MXPA06009006A

MXPA06009006A - Piperidine derivatives useful as ccr3 antagonists

Info

Publication number: MXPA06009006A
Application number: MXPA/A/2006/009006A
Authority: MX
Inventors: Nengyang Shih; Joseph M Kelly; Pauline C Ting; Jianhua Cao; Youhao Dong; Eric J Gilbert; Ying Huang; Stuart Mccombie
Original assignee: Schering Corporation
Priority date: 2004-02-05
Filing date: 2006-08-07
Publication date: 2007-04-10

Abstract

The use of CCR3 antagonists of the formula (I) or a pharmaceutically acceptable salt thereof for the treatment of asthma is disclosed, as well as novel compounds of the formula (II), pharmaceutical compositions comprising them, and their use in the treatment of asthma, wherein R, Ra, X, Xa, R1, R2, R2a, R14, R14a, R16 and n are as defined in the specification.

Description

PIPERID DERIVATIVES USEFUL AS CCR3 ANTAGONISTS BACKGROUND OF THE INVENTION The present invention relates to the use of piperidine derivatives as selective CCR3 antagonists for the treatment of asthma. The invention also relates to a genus of novel compounds useful as selective CCR3 antagonists, pharmaceutical compositions comprising the novel compounds, and methods of treatment using the novel compounds. The new additional compounds that have CCR3 antagonist activity are also claimed. Piperidine derivatives useful as CCR3 antagonists are described in WO 01/77101. Piperidine derivative muscarinic antagonists useful in the treatment of cognitive disorders such as Alzheimer's disease are described in U.S. Patent 5,883,096; 6,037,352; 5,889,006; 5,952,349; and 5,977,138. CCR5 antagonists of piperidine derivatives useful in the treatment of HIV and inflammatory diseases such as rheumatoid arthritis are described in U.S. Patent 6,387,930.

BRIEF DESCRIPTION OF THE INVENTION The present invention relates to the treatment of asthma which comprises administering to a mammal in need of such treatment an effective amount of a CCR3 antagonist represented by structural formula I: or a pharmaceutically acceptable salt thereof, wherein n is 0 or 1; X is -C (R13) 2-, -C (R13) (R19) -, -C (O) -, -O-, -NH-, -N (alkyl) -, OR CH2- (CrC5.}. 3iquiler, fc-R3 NQR4 0.alpha CH-alkyl -CR, 3-, _R13_ »l >) ^", ~ C-, -QR I3"(- C- C (O > - - fvl - alkyl ', R is R6-phenyl, R6-pyridyl, R6-thiophenyl or R6-naphthyl; R1 is hydrogen, halogen, -OH, alkyl, hydroxyalkyl, alkoxy or alkoxyalkyl; R2 is 6-membered heteroaryl substituted with R7, R8, R9; 6-membered heteroaryl N-oxide substituted with R7, R8, R9; 5-membered heteroaryl substituted with R10, R11; R 2-naphthyl; fluorenyl; diphenylmethyl, x is 0-2 and y is 0-2, provided the sum of x and y is 1-4; R3 is R6-phenyl, R6-heteroaryl or R6-naphthyl; R 4 is hydrogen, alkyl, fluoroalkyl, cyclopropylmethyl, -CH 2 CH 2 OH, -CH 2 CH 2 -O-alkyl, -CH 2 C (O) -O-alkyl, -CH 2 C (O) NH 2, -CH 2 C (O) -NH-alkyl or -CH 2 C ( O) -N (alkyl) 2; R6 and R11 are independently selected from the group consisting of hydrogen and alkyl; R6 is 1 to 3 substituents independently selected from the group consisting of hydrogen, halogen, alkyl, alkoxy, -CF3, CF30-, CH3C (O) -, -CN, CH3SO2-, CF3SO2-, R16-phenyl, R16-benzyl, CH3C (= NOCH3) -, CH3C (= NOCH2CH3) -, < xx so, -NH2, -NHCOCF3, -NHCONH-alkyl, -O NHCO-alkyl, -NHSO2-alkyl, 5-membered heteroaryl and? X- ', wherein Z is -O-, -NH- or -N (CH3) -; R7 and R8 are independently selected from the group consisting of alkyl, halogen, -NR20R21, -OH, -CF3, -OCH3, -O-acyl, and -OCF3; R9 is R7, hydrogen, phenyl, -N02, -CN, -CH2F, -CHF2, -CHO, -CH = NOR20, pyridyl, pyridyl N-oxide, pyrimidinyl, pyrazinyl, N (R20) CONR21R22, -NHCONH (chloroalkyl ), -NHCONH-cycloalkylalkyl, -NHCO-alkyl, -NHCOCF3, -NHS02N (alkyl) 2, -NHS02-alkyl, -N (S02CF3) 2, -NHC02-alkyl, cycloalkyl, -SR23, -SOR23, -S02R23, -S02NH-alkyl, -OS02-alkyl, -OS02CF3, hydroxyalkyl, -CONR20R21, -CON (CH2CH2-0-CH3) 2l -OCONH-alkyl, -C02R2 °, -Si (CH3) 3 or -B (0C (CH3 2) 2; R10 is alkyl, -NH2 or R12-phenyl; R12 is 1 to 3 substituents independently selected from the group consisting of hydrogen, alkyl, -CF3, -C02R20, -CN, alkoxy and halogen; R13 and R16 are independently selected from the group consisting of hydrogen and alkyl; R14 is alkyl, alkenyl, haloalkyl, hydroxy, hydroxyalkyl, -CN, - (CR20R21) qO-alkyl, - (CR20R21) q-NR20R24, - (CR20R21) q-N3, - (CR20R21) qC (O) -alkyl, - (CR20R2) qC (O) -phenyl, - (CR20R21) q-COOR20, - (CR20R21) qC (O) NR20R24, - (CR20R21) qS (O) 0-2-R23, - (CR20R21) qN (R20 ) -C (O) NR20R24, - (CR20R21) qN (R20) -C (O) OR23 or - (CR20R1) qOC (O) R23; R14a is hydrogen or alkyl; or R14 and R14a together form = 0 or = NOR20; or R14 and R14a, together with the ring carbon to which they are attached, form a spirocycle ring of 3 to 6 carbon atoms; q is 0, 1, 2, or 3; R15 and R15a are each 1 or 2 substituents independently selected from the group consisting of H, halogen, OH, alkyl, alkoxy, -CF3, -OCF3, -CN, -C (0) R25, -COOR25, -S (O ) 0-2R25, -S (O) 0-2CF3, -NR20R24, phenyl and heterocycloalkyl; or two substituents R15 or two R15a on the carbon atoms of the adjacent ring, together with the carbons to which they are attached, form a fused cycloalkyl ring of 5-6 members; R17 and R18 are independently selected from the group consisting of hydrogen and alkyl, or R17 and R18 together are an alkylene group of C-C5 and with the carbon to which they are attached form a cycloalkyl ring of 3 to 6 carbon atoms; R19 is R6-phenyl, R6-heteroaryl, R6-naphthyl, cycloalkyl, cycloalkylalkyl or alkoxyalkyl; R20, R21 and R22 are independently selected from the group consisting of H and alkyl; R23 is alkyl or phenyl; and R24 is H, alkyl or R12-phenyl. New antagonist compounds are also claimed CCR3 represented by structural formula II or a pharmaceutically acceptable salt thereof, wherein n is 0 or 1; OR13 Xa is -C (R13) 2-, -C (O) - or -CR13-; Ra is R6a-phenyl, R6a-pyridyl, R6a-thiophenyl or R6a-naphthyl; R1 is hydrogen, halogen, -OH, alkyl, hydroxyalkyl, alkoxy or alkoxyalkyl; R2a is x is 0-2 and y is 0-2, provided that the sum of x and y is 1 -4; R6a is 1 to 3 substituents independently selected from the group consisting of hydrogen, halogen, -CF3, CF30-, -CN, CF3S02-, - or -NAZ NHCOCF3, CH3S02-, 5-membered heteroaryl and - ', wherein Z is -O-, -NH- or -N (CH3) -; R13 and R16 are independently selected from the group consisting of hydrogen and alkyl; R 4 is alkyl, alkenyl, haloalkyl, hydroxy, hydroxyalkyl, -CN, - (CR20R21) qO-aIquio, - (CR20R21) q-NR20R24, - (CR20R21) q-N3, - (CR20R21) qC (O) - alkyl, - (CR20R21) qC (O) -phenyl, - (CR20R21) q-COOR20, - (CR20R21) q- C (O) NR20R24, - (CR20R21) qS (O) 0-2-R23, - ( CR20R21) qN (R20) -C (O) NR20R24, - (CR20R21) qN (R20) -C (O) OR23 or - (CR20R21) qOC (O) R23; R14a is hydrogen or alkyl; or R14 and R14a together form = 0 or = NOR20; or R14 and R14a, together with the ring carbon to which they are attached, form a spirocycle ring of 3 to 6 carbon atoms; q is O, 1, 2 or 3; R15 and R15a are each 1 or 2 substituents independently selected from the group consisting of H, halogen, OH, alkyl, alkoxy, -CF3, -OCF3, -CN, -C (0) R25, -COOR25, -S (0 o.2R25, -S (O) 0.2CF3, -NR20R24, phenyl and heterocycloalkyl; or two substituents R15 or two R5a on the carbon atoms of the adjacent ring, together with the carbons to which they are attached, form a fused cycloalkyl ring of 5-6 members; R20 and R21 are independently selected from the group consisting of H and alkyl; R23 is alkyl or phenyl; and R24 is H, alkyl or R12-phenyl; provided that when R14 is - (CR20R1) q-NR20R24 and R24 is H, R20 is alkyl. Another aspect of the invention is the method of treating asthma which comprises administering to a mammal in need of such treatment an effective amount of a compound of formula II.

Another aspect of the invention is a pharmaceutical composition for the treatment of asthma comprising an effective amount of a CCR3 antagonist of formula II in combination with a pharmaceutically acceptable carrier. The following new compounds are also claimed in the genus of formula I: DETAILED DESCRIPTION OF THE INVENTION Preferred are compounds of formula I wherein n is 1. Preferably, R is R6-phenyl, especially wherein R6 is one or two, preferably two halogen substituents. A preferred halogen substituent is chlorine. Also preferred are compounds of formula I wherein R16 is hydrogen. Also preferred are compounds of the formula I wherein X is -C (R13) (R13) -, especially where R13 is hydrogen. For the compounds of the formula I, R1 is preferably hydrogen. R 14 is preferably alkyl, haloalkyl, hydroxy, hydroxyalkyl, alkoxy or alkoxyalkyl and R 14a is preferably hydrogen. In the compounds of the formula I, R 2 is preferably 6-membered heteroaryl substituted with R 7, R 8 R 9 R 12 -naphthyl, wherein R7, R8, R9, R12, R17 and R18 are as defined above and wherein the sum of x and y is 1 or 2, more preferably 1 (ie, R2 is optionally substituted quinolyl). R15 and R15a are preferably unique substituents, both of which are hydrogen, or R15 and R15a are unique substituents independently selected from the group consisting of hydrogen, halogen, methyl, methoxy and CF3.

Preferred are compounds of formula II wherein n is 1. Preferred are compounds of formula II wherein Ra is R6a-phenyl, especially where R6a is one or two, preferably two halogen substituents. A preferred halogen substituent is chlorine. Also preferred are compounds of formula II wherein R16 is hydrogen. Also preferred are compounds of the formula II wherein Xa is -C (R13) (R13) -, especially wherein each R13 is hydrogen. For the compounds of the formula II, R1 is preferably hydrogen. In another embodiment of formula II, R14 is preferably alkyl, haloalkyl, hydroxy, hydroxyalkyl, alkoxy or alkoxyalkyl and R14a is hydrogen. In the compounds of formula II, R a is preferably wherein the sum of x and y is 1 or 2, more preferably 1 (ie, R2a is optionally substituted quinolyl). The most preferred substituents R2a are selected from the group consisting of R15 and R15a are preferably unique substituents, both of which are hydrogen, or R15 and R15a are unique substituents independently selected from the group consisting of hydrogen, halogen, methyl, methoxy and CF3. For the compounds of both formulas I and II, R14 is preferably in the "cis" form relative to R-X or Ra-Xa, ie the compounds of the formulas I and II are preferably as shown in the following partial structures (where R14 and R1 are exemplified as hydrogen). As used herein, the following terms are used as defined below unless otherwise indicated. "Alkyl" means an aliphatic hydrocarbon group which may be straight or branched and comprising about 1 to about 20 carbon atoms in the chain. Preferred alkyl groups contain about 1 to about 12 carbon atoms in the chain. More preferred alkyl groups contain about 1 to about 6 carbon atoms in the chain. Branched means that one or more lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkyl chain. Non-limiting examples of suitable alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, n-pentyl, heptyl, nonyl, and decyl. "Alkenyl" means an aliphatic hydrocarbon group containing at least one carbon-carbon double bond and which may be straight or branched and comprising about 2 to about 15 carbon atoms in the chain. Preferred alkenyl groups have from about 2 to about 12 carbon atoms in the chain; and more preferably about 2 to about 6 carbon atoms in the chain. Branched means that one or more lower alkyl group such as methyl, ethyl or propyl, are attached to a linear alkenyl chain. Non-limiting examples of suitable alkenyl groups include ethenyl, propenyl, n-butenyl, 3-methylbut-2-enyl, n-pentenyl, octenyl and decenyl. "Alkylene" means a difunctional group obtained by removal of a hydrogen atom from an alkyl group defined above. Non-limiting examples of alkylene include methylene, ethylene and propylene. "Cycloalkyl" means a non-aromatic mono- or multicyclic ring system comprising about 3 to about 10 carbon atoms, preferably about 5 to about carbon atoms. Preferred cycloalkyl rings contain about 5 to about 7 ring atoms. Non-limiting examples of suitable monocyclic cycloalkyl include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl and the like. Suitable non-limiting examples of cycloalkyl multicyclic include 1-decalin, norbornyl, adamantyl and the like. "Halogen" or "halo" represents fluoro, chloro, bromo and iodo. "Haloalkyl" means an alkyl as defined above wherein one or more hydrogen atoms in the alkyl are replaced by a halo group defined above. Chloroalkyl and fluoroalkyl refer to alkyl groups substituted either by chloro or fluoro groups, respectively, for example fluoroalkyl represents a straight or branched alkyl chain substituted by 1 to 5 fluoro atoms, which can be attached to the same or different atoms of carbon, for example, -CH2F, -CHF2, -CF3, -CH2CF3 and -CF2CF3. "Hydroxyalkyl" means an HO-alkyl- group in which alkyl is as previously defined. Non-limiting examples of suitable hydroxyalkyl groups include hydroxymethyl, 2-hydroxyethyl and 1-hydroxyethyl. "Alkoxy" means an alkyl-O- group in which the alkyl group is as previously described. Non-limiting examples of suitable alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy and heptoxy. The link to the source portion is through the ether oxygen. Acyl means a radical of a carboxylic acid having the formula alkyl-C (O) -, aryl-C (O) -, aralkyl-C (O) -, cycloalkyl-C (O) -, cycloalkylalkyl-C (O) , and heteroaryl-C (O) -, wherein alkyl and heteroaryl are as defined herein; aryl is R12-phenyl or R12-naphthyl; and aralkyl is arylalkyl, wherein aryl is as defined above. "Heterocycloalkyl" means a saturated, non-aromatic monocyclic or multicyclic ring system comprising about 3 to about 10 ring atoms, preferably about 5 to about 10 ring atoms, in which one or more of the atoms in the system of ring is a different element of carbon, for example nitrogen, oxygen or sulfur, alone or in combination. There are no adjacent oxygen and / or sulfur atoms present in the ring system. Preferred heterocyclyls contain about 5 to about 6 ring atoms. The prefix aza, oxa or thia before the heterocyclic root name means that at least one nitrogen, oxygen or sulfur atom respectively is present as a ring atom. The nitrogen or sulfur atom of the heterocyclyl may optionally be oxidized to the corresponding N-oxide, S-oxide or S, S-dioxide. Non-limiting examples of suitable monocyclic heterocyclic rings include piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, 1,3-dioxolanyl, 1,4-dioxanyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like. Heteroaryl represents cyclic aromatic groups of 5 or 6 atoms or bicyclic groups of 11 to 12 atoms having 1 or 2 heteroatoms independently selected from O, S or N, heteroatoms that disrupt a carbocyclic ring structure and have a sufficient number of electrons pi delocalised to provide aromatic character, as long as the rings do not contain adjacent oxygen and / or sulfur atoms. For 6-membered heteroaryl rings, the carbon atoms can be substituted by groups R7, R8 or R9. Nitrogen atoms can form an N-oxide. All regioisomers are contemplated, for example, 2-pyridyl, 3-pyridyl and 4-pyridyl. Typical 6-membered heteroaryl groups are pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl and the N-oxides thereof. For 5-membered heteroaryl rings, the carbon atoms can be substituted by groups R10 or R11. Typical 5-membered heteroaryl rings are furyl, thienyl, pyrrolyl, thiazolyl, isothiazolyl, imidazolyl, pyrazolyl and isoxazolyl. The 5-membered rings having a heteroatom can be linked through position 2 or 3; the 5-membered rings having two heteroatoms are preferably linked through the 4-position. The bicyclic groups are typically benzo-fused ring systems derived from the heteroaryl groups named above, for example quinolyl, phthalazinyl, quinazolinyl, benzofuranyl, benzothienyl and indolyl As used herein, the term "composition" is intended to include a product that comprises the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from the combination of the ingredients specified in the amounts specified. The wavy line ru-u-, as a bond generally indicates a mixture of, or any of, the possible isomers, for example, containing stereochemistry (R) and (S). For example, The lines drawn in the ring systems, such as, for example: indicate that the indicated line (link) can be attached to any of the carbon atoms of the substitutable ring. When reference is made to a substituted ring, for example, phenyl, the substitution may be in any available position in the phenyl ring. As is well known in the art, a binding drawing of a particular atom in which no portion is represented at the terminal end of the bond indicates a methyl group linked through this bond to the atom, unless otherwise stated. For example: It should also be noted that any carbon or heteroatom with unsatisfied valencies in the text, schemes, examples, structural formulas, and any of the tables in the present is assumed to have the hydrogen atom or atoms to satisfy the valences. Prodrugs and solvates of the compounds of the invention are also contemplated herein. The term "prodrug", as used herein, denotes a compound that is a drug precursor which, upon administration to a subject, undergoes chemical conversion by metabolic or chemical processes to produce a compound of formula I or a salt and / or solvate thereof. A discussion of prodrugs is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems (1987) Volume 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, (1987) Edward B. Roche, ed., American Pharmaceutical Association and Pergamon Press, both are incorporated herein by reference. "Solvate" means a physical association of a compound of this invention with one or more solvent molecules. The physical association involves the variation of ionic and covalent bonding degrees, including hydrogen bonding. In certain cases the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated into the crystal lattice of the crystalline solid. "Solvate" includes both solvates of solution phase and isolators. Non-limiting examples of suitable solvates include ethanolates, methanolates, and the like. "Hydrate" is a solvate in which the solvent molecule is H20. "Effective amount" or "therapeutically effective amount" is understood to describe an amount of compound or composition of the present invention effective in inhibiting CCR3 receptors and thereby producing the desired therapeutic effect in a suitable patient. When R2 is R7, R8, R9-phenyl; 6-membered heteroaryl substituted with R7, R8, R9; or 6-membered heteroaryl N-oxide substituted with R7, R8, R9, the substituents R7 and R8 are preferably attached to the carbon ring members adjacent to the carbon joining the ring to the remainder of the molecule and the substituent R9 can be join any of the remaining unsubstituted carbon ring members, for example as shown in the following structures: When R2 is a 5-membered heteroaryl group, substituents R10 and R11 are preferably attached to the carbon ring members adjacent to the carbon joining the ring to the remainder of the molecule, and R11 is preferably alkyl; however, if a heteroatom is adjacent to the carbon joining the ring to the rest of the molecule (i.e., as in 2-pyrrolyl), R10 is preferably attached to a member of the carbon ring adjacent to the carbon joining the ring to the remainder of the molecule. When R2 or R2a is of the formula in addition to the quinolyl groups described above as preferred, the typical rings include isoquinolyl, phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, cinolinyl, pteridinyl, pyridopyridazine or pyridopyridine. An example of two R 5 substituents or two R 15a forming a fused ring is shown by the following structure: Certain CCR3 antagonist compounds of the invention may exist in different isomeric forms (e.g., enantiomers, diastereoisomers and atropisomers). The invention contemplates all isomers in both pure form and in mixture, including racemic mixtures. Certain compounds will be acidic in nature, for example those compounds which possess a phenolic or carboxyl hydroxyl group. These compounds can form pharmaceutically acceptable salts. Examples of such salts may include sodium, potassium, calcium, aluminum, gold and silver salts. Also contemplated are salts formed with pharmaceutically acceptable amines such as ammonia, alkyl amines, hydroxyalkylamines, N-methylglucamine and the like. Certain basic compounds also form pharmaceutically acceptable salts, for example, acid addition salts. For example, pyrido-nitrogen atoms can form salts with strong acid, while compounds having basic substituents such as amino groups also form salts with weaker acids. Examples of suitable acids for salt formation are hydrochloric, sulfuric, phosphoric, acetic, citric, oxalic, malonic, slicic, malic, fumaric, succinic, ascorbic, maleic, methanesulfonic and other minerals and carboxylic acids well known to those skilled in the art. technique. The salts are prepared by contacting the free base form with a sufficient amount of the desired acid to produce a salt in the conventional manner. The free base forms can be regenerated by treating the salt with the appropriate dilute aqueous base solution such as dilute aqueous NaOH, potassium carbonate, ammonia and sodium bicarbonate. The free base forms differ from their respective salt forms somewhat in certain physical properties, such as solubility in polar solvents, but the acid and base salts are otherwise equivalent to their respective free base forms for purposes of the invention . All acid and base salts are proposed to be pharmaceutically acceptable salts within the scope of the invention and all acid and base salts are considered equivalent to the free forms of the corresponding compounds for purposes of the invention. The polymorphic forms of the compounds of the formulas I and II, and of the salts, solvates and prodrugs of the compounds of the formula I and II, are proposed to be included in the present invention. The compounds of the invention can be made by methods known in the art, in particular by the methods described in US Pat. Nos. 6,387,930, hereby incorporated by reference, as well as by the method described in the patents of US Pat. United States 5,883,096; 6,037,352; 5,889,006; 5,952,349; and 5,977,138, also incorporated herein by reference. The compounds useful in this invention are exemplified by the following preparative examples, which should not be constructed to limit the scope of the description. Alternative mechanical trajectories and analogous structures within the scope of the invention may be apparent to those skilled in the art. The following solvents and reagents can be referred to in the examples by these abbreviations: tetrahydrofuran (THF); Ethanol (EtOH); methanol (MeOH); acetic acid (HOAC or AcOH); ethyl acetate (EtOAc); N, N-dimethylformamide (DMF); trifluoroacetic acid (TFA); trifluoroacetic anhydride (TFAA); 1-hydroxy-benzotriazole (HOBT); m-chloroperbenzoic acid (MCPBA); triethylamine (Et3N); diethyl ether (Et20); tert-butoxycarbonyl (BOC); 9-boraciclo [3.3.1] nonane (9-BBN); 1,8-diazabicyclo [5.4.0] undec-7-ene (DBU); dimethyl sulfoxide (DMSO); p-toluene sulfonic acid (p-TSA); 4-dimethylaminopyridine (DMAP); N, N, N-diisopropylethylamine (Dipea); and tert-butyldiphenylsilyl chloride (TBDPS-CI). TA is room temperature. The enantiomers are separated in Chiral Technology AD columns using 10% isopropanol-hexane with 0.5% diethylamine at 30%, sopropanol-hexane with 0.5% diethylamine as eluent. In the following examples, the "Stage" numbers refer to certain procedures, and each time the procedure is used, the same stage number is repeated; this results in a non-consecutive numbering of stages in the last examples.

EXAMPLE 1 General procedure A Üü A etaj32L. ™ ^ ü -8 Yes [tBx?) Mc2 iaoc and c? 8oc BOC i i? 4 Stage 1 : or JT stage 1 J IBOC 'tBOC 2 Methyl triphenylphosphonium bromide (38.0 g, 0.106 mol) was suspended (250 ml) and cooled to -78 ° C under nitrogen. N-butyl lithium (50.0 ml, 0.100 mol, 2.0 M in cyclohexane) was added dropwise via addition funnel. The cooling bath was stirred and the mixture was stirred at 23 ° C for 45 minutes. 1-t-Butoxycarbonyl-4-piperidone 1 (17.5 g, 0.088 mol) in dry THF (20 ml) dissolved in dry THF (20 ml) was added dropwise via addition funnel. The mixture was stirred at 23 ° C for 3 h, refluxed for 16 h, then cooled to 23 ° C. The solid was separated by filtration, washed with Et20, and the solvent was evaporated. The resulting residue was triturated 3X with Et0, the solid was filtered off, washed with Et20, and the solvent was evaporated. The crude product was purified by silica gel chromatography (eluent: 1: 8 EtOAs.hexane) to yield 14.0 g (0.071 mol, 79%) of product 2 as a colorless oil. MS (ES for M + 1): m / e 198.

Stage 2: stage 2 Selenium dioxide (2.81 g, 0.0253 mol) was suspended in CH2CI2 (150 ml) and cooled to 0 ° C. T-butylhydroperoxide (13.9 ml, 0.101 mol, 70% in water) was added and the mixture was stirred at 0 ° C for 30 minutes. The compound 2 (10.0 g, 0.0507 mol) in CH2Cl2 (20 ml) was added and the mixture was stirred at 0 ° C for 60 minutes, then at 23 ° C for 16 h. Ice chips and 10% by weight of sodium bisulfite in water (150 ml) were added. The layers were separated and the aqueous layer was extracted with CH2Cl2- The combined organic extracts were dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: 25% EtOAc-hexane to 50% EtOAc-hexane) provided 5.26 g (0.0247 mol, 49%) of product 3 as a yellow oil. MS (FAB for M + 1): m / e 214.

Stage 3: Compound 3 (5.25 g, 24.6 mmol) was dissolved in CH2Cl2, and Et3N (3.74 g, 5.1 mL, 36.9 mmol), DMAP (0.75 g, 6.15 mmol), and t-butyldimethylsilyl chloride (4.64 g, 30.77 mmol) they were added. The mixture was stirred at 23 ° C for 16 h. Water (150 ml) was added and the layers separated. The aqueous layer was extracted with CH2Cl2. The combined organic extracts were dried (MgSO), filtered, and concentrated. Purification by silica gel chromatography (eluent: 1: 4 EtOAc: hexane) provided 7.43g (22.7 mmol, 92%) of product 4 as a light yellow oil. MS (FAB for M + 1): m / e 328.

Stage 4: Apa 4 Compound 4 (7.42 g, 22.65 mmol) was dissolved in 9-BBN in THF solution (0.5 N, 68.0 mL, 34.0 mmol) and refluxed under nitrogen for 4 h, then cooled to 23 ° C. C. 1,2-Dichloro-4-iodobenzene (9.27 g, 34.0 mmol), K2CO3 (4.70 g, 34.0 mmol) and 1,1'-bis (diphenylphosphino) ferrocene) dichloropalladium II (1.85 g, 2.27 mmol), DMF were added. (60 ml), and water (4 ml) and the mixture was heated in an oil bath at 90 ° C for 16 h. The reaction mixture was concentrated, 0.5N NaOH (100 ml) was added, and the resulting mixture was extracted with CH2Cl2. The combined organic extracts were dried (MgSO), filtered and concentrated. Purification by silica gel chromatography (eluent: 5% EtOAc-hexane to 10% EtOAc-hexane) provided 4.13 g (8.72 mmol, 38%) of the trans isomer of product 5 as a white waxy solid and 3.83 g (8.09 mmol) , 36%) of the cis isomer of product 5 as a colorless oil. MS (FAB for M + 1): m / e 474.

Stage 5 Compound 5 (8.00 g, 16.9 mmol) was dissolved in THF (60 ml) and tetra-n-butylammonium fluoride in THF (1.0 M, 25.4 ml, 25.4 mmol) was added. The mixture was stirred at 23 ° C for 16 h, then concentrated. Water (100 ml) was added and the mixture was extracted with CH2Cl2. The combined organic extracts were dried (MgSO), filtered, and concentrated. Purification by silica gel chromatography (eluent: 5% EtOAc: hexane to 20% EtOAc-hexane) provided 5.80 g (16.1 mmol, 95%) of product 6 as a colorless oil. MS (FAB for M + 1): m / e 360.

Stage 6: Compound 6 (3.80 g, 10.5 mmol) was dissolved in dry DMF (70 ml) and cooled to 0 ° C under nitrogen. Potassium bis (trimethylsilyl) amide (27.4 ml, 13.7 mmol, 0.5 M in toluene) was added and the mixture was stirred at 0 ° C. minutes. CH3I (2.25 g, 0.98 ml, 15.8 mmol) was added, the mixture was allowed to warm slowly, and stirred at 23 ° C for 16 h. The reaction mixture was concentrated, 0.5N NaOH (70 ml) was added, and the mixture was extracted with CH2Cl2. The combined organic extracts were dried (MgSO), filtered, and concentrated. Purification by silica gel chromatography (eluent: 5% EtOAc: hexane to 10% EtOAc-hexane) provided 3.20 g (8.55 mmol, 81%) of product 7A as a colorless oil. MS (FAB for M + 1): m / e 374.

Stage 7: X O e ^ J Ofcte A ßOC »AH Compound 7A (5.90 g, 0.0158 mol) was dissolved in 1: 1 CH2Cl2: MeOH (150 ml), cooled to 0 ° C, and 4N HCl in dioxane (31.5 ml, 0.126 mol) was added. . The mixture was stirred at 23 ° C for 16 h, and concentrated. MeOH (200 ml) was added and the mixture made basic with diethylaminomethylpolystyrene resin. The resulting mixture was filtered, the resin was washed with MeOH, and the solvent was removed under vacuum to yield 4.30 g (0.0158 mol, 100%) of the 5A product as a cinnamon foam. MS (FAB for M + 1): m / e 274.

Stage 8: Compound 9 (4.70 g, 17.1 mmol) was dissolved in dry CH2Cl2 (250 mL), 1-t-butoxycarbonyl-4-piperidone (5.10 g, 25.7 mmol), sieves of 3 (3 g), sodium triacetoxyborohydride (7.30 g). , 34.3 mmol), and AcOH (1.02 g, 0.94 mL, 17.1 mmol) were added, and the mixture was stirred at 23 ° C for 16 h. 0.5 N NaOH (200 ml) was added, the mixture was filtered and then extracted with CH2Cl2. The combined organic extracts were dried (MgSO), filtered, and concentrated. To remove 1-t-butoxycarbonyl-4-hydroxypiperidine, the crude product was dissolved in CH2Cl2 (200 ml) and Et3N (3.47 g, 4.8 ml, 34.3 mmol), DMAP (1.05 g, 8.57 mmol), and chloride of acetyl (1.35 g, 1.2 ml, 17.1 mmol) were added. The mixture was stirred at 23 ° C for 16 hours. Water (150 ml) was added and the mixture was extracted with CH2Cl2. The combined organic extracts were dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: EtOAc then 5% MeOH with NH3-CH2Cl2) gave 5.20 g (11.4 mmol, 66%) of the product 10 as a brown oil. MS (FAB for M + 1): m / e 457.

Stage 7a: The same procedure as used above for Step 7 was used to prepare compound 11 from compound 10. MS (ES for M + 1): m / e 357.

Stage 9: Compound 11 (0.15 g, 0.42 mmol) was combined with 3-quinolinecarboxylic acid (0.11 g, 0.63 mmol), HOBT (0.085 g, 0.63 mmol), 1-ethyl-3- (3-dimethylamino-propyl) carbodiimide (0.16 g). g, 0.84 mmol), and Dipea (0.11 g, 0.84 mmol) in 1: 1 CH2Cl2: DMF (8 mL) and the mixture was refluxed for 16 h, then cooled to 23 ° C. 0.5 N NaOH (10 mL) was added, and the mixture was filtered and extracted with CH2Cl2. The combined organic extracts were dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: 3% MeOH with NH3-CH2CI2) afforded 0.13 g (0.25 mmol, 62%) of the product of Example 1A as a yellow foam. MS (FAB for M + 1): m / e 512. The following compounds were prepared according to a similar procedure: EXAMPLE 2 General procedure O O OH OH N N H "Cl N H 13 15 1« OC H i * tßOC tDOC e3 Stage 10: Compound 13 (25.0 g, 0.129 mol), di-t-butyl dicarbonate (33.8 g, 0.155 mol), and Et3N (26.1 g, 36.0 ml, 0.258 mol) were combined in CH2Cl2 (240 ml) and MeOH (60 ml), stirred at 23 ° C for 16 h and concentrated. 1.0 N NaOH (100 ml) was added and the mixture was extracted with CH2Cl2. The combined organic extracts were dried (MgSO), filtered, and concentrated. Purification by silica gel chromatography (eluent: 5% EtOAc: hexane to 10% EtOAc-hexane) provided 33.2 g (0.129 mol, 100%) of product 14 as a colorless oil. MS (FAB for M + 1): m / e 258.

Stage 11: tBOC I5 Compound 14 (10.0 g, 0.0389 mol) and NaBH 4 (3.69 g, 0.0973 mol) were dissolved in dry THF (150 ml) and refluxed under nitrogen. MeOH (30 ml) was added dropwise via syringe pump at a rate of 8.4 ml / hour. After the addition of MeOH was complete, the reaction mixture was refluxed for 3 h, then cooled to 23 ° and concentrated. Water (150 ml) was added and the mixture was extracted with CH2Cl2. The combined organic extracts were dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: 5% MeOH-CH 2 Cl 2 to 10% MeOH-CH 2 Cl 2) provided 8.27 g (0.0358 mol, 92%) of product 15 as a colorless oil. MS (ES for M + 1): m / e 232.

Stage 3: Using the same procedure as in Example 1, Step 3, compound 16 was prepared. MS (ES for M + 1): m / e 346.

Stage 12: Oxalyl chloride (5.71 g, 0.0450 mol) was dissolved in dry CH2Cl2 (125 ml) and cooled to -78 ° C under nitrogen. DMSO (7.03 g, 6.4 ml, 0.0900 mol) dissolved in dry CH2Cl2 (25 ml) was added dropwise via addition funnel. The mixture was stirred at -78 ° C for 15 minutes, then compound 16 (12.40 g, 0.0360 mol) dissolved in dry CH2Cl2 (50 ml) was added dropwise via addition funnel. The mixture was stirred at -78 ° C for 60 minutes, then Et3N (10.92 g, 15.0 ml, 0.108 mol) was added and the mixture was stirred again at -78 ° C for 20 minutes, then at 0 ° C for 30 minutes. minutes Water (200 ml) was added, the layers were separated, and the aqueous solution was extracted with CH2Cl2. The combined organic extracts were dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: 1: 4 EtOAc: hexane) provided 11.90 g (0.0347 mol, 97%) of the product 17 as a light yellow oil. MS (FAB for M + 1): m / e 344.

Stage 1 : Using the same procedure as for Example 1, Step, compound 18 was prepared. MS (ESparaM + 1): m / e342.

Stage 4: Using the same procedure as for Example 1, Step, compound 19 was prepared. MS (ES for M + 1): m / e 488.

Stage 5: Cl stage 5 XX = V 20 I tBOC Using the same procedure as for Example 1, Step, compound 20 was prepared.

MS (ES for M + 1): m / e 374.

Stage 13: Compound 20 (0.80 g, 2.13 mmol) was dissolved in dry DMF (25 ml) and cooled to 0 ° C. NaH (0.13 g of 60% by weight in oil, 3.20 mmol) was added and the mixture was stirred at 0 ° C for 15 minutes, then at 23 ° C for 30 minutes. CH3I (0.60 g, 0.27 mL, 4.26 mmol) was added, the mixture was stirred at 23 ° C for 16 h, then concentrated. Water (40 ml) was added and the mixture was extracted with CH CI2. The combined organic extracts were dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: 20% EtOAc: hexane to 30% EtOAc-hexane) provided 0.82 g (2.13 mmol, 100%) of the product 21 as a colorless oil. MS (FAB for M + 1): m / e 388.

Stage 12a: Using the same procedure as for Step 12, above, compound 23 was prepared. MS (ES for M + 1): m / e 372.

Stage 14 Compound 23 (0.95 g, 2.56 mmol) was dissolved in dry THF (20 ml) and CH3MgBr (1.30 ml, 3.84 mmol, 2.95 M in THF) was added. The mixture was refluxed for 3 h, then cooled to 0 ° C and concentrated. Saturated NH 4 Cl (50 mL) was added and the mixture was extracted with CH 2 Cl 2. The combined organic extracts were dried (MgSO), filtered and concentrated. Purification by silica gel chromatography (eluent: 1: 1 EtOAc: hexane) provided 0.39 g (1.01 mmol, 40%) of the product 24 as a white foam. MS (FAB for M + 1): m / e 388.

Stage 7: Using the same procedure as for Example 1, Step 7, compound 22 was prepared. MS (ESparaM + 1): m / e288. Following a similar procedure, the additional intermediaries were prepared: Stage 8: Using the same procedure as for Example 1, Step, the compound of Example 2-A was prepared.

MS (ES for M + 1): m / e 526.

Stage 15: Compound 22 (0.36 g, 1.42 mmol), N- (6-quinolinylcarboxyl) -4-piperidone (0.39 g, 1.42 mmol), and titanium isopropoxide (0.51 g, 1.78 mmol) were combined in dry CH 2 Cl 2 (2 mL) and were stirred at 23 ° C under nitrogen for 8 h. NaBH3CN (0.11 g, 1.78 mmol) and EtOH (5 mL) were added and the mixture was filtered through celite. The celite was washed with CH2Cl2 and water, the filtrate layers were separated, and the aqueous solution was extracted with CH2Cl2. The combined organic extracts were dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: 5% MeOH-CH 2 Cl 2 to 10% MeOH-CH 2 Cl 2) afforded 0.46 g (0.898 mmol, 63%) of Example 2-B as a white foam. MS (FAB for M + 1): m / e 512. The following compounds were prepared according to a similar procedure: EXAMPLE 3 General Procedure and Stage 13: Using the same procedure as for Example 2, Step 13, compound 26 was prepared. MS (ES for M + 1): m / e 286.

Stage 16: Compound 26 (2.90, 10.2 mmol) was suspended in 6N HCl (85 ml) and refluxed for 16 h. The product was concentrated and azeotroped three times with isopropanol to produce a yellow solid. The solid was dissolved in 1: 1 CH2Cl2: MeOH (50 mL) and Et3N (3.08 g, 4.3 mL, 30.5 mmol) and t-butyl dicarbonate (3.32 g, 15.2 mmol) were added. The mixture was stirred at 23 ° C for 16 h, then concentrated. 0.5 N NaOH (50 ml) was added and the mixture was extracted with CH2Cl2. The combined organic extracts were dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: 10% EtOAc: hexane) gave 1.92g (8.46 mmol, 83%) of product 27A as a colorless oil. MS (ES for M + 1): m / e 228.

Stage 17: o Me stage 17 Me * N N I 27B ^ Ph 28 tBOC Compound 28 (15.0 g, 0.0738 mol) and di-t-butyl dicarbonate (17.7 g, 0.0812 mol) were dissolved in EtOAc (400 mL). Palladium hydroxide catalyst (4 g) was added and the mixture was shaken on a Paar shaker under 40 psi hydrogen pressure for 24 h. The resulting mixture was filtered through celite and the celite was washed with EtOAc. The filtrate was concentrated to yield 15.73 g (0.0738 mol, 100%) of product 27B as a colorless oil. MS (ES for M-tBOC + 1): m / e 114.

Stage 1 : XOR14 stage 1 JL.R14 R 4 = Me, Et tBOC 27 tBOC 9 Using the same procedure as for Example 1, Step 1, compounds 29a and 29b were prepared: Stage 4: Using the same procedure as for Example 1, Stage 4, compounds 30a and 30b were prepared. The cis and trans isomers of compound 30b (R14 = Me) were separated by silica gel chromatography (eluent 3% EtOAc: hexane to 10% EtOAc-hexane) at 90% purity.

Stage 7: cc; Using the same procedure as for Example 1, Stage , compounds 31a and 31b were prepared.

Stage 8 or Stage 15: Using the same procedure as for Example 1, Stage , or Example 2, Step 15, the following compounds were prepared: EXAMPLE 4-A Stage A: Stage A to c ?? x 23 * "" N 10th tBOC A solution of 0.5 M potassium bis (trimethylsilyl) amide in toluene (2.68 ml, 1.34 mmol) was added to a suspension of methyl (triphenyl) phosphonium bromide (0.70 g, 1.95 mmol) in Et20 (anhydrous, 10 ml) at -78 ° C. The mixture was stirred for 1 h at -30 ° C, then cooled to -40 ° C, and a solution of the aldehyde 23 (0.50 g, 1.34 mmol) in Et20 (anhydrous, 5 ml) was added dropwise. The reaction mixture was stirred at -30 ° C for 30 min, then quenched with acetone (1 ml). Et2O (100 ml) and water (50 ml) were added, and the organic layer was separated, then dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: 10% v / v acetone / hexane) provided 0.37 g (1.00 mmol, 75%) of the "cis" 100 isomer product as a colorless oil. MS (FAB for M + 1): m / e 370.

Stage B: A 1: 1 solution of TFA: CH 2 Cl 2 (3 mL) was added to a solution of compound 100 (0.37 g, 1.0 mmol) in CH 2 Cl 2 (3 mL) at 23 ° C and stirred for 16 h. The solvent was evaporated, and water (15 ml) and 10% NaOH (3 ml) were added. The mixture was extracted with CH2Cl2 (2 x 25 mL), dried (MgSO), filtered and concentrated to yield 0.24 g (0.89 mmol, 89%) of product 101. MS (FAB for M + 1): m / 270 Stage C: Using the same procedure as for Example 1, Step 8, compound 102 was prepared. MS (FAB for M + 1): m / e 508.

Step D: A solution of compound 102 (70 mg, 0.137 mmol) and tris (triphenylphosphine) -rodio (I) chloride (10 mg) in EtOH (10 mL) was hydrogenated using a Parr shaker overnight. The reaction was filtered through celite and the celite was washed with EtOH. The filtrate was concentrated. Purification by silica gel chromatography (eluent: 10% v / v MeOH with NH3 / EtOAc) gave 36 mg (0.076 mmol, 55%) of the title compound (Example 4-A) as a white foam. MS (FAB for M + 1) = 510.

EXAMPLE 4-B General Procedure Stage 18: 33 Compound 29b (7.92 g, 0.0375 mol) was dissolved in CH2Cl2 (250 ml) and cooled to 0 ° C. NaHCO3 (4.73 g, 0.0563 mol) and MCPBA (12.95 g of 75% purity, 0.0563 mol) were added and the mixture was stirred at 0 ° C for 20 minutes, then at 23 ° C for 16 h. CH2Cl2 (500 ml) was added and the mixture was washed with 0.5 N NaOH (300 ml) and saturated sodium bisulfite (300 ml). The combined organic extracts were dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: 1: 3 EtOAc-hexane) yielded 6.83 g (0.030 mol, 80%) of the product 33 as a colorless oil. MS (ES for M + 1): m / e 228.

Stage 19: Magnesium (1.46 g, 0.060 mol) was suspended in dry THF (10 ml) and 1-bromo-3,4-dichlorobenzene (13.56 g, 0.060 mol) was added via addition funnel. A crystal of iodine and a few drops of 1,2-dibromoethane were added to initiate the grignard reaction. Dry THF (10 ml) was added and the mixture was refluxed for 45 minutes. Dry THF (80 ml) was added and cooled to -30 ° C. Copper iodide (5.71 g, 0.030 mol) and dimethisulfide (10 ml) were added and the mixture was stirred at -30 ° C for 2 h. Compound 33 (6.82 g, 0.030 mol) dissolved in dry THF (25 ml) was added dropwise via addition funnel and the mixture was stirred at -15 ° C for 16 h, then at 23 ° C for 24 h. Saturated NH 4 Cl (200 mL) was added and the mixture was extracted with EtOAc. The combined organic extracts were dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: 1: 2 Et20-hexane, then 1: 1 Et20-hexane, then 1: 2 EtOAc-hexane) yielded 3.29 g (0.0088 mol, 29%) of the product 34 as a foam yellow. MS (ES for M + 1): m / e 374.

Stage 20: Compound 34 (2.38 g, 6.36 mmol) was dissolved in dry CH 2 Cl 2 (40 ml) and cooled to -78 ° C. Diethylaminosulfur trifluoride (1.28 g, 7.95 mmol) was added and the mixture was stirred at -78 ° C for 15 minutes, then slowly warmed to 0 ° C. 1 N NaOH (75 ml) was added and the mixture was extracted with CH2Cl2. The combined organic extracts were dried (MgSO 4), filtered, and concentrated to yield a yellow oil. The oil was dissolved in CH2Cl2 (40 mL) and NaHCO3 (0.53 g, 6.36 mmol) and MCPBA (1.46 g of 75% purity, 6.36 mmol) were added to epoxidize the alkene by-product for purification. The mixture was stirred at 23 ° C for 16 h. 0.5 N NaOH (50 ml) was added and the mixture was extracted with CH2Cl2. The combined organic extracts were dried (MgSO4), filtered, and combined. Purification by silica gel chromatography (eluent: 10% EtOAc-hexane to 25% EtOAc-hexane) yielded 1.20 g (3.19 mmol, 50%) of product 35 as a colorless oil. MS (ES for M + tBu + 1): m / e 319.

Stage 7: Using the same procedure as for Example 1, Step, the following compounds were prepared.

Stage 8: Using the same procedure as for Example 1, Step, the following compounds were prepared: EXAMPLE 5-A Top 21 Stage 21: NaBH 4 (2.18 g, 57.7 mmol) was added to a solution of compound 27B (6.10 g, 28.8 mmol) in ethanol (200 ml) at 0 ° C for a period of 20 minutes. After 30 minutes, the reaction mixture was concentrated. EtOAc (150 ml) was added, and the mixture was washed with brine, dried (Na 2 SO), filtered, and concentrated to provide 5.45 g (25.5 mmol, 88%) of compound 38 as a colorless oil. MS: m / e 160 (M-56).

Stage 22: Diethyl azodicarboxylate (3.7 g, 21 mmol) was added to a solution of compound 38 (4.0 g, 16.7 mmol) and 3,4-dichlorophenol (2.3 g, 14.0 mmol) in THF (65 mL). The mixture was stirred at 23 ° C for 3 days, then at 50 ° C for 27 h. Et20 was added and the mixture was washed with water, 0.4 N NaOH, water and saturated NaCl. The organic extract was dried (Na2SO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: 1: 9 EtOAc: hexane) yielded 2.0 g (5.5 mmol, 40%) of product 39 as a colorless oil. MS: m / e 360.

Stage 23: stage 23 39 TFA (14 ml) was slowly added to a solution of compound 39 (2.0 g, 5.6 mmol) in CH2Cl2 (55 ml) and stirred at 23 ° C for 30 minutes. The mixture was concentrated, 1N NaOH (40 ml) was added and the mixture was extracted with CH2Cl2. The combined organic extracts were dried (Na2SO), filtered, and concentrated. Purification by preparative TLC from silica gel (solvent: 1: MeOH with NH3-CH2CI2) yielded 0.49 g (1.9 mmol, 34%) of the cis isomer of compound 40 as a colorless oil and 0.23 g (0.88 mmol, 16%) of the trans isomer of compound 40 as a colorless oil. MS: m / e 260.

Step 8: Using the same procedure as for Example 1, Step, compound 41 (Ex. 5-A) was prepared from cis compound 40. MS: m / e 498.

Using a similar procedure, Example 5-B was prepared from the trans 40 compound.

MS: m / e: 498 EXAMPLE 6 Procedure for compounds where R14 is -NH2, -NHCOCF3, -NHS02CH3, -NHCOCH3 Stage 24: stage 24 Compound 6 (1.02 g, 2.84 mmol) was dissolved in dry CH2Cl2 (8 mL) and cooled to 0 ° C. Et3N (0.43 g, 0.59 ml, 4.25 mmol) was added, then CH3S02CI (0.27 ml, 3.49 mmol) and the mixture was stirred at 23 ° C for 2 h. EtOAc (40 ml) was added and the mixture was washed with 1 N HCl (25 ml). The aqueous layer was separated and extracted with EtOAc. The combined organic extracts were dried (MgSO), filtered and concentrated to yield the mesylate (1.22 g, 2.77 mmol). The mesylate was dissolved in DMF (10 ml) and sodium azide (0.45 g, 6.92 mmol) was added. The mixture was heated at 65 ° C for 12 days, then cooled to 23 ° C. Et20 (75 ml) was added and the mixture was washed with water (5 x 10 ml). The aqueous layers were combined and extracted with Et20. The combined organic extracts were dried (MgSO), filtered and concentrated. Purification by silica gel chromatography (eluent: 1: 8 EtOAc-hexane to 1: 5 EtOAc-hexane) yielded 0.61 g (1.59 mmol, 56%) of product 42 as a light yellow oil. MS (ES for M + 1): m / e 385.

Stage 25: Compound 42 (0.29 g, 0.76 mmol) was dissolved in THF (5 mL) and water (0.5 mL) and triphenylphosphine (0.41 g, 1.56 mmol). The mixture was refluxed for 16 h. it was cooled to 23 ° C and concentrated. Saturated NaCl was added and the mixture was extracted with CH2Cl2. The combined organic extracts were dried (MgSO), filtered, and concentrated. The crude product was dissolved in dry CH 2 Cl 2 (5 mL), cooled to 0 ° C, and Et 3 N (0.26 mL, 1.87 mmol) and TFAA (0.13 mL, 0.92 mmol) were added. The mixture was stirred at 23 ° C for 4 h, then concentrated. Purification by silica gel chromatography (eluent: 1: 10 EtOAc-hexane to 1: 3 EtOAc-hexane) afforded 0.25 g (0.55 mmol, 73%) of product 43 as a white foam. MS (ES for M + 1 -tBoc): m / e 355.

Stage 23: step 23 NHC0CF3 43 O V H '"44 The same procedure was used as for Example 5, Step 23, to obtain compound 44. MS (ES for M + 1): m / e 355.

Stage 8: The same procedure was used as for Example 1, Step 8, to obtain compound 45. MS (ES for M + 1): m / e 539.

Stage 26: Compound 45 (0.080 g, 0.13 mmol) and K2C03 (0.080 g, 0.58 mmol) were suspended in MeOH (1.5 mL) and water (0.5 mL), stirred at 23 ° C for 3 days and concentrated. CH2Cl2 (15 ml) and water (10 ml) were added, the aqueous layer was separated and extracted with CH CI2 (3 x 15 ml). The organic extracts were combined, dried (MgSO4), filtered and concentrated. Purification by silica gel chromatography (eluent: 1:10 MeOH-CH 2 Cl 2 then 1: 4 4% MeOH with NH 3 -CH 2 Cl 2) yielded 0.052 g (0.10 mmol, 77%) of product 46 as a white foam. MS (ES for +1): m / e 477.

Stage 27: Cl Cl 'XX Compound 46 (0.027 g, 0.054 mmol) was dissolved in CH2Cl2 (2 mL), and Et3N (0.050 mL, 0.36 mmol) and CH3S02CI (0.015 mL, 0.19 mmol) were added. The mixture was stirred at 23 ° C for 16 h. 1N NaOH (10 ml) was added and the mixture was extracted with CH2Cl2. The combined organic extracts were washed with brine, dried (MgSO4), filtered and concentrated. Purification by preparative thin layer chromatography of silica gel (eluent: 1:10 MeOH-CH 2 Cl 2) yielded 0.019 g (0.032 mmol, 60%) of compound 47 as a white foam. MS (ES for M + 1): m / e 575.

Example 6-D was prepared according to the same procedure, using acetyl chloride in place of mesyl chloride; Cl c? XX EM: m / e 539 EXAMPLE 7 Stage 24: Using the same procedure as for Example 6, Step, compound 49 was prepared. MS (ESparaM + 1): m / e537.

Stage 25: Using the same procedure as for Example 6, Step, but without adding the TFAA, compound 50 was prepared. MS (ES for M + 1): m / e 511.

Compound 50 (67 mg, 0.13 mmol) was dissolved in CH2Cl2 (2 mL) and Et3N (0.12 mL, 0.86 mmol) and ethyl chloroformate (0.04 mL, 0.42 mmol) were added. The mixture was stirred at 23 ° C for 16 h. 1N NaOH (5 ml) was added and the mixture was extracted with CH2Cl2. The combined organic layers were dried (MgSO 4), filtered and concentrated. Purification by preparative CCD of silica gel (eluent: 10% MeOH-CH 2 Cl 2) yielded 41 mg (51%) of the product 50a (Ex. 7-C) as a white foam. MS (for M + 1): m / e 583.

Compound 50 (73.4 mg, 0.20 mmol) was dissolved in CH2Cl2 (2 mL) and Et3N (0.17 mL, 1.22 mmol) and trimethylsilylisocyanate (0.08 mL, 0.59 mmol) were added. The mixture was stirred at 23 ° C for 16 h. 1N NaOH (5 ml) was added and the mixture was extracted with CH2Cl2. The combined organic layers were dried (MgSO), filtered and concentrated. Purification by preparative CCD of silica gel (eluent: 10% MeOH with NH3-CH2CI2) afforded 42 mg (38%) of the product 50b (Example 7-D) as a white foam. MS (for M + 1): m / e 554. The following compound was prepared according to a similar procedure: EM (for M + 1): m / e 582 Using the same procedure as for Example 6, Step 27, with acetyl chloride in place of mesyl chloride, compound 50d was prepared (Ex. 7-F).

MS (for M + 1): m / e 553.

Using the same procedure as for Example 6, Step, compound 50e was prepared (Ex. 7-G). MS (for M + 1): m / e 589.

EXAMPLE 8 Stage 28 Compound 51 (0.20 g, 0.39 mmol) was dissolved in CH2Cl2 (15 mL) and Et3N (0.060 g, 0.083 mL, 0.59 mmol) and acetyl chloride (0.037 g, 0.033 mL, 0.47 mmol) were added. The mixture was stirred at 23 ° C for 16 h, water (20 ml) was added and the mixture was extracted with CH 2 Cl 2. The combined organic extracts were dried (MgSO4), filtered and concentrated. Purification by silica gel chromatography (eluent: 5% MeOH with NH3-CH2Cl2) yielded 0.20 g (0.36 mmol, 93%) of the product 52 as a white foam. MS (FAB for M + 1): m / e 554.

Stage 29: Compound 51 (0.20 g, 0.39 mmol) was dissolved in dry THF (10 mL) and Et3N (0.12 g, 0.17 mL, 1.17 mmol) and isopropyl isocyanate (0.075 g, 0.086 mL, 0.89 mmol) were added. The mixture was refluxed for 16 h, then 0.5 N NaOH (20 ml) was added and the mixture was extracted with CH2Cl2. The combined organic extracts were dried (MgSO4), filtered and concentrated. Purification by silica gel chromatography (eluent: 3% MeOH with NH3-CH2CI2) afforded 0.22 g (0.37 mmol, 95%) of the product 53 (Ex. 8-B) as a white foam. MS (FAB for M + 1): m / e 597. The following compounds were prepared according to a similar procedure: EXAMPLE9 Stage 36 Stage 30: A solution of 1-bromo-3,4-dichlorobenzene (16.92 g, 0.074 mol) in Et20 (100 ml) was added to a suspension of magnesium shavings in Et20 (60 ml) and the mixture was refluxed for 1 hour . The reaction was cooled to 23 ° C and a solution of 4-cyanopyridine 54 (7.80 g, 0.075 mol) in 1: 1 Et20: THF (150 ml) was added rapidly with vigorous stirring. The mixture was refluxed for 24 hours, then cooled to 23 ° C. Ice (100 g) was added, followed by 50% H2SO4 (50 ml), and the mixture was stirred for 1 hour. Et20 (100 ml) was added, the aqueous layer was separated, made basic with 10% NaOH, and extracted with EtOAc (2x250 ml). The combined organic layer was dried (Na2SO), filtered and concentrated. Purification by silica gel chromatography (eluent: 30% EtOAc: CH 2 Cl 2) yielded 12 g (0.047 mol, 64%) of product 55 as a colorless oil. MS (FAB for M + 1): m / e 252.

Stage 31: A mixture of compound 55 (10 g, 40 mmol), ethylene glycol (25 ml), and paratoluenesulfonic acid (8.56 g, 45 mmol) in toluene (200 ml) was refluxed for 36 hours, using a Dean-Stark trap. to remove water. The reaction was cooled to 23 ° C and basified with 10% Na 2 CO 3. The reaction was extracted with Et20, and the organic layer was dried (MgSO4), filtered and concentrated. Purification by silica gel chromatography (eluent: 30% EtOAc: CH 2 Cl 2) yielded 11 g (37 mmol, 100% of product 56 as a colorless oil, MS (FAB for M + 1): m / e 296.

Stage 32: Benzyl bromide (7 ml, 58.5 mmol) was added to a solution of compound 56 (9 g, 30.5 mmol) in acetone (250 ml) at 23 ° C then stirred for 16 h. Et20 (300 ml) was added, and the filtered precipitate was washed with Et20 (100 ml), and dried to yield 9 g (23.3 mmol, 76%) of the product 57 as a white solid. MS (FAB for M + 1): m / e 386.

Stage 33: NaBH (1 g, 27 mmol) was added to a solution of compound 57 (4 g, 10.33 mmol) in MeOH (80 ml) at 0 ° C, then warmed to 23 ° C, and stirred for 4 hours. The solvent was evaporated, NaHCO3 (100 mL) was added, and the mixture was extracted with CH2Cl2 (300 mL). The organic layer was separated, dried (MgSO), filtered, and concentrated. Purification by silica gel chromatography (eluent: 20% acetone-CH 2 Cl 2) yielded 3.5 g (8.97 mmol, 87%) of the product 58 as an oil. MS (FAB for M + 1): m / e 391.

Stage 34: Phenyl chloroformate (3 ml, 23.8 mmol) was added to a solution of compound 58 (4.0 g, 10.2 mmol) and Et3N (2 ml, 14.4 mmol) in 1,2-dichloroethane (40 ml) at 23 ° C then the refluxed for 1 hour. The reaction was cooled, concentrated, water was added, the mixture was extracted with CH2Cl2, dried (MgSO), filtered and concentrated. Purification by silica gel chromatography (eluent: 30% acetone-hexane) yielded 4.1 g (9.78 mmol, 76%) of product 59 as a white oil. MS (FAB for M + 1): m / e 420.

Stage 35: P-TSA (0.70 g, 3.67 mmol) was added to a solution of compound 59 (1.0 g, 2.38 mmol) in 3: 1 acetone: water (20 ml) then refluxed for 3 hours. The mixture was cooled and 5% Na 2 CO 3 (50 ml) was added. The mixture was extracted with EtOAc, added (Na2SO4), filtered, and concentrated to yield 0.80 g (2.13 mmol, 89%) of product 60. MS (FAB for M + 1): m / e 376 Stage 36: CH3Li (1.6 mL, 1.6 mmol) was added to a suspension of Cul (99%, 304 mg, 1.6 mmol) in dry Et2O (5 mL) at 10 ° C, then stirred at this temperature for 15 minutes. The reaction was cooled to -50 ° C, and a solution of compound 60 (200 mg, 0.533 mmol) in Et20 (5 mL) was added. The reaction was stirred at -50 ° C for 1 h, then heated to 0 ° C for a period of 1 h. The reaction was quenched by the addition of water (3 ml) and 10% Na 2 CO 3 (10 ml). The organic layer was separated, dried (Na2SO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: 25% acetone-hexane) yielded 100 mg (0.256 mmol, 48%) of compound 61 A as a white solid (FAB for M + 1): m / e 391 and 45 mg (0.115 mmol, 22%) of compound 61 B as a white solid. (FAB for M + 1): m / e 391.

Stage 37: A solution of compounds 61 A and 61B (mixture 2: 1, 500 mg, 1. 28 mmol) in 50% H 2 SO 4 (20 ml) was refluxed for 4 hours, then cooled to 23 ° C, diluted with water, and washed with CH 2 Cl 2 (30 ml). The aqueous layer was basified with 10% NaOH, and extracted with CH2Cl2 (2x50 ml). The combined organic extracts were dried (MgSO4), filtered and concentrated to yield 345 mg (1.27 mmol, 99%) of the products 62A: 62B (ratio 1: 9) as a white solid. MS (FAB for M + 1): m / e 272.

Stage 8: Using the same procedure as for Example 1, Step, compound 63 was prepared from compound 62B. MS (FAB for M + 1): m / e 510.

Step 38: NaBH 4 (15 mg, 0.40 mmol) was added to a solution of compound 63 (150 mg, 0.29 mmol) in EtOH (5 mL) at 0 ° C, then stirred at 23 ° C for 16 hours. The resulting mixture was concentrated and CH CI2 (100 mL), water (50 mL) and 10% NaOH (2 mL) were added. The organic layer was separated, dried (Na 2 SO), filtered, and concentrated. Purification by silica gel chromatography (eluent: 5% MeOH with NH3-CH2CI2) yielded 130 mg (0.25 mmol, 86%) of the product 64 as a mixture of isomers. MS (FAB for M + 1): m / e 512.

EXAMPLE 10 Stage 39: 65 66 NaH (5.26 g, 60%, 0.131 mol) in THF (300 ml) was suspended and cooled to 0 ° C. Compound 65 (10 g, 0.131 mol) was slowly added at 0 ° C, then stirred for 30 minutes. TBDPS-CI (17.0 ml, 0.065 mol) was dissolved in THF (100 ml) and added dropwise to the reaction. The reaction was stirred at 23 ° C for 16 hours, quenched with water, then the organic layer was washed with aqueous NaHCO 3, brine, dried (Na 2 SO 4), filtered and concentrated. Purification by silica gel chromatography (eluent: 10% EtOAc-hexane) yielded 18.31 g (0.058 mol, 89%) of product 66. MS (M + 1): 315.

Stage 40: Oxalyl chloride (0.416 ml, 4.77 mmol) was dissolved in CH2Cl2 (30 ml), and cooled to -78 ° C. DMSO (0.338 ml, 4.77 mmol) was added dropwise and stirred at -78 ° C for 15 minutes. Compound 66 (1.0 g, 3.18 mmol) was dissolved in CH2Cl2 (8 mL), and added dropwise. The mixture was stirred -78 ° C for 20 minutes. Et3N (1.33 mL, 9.54 mmol) was added dropwise, and stirred at -78 ° C for 40 minutes. Methyl (triphenylphosphoranylidene) acetate was dissolved (1.59 g, 4.77 mmol) in CH2Cl2 (8 mL), and added dropwise, and the mixture was stirred at -78 ° C for 10 minutes, then at 23 ° C for 16 hours. The reaction mixture was diluted with Et20 (100 mL) and the precipitate was filtered. The filtrate was washed with water, then brine, dried (Na2SO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: 3% EtOAc-hexane) yielded 0.65 g (55%) of product 67.

MS (M + 1): 369.

Stage 41 Stage 41 TBDPSO ^ ^^. ^^ nu 67"^^ = 0- ^ 68 Compound 67 (0.65 g, 1.67 mmol) was dissolved in CH2Cl2 (10 mL) under N2, and cooled to 0 ° C. DIBAL-H (4.53 ml 1.0 M in hexane, 4.53 mmol) it was added dropwise and the mixture was stirred at 0 ° C for 2 hours. Saturated aqueous Na 2 SO 4 (10 ml) was added and the mixture was filtered through a Celite pad The filtrate was washed with brine, dried (Na2SO4), filtered and concentrated. Purification by silica gel chromatography (eluent: 15% EtOAc-hexane) yielded 420 mg (1.23 mmol, 70%) of product 68.

MS (LCMS for M + 1) = 341.

Stage 42: Compound 68 (6.0 g, 17.6 mmol) was dissolved in CH2Cl2 (150 mL) under N2. Molecular sieves (7 g, 4) were added, and cooled to -20 ° C. Titanium (IV) isopropoxide (5.2 ml, 17.6 mmol) was added, followed by D-diisopropyl tartrate (4.49 ml, 21.1 mmol), and the mixture was stirred at -20 ° C for 40 minutes. Tert-butyl hydroperoxide (5.3 ml, . 0-6.0 M in decane, 26.5 - 31.8 mmol), then stirred at -10 ° C for 4 days. A fresh solution of FeS04 * 7H20 / citric acid (18 ml of 16.5 g / 5.5 g in 50 ml of water) was added dropwise and stirred at -10 ° C for 1 hour. The mixture was heated to 23 ° C and filtered through a pad of celite. The filtrate was washed with brine twice, dried (Na 2 SO), filtered and concentrated. Purification by silica gel chromatography (eluent: 10% EtOAc-hexane to 20% EtOAc-hexane) yielded 1.49 g (4.18 mmol, pure) and 8.08 g (10.7 mmo, 47% mixture with D-tartrate). diisorporpyl) of product 69 (84% combined yield). MS (M + 1): 357.

Stage 43: Compound 69 was dissolved (6.62 g, 47%, 8.73 mmol) in CH 2 Cl 2 (300 ml). Et3N (7.3 ml, 52.4 mmol) was added followed by triphenylmethyl chloride (7.3 g, 26.2 mmol), and DMAP (catalytic amount). The mixture was stirred at 23 ° C for 16 hours, then the solution was washed with water twice, then brine, dried (Na 2 SO), filtered and concentrated. Purification by silica gel chromatography (eluent: 3% EtOAc-hexane) yielded 4.83 g (8.1 mmol, 92%) of product 70. MS (M + 1): 599.

Stage 44: Mg (1.93 g, 79.4 mmol) in Et20 (50 mL) was suspended. A piece of l2 was added, then heated to 45 ° C. 3,4-Dichlorobenzyl chloride (5.50 ml, 39.7 mmol) in Et20 (50 ml) was added slowly and the mixture was stirred at 45CC for 2 hours, then cooled to 23 ° C. In a second flask, CuCN (71 mg, 0.79 mmol) was suspended in Et20 (20 ml), then cooled to -30 ° C under N2. The Grignard reagent was cannulated in the second flask, then heated to -15 ° C. Compound 70 (4.75 g, 7.94 mmol) in Et20 (50 mL) was added dropwise. The mixture was stirred at -15 ° C overnight, then warmed to 23 ° C and stirred for 24 hours. The reaction was quenched with 25% aqueous sodium citrate at 0 ° C and the product was extracted with EtOAc. The combined organic extracts were washed with brine, dried (Na2SO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: 3% EtOAc-hexane) yielded 3.73 g (4.9 mmol, 62%) of product 71. MS (M + 1): 759.

Stage 45: Compound 71 (1.5 g, 2.0 mmol) was dissolved in CH2Cl2 (5 mL) and cooled to 0 ° C. 2,6-Di-tert-butyl pyridine (1.35 ml, 6 mmol) was added, followed by methyltriflate and the mixture was stirred at 23 ° C for 16 h. The mixture was diluted with CH2Cl2, washed with water, brine, dried (Na2SO4), filtered, and concentrated. Purification by silica gel chromatography (eluent: 3% EtOAc-hexane) yielded 1.35 g (87%) of product 72. MS (M + 23): 795.

Stage 46: Compound 72 (1.35 g, 1.7 mmol) was dissolved in MeOH (10 mL) and CH 2 Cl 2 (10 mL) and HCl in 1,4-dioxane (20 mL, 4M) was added. The mixture was stirred at 23 ° C for 30 minutes, then concentrated, redissolved in EtOAc, washed with aqueous NaHCO 3, brine, dried (Na 2 SO), filtered, and concentrated. Purification by silica gel chromatography (eluent: 1: 1 EtOAc-hexane, then 5% MeOH-CH 2 Cl 2) yielded 0.261 g (0.89 mmol, 52%) of product 73. MS (M + 1): 293.

Stage 47: Compound 73 (0.261 g, 0.89 mmol) was dissolved in CH2CI (10 mL), Et3N (0.5 mL, 3.56 mmol) was added, and the mixture was cooled to 0 ° C. CH3S02CI (0.21 ml, 2.67 mmol) was added dropwise. The mixture was stirred at 0 ° C for 1 h, then warmed to 23 ° C, diluted with CH 2 Cl 2, washed with 1 N HCl twice, brine, dried (Na 2 SO), filtered and concentrated to yield 0.391 g (0.87 mmol, 98%) of product 74. MS (M + 23): 471.

Stage 48: Compound 74 (0.391 g, 0.87 mmol) in CH3CN (15 ml) was dissolved in a sealed tube. NH3 / H20 (15 ml, 30%) was added. The tube was sealed and the mixture was stirred at 23 ° C for 3 days. CH3CN was removed under vacuum, the resulting mixture was extracted with CH2Cl2, dried (Na2SO4), filtered, and concentrated. Purification by preparative TLC from silica gel (eluent: 10% MeOH with NH3-CH2CI2) afforded 76 mg (0.28 mmol, 32%) of product 75. MS (M + 1): 274.

Stage 8: Using the same procedure as for Example 1, Step, compound 76 was prepared from compound 75. MS (M + 1): m / e513.

EXAMPLE 11-A Stage 49: i (tBu) Me2 78 Et3N (4 mL, 28.8 mmol) was added to a solution of compound 77 (4.0 g, 10.3 mmol) in CH2Cl2 (50 mL) at 23 ° C. Trichloroethyl chloroformate (4 ml, 29.1 mmol) was added, and the reaction was stirred at 23 ° C for 16 h. Water (130 ml) and CH2Cl2 (200 ml) were added. The organic layer was separated, dried (MgSO), filtered, and concentrated. Purification by silica gel chromatography (eluent: 20% v / v acetone: CH 2 Cl 2) yielded 4.0 g (68%) of the product 78 as an oil. MS (FAB for M + 1): m / e 564.

Stage 50: Tetrabutylammonium fluoride (1.0 M in THF, 15 ml, 15 mmol) was added to a solution of compound 78 (4.0 g, 7.10 mmol) in CH 2 Cl 2 (20 ml) and stirred at 23 ° C for 16 h. Water (100 ml) and CH2Cl2 (200 ml) were added.

The organic layer was separated, dried (MgSO), filtered, and concentrated. Purification by silica gel chromatography (eluent: 20% v / v acetone: CH 2 Cl 2) yielded 2.5 g (94%) of the product 79 as an oil.

Stage 51: Bis- (2-methoxyethyl) amino-sulfur trifluoride (1.0 ml, . 4 mmol) was added to a solution of compound 79 (1.0 g, 2.65 mmol) in CH 2 Cl 2 (10 mL) at 0 ° C and the mixture was then stirred at 23 ° C for 16 h. Water (10 ml) and 10% Na 2 CO 3 (10 ml) were added and the mixture was extracted with CH 2 Cl 2 (100 ml). The organic layer was separated, dried (MgSO 4), filtered and concentrated. Purification by silica gel chromatography (eluent: 1: 5: 5 acetone: CH2Cl2: hexanes) yielded 0.60 g (1.33 mmol, 50%) of product 80 as an oil. MS (FAB for M + 1): m / e 450.

Stage 52: Step 52 - A suspension of compound 80 (300 mg, 0.668 mmol) and zinc metal (250 mg, 3.83 mmol) in glacial AcOH (5 ml) was stirred at 70 ° C for 1 h. The reaction was cooled and concentrated. Water (50 ml) and 10% of NaOH (5 ml) and the mixture was extracted with CH2Cl2 (100 ml). The organic layer was separated, dried (MgSO 4), filtered, and concentrated to yield 150 mg (0.54 mmol, 81%) of product 81. MS (FAB for M + 1): m / e 276.

Stage 8: Using the same procedure as for Example 1, Step, the compound of Example 11 -A was prepared. MS: m / e 514.

Using a similar procedure, the compound of the Example 11-B: MS: m / e 514.

EXAMPLE 12 OvjN? V? Hri, 2 stage 7 Stage 53: Compound 23 (5.00 g, 13.5 mmol) was dissolved in acetone (60 ml) and potassium permanganate (4.50 g, 28.5 mmol) in water (200 ml) was added. The reaction mixture was stirred at 23 ° C for 3 h. Sodium bisulfate (50 g) was added until a colorless solution was obtained. The resulting mixture was filtered, the solid was washed with water, and the filtrate was extracted with CH2Cl2. The combined organic extracts were dried (MgSO4), filtered, and concentrated to yield 4.60 g (11.9 mmol, 88%) of product 82 as a white solid. MS (ES for M + 1): m / e 388.

Stage 9: | Using the same procedure as for Example 1, Step, compound 83 was prepared. MS (ESparaM + 1): m / e387.

Stage 7: Using the same procedure as for Example 1, Step, compound 84 was prepared. MS (ES for M + 1): m / e 287.

Stage 8: Using the same procedure as for Example 1, Step 8, compound 85 was prepared. MS (ES for M + 1): m / e 470.

Stage 54: Compound 85 (1.00 g, 2.13 mmol) was dissolved in CH2Cl2 (10 mL) and the Burgess reagent (1.78 g, 7.45 mmol) was added dropwise. The reaction mixture was stirred at 23 ° C for 16 h. The solvent was evaporated, and the crude product was purified by silica gel chromatography (eluent: 3% MeOH-CH 2 Cl 2) to yield the product 86 as an oil. MS (M + 1): m / e 452.

Stage 7: Using the same procedure as for Example 1, Step, the following intermediates were prepared.

Stage 9: Using the same procedure as for Example 1, Stage , the following compounds were prepared.

EXAMPLE 13 Step 55: cyclopentyl Compound 1 (15.95 g, 0.080 mol) was dissolved in THF (400 ml) under an N atmosphere and cooled to 0 ° C. NaH (6.72 g, 60% in oil, 0.168 mol) was added in portions, and the reaction mixture was stirred at 0 ° C for 30 minutes. CH3I (28.5 g, 12.5 ml, 0.201 mol) was added, and the reaction mixture was stirred at 23 ° C for 48 h. The solvent was removed, water was added, and the resulting mixture was extracted with Et20. The combined organic extracts were dried (MgSO4), filtered, and concentrated. The crude product was recrystallized twice from pentane to yield 8.97 g (0.039 mol, 49%) of product 89 as a white solid. Purification of the mother liquor by silica gel chromatography (eluent: 5% EtOAc-hexane to 20% EtOAc-hexane) yielded an additional 5.00 g (0.022 mol, 27%) of product 89 as a white solid. MS (M + 1): m / e 228.

Using a similar procedure, the following intermediary was prepared: MS (M + 1): m / e254 Stage 1 : Using the same procedure as for Example 1, Step 1, the following intermediates were prepared.

Stage 4: Using the same procedure as for Example 1, Step, the following intermediates were prepared: Stage 7: 91 92 Using the same procedure as for Example 1, Step, the following intermediates were prepared.

Stage 8: Using the same procedure as for Example 1, Step, the following intermediates were prepared.

Step 7: Using the same procedure as for Example 1, Step, the following intermediates were prepared.

Stage 9: Using the same procedure as for Example 1, Step, the following compounds were prepared.

EXAMPLE 14 Stage 56: Compound 96 (7.28 g, 43.0 mmol) was dissolved in CH2Cl2 (170 mL) and cooled to 0 ° C. MCPBA was added in portions (11.87 g of 75% of MCPBA, 51.6 mmol). The reaction mixture was stirred at 23 ° C for 16 h. CH2Cl2 (150 ml) was added and the organic solution was washed three times with NaOH 1 N, once with water, and once with brine. The organic solution was dried (MgSO), filtered, and concentrated. Purification by silica gel chromatography (eluent: 1: 5 EtOAc: PHCH3) afforded 3.89 g (20.4 mmol, 47%) of product 97 as a colorless oil.

Stage 19: 7 98 Using the same procedure as for Example 4-B, Step 19, the following intermediates were prepared.

Stage 6: 98 Using the same procedure as for Example 1, the following intermediaries were staged.

Stage 7: 99 100 Using the same procedure as for Example 1, Stage , the following intermediaries were prepared.

Stage 8: Using the same procedure as for Example 1, Stage , the following intermediaries were prepared.

Stage 9: Using the same procedure as for Example 1, Step, the following compounds were prepared: The following assays can be used to determine the CCR3 inhibitor and antagonist activity of the compounds of the invention.

Spa-based ccr3 receptor binding assay: The procedure used is described in Dairaghi et al, J. Biol. Chem., 272 (1997), pages 28206-28209, and is described in detail below. The source of the CCR3 receptors for this assay were membranes purified from a Y3 cell line transfected with human CCR3. The membranes were prepared by MDS Pharma Services (Bothell, WA) as follows: The cells grew under selection (1 mg / ml G418) in 50-cell bucket bio reactors with minimal media recirculation. The cells were harvested with trypsin and washed once with PBS. The cells were resuspended in 20 volumes of ice-cold 2.5 mM Tris-HCl, pH 7.4 containing protease inhibitors (0.3 mM PMSF, 3 μg / ml aprotinin, 3 μg. / ml luepeptin) and incubated on ice for 15 minutes. The expanded cells were subjected to vigorous vortex and the large debris was removed by centrifugation (1200 rpm, 5 minutes, 4 ° C.) The clarified lysate was subjected to ultra-centrifugation (100,000 X g, 1 hr., 4 ° C) to pellet the membrane fraction and the membrane fraction was resuspended in 25 mM HEPES, pH 7.6, 75 mM NaCl, 1 mM EDTA The protease inhibitors were added as described above after a sample it was removed for protein determination, the membrane preparation was made to a final concentration of 6 to 10 mg / ml and BSA was added to 0.5% The membranes were stored in 1 ml aliquots (-70 ° C).

All compounds to be tested were dissolved in 100% DMSO at a concentration of 10 mM with sonication if necessary. The unlabeled eotaxin (R &D Systems, Minneapolis, MN) was used to generate the standard curves. A primary stock solution (100 nM) was prepared in binding buffer (BB, 25 mM HEPES, pH 7.6, 75 mM NaCl, 1 mM CaCl 2, 5 mM MgCl 2, 0.5% BSA, adjusted to pH 7.6 final) and stored in small aliquots (-70 ° C). [12dl] -Eotaxin (NEN Life Science Products, Boston, MA) was resuspended in water (25 μCi / ml) and stored at -20 ° C. Pearls of PVT-agglutinin wheat germ-SPA (pearls of PVT-WGA-SPA; Amersham Biosciences, Piscataway, NJ) were resuspended in BB (20 mg / ml). The PVT-WGA-SPA beads and membranes were combined in BB and slowly turned at 4 ° C for 1 hr. For 100 binding reactions, 1 mg of membranes was combined with 20 mg of PVT-WGA-SPA beads in 10 ml of BB. For further reactions, the appropriate amounts of membranes and beads were combined in 10 ml of BB. The membranes linked to beads were collected by centrifugation (1500 rpm, 10 minutes, 4 ° C), resuspended in BB and combined with [125 l] -eotaxin. For 100 reactions, [125 L] -eotaxin 5 X 106 cpm was added to 1 mg of membranes in 20 mg of beads in a total of 18 ml of BB. The competition binding reactions were initiated by adding to each well of a 96-well plate (Wallac Oy, Turku, Finland) 180 μl of [125l] -eotaxin / membranes / beads. For compound tests, 20 μl was added to each of the triplicate cavities (final concentration 100 μM to 10 pM). For standard curves, 20 μl of unlabeled eotaxin were added to each of the triplicated cavities (final concentration of 3 nM to 1 pM). Each well contained [125 l] -eotaxin 0.05 nM, 10 μg of membranes and 200 μg of PVT-WGA-SPA beads in a total volume of 200 μl. The plates were shaken briefly and incubated at room temperature for 5 hours before counting in a scintillation counter. An IC5o value was interpolated from the dilution series for each compound and used to calculate a value K1 using the following equation: K1 = IC50 / (1 + [Liigand] / Kd).

Intracellular calcium assay: Cell Culture: CREM3 cells, in which the human CCR3 receptor is stably expressed in the rat Y3 cell line, were cultured in high glucose DMEM containing 10% FBS, 50 U / ml penicillin, 50 μg / ml streptomycin, 2 mM L-glutamine, 1 mg / ml of G418 (Gemini Bioproducts). The cells were cultured in fresh medium every 2 days.

Assay Procedure: The intracellular calcium levels were measured using a fluorometric imaging plate reader (FLIPR), (Molecular Devices, Sunnyvale CA) as described by the manufacturer. CREM3 cells were grown overnight at 20,000 cells / well in 96-well black-walled clear-bottom plates (Packard), pre-coated with 100 μg / ml poly D-lysine hydrobromide (Sigma). Adherent CREM3 cells were loaded with 4 μM Fluo-3 AM (Molecular Probes, Eugene, OR), in HBBS pH 7.4 without phenol red, containing 20 mM HEPES, 0.5% FBS, 2.5 mM probenecid (Sigma), 0.04% acid pluronic (Molecular Probes) for 1 h at 37 ° C. Adherent cells were washed with buffer / diluent (HBBS pH 7.4 without phenol red, containing 20 mM HEPES, 0.5% BSA, 2.5 mM probenecid) by an automated Denley Cell Washer (Labsystemes Oy, Helsinki, Finland); after the final wash, the fluid was aspirated at a level of 100 μl. The chemokines were diluted in wash buffer / diluent to 4 times the final concentration and added in a volume of 50 μl / well. The compounds were resuspended in dimethyl sulfoxide (DMSO) and diluted in wash buffer / diluent to 3 times the final concentration. Each compound was titrated in half logarithm dilutions over 3.5 logarithms. The DMSO control groups, which contain solvent concentrations equivalent to that of the highest concentration of compounds, were included. Cells and chemokines were maintained at 37 ° C during all calcium measurements. Fluid additions were made in accordance with FLIPR recommendations for adherent cells (CREM3). Fluorescence data were collected at 1 second interval for 60 seconds, followed by collection at 2 second intervals for 60 seconds. Background fluorescence was quantified in the cavities containing cells but not chemokines and subtracted from all experimental samples. All conditions were given in quadruplicate. The non-linear regression analysis using Prima GraphPad (Graphpad Software Inc., San Diego, CA) was used to calculate EC50 values for chemokines or IC50 values for compounds. In the assay for determining the CCR3 receptor binding, the compounds of the invention vary in activity from a Ki of about 1 to about 500 nM, with preferred compounds having an activity scale of from about 1 to about 100 nM, more preferably about 1 to about 50 nM. In the calcium flux assay, the preferred isomers of the invention vary in activity from an IC 50 of about 3 nM to about 500 nM, with preferred compounds having an activity scale of from about 3 to about 100 nM, more preferably about 3 at approximately 50 nM. Example 2-EE has a Ki of 2.8 nM an IC50 of 3.6 nM. Compared with the CCR3 antagonists described in WO 01/77101, the compounds of the present invention exhibit lower Ki values in the binding assay, and lower IC50 values in the calcium flux assay. To prepare pharmaceutical compositions from CCR3 antagonist compounds described by this invention, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, wafers and suppositories. The powders and tablets may be comprised of from about 5 to about 95 percent active ingredient. Suitable solid carriers are known in the art, for example, magnesium carbonate, magnesium stearate, talc, sugar or lactose. Tablets, powders, wafers and capsules can be used as solid dosage forms suitable for oral administration. Examples of pharmaceutically acceptable carriers and manufacturing methods for various compositions can be found in A. Gennaro (ed.), Remington's Pharmaceutical Sciences, 18th Edition, (1990), Mack Publishing Co., Easton, Pennsylvania. Liquid form preparations include solutions, suspensions and emulsions. As an example there may be mentioned water or water-propylene glycol solutions for parenteral injection or addition of sweeteners and opacifiers for solutions, suspensions and oral emulsions. Liquid form preparations may also include solutions for intranasal administration. Aerosol preparations suitable for inhalation may include solutions and solids in powder form, which may be in combination with a pharmaceutically acceptable carrier, such as an inert compressed gas, for example nitrogen.

Also included are solid form preparations which are intended to be converted, shortly before use, into liquid form preparations for either oral or parenteral administration. Such liquid forms include solutions, suspensions or emulsions. The compounds of the invention can also be transdermally available. The transdermal compositions may take the form of creams, lotions, aerosols and / or emulsions and may be included in a transdermal patch of the matrix or reservoir type as are conventional in the art for this purpose. Preferably, the compound is orally administered. Preferably, the pharmaceutical preparation is in a unit dosage form. In such form, the preparation is subdivided into suitably sized unit doses containing appropriate quantities of the active component, for example, an effective amount to achieve the desired purposes. The amount of the active compound in a unit dose of preparation can be varied or adjusted from about 10 mg to about 500 mg, preferably from about 25 mg to about 300 mg, more preferably from about 50 mg to about 250 mg, and most preferably from about 55 mg to about 200 mg, according to the particular application. The current dosage of the CCR3 compound used can be varied depending on the requirements of the patient and the severity of the condition being treated. The determination of the appropriate dosage regimen for a particular situation is within the skill of the art. For convenience, the total daily dosage can be divided and administered in portions during the day as required. The amount and frequency of administration of the CCR3 compounds of the invention and / or pharmaceutically acceptable salts thereof will be regulated according to the judgment of the attending clinician considering such factors as age, condition and size of the patient as well as severity of the symptoms to be treated. A typical recommended daily dosage regimen for oral administration can range from about 100 mg / day to about 300 mg / day, preferably 150 mg / day to 250 mg / day, more preferably about 200 mg / day, in two to four divided doses . While the present invention has been described in conjunction with the specific embodiments described above, many alternatives, modifications and variations thereof will be apparent to those of ordinary skill in the art. All alternatives, modifications and variations are proposed to fall within the spirit and scope of the present invention.

Claims

NOVELTY OF THE INVENTION CLAIMS

1. - A compound represented by structural formula II or a pharmaceutically acceptable salt thereof, further characterized OR 13 because; n is 0 or 1; Xa is -C (R13) 2-, -C (O) - or -CR13-; Ra is R6a-phenyl, R6a-pyridyl, R6a-thiophenyl or R6a-naphthyl; R1 is hydrogen, halogen, -OH, alkyl, hydroxyalkyl, alkoxy or alkoxyalkyl; R2a is x is 0-2 and y is 0-2, provided the sum of x and y is 1-4; R6a is 1 to 3 substituents independently selected from the group consisting of hydrogen, halogen, -CF3, CF30-, -CN, CF3S02-, -NHCOCF3, CH3S02-, Or 5-membered heteroaryl and ?? iz t where Z is -O-, -NH- or -N (CH3) -; R, 13 and R > 16 are independently selected from the group consisting of hydrogen and alkyl; R14 is alkyl, alkenyl, haloalkyl, hydroxy, hydroxyalkyl, -CN, - (CR20R21) qO-alkyl, - (CR20R21) q-NR20R24, - (CR20R1) q-N3) - (CR20R21) qC (O) -alkyl , - (CR20R21) qC (O) -phenyl, - (CR20R21) q-COOR20, - (CR20R21) qC (O) NR20R24, - (CR20R21) qS (0) or-2-R23I - (CR20R21) qN (R20 ) -C (O) NR20R24, - (CR20R21) qN (R20) -C (O) OR23 or - (CR20R21) qOC (O) R23; R14a is hydrogen or alkyl; or R14 and R14a together form = 0 or = NOR20; or R14 and R14a, together with the ring carbon to which they are attached, form a spirocycle ring of 3 to 6 carbon atoms; q is O, 1, 2 or 3; R15 and R5a are each 1 or 2 substituents independently selected from the group consisting of H, halogen, OH, alkyl, alkoxy, -CF3, -OCF3, -CN, -C (0) R25, -COOR25, -S ( 0) or-R25, -S (O) 0-2CF3, -NR20R24, phenyl and heterocycloalkyl; or two substituents R15 or two R15a on the carbon atoms of the adjacent ring, together with the carbons to which they are attached, form a fused cycloalkyl ring of 5-6 members; R20 and R21 are independently selected from the group consisting of H and alkyl; R23 is alkyl or phenyl; and R24 is H, alkyl or R12-phenyl; provided that when R14 is - (CR20R21) q-NR20R24 and R24 is H, R20 is alkyl.

2. The compound according to claim 1, further characterized in that n is 1, R1 is hydrogen, R16 is hydrogen, Ra is R6a-phenyl, R6a is one or two halogen substituents, and Xa is -C (R13) ( R13) -.

3. The compound according to claim 1, further characterized in that R14 is alkyl, haloalkyl, hydroxy, hydroxyalkyl, alkoxy or alkoxyalkyl and R14a is hydrogen.

4. - The compound according to claim 1, further characterized in that R2a is wherein the sum of x and y is 1 or 2.

5. The compound according to claim 4, further characterized in that R2a is selected from the group consisting of s independently selected from the group consisting of hydrogen, halogen, methyl, methoxy and CF3.

6. A pharmaceutical composition, comprising an effective amount of a CCR3 antagonist of claim 1 in combination with a pharmaceutically acceptable carrier.

7. The use of a compound as defined in claim 1, for the preparation of a medicament for the treatment of asthma.

8. The use of a compound of the formula or a pharmaceutically acceptable salt thereof, wherein n is 0 or 1; X is C (R13) 2-, -C (R13) (R19) -, -C (0) -, -O-, -NH-, -N (alkyl) -, I rent O-C (O) - alkyl 0-C (0) -0- alkyl 0-C (0) -NH- alkyl Q-C (O) -N (alkyl) -CR 13- -CR 13-. -CR13-. -CR13- (alkyl) 2 C (O) - or -N-alkyl; R is R6-phenyl, R6-pyridyl, R6-thiophenyl or R6-naphthyl; R1 is hydrogen, halogen, -OH, alkyl, hydroxyalkyl, alkoxy or alkoxyalkyl; R2 is 6-membered heteroaryl substituted with R7, R8, R9; Heteroaryl N-oxide 6 members substituted with R7, R8, R9; 5-membered heteroaryl substituted with R10, R11; R12-naphthyl; fluorenyl; diphenylmethyl, and be 1-4; R3 is R6-phenyl, R6-heteroaryl or R6-naphthyl; R 4 is hydrogen, alkyl, fluoroalkyl, cyclopropylmethyl, -CH 2 CH 2 OH, -CH 2 CH 2 -O-alkyl, -CH 2 C (O) -O-alkyl, -CH 2 C (0) NH 2, -CH 2 C (0) -NH-alkyl or -CH 2 C ( 0) -N (alkyl) 2; R5 and R11 are independently selected from the group consisting of hydrogen and alkyl; R6 is 1 to 3 substituents independently selected from the group consisting of hydrogen, halogen, alkyl, alkoxy, -CF3, CF30-, CH3C (O) -, - CN, CH3S02-, CF3S02-, R16-phenyl, R16-benzyl, CH3C (= N0CH3) -, -NH2, -NHCOCF3, -NHCONH-alkyl, O-NAZ-NHCO-alkyl, -NHS02-alkyl, 5-membered heteroaryl and \ -f > wherein Z is -O-, -NH- or -N (CH3) -; R7 and R8 are independently selected from the group consisting of alkyl, halogen, -NR20R21, -OH, -FC3, -OCH3, -O-acyl, and -OCF3; R9 is R7, hydrogen, phenyl, -N02, -CN, -CH2F, -CHF2, -CHO, -CH = NOR20, pyridyl, pyridyl N-oxide, pyrimidinyl, pyrazinyl, -N (R20) CONR21R22, -NHCONH (chloroalkyl), -NHCONH-cycloalkylalkyl, -NHCO-alkyl, -NHCOCF3, - NHS02N (alkyl) 2, -NHS02-alkyl, -N (S02CF3) 2, - NHC02-alkyl, cycloalkyl, -SR23, -SOR23, -S02R23, -S02NH-alkyl, -OS02-alkyl, -OS02CF3, hydroxyalkyl, - CONR20R21, -CON (CH2CH2-0-CH3) 2, - OCONH-alkyl, -C02R20, -Si (CH3) 3 or -B (OC (CH3) 2) 2; R10 is alkyl, -NH2 or R12-phenyl; R 2 is 1 to 3 substituents independently selected from a group consisting of hydrogen, alkyl, -CF3, -C02R2 °, -CN, alkoxy and halogen; R13 and R16 are independently selected from the group consisting of hydrogen and alkyl; R14 is alkyl, alkenyl, haloalkyl, hydroxy, hydroxyalkyl, -CN, - (CR20R21) qO-alkyl, - (CR20R21) q-NR20R24, - (CR20R21) q-N3, - (CR20R21) qC (O) -alkyl, - (CR20R21) qC (O) -phenyl, - (CR20R21) q-COOR20, - (CR20R21) qC (O) NR20R24, - (CR20R21) qS (O) 0-2-R23, - (CR20R2) qN (R20 ) -C (O) NR20R24, - (CR20R21) qN (R20) -C (O) OR23 or - (CR 0R2) qOC (O) R23; R 4a is hydrogen or alkyl; or R14 and R14a together form = 0 or = NOR20; or R14 and R14a, together with the ring carbon to which they are attached, form a spirocycle ring of 3 to 6 carbon atoms; q is O, 1, 2, or 3; R15 and R15a are each 1 or 2 substituents independently selected from the group consisting of H, halogen, OH, alkyl, alkoxy, -CF3, -OCF3, -CN, -C (0) R25, -COOR25, -S (O ) 0-2R25, -S (0) or-2CF3I-NR20R24, phenyl and heterocycloalkyl; or two substituents R15 or two R15a on the carbon atoms of the adjacent ring, together with the carbons to which they are attached, form a fused cycloalkyl ring of 5-6 members; R17 and R18 are independently selected from the group consisting of hydrogen and alkyl, or R17 and R18 together are an alkylene group of C2-C5 and with the carbon to which they are attached form a cycloalkyl ring of 3 to 6 carbon atoms; R19 is R6-phenyl, R6-heteroaryl, R6-naphthyl, cycloalkyl, cycloalkylalkyl or alkoxyalkyl; R20, R21 and R22 are independently selected from the group consisting of H and alkyl; R23 is alkyl or phenyl; and R24 is H, alkyl or R12-phenyl; for the preparation of a medication for the treatment of asthma.

9. The use as claimed in claim 8, wherein n is 1; R1 is hydrogen; R16 is hydrogen; R is R6-phenyl, wherein R6 is one or two halogen substituents; X is -C (R13) (R13) -; R 14 is alkyl, haloalkyl, hydroxy, hydroxyalkyl, alkoxy or alkoxyalkyl; R14a is hydrogen; R2 is selected from the group consisting of uniquely independently selected from the group consisting of hydrogen, halogen, methyl, methoxy and CF3.

10. A compound selected from the group consisting of