuran-l ⁇ '-indololl ⁇ - ⁇ ll ⁇ lbenzoxazocinel-ll'-carboxylic acid
• Example 7b (2R,4S) or (2S,4R)-14'-cyclohexyl-4-(dimethylamino)-4,5-dihydro-3H- spiro[furan-2,7'-indolo[l,2-e] [l,5]benzoxazocine]-ll'-carboxylic acid
• the first eluting racemate (hplc-A from example 7 step 6) was further separated into its respective enantiomers by chiral SFC (flow lOml/min; column chiralpack AD- ⁇ ; isocratic 30 % MeO ⁇ /0.2% diethylamine) to afford with 39 % recovery the first eluting enantiomer SFC-I (retention time 5.13 min; example 7a) and second eluting enantiomer SFC-2 (retention time 7.05 min; example 7b).
• 1 H nmr and MS data as for example 7.
• Example 8 (2S,45) and (2R,4R)-14'-cyclohexyl-4-(dimethylamino)-4,5-dihydro-3H- spiro[furan-2,7'-indolo[l,2-e] [l,5]benzoxazocine]-ll'-carboxylic acid
• steps 1-6 afforded 2 racemic mixtures upon hplc purification.
• the second eluting racemate from hplc was prepared as described in example 7.
• Example 8a (2S,4S) or (2R,4R)-14'-cyclohexyl-4-(dimethylamino)-4,5-dihydro-3H- spirolfuran-l ⁇ '-indololl ⁇ - ⁇ ll ⁇ lbenzoxazocinel-ll'-carboxylic acid
• Example 8b (2R,4R) or (2S,4S)-14'-cyclohexyl-4-(dimethylamino)-4,5-dihydro-3H- spiro[furan-2,7'-indolo[l,2-e] [l,5]benzoxazocine]-ll '-carboxylic acid
• the second eluting racemate (hplc-B; example 8) was further separated into its respective enantiomers by chiral SFC (flow 10 ml/min; column chiralpack AD- ⁇ ; isocratic 35 % MeO ⁇ /0.2% diethylamine for 6 mins, then 50 % MeOH/0.2% diethylamine) to afford with 36 % and 33 % recovery respectively the first eluting enantiomer SFC-I (retention time 3.63 min; example 2a) and second eluting enantiomer SFC-2 (retention time 9.40 min; example 2b).
• Step 1 methyl 14'-cvclohexyl-4-(methylamino)-4 ,5-dihvdro-3H-spiro [furan-2 ', 7'-indolo[l,2- e] [l,5]benzoxazocine]-l l '-carboxylate
• 2,2,2-trifluoroethyl formate (2.45 eq) was added to a solution (0.1 M) of methyl 4-amino-14'- cyclohexyl-4,5-dihydro-3H-spiro[furan-2,7'-indolo[l,2-e][l,5]benzoxazocine]-H'-carboxylate (prepared as described in example 7, step 4) in dry T ⁇ F.
• the reaction was stirred at RT for 3 h, at which point JV-formylation was complete. Volatiles were then removed in vacuo, and the resultant yellow foam re-solvated in T ⁇ F to give a 0.05 M solution.
• Step 2 methyl 4- / [N-(tert-butoxycarbonyl)-N-methyl-beta-alanyl) '(methyl) amino/ '- 14'- cyclohexyl-4,5-dihydro-3H-spiro[furan-2, 7'-indolo[l ,2-e] [1 ,5]benzoxazocine]-l l '-carboxylate ⁇ /-(tert-butoxycarbonyl)- ⁇ /-methyl- ⁇ -alanine (1.1 eq) was added to a solution of the foregoing compound (0.03 M) in dry DCM under N 2 .
• Step 3 methyl 14'-cvclohexyl-4- ⁇ methyl[3-(methylamino)propyl]amino ⁇ -4, 5-dihvdro-3H- spiro[furan-2, 7'-indolo[l,2-e][l,5]benzoxazocine]-l l '-carboxylate Borane DMS complex (2M solution in THF; 20 eq) was added to a stirred solution of the foregoing compound (0.05 M) in dry THF at RT under N 2 . The reaction was stirred at RT overnight before introducing a further 20 eq of borane DMS complex and stirring an additional 8 h.
• Step 4 methyl 4-[ ⁇ 3-[ ⁇ [(tert- butoxycarbonyl) amino] sulfonyl) (methyl) amino] propyl ⁇ (methyl) amino) '- 14'-cvclohexyl-4 ' , 5- dihvdro-3H-spiro [furan-2 , 7'-indolo [ 1 ,2-e] [ 1 ,5]benzoxazocine] - 1 l '-carboxylate
• Step 5 4- f ⁇ 3- [ ⁇ [(tert-butoxycarbonyl) amino) ] sulfonyl ⁇ (methyl) amino] propyl ⁇ (methyl) amino] - 14'-cvclohexyl-4,5-dihvdro-3H-spiro [furan-2, 7'-indolo / 1, 2-e] Jl, 5]benzoxazocine]- IV- carboxylic acid 2M NaOH (aq) (22 eq) was added to a solution of the foregoing compound (0.03 M) in a 2/1 mixture of MeOH/THF. The reaction was stirred with heating at 60 0 C for 2 h before being allowed to cool to RT and the volatiles reduced in vacuo.
• Step 6 4-[ ⁇ 3-[(aminosulfonyl) (methyl) amino] propyl ⁇ (methyl) amino) '- 14'-cvclohexyl-4 ' , 5- dihvdro-3H-spiroffuran-2, 7'-indolofl,2-eJfl,5JbenzoxazocineJ-ll '-carboxylic acid
• Step 1 methyl 4-cvano-14'-cvclohexyl-4,5-dihvdro-3H-spiro[furan-2, 7'-indolo[l,2- e] (1 ,5] benzoxazocine] - 1 l '-carboxylate
• Step 3 methyl 14'-cvclohexyl-4-[(dimethylamino)methyl]-4,5-dihvdro-3H-spiro[furan-2, 7 - indolofl ,2-e] [1 ,5] benzoxazocine]-! l '-carboxylate
• Aqueous formaldehyde solution (37 wt %; 6 eq) was added to a solution in methanol (0.05 M) of the preceding compound.
• the pH of the reaction medium was adjusted to circa pH 5 - 6 by the dropwise addition of acetic acid.
• Sodium cyanoborohydride (6 eq) was introduced and the reaction mixture left to stir at RT for 30 min.
• the reaction mixture was partitioned between EtOAc and sat. aq. NaHCO 3 .
• the organics were washed with brine, dried (Na 2 SO 4 ), filtered and concentrated in vacuo to afford the product as a clear glass that was taken on without further purification.
• (ES + ) m/z 503 (M+H) + .
• Step 4 14'-cvclohexyl-4-ffdimethylamino)methyl]-4,5-dihvdro-3H-spiroffuran-2, 7'-indolofl,2- e] [ 1 ,5] benzoxazocine] - 11 '-carboxylic acid
• Example 10a (2S,4S) or (2R,4R)-14'-cyclohexyl-4-[(dimethylamino)methyl]-4,5-dihy(iro-
• Example 10b (2R,4R) or (2S,4S)-14'-cyclohexyl-4-[(dimethylamino)methyl]-4,5-dihydro- 3H-spiro[furan-2,7'-indolo[l,2-e] [l,5]benzoxazocine]-ll'-carboxylic acid
• the first eluting racemate (hplc-A from example 10, step 4) was further separated into its respective enantiomers by chiral SFC (flow 10 ml/min; column chiralpack AD- ⁇ ; isocratic 30 % EtO ⁇ /0.4% diethylamine) to afford the first eluting enantiomer SFC-I (retention time 5.20 min; example 4a) and second eluting enantiomer SFC-2 (retention time 10.10 min; example 4b).
• 1 H nmr and MS data as for example 10.
• Example 11a (2S,4R) or (2R,4S)-14'-cyclohexyl-4-[(dimethylamino)methyl]-4,5-dihydro- SH-spirotfuran ⁇ '-indolotl ⁇ -eHl ⁇ benzoxazocinej-ir-carboxylic acid
• Example lib (2R,4S) or (2S,4R)-14'-cyclohexyl-4-[(dimethylamino)methyl]-4,5-dihydro- 3H-spiro[furan-2,7'-indolo[l,2-e] [l,5]benzoxazocine]-ll'-carboxylic acid
• the second eluting racemate (hplc-B; example 11) was further separated into its respective enantiomers by chiral SFC (flow 10 ml/min; column chiralpack AD- ⁇ ; isocratic 30 % EtOH/0.4% diethylamine) to afford the first e
• Step 1 ethyl 14'-cvclohexyl-4-(morpholin-4-ylmethyl)-4,5-dihvdro-3H-spiroffuran-2, 7 - indolo[l,2-e] [l,5]benzoxazocine]-l l '-carboxylate
• Methyl 4-(aminomethyl)-14'-cyclohexyl-4,5-dihydro-3H-spiro[furan-2,7'-indolo[l,2- e][l,5]benzoxazocine]-l l'-carboxylate was dissolved in dry ethanol (0.02 M).
• Step 2 14'-cvclohexyl-4-fmorpholin-4-ylmethyl)-4,5-dihvdro-3H-spiro [furan-2 J'-indolo / 1 ,2- e] [ 1 ,5] benzoxazocine] - 11 '-carboxylic acid

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