Macrocyclic inhibitors of hepatitis c virus

TECHNICAL MEMORY

MACROCYCLIC INHIBITORS OF HEPATITIS C VIRUS

The present invention relates to macrocyclic compounds that possess inhibitory activity on the replication of the hepatitis C virus (HCV). It further relates to compositions comprising these compounds as active components, as well as processes for preparing these compounds and compositions.

The hepatitis C virus is the leading cause of chronic liver disease worldwide and has become a focus of considerable medical research. HCV is a member of the Flaviviridae family of viruses in the genus hepacivirus, and is closely related to the genus flavivirus, which includes a number of viruses implicated in human disease, such as dengue virus and yellow fever virus, and with the family of animal pesteviruses, which includes bovine viral diarrhea virus (VBDV). HCV is a positive-sense, single-stranded RNA virus with a genome of about 9600 bases. The genome comprises both 5' and 3' untranslated regions that adopt RNA secondary structures and a central open reading frame that encodes a single polyprotein of about 3,010-3,030 amino acids. The polyprotein encodes ten gene products that are generated from the precursor polyprotein by an organized series of co- and post-translational endoproteolytic cleavages mediated by host and viral proteases. Viral structural proteins include the core nucleocapsid protein and two envelope glycoproteins E1 and E2. Non-structural (NS) proteins encode some essential viral enzymatic functions (helicase, polymerase, protease), as well as proteins of unknown function. Viral genome replication is mediated by an RNA-dependent RNA polymerase, encoded by the non-structural protein 5b (NS5B). In addition to polymerase functions, viral helicase and protease functions, both encoded in the bifunctional NS3 protein, were shown to be essential for HCV RNA replication. In addition to the NS3 serine protease, HCV also encodes a metalloproteinase in the NS2 region.

After the initial acute infection, a majority of infected individuals developed chronic hepatitis because HCV replicates preferentially in hepatocytes, but is not directly cytopathic. In particular, the lack of a vigorous T cell response and the high tendency of the virus to mutate seem to promote a high degree of chronic infection. Chronic hepatitis can progress to liver fibrosis leading to cirrhosis, end-stage liver disease and HCC (hepatocellular carcinoma), making it the leading cause of liver transplantation.

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There are 6 main HCV genotypes and more than 50 subtypes, which are geographically distributed differently. HCV type 1 is the predominant genotype in Europe and the United States. The extensive genetic heterogeneity of HCV has important diagnostic and clinical implications, possibly explaining the difficulties in vaccine development and the lack of response to therapy.

HCV transmission can occur through contact with contaminated blood or blood products, for example following blood transfusion or intravenous drug use. The introduction of diagnostic tests used in the evaluation of blood produced a downward trend in the incidence of HCV post-transfusion. However, given the slow progression to end-stage liver disease, existing infections will continue to present a serious medical and economic burden for decades.

Current HCV therapies are based on interferon-alpha (IFN-α) (pegylated) in combination with ribavirin. This combination therapy produces a sustained virologic response in more than 40% of patients infected with genotype 1 viruses and about 80% of those infected with genotypes 2 and 3. In addition to limited efficacy against HCV type 1, this combination therapy has side effects and is poorly tolerated in many patients. Most side effects include influenza-like symptoms, hematologic abnormalities, and neuropsychiatric symptoms. Therefore, there is a need for more effective, convenient and better tolerated treatments.

Recently, two peptimimetic HCV protease inhibitors gained attention as clinical candidates, namely, BILN-2061 disclosed in WOOO/59929 and VX-950 disclosed in W003/87092. A number of similar HCV protease inhibitors have also been disclosed in the academic and patent literature. It is already evident that prolonged administration of BILN-2061 or VX-950 selects for HCV mutants that are resistant to the respective drug, so-called drug escape mutants. These drug escape mutants possess characteristic mutations in the HCV protease genome, notably D168V, D168A and/or A156S. Therefore, additional drugs with different resistance patterns are required to provide non-improving patients with treatment options, and multidrug combination therapy is likely to be the norm in the future, even for first-line treatment.

Experience with anti-HIV drugs, and HIV protease inhibitors in particular, has emphasized that sub-optimal pharmacokinetics and complex dosing regimens rapidly result in inadvertent failures to adhere. This in turn means that the 24-hour trough concentration (trough plasma concentration) for the respective drugs in an HIV regimen often falls below the IC threshold.₉₀ or ED_go for much of the day. It is considered that a minimum level of 24 hours of at least the HF₅₀, and more realistically, the CI₉₀ or ED_go, is essential for slowing the development of drug escape mutants. Achieving the pharmacokinetics and drug metabolism necessary to allow such minimal levels provides a rigorous challenge for drug design. The strong peptimimetic nature of prior art HCV protease inhibitors, with multiple peptide bonds, represents pharmacokinetic obstacles to effective dosing regimens.

There is a need for HCV inhibitors that can overcome the drawbacks of current HCV therapy, such as side effects, limited efficacy, the rise of resistance, and failure to adhere.

The present invention relates to HCV inhibitors that are superior in one or more of the following related pharmacological properties, ie potency, reduced cytotoxicity, improved pharmacokinetics, improved resistance profile, acceptable dosing, and pill loading.

In addition, the compounds of the present invention have relatively low molecular weight and are easy to synthesize from starting materials that are commercially available or readily available through synthetic procedures known in the art.

W005/010029 discloses macrocyclic aza-peptide serine protease inhibitors of hepatitis C, pharmaceutical compositions comprising the aforementioned compounds for administration to a subject suffering from HCV infection, and methods for treating an HCV infection in a patient by administering a pharmaceutical composition comprising the aforementioned compounds.

The present invention relates to inhibitors of HCV replication, which can be represented by Formula (I):

<img file="ECSP088150A_D0001.tif" />

and the n-oxides, salts and stereoisomers thereof, where each dotted line (represented by.....) represents an optional double bond;

X is N, CH and when X has a double bond it is C;

R.¹ is -OR⁷, -NH-SO₂R.⁸;

R.² is hydrogen and when X is C or CH, R² can also be CA alkyl;

R.³ is hydrogen, C alkyl_V6, alkoxyCj₆-alkyl Ο_ν6, C-cycloalkyl₃_₇;

R.⁴ is aril or Het;

n is 3, 4, 5, or 6;

R.⁵ represents halo, CA alkyl, hydroxy, CA alkoxy, polyhalo-Ci_ alkyl₆, phenyl or Het;

R.⁶ represents CA alkoxy, mono- or di-alkylamino Ci__yes;

R.⁷ is hydrogen; aryl; Het; C cycloalkyl₃.₇ optionally substituted with alkyl Ci-₅; or alkyl Ci-₆optionally substituted with C cycloalkyl₃.₇, aril or by Het;

R.⁸ is aril; Het; C cycloalkyl₃.₇ optionally substituted with alkyl Ci-₆; or alkyl Ci-₆ optionally substituted with C cycloalkyl₃.₇, aril or by Het;

Aryl as a group or part of a group is phenyl optionally substituted with one, two, or three substituents selected from halo, hydroxy, nitro, cyano, carboxyl, CA alkyl, CA alkoxy, CA alkoxy CA alkyl, CA alkylcarbonyl, amino, mono- or di-CA alkylamino, azido, mercapto, polyhalo-CA alkyl, polyhalo-CA alkoxy, C cycloalkyl₃.₇ , pyrrolidinyl, piperidinyl, piperazinyl, 4-alkyl CA-piperazinyl, 4-alkylcarbonyl CA-piperazinyl, and morpholinyl; where the morpholinyl and piperidinyl groups can be optionally substituted with one with two CA alkyl radicals;

Het as a group or part of a group is a 5- or 6-membered saturated, partially unsaturated, or fully unsaturated heterocyclic ring containing 1 to 4 heteroatoms each independently selected from nitrogen, oxygen, and sulfur, said heterocyclic ring optionally being fused with a benzene ring; and Het as a full ring optionally substituted with one, two, or three substituents each independently selected from the group consisting of halo, hydroxy, nitro, cyano, carboxyl, Ci_ alkyl₆, alkoxy Ci_₅, alkoxy Ci.₆-Ci-alkyl₆, alkylcarbonyl Ci_₆, amino, mono- or di-alkylamino Ci_₆, azido, mercapto, polyhaloalkyl Ci_₆, polyhaloalkoxy Ci_₆, C-cycloalkyl_{3 7}, pyrrolidinyl, piperidinyl, piperazinyl, 4-alkyl Ci-₆-piperazinyl, 4-alkylcarbonylCi-₆-piperazinyl and morpholinyl; where the morpholinyl and piperidinyl groups can be optionally substituted with one or two Ci-6 alkyl radicals. The invention also relates to methods for the preparation of the compounds of formula (I), the /V-oxides, addition salts, quaternary amines , metal complexes and stereochemically isomeric forms thereof, their intermediates and the use of the intermediates in the preparation of the compounds of formula (I).

The invention relates to the compounds of formula (I) per se, the Μ-oxides, addition salts, quaternary amines, metal complexes and stereochemically isomeric forms thereof, for use as a medicament. The invention further relates to pharmaceutical compositions comprising a carrier and a filler effective as an antiviral agent of a compound of formula (I), as specified herein. Pharmaceutical compositions may comprise combinations of the aforementioned compounds with other anti-HCV agents. The invention further relates to the aforementioned pharmaceutical compositions for administration to a subject suffering from HCV infection.

The invention also relates to the use of a compound of formula (I), or an N-oxide, addition salt, quaternary amine, metal complex, or stereochemically isomeric forms thereof, for the manufacture of a medicament for inhibiting replication of HCV or the invention relates to a method for inhibiting HCV replication in a warm-blooded animal said method comprises the administration of an effective amount of a compound of formula (I), or a pro-drug, /V-oxide , addition salt, quaternary amine, metal complex, or stereochemically isomeric forms thereof.

As used hereinafter and hereinbefore, the following definitions apply unless otherwise specified.

The term halo is generic to fluoro, chloro, bromo, and iodo.

The term "polyhaloC4alkyl" as a group or part of a group, for example in polyhalo-alkoxy Ci-₆, is defined as mono- or polyhalo alkyl Ci.₅ substituted, especially Ci alkyl.₅ substituted by up to one, two, three, four, five, six or more halo atoms, such as methyl or ethyl by one or more fluoro atoms, eg, difluoromethyl, trifluoromethyl, trifluoroethyl. Trifluoromethyl is preferred. Also included are perfluoro-alkyl Ci groups.₆, which are Ci_ alkyl groups₆ where all hydrogen atoms are replaced by fluoro atoms, eg pentafluoroethyl. In the case that more than one halogen atom is attached to an alkyl group in the definition of polyhalo-alkyl (0₆, the halogen atoms can be the same or different.

As used herein, “C alkyl_m” as a group or part of a group defines straight or branched chain saturated hydrocarbon radicals having 1 to 4 carbon atoms, such as for example methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2- butyl, 2-methyl-1 -propyl; “alkyl Ci-₆”comprises alkyl radicals and higher homologues thereof having 5 or 6 carbon atoms such as, for example, 1-pentyl, 2-pentyl, 3-pentyl, 1-hexyl, 2-hexyl, 2-methyl-1 -butyl, 2-methyl-1-pentyl, 2-ethyl-1-butyl, 3-methyl-2-pentyl and the like. It is of interest among renters (0₆ the rent (0₄.

The term “alkenyl C₂.₆” as a group or part of a group defines straight and branched chain hydrocarbon radicals possessing saturated carbon-carbon bonds and at least one double bond and possessing from 2 to 6 carbon atoms, such as, for example, ethenyl ( or vinyl), 1-propenyl, 2-propenyl (or allyl), 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-2-propenyl, 2-pentenyl, 3-pentenyl, 2-hexenyl, 3-hexenyl, 4- hexenyl, 2-methyl-2-butenyl, 2-methyl-2-pentenyl and the like. It is of interest between the alkenyls C₂.₃ alkenyl C₂.₄,

The term “alkynyl C₂.₃” as a group or part of a group defines straight and branched chain hydrocarbon radicals possessing saturated carbon-carbon bonds and at least one triple bond and possessing 2 to 6 carbon atoms, such as, for example, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 2-pentynyl, 3-pentynyl, 2-hexynyl, 3-hexynyl and the like. It is of interest among the alkynyls (0₃ the alkynyl (0₄.

C cycloalkyl₃.₇ is generic to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

Alkanediyl C₁₅ defines straight and branched bivalent chain hydrocarbon radicals having 1 to 6 carbon atoms such as, for example, methylene, ethylene, 1,3-propanediyl,

1,4-butanediyl, 1,2-propanediyl, 2,3-butanediyl, 1,5-pentanediyl, 1,6-hexanediyl and the like. It is of interest among the alkanediyls C₁₆alkanediyl C₁₄.

Alkoxy Ci-₆ means Ci alkyloxy.₆where I rent Ci.₆ is as defined above.

As used herein, above, the term (=0) or oxo forms a carbonyl moiety when attached to a carbon atom, a sulfoxide moiety when attached to a sulfur atom, and a sulfonyl moiety when two of said terms bond to a sulfur atom. Whenever an oxo group is substituted for a ring or ring system, the carbon atom to which the oxo is attached is a saturated carbon.

The radical Het is a heterocycle, as specified in the present specification and claims. Preferred among the Het radicals are those that are monocyclic,

Examples of Het comprise, for example, pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazinolyl, isothiazinolyl, thiazolyl, isothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl (including 1,2,3-triazolyl , 1,2,4-triazolyl), tetrazolyl, furanyl, thienyl, pyridyl, pyrimidyl, pyridazinyl, triazinyl, and the like. Of interest among the Het radicals are those that are unsaturated, especially those that possess an aromatic character. Of additional interest are those Het radicals that possess one or two nitrogens.

Each of the Het radicals mentioned in this and in the following paragraphs can be optionally substituted with the number and type of substituents mentioned in the definitions of the compounds of formula (I) or any of the subgroups of compounds of formula (I). Some of the Het radicals mentioned in this and the following paragraphs may be substituted by one, two or three hydroxy substituents. Such substituted hydroxy rings can be produced as their tautomeric forms bearing keto groups. For example, a 3-hydroxypyridazine moiety can occur in its tautomeric form, 2H-pyridazin-3-one. When Het is piperazinyl, it is preferably substituted in the 4-position by a substituent attached to the 4-nitrogen with a carbon atom, for example 4-alkyl Ci_₆,

4-polyhalo-alkyl Ci-₆, alkoxy Ci-₆-Ci-alkyl₆, alkylcarbonyl Ci-₆, C-cycloalkyl₃-₇·

Het radicals of interest comprise, for example, pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl (including 1,2,3-triazolyl, 1 ,2,4-triazolyl), tetrazolyl, furanyl, thienyl, pyridyl, pyrimidyl, pyridazinyl, pyrazolyl, triazinyl, or any such heterocycle fused with a benzene ring, such as indolyl, indazolyl (especially 1H-indazolyl), indolinyl, quinolinyl, tetrahydroquinolinyl (especially 1,2,3,4-tetrahydroquinolinyl), isoquinolinyl, tetrahydroisoquinolinyl (especially 1,2,3,4-tetrahydroisoquinolinyl), quinazolinyl, phthalazinyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzoxadiazolyl, benzothiadiazolyl, benzofuranyl, benzothienyl.

The radicals Het pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, 4-substituted piperazinyl are preferably attached via their nitrogen atom (i.e. 1-pyrrolidinyl, 1-piperidinyl, 4-thiomorpholinyl, 4-morpholinyl, 1-piperazinyl, substituted piperazinyl in position 4).

It should be noted that the locations of the radicals in any molecular moiety used in the definitions can be anywhere on said moiety, as long as it is chemically stable.

The radicals used in the definitions of the variables include all possible isomers, unless otherwise indicated. For example, pyridyl includes 2-pyridyl, 3-pyridyl, and 4-pyridyl; pentyl includes 1-pentyl, 2-pentyl and 3-pentyl.

When any variable occurs more than once in any constituent, each definition is independent.

Whenever used hereinafter, the term "compounds of formula (I)", or the present compounds" or similar terms, is intended to include the compounds of formula (I), their prodrugs, /V-oxides, addition salts, quaternary amines, metal complexes and stereochemical isomeric forms. One embodiment comprises the compounds of formula (I) or any subgroup of compounds of formula (I) specified herein, as well as /V-oxides, salts, as well as possible stereoisomeric forms thereof. Another embodiment comprises the compounds of formula (I) or any subgroup of compounds of formula (I) specified herein, as well as salts as their possible stereoisomeric forms.

Compounds of formula (I) possess several centers of chirality and exist as stereochemical isomeric forms. The term "stereochemical isomeric forms" as used herein defines all possible compounds prepared from the same atoms linked by the same sequence of bonds, but possessing different non-interchangeable three-dimensional structures, that compounds of formula (YO).

In reference to the instances in which (ñ) or (S) is used to designate the absolute configuration of a chiral atom in a substituent, the designation is carried out considering the complete compound and not the isolated substituent.

Unless otherwise mentioned or indicated, the chemical designation of a compound includes the mixture of all possible stereochemical isomeric forms that said compound may possess. Said mixture may contain all diastereomers and/or enantiomers of the basic molecular structure of said compound. All stereochemical isomeric forms of the compounds of the present invention which require both the pure form or in combination with each other are intended to be within the scope of the present invention.

Stereoisomeric pure forms of compounds and intermediates as mentioned herein are defined as essentially free isomers of other enantiomeric or diastereomeric forms of the same basic molecular structure of said compounds or intermediates. In particular, the term "stereoisomerically pure" refers to compounds or intermediates that have a stereoisomeric excess of at least 80% (ie 90% minimum of one isomer and a maximum of 10% of other possible isomers) up to a stereoisomeric excess. 100% (ie 100% of one isomer and none of the others), more especially, compounds and intermediates having a stereoisomeric excess of 90% to 100%, even more especially that they have a stereoisomeric excess of 94% to 100% and even more especially that they have a stereoisomeric excess of 97% to 100%. The terms "enantiomerically pure" and "diastereomerically pure" should be understood in a similar way, but considering the enantiomeric excess and the diastereomeric excess, respectively, of the mixture in question.

Pure stereoisomeric forms of the compounds and Intermediates of the present invention can be obtained by applying procedures known in the art. For example, enantiomers can be separated from one another by selective crystallization of their diastereomeric salts with optimally active acids or bases. Examples thereof are tartaric acid, dibenzoyltartaric acid, ditoluoyltartaric acid and camphorsulfonic acid. Alternatively, enantiomers can be separated by chromatographic techniques using chiral stationary phases.

Said pure stereochemical isomeric forms may also be derived from the corresponding pure stereochemical isomeric forms of the appropriate starting materials, provided that the reaction occurs stereospecifically. Preferably, if a specific stereoisomer is desired, said compound will be synthesized by specific methods of preparation. These methods will advantageously use enantiomerically pure starting materials.

The diastereomeric racemates of the compounds of formula (I) can be obtained separately by conventional methods. Appropriate physical separation methods that can be used to advantage are, for example, selective crystallization and chromatography, for example column chromatography.

For some of the compounds of formula (I), their pro-drugs, /V-oxides, salts, solvates, quaternary amines, or metal complexes and the intermediates used in their preparation, the absolute stereochemical configuration was not determined from experimental way. A person skilled in the art is able to determine the absolute configuration of such compounds using methods known in the art, such as, for example, X-ray diffraction.

It is also intended that the present invention include all isotopes of atoms that occur in the present compounds. Isotopes include those atoms that have the same atomic quantity but different mass numbers.

By way of general example and without limitation, isotopes of hydrogen include tritium and deuterium. Carbon isotopes include C-13 and C-14.

The term "pro-drug", as used throughout this text, means pharmaceutically acceptable derivatives such as esters, amides and phosphates, such that the product resulting from in vivo biotransformation of the derivative is the active drug, as defined in the compounds of formula (I). The reference of Goodman and Gilman (The Pharmacological Basis of Therapeutics, 8^{1 hour} ed, McGraw-Hill, Int. Ed. 1992, "Biotransformation of Drugs", p 13-15) which generally describes prodrugs. Prodrugs preferably possess excellent aqueous solubility, increased bioavailability, and are readily metabolized to active inhibitors in vivo. Prodrugs of a compound of the present invention can be prepared by modifying functional groups present in the compound such that the modifications are cleaved, either by routine manipulation or in vivo, to the parent compound.

Preferred are pharmaceutically acceptable ester prodrugs which are hydrolyzable in vivo and which are derived from those compounds of formula (I) which possess a hydroxy or carboxyl group. An in vivo hydrolyzable ester is an ester, which is hydrolyzed in the human or animal body to produce the parent acid or alcohol. Suitable pharmaceutically acceptable esters for carboxy include Ci alkoxymethyl esters.₆, for example methoxymethyl, alkanoyloxymethyl esters Ci.₅ for example pivaloyloxymethyl, phthalidyl esters, cycloalkoxycarbonyloxy C esters₃.₈-Ci_ alkyl₆ for example 1-cyclohexylcarbonyloxyethyl; 1,3-dioxolen-2-onylmethyl esters, for example 5-methyl-1,3-dioxolen-2-onylmethyl; and Ci_ alkoxycarbonyloxyethyl esters₆ for example 1-methoxycarbonyloxyethyl, which can be formed at any carboxy group in the compounds of this invention.

An in vivo hydrolyzable ester of a compound of formula (I) containing a hydroxy group includes organic esters such as phosphate esters and α-acyloxyalkyl ethers and related compounds which as a result of in vivo hydrolysis of ester cleavage cleave to give the parent hydroxy group. Examples of α-acyloxyalkyl ethers include acetoxymethoxy and 2,2-dimethylpropionyloxy-methoxy. A selection of in vivo hydrolyzable esters that form para hydroxy groups include alkanoyl, benzoyl, phenylacetyl, and substituted benzoyl and phenylacetyl, alkoxycarbonyl (to give alkyl carbonate esters), dialkylcarbamoyl, and N-(dialkylam¡noethyl)-N-alkyl lcarbamoyl (to give carbamates), dialkylaminoacetyl and carboxyacetyl. Examples of substituents on benzoyl include morpholino and piperazino attached from a ring nitrogen atom through a methylene group to the 3- or 4-position of the benzoyl ring.

For therapeutic use, the salts of the compounds of formula (I) are those in which the counter-ion is acceptable for pharmaceutical use. However, salts of acids and bases that are not pharmaceutically acceptable may also be used, for example, in the preparation or purification of a pharmaceutically acceptable compound. All salts, whether acceptable for pharmaceutical use or not, are included within the scope of the present invention.

Acid and base addition salts are acceptable for pharmaceutical use as mentioned above are herein intended to comprise the therapeutically active non-toxic acid and base addition salt forms that the compounds of formula (I) are capable of forming. Pharmaceutically acceptable acid addition salts may conveniently be obtained by treating the base form with said appropriate acid. Appropriate acids comprise, for example, inorganic acids such as hydrocids, for example hydrochloric or hydrobromic, sulfuric, nitric, phosphoric and the like acids; or organic acids such as, for example, acetic, propanoic, hydroxyacetic, lactic, pyruvic, oxalic (ie ethanedioic), malonic, succinic (ie butanedioic), maleic, fumaric, malic (ie hydroxybutanedioic), tartaric acids , citric, methanesulfonic, ethanesulfonic, benzenesulfonic, p-toluenesulfonic, cyclamic, salicylic, p-aminosalicylic, pamoic, and similar acids.

Conversely, such salt forms can be transformed by treatment with an appropriate base into the free base form.

Compounds of formula (I) containing an acidic proton can also be transformed into their non-toxic amine or metal addition salt forms by treatment with appropriate organic and inorganic bases. Salt forms with bases include, for example, ammonium salts, alkali and alkaline earth metal salts, for example lithium, sodium, potassium, magnesium, calcium salts and the like, salts with organic bases, for example benzathine , /V-methyl-D-glucamine, hydrabamine salts and salts with amino acids such as, for example, arginine, lysine and the like.

The term addition salt, as used herein above, also encompasses the solvates that the compounds of formula (I) are capable of forming, as well as the salts thereof. Such solvates are, for example, hydrates, alcoholates and the like.

The term "quaternary amine" as used herein above, defines the quaternary ammonium salts that the compounds of formula (I) are capable of forming through the reaction between a basic nitrogen of a compound of formula (I) and an agent quaternization agent, such as, for example, an alkyl halide, aryl halide, or optionally substituted arylalkyl halide, for example, methyl iodide or benzyl iodide. Other reagents with good leaving groups may also be used, such as alkyl trifluoromethanesulfonates, alkyl methanesulfonates, and alkyl p-toluenesulfonates. A quaternary amine has a positively charged nitrogen. Counter ions acceptable for pharmaceutical use include chlorine, bromine, iodine, trifluoroacetate and acetate. The contract of choice can be introduced using ion exchange resins.

The A/-oxide forms of the present compounds are intended to comprise compounds of formula (I) wherein one or more nitrogen atoms are oxidized to the so-called /V-oxide.

It will be appreciated that compounds of formula (I) may possess complex, chelating, metal bond-forming properties and can therefore exist as metal complexes or metal chelates. Such metal derivatives of the compounds of formula (I) are intended to be included within the scope of the present invention.

Some of the compounds of formula (I) may also exist in their tautomeric form. Such forms, although not explicitly indicated in the above formula, are intended to be included within the scope of the present invention.

As mentioned above, the compounds of formula (I) possess several asymmetric centers. To refer more efficiently to each of these asymmetric centers, the numbering system will be used, as indicated in the following structural formula.

<img file="ECSP088150A_D0002.tif" />

Asymmetric centers are present at the 1,4 and 6 positions of the macrocycle, as well as at the 3' carbon atom in the 5-membered ring, the 2' carbon atom where the R substituent is² is C-alkyl._ñ and at the Γ position of the carbon atoms, where X is CH. Each of these asymmetric centers can be presented in its R or S configuration.

The stereochemistry at position 1 preferably corresponds to that of an L-amino acid configuration, ie, that of L-proline.

When X is CH, the 2 substituted carbonite groups at the 1' and 5' positions of the cyclopentane ring are preferably in a trans configuration. The carbonite substituent in the 5' position is preferably in that configuration that corresponds to an L-proline configuration. The carbonite groups substituted at the 1' and 5' positions are preferably as described below in the structure of the following formula.

<img file="ECSP088150A_D0003.tif" />

“S.

Compounds of formula (I) include a cyclopropyl group, as represented in the structural fragment below:

Or where C₇ represents the carbon at position 7 and the carbons at position 4 and 6 are asymmetric carbon atoms of the cyclopropane ring.

Regardless of other possible asymmetric centers in other segments of the compounds of formula (I), the presence of these two asymmetric centers means that the compounds can exist as mixtures of diastereomers, such as the diastereomers of the compounds of formula (I) where the carbon at position 7 is set to either syn for the carbonyl or syn for the amide, as shown below.

ooh

<img file="ECSP088150A_D0004.tif" />

¹¹ 5&lt;LH

O ^6O c₇H=c

C7 syn to carbonyl^{07 yes}AND^no P^now carbonyl C7 syn for amide

ooh

<img file="ECSP088150A_D0005.tif" />

C7 syn to carbonyl C7 syn to carbonyl

C7 syn for amide

One embodiment relates to compounds of formula (I) where the carbon at position 7 is configured syn for the carbonyl. Another embodiment relates to compounds of formula (I) where the configuration at the carbon at position 4 is R. A specific subgroup of compounds of formula (I) are those in which the carbon at position 7 is syn configured for the carbonyl and where the configuration on the carbon at position 4 is R.

Compounds of formula (I) may also include a proline residue (when X is N) or a cyclopentyl or cyclopentenyl residue (when X is CH or C). Preferred are compounds of formula (I) where the substituent at the (5') position and the substituent at the 3' position are in a trans configuration. Of particular interest are the compounds of formula (I) where position 1 has the configuration corresponding to L-proline and the substituent in position 3' is in a trans configuration with respect to position 1. Preferably, compounds of formula (I) possess the stereochemistry as indicated by the structures of formulas (la) and (lb) below:

<img file="ECSP088150A_D0006.tif" />

(the)

<img file="ECSP088150A_D0007.tif" />

(lbs)

One embodiment of the present invention relates to compounds of formula (I) or formula (la) or any subgroup of compounds of formula (I), where one or more of the following conditions apply:

(a) R² is hydrogen;

(b) X is nitrogen;

(c) A double bond is present between carbon atoms 7 and 8.

One embodiment of the present invention relates to compounds of formula (I) or of formulas (la), (lb), or any subgroup of compounds of formula (I), where one or more of the following conditions apply:

(a) R² is hydrogen;

(b) XesCH;

(c) A double bond is present between carbon atoms 7 and 8.

Special subgroups of compounds of formula (I) are those represented by the following structural formulas:

<img file="ECSP088150A_D0008.tif" />

(lc)

<img file="ECSP088150A_D0009.tif" />

(ld)

Among the compounds of formula (lc) and (ld), those having the stereochemical configuration of the compounds of formulas (la) and (lb), respectively, are of particular interest.

The double bond between carbon atoms 7 and 8 in compounds of formula (I), or in any subgroup of compounds of formula (I), may be in either a cis or a trans configuration. Preferably, the double bond between carbon atoms 7 and 8 is in a cis configuration, as described in formulas (lc) and (ld).

A double bond between the 1' and 2' carbon atoms may be present in compounds of formula (I), or in any subgroup of compounds of formula (I), as described in formula (le) below.

R.⁵

<img file="ECSP088150A_D0010.tif" />

EITHER

<img file="ECSP088150A_D0011.tif" />

Still another particular subgroup of compounds of formula (I) are those represented by the following structural formulas:

<img file="ECSP088150A_D0012.tif" />

<img file="ECSP088150A_D0013.tif" />

(lf) (ig)

<img file="ECSP088150A_D0014.tif" />

I\k Y (

<img file="ECSP088150A_D0015.tif" />

(lh)

Among the compounds of formulas (lf), (lg) or (lh), those having the stereochemical configuration of the compounds of formulas (la) and (lb) are of particular interest.

In (la), (lb), (lc), (ld), (le), (lf), (lg) and (lh), where applicable, X, n, R¹,R²,R³,R⁴,R⁵ and R⁶are as specified in the definitions of the compounds of formula (I) or in any of the subgroups of compounds of formula (I) specified herein.

It is to be understood that the above defined subgroups of compounds of formulas (la), (lb), (lc), (ld), (le), (lf), (lg) or (lh), as well as any other subgroups defined herein, also include any /V-oxides, addition salts, quaternary amines, metal complexes, and stereochemical isomeric forms of such compounds.

When n is 2, the remainder -CH₂- grouped by "n" corresponds to ethanediyl in the compounds of formula (I) or in any subgroup of compounds of formula (I). When n is 3, the remainder -CH₂- grouped by "n" corresponds to propanediyl in the compounds of formula (I) or in any subgroup of compounds of formula (I). When n is 4, the remainder -CH₂- grouped by "n" corresponds to butanediyl in the compounds of formula (I) or in any subgroup of compounds of formula (I). When n is 5, the remainder —CH₂- grouped by "n" corresponds to pentanediyl in the compounds of formula (I) or in any subgroup of compounds of formula (I). When n is 6, the remainder -CH₂- grouped by "n" corresponds to hexanediyl in the compounds of formula (I) or in any subgroup of compounds of formula (I). Particular subgroups of the compounds of formula (I) are those compounds where n is 4 or 5.

Embodiments of the invention are compounds of formula (I) or any of the subgroups of compounds of formula (I), in which (a) R¹ is -OR⁷, especially where R⁷ is alkyl Ci-₆, such as methyl, ethyl, or tert-butyl (or t.butyl) and more preferably where R⁷ is hydrogen;

(b) R¹ is -NHS(=O)2R⁸, especially where R⁸ is C1.6 alkyl, C3-C7 cycloalkyl, or aryl, for example where R⁸is methyl, cyclopropyl, or phenyl; or (c) R.¹ is -NHS(=O)2R⁸, especially where R⁸ is C3-cycloalkyl₇ substituted with Ci-alkyl₅, preferably where R⁸is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, any of which is replaced by alkyl Ci-₄, that is to say by methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, or isobutyl.

Furthermore, embodiments of the invention are compounds of formula (I) or any of the subgroups of compounds of formula (I) where R¹ is -NHS(=O)₂R.⁸, especially where R⁸ is cyclopropyl substituted by C1.4alkyl, ie by methyl, ethyl, propyl, or isopropyl.

Furthermore, embodiments of the invention are compounds of formula (I) or any of the subgroups of compounds of formula (I) where R¹ is -NHS(=O)₂R.⁸, especially where R⁸ is 1-methylcyclopropyl.

Furthermore, embodiments of the invention are compounds of formula (I) or any of the subgroups of compounds of formula (I) where (a) R² is hydrogen;

(b) R² is alkyl Ci_₆, preferably methyl.

Embodiments of the invention are compounds of formula (I) or any of the subgroups of compounds of formula (I) where (a) X is N, C (X being linked by a double bond) or CH (X being linked by a double bond). single bond) and R² is hydrogen;

(b) X is C (X being joined by a double bond) and R² is alkyl Ci_₆, preferably methyl.

Other embodiments of the invention are compounds of formula (I) or any of the subgroups of compounds of formula (I) where (a) R³ is hydrogen;

(b) R³ is alkyl Ci_₆;

(c) R³ is Ci_g alkoxy-Ci_ alkyl₆ or C cycloalkyl₃_₇.

Preferred embodiments of the invention are compounds of formula (I) or any of the subgroups of compounds of formula (I) where R³ is hydrogen, or alkyl Ci.₆, more preferably hydrogen or methyl.

Embodiments of the invention are compounds of formula (I) or any of the subgroups of compounds of formula (I) where R⁴ is aryl or Het, each independently, optionally substituted with any of the Het or aryl substituents mentioned in the definitions of the compounds of formula (I) or of any of the subgroups of compounds of formula (I); or specifically, said aryl or Het each independently being optionally substituted with alkyl Ci_₆, halo, amino, mono- or di-alkylamino Ci_₆, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl,

4-alkyl Ci.₆-piperaz¡n¡lo; and where the morpholinyl and piperidinyl groups can be optionally substituted by one or two alkyl radicals Ci-₆.

Embodiments of the invention are compounds of formula (I) or any of the subgroups of compounds of formula (I) where R⁴is a radical

<img file="ECSP088150A_D0016.tif" />

or, especially, where R⁴ is selected from the group consisting of:

<img file="ECSP088150A_D0017.tif" />

<img file="ECSP088150A_D0018.tif" />

where, when possible a nitrogen may possess an R substituent^4a or a binding to the rest of the molecule; each R^4a on any of the R substituents⁴ it can be selected from those mentioned as possible substituents on Het, as specified in the definitions of the compounds of formula (I) or of any of the subgroups of compounds of formula (I);

more specifically each R^4a can be hydrogen, halo, alkyl Ci_₆, amino, or mono- or di-alkylamino Ci_₆, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, 4-alkyl Ci-₆-piperazino; and where the morpholinyl and piperidinyl groups can be optionally substituted by one or two C^ alkyl radicals;

more specifically each R^4a is, each independently, hydrogen, halo, alkyl Ci_₅, amino, or mono- or di-C alkylamino₁₆;

and where R^4a is substituted on a nitrogen atom, preferably it is a carbon containing the substituent that is connected to nitrogen through a carbon atom or one of its carbon atoms; and in which in this instance R^4a preferably it is alkyl Ci.₆.

Embodiments of the invention are compounds of formula (I) or any of the subgroups of compounds of formula (I) where R⁴ is phenyl or pyridiyl (especially 4-pyridyl) which may each be substituted by 1, 2 or 3 substituents selected from those mentioned for aryl in the definitions of the compounds of formula (I) or any of its subgroups. In particular, said phenyl or pyridyl is substituted by 1-3 (or by 1-2, or by one) substituent or substituents selected from halo, Ο-alkyl_Ί.₆ or alkoxy Ci_₆.

Embodiments of the invention are compounds of formula (I) or any of the subgroups of compounds of formula (I) where R⁵ is halo, or alkyl Ci-₆, preferably methyl, ethyl, isopropyl, tertbutyl, fluoro, chloro, or bromo. They include polyhalo-alkyl Ci-₆.

Embodiments of the invention are compounds of formula (I) or any of the subgroups of compounds of formula (I) where R⁶ is Ci-alkoxy₈ or di-alkylamino Ci-₈; preferably R⁶ is methoxy or dimethylamino; more preferably R⁶ it's methoxy.

Compounds of formula (I) consist of three building blocks P1, P2, P3. Building block P1 further contains a tail PT. The carbonyl group marked with an asterisk in compound (lc) below, can be part of either the P2 building block or the P3 building block. Due to chemical reasons, the P2 building block of compounds of formula (I) where X is C incorporates the carbonyl group attached to the 1' position.

The union of the building blocks P1 with P2, P2 with P3 and P1 with P1' (when R¹ is -NHSO₂R.⁸ or -OR⁷) comprises forming an amide bond. The union of the P1 and P3 blocks comprises the formation of the double bond. The ligation of the building blocks Ρ1, P2 and P3 to prepare compounds (li) or (lj) can be carried out in any given sequence. One of the steps comprises the cyclization by which the macrocycle is formed.

Presently represented below are the compounds (l-¡) which are compounds of formula (I) where the C7 and 08 carbon atoms are linked by a double bond and the compounds (lj) which are compounds of formula (I ) where C7 and C8 carbon atoms are linked by a single bond. Compounds of formula (lj) can be prepared from the corresponding compounds of formula (ll) by reduction of the double bond in the macrocycle.

<img file="ECSP088150A_D0019.tif" />

<img file="ECSP088150A_D0020.tif" />

It should be noted that in compounds of formula (lc), the formation of the amide bond between the P2 and P3 blocks can be achieved at two different positions on the urea fragment. A first amide bond comprises the nitrogen of the pyrrolidine ring and the adjacent carbonyl (marked with an asterisk). A second alternative amide bond formation involves the reaction of an asterisked carbonyl with a -NHR group.³. Both amide bond formations between building blocks P2 and P3 are feasible.

The synthetic procedures described herein below are intended to be applicable also for the racemates, stereochemically pure intermediates or final products, or any stereochemical mixtures. Racemates or stereochemical mixtures can be separated into stereoisomeric forms at any stage of the synthetic procedures. In one embodiment, the intermediates and final products possess the stereochemistry as specified above in the compounds of formula (la) and (lb).

To simplify the structural representation of the compounds of formula (I) or the intermediates, the group

<img file="ECSP088150A_D0021.tif" />

is represented by R⁹ and the dotted line represents the bond that unites said group represented by R⁹ for the rest of the molecule.

In one embodiment, compounds (1-¡) are prepared by first forming the amide bonds and subsequently forming the double bond junction between P3 and P1 with concomitant cyclization to the macrocycle.

In a preferred embodiment, the compounds (I) in which the bond between C₇ and C.₈ is a double bond, which are compounds of formula (1-¡), as defined above, can be prepared as indicated in the following reaction scheme:

<img file="ECSP088150A_D0022.tif" />

The formation of the macrocycle can be carried out by an olefin metathesis reaction in the presence of a suitable metal catalyst, such as for example the Ru-based catalyst reported by Miller, SJ, Blackwell, HE, Grubbs, RHJ Am 118, (1996), 9606-9614; Kingsbury, JS, Harrity, JPA, Bonitatebus, PJ, Hoveyda, AH, J. Am. Chem. Soc. 121, (1999), 791-799; and Huang et al., J. Am. Chem. Soc. 121, (1999), 2674-2678; for example a Hoveyda Grubbs catalyst.

Air-stable ruthenium catalysts such as bis(tricyclohexylphosphine)-3-phenyl-1H-indene-1-ylidene ruthenium chloride (Neolyst M1®) or bis(tricyclohexylphosphine) dichloride can be used. -[(phenylthio)methylene]ruthenium (IV). Other catalysts that can be used are Grubbs' first and second generation catalysts, ie, benzylidenebis(tricyclohex¡lf osfinajdichlororuthenium and (1,3-b¡s-(2,4,6-trimethylphenyl)- 2-imidazolidinylidene)dichloro(phenylmethylene)-(tricyclohexylphosphine)ruthenium, respectively. Of particular interest are the first and second generation Hoveyda-Grubbs catalysts, which are dichloro(oisopropoxyphenylmethylene)(tricyclohexylphosphine)-ruthenium(l I) and 1,3-bis-(2,4,6-trimethylphene¡ l )-2-imidazolidinylidene)dichloro(o-¡sopropoxy¡phenylmethylene)ruthen¡or respectively. Also, other catalysts containing other transition metals, such as Mo, can be used for this reaction.

Metathesis reactions can be carried out in a suitable solvent such as, for example, ethers, eg THF, dioxane; halogenated hydrocarbons, eg dichloromethane, CHCL, 1,2-dichloroethane and the like, hydrocarbons, eg toluene. In a preferred embodiment, the metathesis reaction is carried out in toluene. These reactions are carried out at increased temperatures under a nitrogen atmosphere.

Compounds of formula (I) where the link between C7 and C8 in the macrocycle is a single bond, i.e. compounds of formula (lj), can be prepared from compounds of formula (li) by reduction of the double bond C7-C8 in the compounds of formula (1-¡). this reduction can be carried out by catalytic hydrogenation with hydrogen in the presence of a noble metal catalyst, such as, for example, Pt, Pd, Rh, Ru or Raney nickel. It is of interest Rh in alumina. The hydrogenation reaction is preferably carried out in a solvent, such as, for example, an alcohol such as methanol, ethanol, or an ether such as THF, or mixtures thereof. Water can also be added to these solvents and solvent mixtures.

The R group¹ it may be found connected to the P1 building block at any stage of the synthesis, ie, before or after deletion or before or after deletion and reduction, as described herein above. Compounds of formula (I) in which R¹ represents -NHSO2R⁸, said compounds being represented by the formula (lk-1), can be prepared by joining the group R¹ to P1 by forming an amide bond between both residues. Similarly, compounds of formula (I), in which R¹ represents -OR⁷, i.e. compounds (lk-2), can be prepared by joining the R group¹ to P1 by forming an ester bond. In one embodiment, the -OR groups⁵ In the last step of the synthesis of the compounds, the compounds (I) are introduced as indicated in the following reaction schemes in which G represents a group:

<img file="ECSP088150A_D0023.tif" />

<img file="ECSP088150A_D0024.tif" />

(2a)

G-COOH + (2a)

Intermediate (2a) can be coupled with amine (2b) via an amine formation reaction such as any of the procedures for amide bond formation described herein below. In particular, (2a) can be treated with the coupling agent, for example A/,A/-carbonyldiimidazole (CDI), EEDQ, IIDQ, EDCI or benzotriazol-1-yl-oxy-trispirrolidinophosphonium hexafluorophosphate (commercially available as PyBOP ®), in a solvent such as ether, for example THF, or a halogenated hydrocarbon, for example dichloromethane, chloroform, dichloroethane and can be reacted with the desired sulfonamide (2b), preferably after reaction (2a) with the agent coupling. The reactions of (2a) with (2b) are preferably carried out in the presence of a base, for example a trialkylamine such as triethylamine or diisopropylethylamine, or 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). Intermediate (2a) can also be transformed into an active form, for example an active form of the general formula G-CO-Z, where Z represents halo, or the remaining part of an active ester, for example Z is an aryloxy group. such as phenoxy, p.nitrophenoxy, pentafluorophenoxy, trichlorophenoxy, pentachlorophenoxy and the like; or Z can be the residue of a combined anhydride. In one embodiment, G-CO-Z is an acid chloride (G-CO-CI) or a mixed acid anhydride (G-CO-O-CO-R or G

CO-O-CO-OR, R in the latter being, for example, C1.4 alkyl, such as methyl, ethyl, propyl, i-propyl, butyl, t-butyl, i-butyl or benzyl). The active form G-CO-Z is reacted with the sulfonamide (2b).

Activation of the carboxylic acid in (2a) as described in the above reactions can lead to an internal cleavage reaction to an azalactone intermediate of formula

<img file="ECSP088150A_D0025.tif" />

where X,R²,R³,R⁹, n are as specified above and where the stereogenic centers may possess the stereochemical configuration as specified above, eg as in (la) or (Ιό). Intermediates (2a-1) can be isolated from the reaction mixture, using conventional methodology and the isolated intermediate (2a-1) is then reacted with (2b), or the reaction mixture containing (2a-1). it can be further reacted with (2b) without isolation of (2a-1). In one embodiment, where the reaction with the coupling agent is carried out in a water-immiscible solvent, the reaction mixture containing (2a-1) can be washed with water or slightly basic water to remove all by-products. soluble in water. The washed solution obtained in this way can then be reacted with (2b), without further purification steps. Isolation of intermediates (2a-1), on the other hand, can provide certain advantages in that the isolated product, after optional further purification, can be reacted with (2b), giving rise to fewer by-products and less processing. easier to react.

Intermediate (2a) can be coupled with alcohol (2c) via an ester formation reaction. For example, (2a) and (2c) are reacted along with the removal of water, either physically, eg, by azeotropic removal of water, or chemically, by use of a dehydrating agent. Intermediate (2a) can also be converted to an active form of G-CO-Z, such as the active forms mentioned above and subsequently reacted with the alcohol (2c).

The ester formation reactions are preferably carried out in the presence of a base such as an alkali metal carbonate or hydrogen carbonate, for example sodium or potassium hydrogen carbonate, or a tertiary amine, such as the amines mentioned. herein in connection with amide formation reactions, especially a trialkylamine, eg triethylamine. Solvents that can be used in ester-forming reactions include ethers such as THF; halogenated hydrocarbons, such as dichloromethane, CH₂IC₂; hydrocarbons such as toluene; polar aprotic solvents such as DMF, DMSO, DMA; and the like solvents.

Compounds of formula (I) where R³ is hydrogen, said compounds being represented by (ll), can also be prepared by removing a protecting group PG, from a corresponding intermediate protected by nitrogen (3a), as in the following reaction scheme. The protecting group PG in particular is any of the nitrogen protecting groups mentioned herein below and can be removed using procedures also mentioned herein below:

<img file="ECSP088150A_D0026.tif" />

<img file="ECSP088150A_D0027.tif" />

The starting materials (3a) in the above reaction can be prepared by following the procedures for the preparation of compounds of formula (I), but using intermediates where the group R³ it's PG.

Compounds of formula (I) can also be prepared by reacting intermediate (4a) with intermediate (4b) as indicated in the following reaction scheme where the various radicals have the meanings specified above:

ooh

<img file="ECSP088150A_D0028.tif" />

YR⁹ (4b)

---------►

<img file="ECSP088150A_D0029.tif" />

(YO)

Y in (4b) represents hydroxy or a leaving group LG such as halide, eg bromide or chloride, or an arylsulfonyl group, eg mesylate, triflate or tosylate and the like.

In one embodiment, the reaction of (4a) with (4b) is an O-arylation reaction and Y represents a leaving group. This reaction can be carried out following the procedures described by EM Smith et al. (J. Med. Chem. (1988), 31, 875-885). In particular, this reaction is carried out in the presence of a base, preferably a strong base, in a reaction-inert solvent, for example one of the solvents mentioned for the formation of an amide bond.

In a particular embodiment, starting material (4a) is reacted with (4b) in the presence of a base that is strong enough to reduce a hydrogen from the hydroxy group, for example, an alkali metal hydride alkali such as LiH or sodium hydride, or alkali metal alkoxide, such as sodium or potassium methoxide or ethoxide, potassium tert-butoxide, in a reaction inert solvent such as a dipolar aprotic solvent, for example DMA, DMF and the like. The resulting alcoholate is reacted with an arylating agent (4b), where Y is a suitable leaving group, mentioned above. The conversion of (4a) to (I) using this type of O-arylation reaction does not change the stereochemical configuration on the carbon that has the hydroxy group.

Alternatively, the reaction of (4a) with (4b) can also be carried out by a Mitsunobu reaction (Mitsunobu, 1981, Synthesis, January, 1-28; Rano et al., Tetrahedron Lett., 1995, 36, 22, 3779-3792; Krchnak et al., Tetrahedron Lett., 1995, 36, 5, 6193-6196; Richter et al., Tetrahedron Lett., 1994, 35, 27, 4705-4706). This reaction comprises treatment of intermediate (4a) with (4b) where Y is hydroxyl, in the presence of triphenylphosphine and an activating agent such as a dialkyl azocarboxylate, for example, diethyl azodicarboxylate (DEAD), diethyl azodicarboxylate. diisopropyl (DIAD) or the like. The Mitsunobu reaction changes the stereochemical configuration at the carbon that holds the hydroxy group.

Alternatively, to prepare compounds of formula (I), an amide bond is first formed between building blocks P2 and P1, followed by coupling of building block P3 to the P1 moiety in P1-P2 and subsequent formation from the carbamate or ester bond between P3 and the P2 moiety in P2-P1-P3 with simultaneous ring closure.

Yet another alternative synthetic methodology is the formation of an amide bond between the P2 and P3 building blocks, followed by the coupling of the P1 building block to the P3 moiety at P3-P2 and a final amide bond formation between P1 and P2 at P1-P3-P2 with simultaneous ring closure.

Building blocks P1 and P3 can be linked to a P1-P3 sequence. If desired, the binding of the P1 and P3 double bond can be reduced. The P1-P3 sequence so formed, whether reduced or not, can be coupled to the P2 building block, thereby forming the P1-P3-P2 sequence, subsequently cyclized, by formation of an amide bond.

Building blocks P1 and P3 in any of the above approaches can be joined by double bond formation, for example, by the olefin metathesis reaction described hereinafter or a Wittig-type reaction. If desired, the double bond so formed can be reduced, in a manner similar to that described above for the conversion from (l-¡) to (lj). The double bond can also be reduced at a later stage, ie after the addition of a third building block or after macrocycle formation. Building blocks P2 and P1 are linked by amide bond formation and P3 and P2 are linked by carbamate or ester formation.

The P1' tail can be found linked to the P1 building block at any stage in the synthesis of the compounds of formula (I), for example before or after the coupling of the P2 and P1 building blocks; before or after the coupling of the building block P3 to P1; or before or after the closure of the ring.

Individual building blocks can be prepared first and then coupled together or alternatively, building block precursors can be coupled together and modified at a later stage to the desired molecular composition.

The functional groups on each of the building blocks can be protected to prevent side reactions.

Amide bond formation can be carried out using standard procedures, such as those used for bond coupling in peptide synthesis. The latter involves the dehydrative coupling of a carboxyl group of one reactant with an amino group of the other reactant to form a linking amide bond. Amide bond formation can be accomplished by reacting the starting materials in the presence of a coupling agent by converting the carboxyl functional group to an active form, such as an active ester, combined anhydride, or a carboxyl acid chloride or bromide. . General descriptions of such coupling reactions and the reagents used therein can be found in general textbooks on peptide chemistry, for example, M. Bodanszky, "Peptide Chemistry", 2nd rev. ed., Springer-Verlag, Berlin, Germany, (1993).

Examples of coupling reactions with amide bond formation include the azide method, the mixed carbonic-carboxylic acid anhydride method (isobutyl chloroformate), the carbodiimide method (dicyclohexylcarbodiimide, diisopropylcarbodiimide, or a soluble carbodiimide). water such as /V-ethyl-A/'-[(3-dimethylamino)propyl]carbodiimide), the active ester method (for example p-nitrophenyl, p-chlorophenyl, trichlorophenyl, pentachlorophenyl, pentafluorophenyl, /V-hydroxysuccinic imido and the like), Woodward's reagent K method, 1,1-carbonyldiimidazole (CDI or Ν,Ν'-carbonyldiimidazole) method, phosphorous or oxidation-reduction reagent methods. Some of these methods can be refined by adding suitable catalysts, for example in the carbodiimide method by adding 1-hydroxybenzotriazole, DBU (1,8-diazabicyclo[5,4,0]undec-7-ene), or 4- DMAP. Other coupling agents are (benzotriazol-1-yloxy)tris-(dimethylamino)phosphonium hexafluorophosphate, either by itself or in the presence of 1-hydroxybenzotriazole or 4-DMAP; or 2-(1/-/benzotriazol-1-yl)-A/,/V,A/',N-tetra-methyluron¡o tetrafluoroborate, or 0-(7-azabenzotriazole-1-hexafluorophosphate -il)/V,/V,/V',/\/'-tetramet¡luron¡o. These coupling reactions can be carried out in either solution (liquid phase) or solid phase.

Preferred amide bond formation is carried out using N-ethyloxycarbonyl-2-ethyloxy¡-1,2-dihydroquinoline (EEDQ) or N-isobutyloxy-carbonyl-2-isobutyloxy¡-1,2-dihydroquinoline (IIDQ). Unlike the classical anhydride procedure, EEDQ and IIDQ do not require base or low reaction temperatures.

Typically, the process comprises reacting equimolar amounts of the carboxyl and amine components in an organic solvent (a wide variety of solvents can be used). Then, excess EEDQ or IIDQ is added and the mixture is allowed to stir at room temperature.

Coupling reactions are preferably carried out in an inert solvent, such as halogenated hydrocarbons, eg dichloromethane, chloroform, dipolar aprotic solvents such as acetonitrile, dimethylformamide, dimethylacetamide, DMSO, HMPT, ethers such as tetrahydrofuran (THF).

In many instances, the coupling reactions are carried out in the presence of a suitable base such as a tertiary amine, for example triethylamine, diisopropylethylamine (DIPEA), /V-methylmorpholino, /V-methylpyrrolidine, 4-DMAP or 1,8 -diazabicyclo[5,4,0]undec-7-ene (DBU). The reaction temperature can range from 0°C to 50°C and the reaction time can range from 15 min to 24 h.

Functional groups on the building blocks that are attached can be deprotected to prevent undesired linkage formation. Appropriate protecting groups that can be used are listed for example in Greene, "Protective Groups in Organic Chemistry", John Wiley &amp; Sons, New York (1999) and "The Peptides: Analysis, Synthesis, Biology", Vol. 3, Academic Press, New York (1987).

Carboxyl groups can be protected as an ester which can be cleaved to carboxylic acid. Protecting groups that can be used include 1) alkyl esters such as methyl, trimethylsilyl, and tert-butyl; 2) arylalkyl esters such as benzyl and substituted benzyl; or 3) esters that can be cleaved by moderate base or mild reducing means, such as trichloroethyl phenacyl esters.

Amino groups can be protected by a variety of protecting groups, such as:

1) acyl groups, such as formyl, trifluoroacetyl, phthalyl, and p-toluenesulfonyl;

2) aromatic carbamate groups, such as benzyloxycarbonyl (Cbz or Z) and substituted benzyloxycarbonyls and 9-fluorenylmethyloxycarbonyl (Fmoc);

3) aliphatic carbamate groups such as tert-butyloxycarbonyl (Boc), ethoxycarbonyl, diisopropylmethoxycarbonyl and allyloxycarbonyl;

4) cyclic alkyl carbamate groups, such as cyclopentyloxycarbonyl and adamantyloxycarbonyl;

5) alkyl groups, such as triphenylmethyl, benzyl, or substituted benzyl such as 4-methoxybenzyl;

6) trialkylsilyl, such as trimethylsilyl or t.Bu dimethylsilyl; and

7) thiol-containing groups, such as phenylthiocarbonyl and dithiasuccinoyl. The amino protecting groups of interest are Boc and Fmoc.

Preferably, the amino protecting group is cleaved before the next coupling step. Removal of the N-protecting groups can be carried out following procedures known in the art. When a Boc group is used, the methods of choice are trifluoroacetic acid, neat or in dichloromethane, or HCI in dioxane or in ethyl acetate. The resulting ammonium salt is then neutralized, either prior to coupling or in situ with basic solutions such as aqueous buffers, or tertiary amines in dichloromethane or acetonitrile or dimethylformamide. When the Fmoc group is used, the reagents of choice are piperidine or substituted piperidine in dimethylformamide, but any secondary amine can be used. The deprotection is carried out at a temperature between 0°C and room temperature, commonly around 15-25Ό or 20-22°C.

Other functional groups that can interfere with the coupling reactions of the building blocks can also be protected. For example, hydroxyl groups can be protected as benzyl or substituted benzyl ethers, eg 4-methoxybenzyl ether, benzoyl or substituted benzoyl esters, eg 4-nitrobenzoyl ester, or with trialkylsilyl groups (eg trimethylsilyl or tert-butyldimethylsilyl). .

Other amino groups can be protected by selectively cleavable protecting groups. For example, when Boc is used as the α-amino protecting group, the following side chain protecting groups are suitable: p-toluenesulfonyl (tosyl) moieties can be used to protect other amino groups; benzyl (Bn) ethers can be used to protect hydroxy groups; and benzyl esters can be used to protect other carboxyl groups. Or when Fmoc is chosen for α-amino protection, tert-butyl based protecting groups are commonly acceptable. For example, Boc can be used for other amino groups; tert-butyl ethers for hydroxyl groups; and tert-butyl esters for other carboxyl groups.

Any of the protecting groups can be removed at any stage of the synthesis procedure, but preferably, the protecting groups of any of the functional groups involved in the reaction steps are removed after completion of the macrocycle preparation. Removal of the protecting groups can be carried out in any manner determined by the choice of protecting groups, which manners are known to those skilled in the art.

Intermediates of formula (1a) where X is N, said Intermediates being represented by formula (1a-1), can be prepared from intermediates (5a) which are reacted with an alkenamine (5b) in the presence of a carbonyl introducing agent as indicated in the following reaction scheme.

<img file="ECSP088150A_D0030.tif" />

<img file="ECSP088150A_D0031.tif" />

Carbonyl (CO) introducing agents include phosgene or phosgene derivatives, such as carbonyl diimidazole (CDI) and the like. In one embodiment (5a) it is reacted with the CO introducing agent in the presence of a suitable base and a solvent, which may be the bases and solvents used in the amide formation reactions, as described above. In a particular embodiment, the base is a hydrogen carbonate, for example NaHCO₃, or a tertiary amine, such as triethylamine and the like, and the solvent is an ether or halogenated hydrocarbon, for example THF, CH₂IC₂, CHCI₃ and the like. Then, the amine (5b) is added, thus obtaining intermediates (1a1) as in the previous scheme. An alternative route using similar reaction conditions comprises first reacting the CO introducing agent with the alkenamine (5b) and then reacting the Intermediate so formed with (5a).

Intermediaries (1-to-1), alternatively, can be prepared as follows:

<img file="ECSP088150A_D0032.tif" />

PG¹ is an O-protecting group, which can be any of the groups mentioned herein, and is especially a benzoyl or substituted benzoyl group, such as 4-nitrobenzoyl. In the latter instance, this group can be removed by reaction with an alkali metal hydroxide (LiOH, NaOH, KOH), especially where PG¹ is 4-nitrobenzoyl, with LiOH, in an aqueous medium comprising water and a water-soluble organic solvent, such as alkanol (methanol, ethanol) and THF.

Intermediates (6a) are reacted with (5b) in the presence of a carbonite introducing agent, similar as described above and this reaction produces intermediates (6c). These are especially deprotected using the reaction conditions mentioned above. The resulting alcohol (6d) is reacted with intermediates (4b), as described above for the reaction of (4a) with (4b), and this reaction produces intermediates (1a-1).

Intermediates of formula (1a) where X is C, said intermediates being represented by formula (1a-2), can be prepared by an amine formation reaction starting from intermediates (7a) which are reacted with an amine (5b), as shown in the following reaction scheme, using the reaction conditions for preparing amides, such as those described above.

<img file="ECSP088150A_D0033.tif" />

Intermediates (1-to-1) can be prepared alternatively, as follows:

<img file="ECSP088150A_D0034.tif" />

the PG¹ is an O-protecting group, as described above. The same reaction conditions can be used, as described above; amide formation, as described above, PG removal¹ as in the description of the protecting groups and the introduction of R⁹, as in the reactions of (4a) with the reactants (4b).

Intermediates of formula (2a) can be prepared by first cyclizing the open amide (9a) to a macrocyclic ester (9b), which in turn is converted to (2a), as follows:

<img file="ECSP088150A_D0035.tif" />

PG² is a carboxyl protecting group, eg one of the above mentioned carboxyl protecting groups, especially a Cm alkyl or benzyl ester, eg a methyl, ethyl or t-butyl ester. The reaction from (9a) to (9b) is a metathesis reaction and is carried out as described above. the PG group² it is removed following the procedures also described above. when PG¹ is a C-|alkyl ester.₄, is removed by alkaline hydrolysis, for example with NaOH or preferably LiOH, in an aqueous solvent, for example an alkanol mixture Ci.₄/water. A benzyl group can be removed by catalytic hydrogenation.

In an alternative synthesis, intermediates (2a) can be prepared as follows:

<img file="ECSP088150A_D0036.tif" />

the PG group¹ is selected such that it can be selectively cleaved with respect to

PG². PG² it can be, for example, methyl or ethyl esters, which can be removed by treatment with an alkali metal hydroxide in an aqueous medium, in which case PG¹, for example, is t.butyl or benzyl. PG² can be t-butyl esters which can be removed under weakly acidic conditions or PG¹ can be benzyl esters that can be removed with strong acid or by catalytic hydrogenation, in the latter two cases, PG¹ for example it is a benzoic ester such as a 4-nitrobenzoic ester.

First, the intermediates (10a) are cyclized to the macrocyclic esters (10b), the latter are deprotected by removal of the PG group.¹ a (10c), which are reacted with intermediates (4b), followed by removal of the carboxyl protecting group PG². The cyclization, deprotection of PG¹ and PG² and coupling with (4b) are as described above.

The R groups¹ groups can be introduced at any stage of the synthesis, either as the last step, as described above, or earlier, before macrocycle formation. In the following scheme, the groups R are introduced¹ where -NH-SO₂R.⁸ or -OR⁷ (which are as specified above):

<img file="ECSP088150A_D0037.tif" />

<img file="ECSP088150A_D0038.tif" />

In the above scheme, PG² is as defined above and L¹ is a P3 group or

R.³ (b), where n and R³ are as defined above and where X is N, L¹ may also be a nitrogen protecting group (PG, as defined above) and where X is C, L¹ can also be a COOPG group^2a, where the group PG^2a is a carboxyl protecting group similar to PG², but where PG^2a can be cleaved selectively with respect to PG². In one embodiment, PG^2a is t-butyl and PG² is it methyl or ethyl.

Intermediates (11c) and (11d) where L¹ represents a group (b) correspond to intermediates (1a) and can be further processed as specified above.

Coupling of building blocks P1 and P2

Building blocks P1 and P2 are joined using an amide formation reaction following the procedures described above. The building block P1 may possess a carboxyl protecting group PG² (as in (12b)) or can already be found attached to the P1' group (as in (12c)). L² is a protecting group N (PG), or a group (b), as specified above. L³ is hydroxy, -OPG¹ or an -OR group⁹ as specified above. When in any of the following reaction schemes, L³ is hydroxy, before each reaction step, it can be protected as a -OPG group¹ and, if desired, can then be deprotected again for a free hydroxy function. Similarly, as described above, the hydroxy function can be converted to a -0- group

<img file="ECSP088150A_D0039.tif" />

In the procedure of the above scheme, a cyclopropyl amino acid (12b) or (12c) is coupled to the acid function of the P2 building block (12a) with the formation of an amide linkage, following the procedures described above. Intermediates (12d) or (12e) were obtained. where in the latter, L² is a group (b), the resulting products are P3-P2-P1 sequences comprising some of the intermediates (11c) or (11d) in the above reaction scheme. Removal of the acidic protecting group in (12d), using the appropriate conditions for the protecting group used, followed by coupling with an amine H₂N-SW₂R.⁸ (2b) or with HOR⁷ (2c), as described above, again gives the intermediaries (12e), where -COR¹ They are amide or ester groups. When I² is an N-protecting group, it can be removed giving intermediates (5a) or (6a). In one embodiment, PG in this reaction is a BOC group and PG² is it methyl or ethyl. When, additionally L³ is hydroxy, the starting material (12a) is Boc-L-hydroxyproline. In a particular embodiment, PG is BOC, PG² is methyl or ethyl and L³ is -OR⁹.

In one embodiment, L.² is a group (b) and these reactions comprise the coupling of P1 to P2P3, which produces the intermediates (1a-1) or (1a) mentioned above. In another embodiment, L² is a protecting group N PG, which is as specified above and the coupling reaction produces intermediates (12d-1) or (12e-1), from which the PG group can be removed, using the reaction conditions mentioned above, obtaining the intermediates (12-f) or respectively (12g), comprising the intermediates (5a) and (6a), as specified above:

<img file="ECSP088150A_D0040.tif" />

<img file="ECSP088150A_D0041.tif" />

<img file="ECSP088150A_D0042.tif" />

<img file="ECSP088150A_D0043.tif" />

<img file="ECSP088150A_D0044.tif" />

In one embodiment, the group L3 in the above schemes represents a group -O-PG1 that can be introduced into a starting material (12a) where L3 is hydroxy. In this instance, PG1 is selected such that it can be selectively cleaved with respect to the L2 group which is PG.

Similarly, building blocks P2 where X is C, which are derivatives of cyclopentane or cyclopentene, can be linked to building blocks P1, as indicated in the following scheme where R¹,R², L³ are as specified above and PG² and PG^2a are carboxyl protecting groups. PG^2a typically selected so that it can be selectively cleaved with respect to the PG group². the removal of the PG group^2a in (13c) gives intermediates (7a) or (8a), which can be reacted with (5b), as described above.

<img file="ECSP088150A_D0045.tif" />

In a particular embodiment, where X is C,

R.² is H and where X and the R² that has carbon are linked by a single bond (P2 being a cyclopentane residue), PG^2a and L³ taken together they form a bond and the building block P2 is represented by the formula:

<img file="ECSP088150A_D0046.tif" />

0'

Bicyclic acid (14a) is reacted with the like (12b) or (12c), as described above for (14b) and (14c) respectively, where the lactone is opened to give intermediates (14c) and (14e). Lactones can be opened using ester hydrolysis procedures, for example using the reaction conditions described above for the alkaline removal of a PG group.¹ in (9b), especially using basic conditions, such as an alkali metal hydroxide, eg NaOH, KOH, especially LiOH.

ooh

<img file="ECSP088150A_D0047.tif" />

(14 to)

<img file="ECSP088150A_D0048.tif" />

Intermediates (14c) and (14e) can be further processed, as described herein below.

Mating of building blocks P3 and P2

For the P2 building blocks that possess a pyrrolidine moiety, the P3 and P2 or P3 and P2-P1 building blocks are linked using a carbamate formation reaction following the procedures previously described for the coupling of (5a) with (5b). A general procedure for the coupling of P2 blocks possessing a pyrrolidine moiety is depicted in the following reaction scheme where L³ is as specified above and L⁴ is a group -O-PG², a group or

---NH

OPG²

<img file="ECSP088150A_D0049.tif" />

or group

<img file="ECSP088150A_D0050.tif" />

<img file="ECSP088150A_D0051.tif" />

In one embodiment, L.⁴ in (15a) is a group -OPG², the PG group² can be removed and the resulting acid coupled with cyclopropyl amino acids (12a) or (12b), giving intermediates (12d) or (12e) where L² is a radical (d) or (e).

A general procedure for the coupling of P3 blocks with a P2 block or with a P2-P1 block where the P2 is a cyclopentane or cyclopentene is shown in the following scheme. L³ and L⁴ are as specified above.

<img file="ECSP088150A_D0052.tif" />

(16a)

<img file="ECSP088150A_D0053.tif" />

In a particular embodiment, L³ and L⁴ taken together they can form a lactone bridge as in (14a) and the coupling of a P3 block with a P2 block is as follows:

ooh

<img file="ECSP088150A_D0054.tif" />

The bicyclic lactone (14a) is reacted with (5b) in an amide to amide formation reaction (16c) in which the lactone bridge is opened to (16d). The reaction conditions for the amide formation and lactone opening reactions are as described above or hereinafter. Intermediate (16d) in turn can be coupled to a P1 group, as described above.

The reactions in the above schemes are carried out using the same procedures as described above for the reactions of (5a), (7a) or (8a) with (5b) and, especially, the above reactions where L⁴ is a group (d) or (e) correspond to the reactions of (5a), (7a) or (8a) with (5b), as described above.

The building blocks P1, P1', P2 and P3 used in the preparation of the compounds of formula (I) can be prepared from intermediates known in the art. A number of such syntheses are described in further detail below.

Individual building blocks can be first prepared and then coupled together or alternatively, building block precursors can be coupled together and modified at a later stage to the desired molecular composition.

The functional groups on each of the building blocks can be protected to prevent side reactions.

Synthesis of the P2 building blocks

The P2 building blocks contain any of a pyrrolidine moiety, a cyclopentane, or cyclopentene substituted by an -OR group.⁴.

The P2 building blocks containing a pyrrolidine moiety can be derived from commercially available hydroxy proline.

Preparation of the P2 building blocks containing a cyclopentane ring can be carried out as shown in the following scheme.

either

<img file="ECSP088150A_D0055.tif" />

ooh

<img file="ECSP088150A_D0056.tif" />

Bicyclic acid (17b) can be prepared, for example, from 3,4-bis(methoxycarbonyl)cyclopentanone (17a), as described by Rosenquist et al. in Acta Chem. Scand. 46 (1992) 11271129. A first step in this procedure involves reduction of a keto group with a reducing agent such as sodium borohydride in a solvent such as methanol, followed by hydrolysis of the esters, and finally ring closure to the bicyclic lactone (17b) using procedures of lactone formation, especially through the use of acetic anhydride in the presence of a weak base, such as pyridine. The carboxylic acid functional group in (17b) can then be protected by introducing a suitable carboxyl protecting group, such as a PG group.², which is as specified above, thereby providing bicyclic ester (17c). the PG group² in particular it is acid unstable, such as a t.butyl group and is introduced, for example, by treatment with isobutene in the presence of a Lewis acid or with di-tert-butyl dicarbonate in the presence of a base such as a tertiary amine, such as dimethylaminopyridine or triethylamine in a solvent such as dichloromethane. Opening of lactone (17c) using the reaction conditions as described above, especially with lithium hydroxide, gives the acid (17d), which can be further used in coupling reactions with the P1 building blocks. The free acid in (17d) can also be protected, preferably with an acid protecting group PG^2a that can be selectively cleaved with respect to PG² and the hydroxy function can be converted to an -OPG group¹ or in an -OR group⁹. The products obtained by removing the PG group² are the intermediaries (17g) and (17¡) that correspond to the intermediaries (13a) or (16a) specified above.

Intermediates with specific stereochemistry can be prepared by resolving the intermediates in the above reaction sequence. For example, (17b) can be resolved following procedures known in the art, for example by action of the salt form with an optically active base or by chiral chromatography and the resulting stereoisomers can also be processed as described. previously. The OH and COOH groups in (17d) are found in this cis position. Trans analogs can be prepared by reversing the stereochemistry at the carbon holding the OH function by using specific reagents in reactions introducing OPG¹ or OR⁹ that reverse the stereochemistry, such as, for example, by applying a Mitsunobu reaction.

In one embodiment, intermediates (17d) are coupled to P1 blocks (12b) or (12c), which coupling reactions correspond to the coupling of (13a) or (16a) with the same P1 blocks, using the same conditions. The subsequent introduction of a replaceme -OR⁹, as described above, followed by removal of the acidic protecting group PG² gives the intermediaries (8a-1), which are a subclass of the intermediaries (7a), or a part of the intermediaries (16a). The reaction products of PG removal² they can also be attached to the building block P3. In one embodiment PG² in (17d) is t-butyl which can be removed under acidic conditions, for example with trifluoroacetic acid.

<img file="ECSP088150A_D0057.tif" />

<img file="ECSP088150A_D0058.tif" />

An unsaturated P2 building block, ie a cyclopentene ring, can be prepared as illustrated in the scheme below.

<img file="ECSP088150A_D0059.tif" />

(17a)

<img file="ECSP088150A_D0060.tif" />

A bromination elimination reaction of 3,4-bis(methoxycarbonyl)cyclopentanone (17a) as described by Dolby et al. in J.Org. Chem. 36 (1971) 1277-1285 followed by reduction of the keto functional group with a reducing agent such as sodium borohydride provides the cyclopentenol (19a).

Selective ester hydrolysis using, for example, lithium hydroxide in a solvent such as a mixture of dioxane and water, provides the hydroxy-substituted cyclopentenol monoester (19b).

An unsaturated building block P2 where R² it can also be other than hydrogen, it can be prepared as illustrated in the scheme below.

<img file="ECSP088150A_D0061.tif" />

Oxidation of commercially available 3-methyl-3-buten-1-ol (20a), especially by an oxidizing agent such as pyridinium chlorochromate, gives (20b), which is converted to the corresponding methyl ester, e.g., by treatment with acetyl chloride in methanol, followed by bromination reaction with bromine to give the bromo α-bromo ester (20c). The latter can be condensed with the alkenyl ester (20e), obtained from (20d) by an ester formation reaction. The ester in (20e) is preferably a t-butyl ester which can be prepared from the corresponding commercially available acid (20d), for example, by treatment with di-fer-butyl dicarbonate in the presence of a base such as dimethylaminopyridine. Intermediate (20e) is treated with a base, such as lithium diisopropyl amide in a solvent such as tetrahydrofuran, and reacted with (20c) to give the alkenyl ester (20f).

Deletion of (20f) by an olefin metathesis reaction, which is carried out as described above, provides cyclopentene derivative (20g). Stereoselective epoxidation of (20g) can be carried out using Jacobsen's asymmetric epoxidation method to obtain epoxide (20h). Finally, an opening reaction of the epoxide under basic conditions, for example by adding a base, especially DBN (1,5-diazabicyclo-[4,3,0]non-5-ene), gives the alcohol (twenty). Optionally, the double bond in intermediate (20¡) can be reduced, for example by catalytic hydrogenation using a catalyst such as palladium on carbon, to give the corresponding cyclopentane compound. The t-butyl ester can be removed to the corresponding acid, which is subsequently coupled to a P1 building block.

The -R group⁹ it can be introduced into the pyrrolidine, cyclopentane or cyclopentene rings at any convenient stage in the synthesis of the compounds according to the present invention. One approach is to first introduce the -R group⁹ to the mentioned rings and subsequently adding the other desired building blocks, ie P1 (optionally with PT tail) and P3, followed by formation of the macrocycle. Another approach is to couple the building blocks P2, which has no -OR substituent⁹, with each P1 and P3 and add the -R group⁹ either before or after macrocycle formation. In this latter procedure, the P2 residues possess a hydroxy group, which can be protected by a PG protecting group.¹.

The R groups⁹ can be introduced into the P2 building blocks by reacting the hydroxy-substituted intermediates (21a) or (21b) with similar intermediates (4b), as described above for the synthesis of (l) from (4a). ). These reactions are represented in the following schemes, where L² is as specified above and L⁵ and L^5a independently of each other, they represent hydroxy, a carboxyl protecting group -OPG² or -OPG^2a, or L⁵ may also represent a group P1 such as a group (d) or (e), as specified above, or L^5a it can also represent a group P3 such as a group (b) as specified above. PG groups²and PG^2a are as specified above. When the groups L⁵ and L^5a they are PG² or PG^2a, are selected such that each group can be selectively cleaved relative to the other. For example, one of L.⁵ and L^5a it may be one methyl or ethyl group and the other a benzyl or t-butyl group.

In one embodiment in (21a), L² is PG and L⁵ is -OPG² or in (21 d), L^5a is -OPG² and L⁵ is -OPG² and the PG groups² are removed as described above.

<img file="ECSP088150A_D0062.tif" />

(21a) (21b)

<img file="ECSP088150A_D0063.tif" />

<img file="ECSP088150A_D0064.tif" />

<img file="ECSP088150A_D0065.tif" />

<img file="ECSP088150A_D0066.tif" />

<img file="ECSP088150A_D0067.tif" />

Alternatively, when handling hydroxy-substituted cyclopentane analogues, the quinoline substituent can be introduced via a similar Mitsunobu reaction by reacting the hydroxy group of a compound (2a') with the desired alcohol (3b) in the presence of triphenylphosphine. and an activating agent such as diethyl azodicarboxylate (DEAD), diisopropyl azodicarboxylate (DIAD) or the like.

In another embodiment, group L² it's BOC, L⁵ is hydroxy and the starting material (21a) is commercially available BOC-hydroxyproline, or any other stereoisomeric form thereof, for example BOC-L-hydroxyproline, especially the trans isomer of the latter. When I⁵ in (21b) is a carboxyl protecting group, it can be removed by following the procedures as described above for (21c). In yet another embodiment, PG in (21 b-1) is Boc and PG² it is a lower alkyl ester, especially a methyl or ethyl ester. Hydrolysis of the latter ester to acid can be carried out by standard procedures, for example, acid hydrolysis with hydrochloric acid in methanol or with an alkali metal hydroxide such as NaOH, especially LiOH. In another embodiment, the hydroxy-substituted cyclopentane or cyclopentene analogues (21 d) are converted to (21 e), which, when L⁵ and L^5a they are OPG² or -OPG^2a, can be converted to the corresponding acids (21 f) by removal of the group

PG². PG removal^2a in (21 e-1) leads to similar intermediaries.

Intermediaries Y-R⁹ (4b) can be prepared following methods known in the art using known starting materials. A number of synthesis routes for such intermediates will be described in further detail hereinafter. For example, the preparation of the aforementioned intermediate quinolines is shown below in the following scheme.

<img file="ECSP088150A_D0068.tif" />

Friedel-Craft acylation of a suitable substituted aniline (22a), either commercially available or by procedures known in the art, using an acylating agent such as acetyl chloride or the like in the presence of one or more Lewis acids, such as boron trichloride and aluminum trichloride in a solvent such as dichloromethane provides (22b). The coupling of (22b) with a carboxylic acid (22c), preferably under basic conditions, such as in pyridine, in the presence of an activating agent for the carboxylate group, for example POCI₃, followed by ring closure and dehydration under basic conditions as potassium ferr-butoxide in ferr-butanol gives the quinoline derivative (22e). The latter can be converted to (22f) where LG is a leaving group, for example by reacting (22e) with a halogenating agent, for example phosphoryl chloride or the like, or with an arylsulfonyl chloride, for example with chloride of tosyl. The quinoline derivative (22e) can be coupled in a Mitsunobu reaction to an alcohol, as described above, or the quinoline (22f) can be reacted with (1a) in an O-arylation reaction, as described above. .

A variety of carboxylic acids with the general structure (22c) can be used in the above synthesis. These acids are either commercially available or can be prepared by procedures known in the art. An example of the preparation of 2(substituted)aminocarboxy-aminothiazole derivatives (23a-1) is shown, following the procedure described by Berdikhina et al. in

Chem. Heterocycl. Compd. (Engl. Transí.) (1991), 427-433, in the following reaction scheme that illustrates the preparation of 2-carboxy-4-isopropílt¡azole (22c-1):

<img file="ECSP088150A_D0069.tif" />

Ethyl thiooxamate (23a) is reacted with the β-bromoketone (23b) to form the carboxylic acid thiazolyl ester (23c), which is hydrolyzed to the corresponding acid (25c-1). The ethyl ester in these intermediates can be replaced by the PG carboxyl protecting groups.², as defined above. In the above scheme, R^4a is as defined above and is especially C-alkyl. 4, more especially i-propyl.

Bromoketone (23b) can be prepared from 3-methyl-butan-2-one (MIK) with a silylating agent (such as TMSCI) in the presence of a suitable base (especially LiHMDS) and bromine.

The synthesis of other carboxylic acids (22c), especially amino substituted thiazole carboxylic acids (25a-2) is illustrated herein below:

<img file="ECSP088150A_D0070.tif" />

<img file="ECSP088150A_D0071.tif" />

Thiourea (24c) with several R substituents^4a, which, in particular, are alkyl Ci.₆, can be formed by reaction of the appropriate amine (24a) with tert-butylisothiocyanate in the presence of a base such as diisopropylethylamine in a solvent such as dichloromethane followed by removal of the tert-butyl group under acidic conditions. Subsequent condensation of the thiourea derivative (24c) with 3-bromopyruvic acid provides the thiazole carboxylic acid (22c-2).

Synthesis of the P1 building blocks

The cyclopropanamino acid used in the preparation of the P1 fragment is commercially available or can be prepared using procedures known in the art.

In particular, the amino-vinyl-cyclopropyl ethyl ester (12b) can be obtained according to the procedure described in WO 00/09543 or as illustrated in the following scheme, where PG² is a carboxyl protecting group as specified above:

<img file="ECSP088150A_D0072.tif" />

<img file="ECSP088150A_D0073.tif" />

h₂Nv ,,'COOPG²

Δ(12b-1)

<img file="ECSP088150A_D0074.tif" />

The imine treatment (25a) available commercially or easily obtainable with

1,4-Dihalobutene in the presence of base produces (25b), which after hydrolysis gives cyclopropyl amino acid (12b), which has the allyl syn substituent for the carboxyl group. Resolution of the enantiomeric mixture (12b) yields (12b-1). Resolution is carried out using procedures known in the art such as enzymatic separation; crystallization with a chiral acid; or chemical derivation; or by chiral column chromatography. Intermediates (12b) or (12b-1) may be coupled to the appropriate P2 derivatives as described above.

The building blocks P1 for the preparation of compounds according to the general formula (I) where R¹ is -OR⁷ or -NH-SO₂R.⁸ they can be prepared by reacting the amino acids (23a) with the appropriate alcohol or amine, respectively, under standard conditions for ester or amide formation. The cyclopropyl amino acids (23a) are prepared by introducing an N PG protecting group and removing PG² and the amino acids (a) are converted to the amides (12c-1) or esters (12c-2), which are subgroups of the intermediates (12c), as indicated in the following reaction scheme, where PG is as specified previously.

<img file="ECSP088150A_D0075.tif" />

The reaction of (26a) with amine (2b) is an amine formation process. The similar reaction with (2c) is an ester formation reaction. Both can be carried out by following the procedures described above. This reaction gives intermediates (26b) or (26c) from which the amino protecting group is removed by standard methods such as those described above. This, in turn, produces the desired intermediate (12c-1). Starting materials (26a) can be prepared from the aforementioned intermediates (12b) by first introducing an N PG protecting group and subsequently removing the PG group.^2,

In one embodiment, the reaction of (26a) with (2b) is carried out by treating the amino acid with the coupling agent, for example Ν,Ν'-carbonyl-diimidazole (CDI) or the like, in a solvent such as THF. , followed by reaction with (2b) in the presence of a base such as 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). Alternatively, the amino acid can be treated with (2b) in the presence of a base such as diisopropylethylamine, followed by treatment with a coupling agent such as benzotriazol-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate. (commercially available as PyBOP®) to effect introduction of the sulfonamide group.

Intermediates (12c-1) or (12c-2) in turn, can be coupled to the appropriate proline, cyclopentane or cyclopentene derivatives as described above.

Synthesis of the P3 building blocks

P3 building blocks are commercially available or can be prepared according to methodologies known to those of skill in the art. One of these methodologies is shown in the scheme below and uses monoacylated amines, such as trifluoroacetamide or a Boc-protected amine.

either

<img file="ECSP088150A_D0076.tif" />

(27a)

<img file="ECSP088150A_D0077.tif" />

(27b) |₃ (27c) *YYY

HN |₃ (5b)

In the above scheme, R together with the group CO forms a protecting group N, in particular R is t-butoxy, trifluoromethyl; R.³ and n are as defined above and LG is a leaving group, in particular halogen, eg. chlorine or bromine

The monoacylated amines (27a) are treated with a strong base such as sodium hydride and subsequently reacted with a reagent LG-alkenyl C₅.₈ (27b), in particular haloalkenyl C₅.

₈, to form the corresponding protected amines (27c). Deprotection of (27c) produces (5b), which are P3 building blocks. Deprotection will depend on the functional group R, so if R is i-butoxy, deprotection of the corresponding Boc-protected amine can be achieved by treatment with an acid, eg. trifluoroacetic acid. Alternatively, when R is eg trifluoromethyl, removal of the R group is accomplished with a base, eg. sodium hydroxide.

The following scheme illustrates yet another method for preparing a P3 building block, ie a Gabriel synthesis of C alkenylamines.₅.₈ primers, which can be carried out by treatment of a phthalimide (28a) with a base, such as NaOH or KOH, and with (27b), which is as specified above, followed by hydrolysis of the intermediate N-alkenylimide to generate a

<td>alkenylamine C₅. 0 //</td><td colspan="2">₈ primary (5b-1).</td>

<td></td><td>1. basis</td><td></td>

<td>í T&gt;</td><td> -------►</td><td>h₂No.</td>

<td></td><td></td><td>(5b-1)</td>

<td>(28a) 0</td><td>(27b)</td><td></td>

In the scheme above, n is as defined above.

Compounds of formula (I) can be converted into each other following art-known functional group transformation reactions. For example, amino groups can be N-alkylated, nitro groups reduced to amino groups, a halo atom can be exchanged for another halo.

Compounds of formula (I) can be converted to the corresponding N-oxide form following art-known procedures for converting a trivalent nitrogen to its /V-oxide form. Said /V-oxidation reaction can generally be carried out by reacting the starting material of formula (I) with an appropriate organic or inorganic peroxide. Suitable inorganic peroxides comprise, for example, hydrogen peroxide, alkali metal or alkaline earth metal peroxides, eg. sodium peroxide, potassium peroxide; suitable organic peroxides may comprise peroxyacids such as, for example, benzenecarboperoxoic acid or halo-substituted benzenecarboperoxoic acid, eg. 3-chlorobenzenecarboperoxoic acid, peroxoalkanoic acids, eg. peroxoacetic acid, alkylhydroperoxides, eg. tert-butyl hydroperoxide. Suitable solvents are, for example, water, lower alcohols, eg. ethanol and the like, hydrocarbons, eg. toluene, ketones, eg. 2-butanone, halogenated hydrocarbons, eg. dichloromethane, and mixtures of said solvents.

The stereochemically pure form of the compounds of formula (I) can be obtained by applying procedures known in the art. Diasteromers can be separated by physical methods such as chromatographic techniques and selective crystallization, eg, countercurrent distribution, liquid chromatography, and the like.

Compounds of formula (I) can be obtained as racemic mixtures of enantiomers which can be separated from one another following art-known resolution procedures. Racemic compounds of formula (I), which are sufficiently alkaline or acidic, can be converted into the corresponding diasteromeric salt form by reaction with an appropriate chiral acid, respectively chiral base. Said diastereomeric salt forms are subsequently separated, for example, by selective or fractional crystallization and the enantiomers are liberated from these by alkali or acid. An alternative way of separating the enantiomeric form of the compounds of formula (I) involves liquid chromatography, in particular liquid chromatography using a chiral fixed phase. Said stereochemically pure isomeric form may also be derived from the corresponding stereochemically pure form of the appropriate starting materials, provided that the reaction occurs stereospecifically. Preferably if a specific stereoisomer is desired, said compound can be synthesized by stereospecific preparation methods. These methods can advantageously employ enantiomerically pure starting materials.

In a further aspect, the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) as specified herein, or a compound from any of the sub-groups of compounds of formula (I). ) as specified herein, and a pharmaceutically acceptable vehicle. A therapeutically effective amount in this context is an amount sufficient to act prophylactically, to stabilize or reduce viral infection, and in particular HCV viral infection, in infected subjects or subjects at risk of infection. In yet a further aspect, this invention relates to a process for preparing a pharmaceutical composition as specified herein, comprising thoroughly mixing a pharmaceutically acceptable carrier with a therapeutically effective amount of a compound of formula (I), as specified herein, or a compound of any of the sub-groups of compounds of formula (I) as specified herein.

Therefore, the compounds of the present invention or any sub-group thereof can be formulated into various dosage forms for administration purposes. Mention may be made as suitable compositions of all the compositions normally used for the systemic administration of drugs. To prepare the pharmaceutical compositions of this invention, an effective amount of the particular compound, optionally in the form of an addition salt or metal complex, as an active component is combined in intimate admixture with a pharmaceutically acceptable carrier, the vehicle can take a wide variety of forms depending on the form of preparation desired for administration. These pharmaceutical compositions are intended in appropriate unit dosage form, in particular, for administration orally, rectally, percutaneously, or by parenteral injection. For example, in the preparation of the compositions in oral dosage form, any of the usual pharmaceutical media may be employed such as, for example, water, glycols, oils, alcohols and the like in the case of oral liquid preparations such as suspensions, syrups , elixirs, emulsions and solutions; or solid carriers such as starches, sugars, kaolin, lubricants, binders, disintegrating agents, and the like in the case of powders, pills, capsules, and tablets. Due to their ease of administration, tablets and capsules represent the most advantageous oral dosage unit form, in which case solid pharmaceutical carriers are obviously employed. For parenteral compositions, the vehicle will usually comprise sterile water, at least in large part, although other components may be included, for example, to aid solubility. Injectable solutions can be prepared, for example, in which the vehicle comprises saline, glucose solution, or a mixture of saline and glucose solution.

Injectable suspensions may also be prepared in which case appropriate liquid carriers, suspending agents and the like may be employed. Also included are solid form preparations that are intended to be converted immediately prior to use into liquid form preparations. In compositions suitable for percutaneous administration, the vehicle optionally comprises a penetration-enhancing agent and/or a suitable wetting agent, optionally combined with suitable additives of any nature in minor proportions, which additives do not introduce an effect significant detrimental to the skin.

The compounds of the present invention may also be administered by oral inhalation or insufflation by means of methods and formulations employed in the art for administration by this route. Thus, in general the compounds of the present invention can be administered to the lungs in the form of a solution, a suspension or a dry powder, with a solution being preferred. Any systems developed for the administration of solutions, suspensions or dry powders by inhalation or oral insufflation are suitable for the administration of the present compounds.

Thus, the present invention further provides a pharmaceutical composition adapted for administration by inhalation or insufflation through the mouth comprising a compound of formula (I) and a pharmaceutically acceptable carrier. Preferably, the compounds of the present invention are administered via inhalation of a solution in nebulized or aerosol doses.

It is especially advantageous to formulate the aforementioned pharmaceutical compositions in individual dosage form for ease of administration and uniformity of dosage. Single dosage form as used herein refers to physically individual units suitable as unit dosages, each unit containing a predetermined amount of active component calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Some examples of such unit dosage forms are tablets (including scored or film-coated tablets), capsules, pills, suppositories, powder packets, wafers, injectable solutions or suspensions and the like, and the like.

Compounds of formula (I) show antiviral properties. Viral infections and their associated diseases that can be treated using the compounds and methods of the present invention include those infections generated by HCV and other pathogenic flaviviruses such as Yellow Fever, Dengue Fever (types 1-4), St. Louis Encephalitis , Japanese Encephalitis, Murray Valley Encephalitis, West Nile Virus, and Kunjin Virus. Diseases associated with HCV include progressive liver fibrosis, inflammation, and necrosis leading to cirrhosis, end-stage liver disease, and HCC; and for the other pathogenic flaviviruses the diseases include yellow fever, dengue fever, hemorrhagic fever and encephalitis. A number of the compounds of this invention are even active against HCV mutated strains. Additionally, many of the compounds of this invention show a favorable pharmacokinetic profile and have attractive bioavailability properties, including acceptable half-life, AUC, and peak values, and are devoid of unfavorable phenomena. such as insufficient rapid onset and tissue retention.

The in vitro antiviral activity against HCV of the compounds of formula (I) was evaluated in a cellular HCV replicon system based on Lohmann et al. (1999) Science 285:110-113, with the additional modifications described by Krieger et al. (2001) Journal of Virology 75: 4614-4624, which is further exemplified in the examples section. This model, while not a complete infection model for HCV, is widely accepted as the most robust and efficient model of autonomous HCV RNA replication currently available. Compounds that exhibit anti-HCV activity in this cell model are considered candidates for further development in the treatment of HCV infections in mammals. It will be appreciated that it is important to distinguish between compounds that specifically interfere with HCV functions from those that exert cytotoxic or cytostatic effects on the HCV replicon model, and as a consequence cause a reduction in HCV RNA or related reporter enzyme concentration. Assays for the evaluation of cellular cytotoxicity based, for example, on the activity of mitochondrial enzymes using fluorogenic redox dyes such as resazurin are known in the art. Additionally, there are counter-cellular screens for the evaluation of non-selective inhibition of the activity of related reporter genes, such as firefly luciferase. Appropriate cell types can be equipped by stable transfection with a luciferase reporter gene whose expression is dependent on a constitutively active promoter, and such cells can be used as a counterscreen to remove non-selective inhibitors.

Due to their antiviral properties, in particular their anti-HCV properties, the compounds of formula (I) or any sub-group thereof, their prodrugs, /V-oxides, addition salts, quaternary amines, metal complexes and stereochemically isomeric forms , are useful in the treatment of individuals experiencing a viral infection, in particular an HCV infection, and for the prophylaxis of these infections. In general, the compounds of the present invention may be useful in the treatment of warm-blooded animals infected with viruses, in particular flaviviruses such as HCV.

The compounds of the present invention or any sub-group thereof can therefore be used as medicaments. Said use as a medicament or method of treatment comprises the systemic administration to subjects infected with the virus or to subjects susceptible to contracting viral infections of an amount effective to combat the conditions associated with the viral infection, in particular HCV infection.

The present invention further relates to the use of the present compounds or any sub-group thereof in the manufacture of a medicament for the treatment or prevention of viral infections, in particular HCV infection.

The present invention additionally relates to a method of treating a warm-blooded animal infected by a virus, or that presents a risk of infection by a virus, in particular by HCV, said method comprises the administration of an effective amount from the antiviral point of view of a compound of formula (I), as specified herein, or of a compound of any of the subgroups of compounds of formula (I), as specified herein.

Additionally, the combination of the previously known anti-HCV compound, such as, for example, interferon-α (IFN-α), pegylated interferon-α and/or ribavirin, and a compound of formula (I) can be used as drug in a combination treatment. The term "combination therapy" refers to a product containing compulsorily (a) a compound of formula (I), and (b) optionally another anti-HCV compound, as a combined preparation for simultaneous, separate or consecutive use in the treatment of infections caused by HCV, in particular, in the treatment of infections with HCV.

Anti-HCV compounds encompass agents selected from an HCV polymerase inhibitor, an HCV protease inhibitor, an inhibitor of another target in the HCV life cycle, and an immunomodulatory agent, an antiviral agent, and combinations thereof.

HCV polymerase inhibitors include, but are not limited to, NM283 (valopicitabine), R803, JTK-109, JTK-003, HCV-371, HCV-086, HCV-796, and R-1479.

HCV protease inhibitors (NS2-NS3 Inhibitors and NS3-NS4A Inhibitors) include, but are not limited to, the compounds of WO02/18369 (see, eg, page 273, lines 9-22 and page 274, line 4 to page 276, line 11); BILN-2061, VX-950, GS-9132 (ACH-806), SCH-503034, and SCH-6. Other additional agents that can be used are those disclosed in WO-98/17679, WO00/056331 (Vertex); WO 98/22496 (Roche); WO 99/07734, (Boehringer Ingelheim), WO 2005/073216, W02005073195 (Medivir) and agents with similar structures.

Inhibitors of other targets in the HCV life cycle, including NS3 helicase; metalloprotease inhibitors; antisense oligonucleotide inhibitors, such as ISIS-14803, AVI-4065, and the like; siRNA such as SIRPLEX-140-N and the like; vector-encoded short hair bulb RNA (shRNA); DNAzymes; HCV-specific ribozymes such as heptazime, RPI,13919 and the like; entry inhibitors such as HepeX-C, HuMax-HepC and the like; alpha glucosidase inhibitors such as celgosivir, UT-231B and the like; KPE-02003002; and BIVN 401.

Immunomodulatory agents include, without limitation; natural and recombinant interferon isoform compounds, including α-interferon, β-interferon, γ-interferon, ω-interferon and the like, such as Intron A®, Roferon-A®, Canferon-A300®, Advaferon®, Infergen ®, Humoferon ® , Sumiferon MP ® , Alfaferone ® , IFN-beta ® , Feron ® and the like; compounds with an interferon structure derived (pegylated) from polyethylene glycol, such as PEG interferon-a-2a (Pegasys®), PEG interferon-a-2b (PEG-Intron®), pegylated IFN-a-con1 and the like; long-acting formulations and derivations of compounds with an interferon structure such as albumin-fused interferon albumiferon a and the like; compounds that stimulate the synthesis of interferon in cells, such as resiquimod and the like; interleukins; compounds that enhance the development of the type 1 helper T cell response, such as SCV-07 and the like; TOLL-like receptor agonists such as CpG-10101 (actilon), isatoribine and the like; thymosin α-1; ANA-245; ANA-246; histamine dihydrochloride; propagermanium; tetrachlorodecaoxide; magnify; IMP-321; KRN-7000; antibodies, such as civacir, XTL6865 and the like; and prophylactic and therapeutic vaccines such as InnoVac C, HCV E1E2/MF59 and the like.

Other antiviral agents include, but are not limited to, ribavirin, amantadine, viramidine, nitazoxanide; telbivudine; NOV-205; taribavirin; inhibitors of internal ribosome entry; broad-spectrum viral inhibitors, such as IMPDH inhibitors (eg, compounds from US5,807,876,

US6,498,178, US6,344,465, US6,054,472, W097/40028, WO98/40381, WOOO/56331, and mycophenolic acid and its derivatives, including, but not limited to, VX-950, merimepodib (VX-497), VX- 148, and/or VX-944); or combinations of any of the above.

Thus, to combat or treat HCV infections, the compounds of formula (I) can be co-administered in combination with, for example, interferon-α (IFN-α), pegylated interferon-α and/or ribavirin, such as also therapeutics based on antibodies directed against HCV epitopes, small interfering RNA (Si RNA), ribozymes, DNAzymes, antisense RNA, small molecule antagonists of eg NS3 protease, NS3 helicase and NS5B polymerase.

Consequently, the present invention relates to the use of a compound of formula (I) or any sub-group thereof as defined above for the manufacture of a medicament useful for inhibiting HCV activity in a mammal infected with HCV virus. HCV, where said medicament is used in combination treatment, said combination treatment preferably comprises a compound of formula (I) and another HCV inhibitor compound, eg. IFN-α (pegylated) and/or ribavirin.

In yet another aspect combinations of a compound of formula (I) as specified herein and an anti-HIV compound are provided. The latter are preferably those HIV inhibitors that have a positive effect on drug metabolism and/or on their pharmacokinetics that improve bioavailability. An example of such an HIV inhibitor is ritonavir.

As such, the present invention further provides a combination comprising (a) an HCV NS3/4a protease inhibitor of formula (I) or a pharmaceutically acceptable salt thereof; and (b) ritonavir or a pharmaceutically acceptable salt thereof.

The compound ritonavir, and its pharmaceutically acceptable salts, and methods for their preparation are described in WO94/14436. For a preferred dosage form of ritonavir, see US6,037,157, and the documents cited therein: US 5,484,801, US 08/402,690, and WO95/07696 and WO95/09614. Ritonavir has the following formula:

ooh

In a further embodiment, the combination comprises (a) an HCV NS3/4a protease inhibitor of formula (I) or a pharmaceutically acceptable salt thereof; and (b) ritonavir or a pharmaceutically acceptable salt thereof; additionally it comprises a further anti-HCV compound selected from compounds as described herein.

In one embodiment of the present invention there is provided a process for preparing a combination as described herein, comprising the step of combining an HCV NS3/4a protease inhibitor of formula (I) or a point of pharmaceutically acceptable salt thereof pharmaceutically, and ritonavir or a pharmaceutically acceptable salt thereof. An alternative embodiment of this invention provides a process in which the combination comprises one or more additional agents as described herein.

The combinations of the present invention can be used as medicaments. Said use as a medicament or method of treatment comprises the systemic administration to HCV-infected subjects of an amount effective to combat conditions associated with HCV and other pathogenic flavi- and pestiviruses. Accordingly, the combinations of the present invention may be used in the manufacture of a medicament useful for treating, preventing or combating infection or disease associated with HCV infection in a mammal, in particular for treating conditions associated with HCV and other flaviy pestiviruses. pathogenic.

In one embodiment of the present invention there is provided a pharmaceutical composition comprising a combination according to any of the embodiments described herein and a pharmaceutically acceptable excipient. In particular, the present invention provides a pharmaceutical composition comprising (a) a therapeutically effective amount of an HCV NS3/4a protease inhibitor of formula (I) or a pharmaceutically acceptable salt thereof, (b ) a therapeutically effective amount of ritonavir or a pharmaceutically acceptable salt thereof, and (c) a pharmaceutically acceptable excipient. Optionally, the pharmaceutical composition further comprises an additional agent selected from an HCV polymerase inhibitor, an HCV protease inhibitor, an inhibitor of another target in the HCV life cycle, and an immunomodulatory agent, an agent antivirals and their combinations.

The compositions can be formulated in appropriate pharmaceutical dosage forms such as the dosage form described above. Each of the active components can be formulated separately and the formulations can be administered concomitantly or a formulation containing both and additional active components can be provided if desired.

As used herein, the term "composition" is intended to encompass a product comprising the specified components, as well as any product that is obtained, directly or indirectly, from the combination of the specified components.

In one embodiment the combinations provided herein may also be formulated as a combination preparation for simultaneous, separate or consecutive use in anti-HIV therapy. In such a case, the compound of general formula (I) or any sub-group thereof, is formulated in a pharmaceutical composition containing other pharmaceutically acceptable excipients, and ritonavir is formulated separately in a pharmaceutical composition containing other pharmaceutically acceptable excipients. Conveniently, these two separate pharmaceutical compositions can be part of a kit for simultaneous, separate or consecutive use.

Thus, the individual components of the combination of the present invention may be administered separately at different times during the course of treatment or concurrently in single or divided combination form. It is to be understood that the present invention, therefore, encompasses all such alternative or concurrent treatment regimens and the term "administer" is to be construed accordingly. In a preferred embodiment, the separate dosage forms are administered approximately simultaneously.

In one embodiment, the combination of the present invention contains an amount of ritonavir, or a pharmaceutically acceptable salt thereof, which is sufficient to clinically improve the bioavailability of the HCV NS3/4a protease inhibitor of formula (I) in relation to bioavailability when said HCV NS3/4a protease inhibitor of formula (I) is administered alone.

In another embodiment, the combination of the present invention contains an amount of ritonavir, or a pharmaceutically acceptable salt thereof, that is sufficient to increase at least one of the NS3/4a protease inhibitor pharmacokinetic variables. of the HCV of formula (I) selected from t_1/2, C_m¡_no, C_max, C_H.H, AUC at 12 hours, or AUC at 24 hours, relative to said at least one pharmacokinetic endpoint when the HCV NS3/4a protease inhibitor of formula (I) is administered alone.

A further embodiment relates to a method of enhancing the bioavailability of an HCV NS3/4a protease inhibitor comprising administering to an individual in need of said enhancement a combination as defined herein comprising a therapeutically effective amount of each component of said combination.

In an additional embodiment, the invention relates to the use of ritonavir or one of its pharmaceutically acceptable salts, as an enhancer of at least one of the pharmacokinetic variables of an HCV NS3/4a protease inhibitor of the formula (I) Selected from t_1/2, C_min, C_max, C_S£, ABC at 12 o'clock, or ABC at 24 o'clock; with the proviso that such use is not practiced on the human or animal body.

The term "individual" as used herein refers to an animal, preferably a mammal, most preferably a human, that has been the subject of treatment, observation, or experimentation.

Bioavailability is defined as the fraction of the administered dose that reaches the systemic circulation. you_1/2 represents the half-life or the time elapsed for the plasma concentration to return to half of its original value. C._H.H is the steady-state concentration, that is, the concentration at which the rate of entry of the drug is equal to the rate of elimination. C._min it is defined as the lowest (minimum) concentration measured during the dosing interval. C._max, represents the highest (maximum) concentration during the dosing interval. AUC is defined as the area under the plasma concentration-time curve for a defined period of time.

The combinations of this invention can be administered to humans in specific dosage ranges for each component included in such combinations. The components comprised in such combinations can be administered together or separately. The NS3/4a protease inhibitors of formula (I) or any sub-group thereof, and ritonavir or a pharmaceutically acceptable salt or ester thereof, may have dosage levels in the order of 0.02 to 5.0 grams per day.

When the HCV NS3/4a protease inhibitor of formula (I) and ritonavir are administered in combination, the weight ratio of the HCV NS3/4a protease inhibitor of formula (I) to ritonavir is appropriately in the range from about 40:1 to about

1:15, or from about 30:1 to about 1:15, or from about 15:1 to about 1:15, typically from about 10:1 to about 1:10, and more typically from about 8:1 to about 1:8. Also useful are weight ratios of the HCV NS3/4a protease inhibitors of formula (I) to ritonavir ranging from about 6:1 to about 1:6, or from about 4:1 to about 1:4, or from about 3:1 to about 1:3, or from about 2:1 to about 1:2, or from about 1.5:1 to about 1:1.5. In one aspect, the amount by weight of the HCV NS3/4a protease inhibitors of formula (I) is equal to or greater than that of ritonavir, where the weight ratio of the HCV NS3/4a protease inhibitor of formula (I ) to ritonavir suitably ranges from about 1:1 to about 15:1, usually from about 1:1 to about 10:1, and more usually from about 1:1 to about 8:1. Also useful are weight ratios of the HCV NS3/4a protease inhibitor of formula (I) to ritonavir ranging from about 1:1 to about 6:1, or from about 1:1 to about 5:1, or from about 1:1 to about 4:1, or about 3:2 to about 3:1, or about 1:1 to about 2:1, or about 1:1 to about 1.5:1.

The term "therapeutically effective amount" as used herein refers to that amount of active compound or pharmaceutical component or agent that produces the intended biological or medicinal response in a tissue, system, animal or human, in view of the present invention. , by a researcher, veterinarian, physician or other clinician, including alleviation of symptoms of the disease being treated. Since the present invention relates to combinations comprising two or more agents, the "therapeutically effective amount" is that amount of agents taken together such that the combined effect produces the desired biological or medicinal response. For example, the therapeutically effective amount of a composition comprising (a) the compound of formula (I) and (b) ritonavir, would be the amount of the compound of formula (I) and the amount of ritonavir that when taken together have a combined effect that is therapeutically effective.

It is generally contemplated that an effective antiviral daily amount would be from 0.01 mg/kg to 500 mg/kg of body weight, more preferably from 0.1 mg/kg to 50 mg/kg of body weight. It may be appropriate to administer the required dose as two, three, four, or more sub-doses at appropriate intervals throughout the day. Said sub-doses may be formulated as a unit dosage form, eg containing 1 to 1000 mg, and in particular 5 to 200 mg of active ingredient per unit dosage form.

The exact dose and frequency of administration depends on the particular compound of formula (I) used, the particular condition treated, the severity of the condition treated, age, weight, sex, degree of disorder and general physical condition. of the particular patient as well as other medication the Individual may be taking, as is known to those of skill in the art. Additionally, it is evident that said effective daily amount can be reduced or increased depending on the response of the treated subject and/or depending on the evaluation of the physician who prescribes the compounds of the present invention. The effective daily amount ranges mentioned above are therefore only guides.

According to one embodiment, the HCV NS3/4a protease inhibitor of formula (I) and ritonavir can be administered concomitantly once or twice daily, preferably orally, where the amount of the compounds of formula (I ) per dose is from about 1 to about 2500 mg, and the amount of ritonavir per dose is from 1 to about 2500 mg. In another embodiment, the amounts per dose for once or twice daily co-administration are from about 50 to about 1500 mg of the compound of formula (I) and from about 50 to about 1500 mg of ritonavir. In yet another embodiment, the amounts per dose for once or twice daily co-administration are from about 100 to about 1000 mg of the compound of formula (I) and from about 100 to about 800 mg of ritonavir. In yet another embodiment, the amounts per dose for once or twice daily co-administration are from about 150 to about 800 mg of the compound of formula (I) and from about 100 to about 600 mg of ritonavir. In yet another embodiment, the amounts per dose for once or twice daily co-administration are from about 200 to about 600 mg of the compound of formula (I) and from about 100 to about 400 mg of ritonavir. In yet another embodiment, the amounts per dose for once or twice daily co-administration are from about 200 to about 600 mg of the compound of formula (I) and from about 20 to about 300 mg of ritonavir. In yet another embodiment, the amounts per dose for once or twice daily co-administration are from about 100 to about 400 mg of the compound of formula (I) and from about 40 to about 100 mg of ritonavir.

Example combinations of the compound of formula (I) (mg)/ritonavir (mg) for once or twice daily dosing 50/100, 100/100, 150/100, 200/100, 250/100, 300 /100, 350/100, 400/100,

450/100, 50/133, 100/133, 150/133, 200/133, 250/133, 300/133, 50/150, 100/150, 150/150, 200/150,

250/150, 50/200, 100/200, 150/200, 200/200, 250/200, 300/200, 50/300, 80/300, 150/300, 200/300,

250/300, 300/300, 200/600, 400/600, 600/600, 800/600, 1000/600, 200/666, 400/666, 600/666, 800/666,

1000/666, 1200/666, 200/800, 400/800, 600/800, 800/800, 1000/800, 1200/800, 200/1200, 400/1200, 600/1200, 800/1200,1000/ 1200, and 1200/1200. Other exemplary combinations of the compound of formula (I) (mg)/ritonavir (mg) for once or twice daily dosing 1200/400, 800/400, 600/400, 400/200, 600/200, 600 /100, 500/100, 400/50, 300/50, and 200/50.

In one embodiment of the present invention there is provided an article of manufacture comprising a composition effective for treating HCV infection or inhibiting HCV NS3 protease; and packaging material comprising a label indicating that the composition can be used to treat infection caused by the hepatitis C virus; wherein the composition comprises a compound of formula (I) or any sub-group thereof, or the combination as described herein.

Another embodiment of the present invention relates to a kit or container comprising a compound of formula (I) or any sub-group thereof, or a combination according to the invention combining an HCV NS3/4a protease inhibitor of formula (I) or a pharmaceutically acceptable salt thereof, and ritonavir or a pharmaceutically acceptable salt thereof, in an effective amount for use as a standard or reagent in a test or assay to determine the ability of potential pharmaceuticals to inhibit HCV NS3/4a protease, HCV growth, or both. This aspect of the invention may find use in pharmaceutical research programs.

The compounds and combinations of the present invention can be used in high resolution analytical blank assays such as those to measure the efficacy of said combination in the treatment of HCV.

examples

The following examples are intended to illustrate the present invention and not to limit it.

Example 1: Preparation of representative intermediates.

Synthesis of 4-hydroxy-7-methoxy-8-methyl-2-(thiazol-2-yl)quinoline (4).

step A

NH.

Added a BCI fix₃ (1.0M in CH₂IC₂, 194 mL) dropwise via cannula over 20 min, under argon pressure, at 0°C, to a solution of 3-methoxy-2-methalanyl-line (25.4 g, 185 mmol) in xylene (300 mi). The temperature was maintained between 0°C and 10°C, until the addition was complete. After an additional 30 min, at 0°C, acetonitrile (12.6 mL, 241 mmol) was added dropwise under argon at 0°C. After 30 min at 0 °C, the resulting suspension was transferred into a dropping funnel and diluted with CH₂IC₂ (40 mi). This mixture was added to 0^yesC under argon for 20 min to a suspension of AICI3 (25.9 g, 194 mmol) in CH2CI2 (40 mL). The resulting orange solution was heated in a 70°C oil bath under a stream of nitrogen for 12h. The reaction mixture was then cooled to room temperature and ice water and CH2CI2 were added. This mixture was refluxed for 6h and then cooled to room temperature. After 12h, the pH was adjusted to 0.^QC to 3 with 6N NaOH. The solution was extracted with CH2CI₂, was subsequently washed with water, 1N NaOH and brine. The organic layer was dried (Na₂SW₄), filtered and concentrated in vacuo. The residue was triturated at room temperature in diisopropyl ether (50 mL) for 0.5 h. The suspension was then cooled to 0^QC, filtered and washed with a small amount of diisopropyl and dried in high vacuum to give 15.4 g (46%) of the desired product 2: m/z = 180 (M+H)⁺.

step B

<img file="ECSP088150A_D0078.tif" />

<img file="ECSP088150A_D0079.tif" />

EDCI (257 mg, 1.34 mmol) and HOAt (152 mg, 1.12 mmol) were added to a stirred solution of 2 (200 mg, 1.12 mmol) in CH₂IC₂ (10 ml) and dry DMF (1 ml). The resulting solution was stirred at room temperature for 3 days. The reaction mixture was then partitioned between CH₂IC₂ and NaHCO₃1N. The organic layer was washed successively with NH₄CI 1N and water, dried (Na₂SW₄) and evaporated. Purification by flash chromatography (AcOEt/heptane gradient, 10:90 to 50:50) gave 62 mg (19%) of the desired product: m/z = 291 (M+H)⁺.

Step C

<img file="ECSP088150A_D0080.tif" />

<img file="ECSP088150A_D0081.tif" />

tBuOK (50 mg, 0.448 mmol) was added to a suspension of acetophenone 3 (62 mg, 0.213 mmol) in fBuOH (5 mL). The resulting mixture was stirred at 80°C overnight, then cooled to room temperature. The reaction mixture was diluted with AcOEt, acidified with KHSO₄ and subsequently washed with water and brine. The organic layer was dried (Na₂SW₄) and evaporated to give 43 mg (74%) of the sought product as a white powder: m/z = 273 (M+H)⁺.

Synthesis of (hex-5-enyl)(methyl)amine (21).

<img file="ECSP088150A_D0082.tif" />

<img file="ECSP088150A_D0083.tif" />

<img file="ECSP088150A_D0084.tif" />

step A

Sodium hydride (1.05 eq) was added slowly at 0^andC to a solution of /V-methyltrifluoro-acetamide (25 g) in DMF (140 ml). The mixture was stirred for 1h at room temperature under nitrogen. Then, a solution of bromohexene (32.1 g) in DMF (25 ml) was added dropwise and the mixture was heated at 70°C for 12 hours. The reaction mixture was poured into water (200 ml) and extracted with ether (4 x 50 ml), dried (MgSO₄), filtered and evaporated to give 35 g of the target product 20 as a yellow oil which was used without further purification in the next step.

step B

A solution of potassium hydroxide (187.7 g) in water (130 ml) was added dropwise to a solution of 20 (35 g) in methanol (200 ml). The mixture was stirred at room temperature for 12 hours. The reaction mixture was then poured into water (100 ml) and extracted with ether (4 x 50 ml), dried (MgSO₄), was filtered and the ether was distilled at atmospheric pressure. The resulting oil was purified by vacuum distillation (13 mm Hg pressure, 50°C) to give 7.4 g (34%) of the title product 21 as a colorless oil:¹H-NMR (CDCI₃): δ 5.8 (m, 1H), 5 (ddd, J= 17.2 Hz, 3.5 Hz, 1.8 Hz, 1H), 4.95 (m, 1H), 2.5 (t , 7.0 Hz, 2H), 2.43 (s, 3H), 2.08 (q, J= 7.0 Hz, 2H), 1.4 (m, 4H), 1.3 (br s, 1 HOUR).

Example 2: Preparation of 17-[7-methoxy-8-methyl-2-(t¡azol-2-¡l)quinolin-4-ylox¡]-13-methyl-2,14-dioxo3,13 acid -diazatricyclo[13.3.0.0^4S]octadec-7-en-4-carboxylic acid (29)

step A

3-Oxo-2-oxa-bicyclo[2.2.1]heptane-5-carboxylic acid 22 (500 mg, 3.2 mmol) in 4 mL DMF was added to 0^QC a HATU (1.34 g, 3.52 mmol) and A/-methylhex-5-enylamine (435 mg, 3.84 mmol) in DMF (3 mL), followed by DIPEA. After stirring for 40 min at 0^andC, the mixture was stirred at room temperature for 5 h. Then the solvent was evaporated, the residue was dissolved in EtOAc (70 ml) and washed with NaHCO₃saturated (10 ml). The aqueous layer was extracted with EtOAc (2 x 25 mL). The organic layers were combined, washed with saturated NaCI (20 mL), dried (Na₂SW₄) and evaporated. Purification by flash chromatography (EtOAc/petroleum ether, 2:1) gave 550 mg (68%) of the target product 23 as a colorless oil: m/z=252 (M+H)⁺.

step B

ooh

ooh

A solution of LiOH (105 mg in 4 mL of water) was added at 0^QC to lactone amide 23. After 1 h, the conversion was complete (HPLC). The mixture was acidified to pH 2-3 with 1N HCI, extracted with

AcOEt, dried (MgSO₄), evaporated, co-evaporated with toluene several times and dried in high vacuum overnight to give 520 mg (88%) of the desired product 24: m/z = 270 (M+H)⁺.

<img file="ECSP088150A_D0085.tif" />

<img file="ECSP088150A_D0086.tif" />

.HCI . COOEt

<img file="ECSP088150A_D0087.tif" />

<img file="ECSP088150A_D0088.tif" />

1-(Amino)-2-(vinyl)cyclopropanecarboxylic acid ethyl ester hydrochloride 25 (4.92 g, 31.7 mmol) and HATU (12.6 g, 33.2 mmol) were added to 24 ( 8.14g, 30.2mmol). The mixture was cooled in an ice bath under argon and then DMF (100 mL) and DIPEA (12.5 mL, 11.5 mmol) were added subsequently. After 30 min at 0 °C, the solution was stirred at room temperature for an additional 3 h. The reaction mixture was then partitioned between EtOAc and water, post-washed with 0.5 N HCI (20 mL) and saturated NaCI (2 x 20 mL) and dried (Na₂SW₄). Purification by flash chromatography (AcOEt/CH₂IC₂/petroleum ether, 1:1:1) gave 7.41 g (60%) of the desired product 26 as a colorless oil: m/z =407 (M+H)⁺.

Step D

<img file="ECSP088150A_D0089.tif" />

DIAD (218 μί, 1.11 mmol) was added at -20°C under a nitrogen atmosphere to a solution of 26 (300 mg, 0.738 mmol), quinoline 4 (420 mg, 1.03 mmol), and triphenylphosphine (271 mg , 1.03 mmol) in dry THF (15 mL). The reaction is then warmed to room temperature. After 1.5 h, the solvent was evaporated and the crude product was purified by flash column chromatography (petroleum/CH gradient).₂IC₂/ether, 3:1.5:0.5 to 1:1:1) to give the desired product 27: m/z = 661 (M+H)⁺.

Step E

<img file="ECSP088150A_D0090.tif" />

A solution of 27 (200 mg, 0.30 mmol) and HoveydaGrubbs 1st generation catalyst (18 mg, 0.030 mmol) dry, dephasified 1,2-dichloroethane (300 mL) was heated at 70 °C under nitrogen for 12 h. . Then, the solvent was evaporated and the residue purified by silica gel chromatography (Petroleum ether/CH₂IC₂/et₂0; 3:1:1) to give the desired product 28: m/z= 633 (M+H)⁺.

<img file="ECSP088150A_D0091.tif" />

<img file="ECSP088150A_D0092.tif" />

A solution of LiOH (327 mg) in water (3 mL) was added to a stirred solution of 28 (150 mg, 0.237 mmol) in THF (15 mL) and MeOH (10 mL). After 48h the solvent was evaporated and the residue was partitioned between water and ether. The aqueous layer was acidified (pH = 3) and extracted with AcOEt, dried (MgSO₄) and evaporated. The residue was crystallized from ether to give the objective compound 29: m/z = 605 (M+H)⁺.

Example 3: Preparation of /V-í17-[7-methox¡-8-methyl-2-(t¡azol-2-¡l)au¡nol¡n-4-¡lox¡1-13-met ¡l-2,14-d¡oxo-3,13d¡azatr¡c¡clo[13.3.0.0⁴’⁶1octadec-7-en-4-carbon¡ll-(cycloprop¡l)sulfonamide (30)

<img file="ECSP088150A_D0093.tif" />

A mixture of 29 (85 mg, 0.14 mmol) and CDI (47 mg, 0.29 mmol) in dry THF (7 mL) was heated under reflux for 2h under nitrogen. LCMS analysis shows a peak of the intermediate (r = 5.37). The reaction mixture was cooled to room temperature and cyclopropylsulfonamide (52 mg, 0.43 mmol) was added. Then, DBU (50 μί, 0.33 mmol) was added and the reaction mixture was stirred at room temperature for 1h and then heated at 55°C for 24h. The solvent was evaporated and the residue was partitioned between AcOEt and acidic water (pH = 3). The crude material was purified by column chromatography (AcOEt/CH₂IC₂/Petroleum ether, 1:1:1). The residue was crystallized from Et₂Or, filtered to give the target compound contaminated with the cyclopropyl sulfonamide. This material was triturated in 3 mL of water, filtered, washed with water, and dried overnight in the high vacuum pump to give the target compound 30 as a white powder: m/z=708 (M+ h)⁺.

Example 4: Preparation of the acid 17-[2-(4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-yloxyl]-13-methyl2,14-dioxo-3,13-diazatricyclo[13.3 .0.0^4,6]octadec-7-en-4-carboxylic acid (46)

Synthesis of 4-h¡drox¡-2-(4-¡soprop¡lt¡azol-2-¡l)-7-methox¡-8-methylgu¡nol¡na (36)

Step 1: Synthesis of /V-(yer-butyloxycarbonyl)-3-methoxy-2-methylaniline (32)

<img file="ECSP088150A_D0094.tif" />

<img file="ECSP088150A_D0095.tif" />

Triethylamine (42.4 mL, 302 mmol) was added to a suspension of 3-methoxy-2-methylbenzoic acid (45.6 g, 274 mmol) in dry toluene (800 mL). A clear solution was obtained. Then dppa (65.4 mL, 302 mmol) in toluene (100 mL) was slowly added. After 1h at room temperature, the reaction mixture was subsequently heated to 50°C for 0.5h, 70°C for 0.5h then 100°C for 1h. To this solution, t-BuOH (30.5 g, 411 mmol) in toluene (40 mL) was added at 100°C and the resulting mixture was refluxed for 7h. The solution was cooled to room temperature then post-washed with water, 0.5 N HCI, 0.5 N NaOH and brine, dried (Na₂SW₄) and evaporated to give 67 g of the sought product: m/z = 237 (M)⁺.

Step 2: Synthesis of 3-methoxy-2-methylaniline (33)

<img file="ECSP088150A_D0096.tif" />

<img file="ECSP088150A_D0097.tif" />

TFA (40.7 mL, 548 mmol) was added to a solution of A/-(tert-butyloxycarbon¡l)-3-methoxy¡-2-methylaniline, in dichloromethane (500 mL). After 2 h at room temperature, TFA (40.7 ml,

548 mmol) and the resulting mixture was stirred at room temperature overnight. The volatiles were then evaporated. The residue was triturated with toluene (100 ml) and diisopropyl ether (250 ml), filtered and washed with diisopropyl ether (100 ml) to give 56.3 g of the title product as a TFA salt: m/z = 138 (M+H)⁺. The TFA salt was transformed into the free aniline by treatment with NaHCO₃.

Step 3: Synthesis of (2-amino-4-methox¡-3-methylphenyl)(methyl)ketone (34)

<img file="ECSP088150A_D0098.tif" />

<img file="ECSP088150A_D0099.tif" />

BCI solution added slowly₃ (1.0 M, 200 mL, 200 mmol) in CH₂IC₂ under nitrogen to a solution of 3-methoxy-2-methylaniline (26.0 g, 190 mmol) in xylene (400 mL). The temperature was monitored during the addition and was kept below 10°C. The reaction mixture was stirred at 5°C for 0.5h. Then, dry acetonitrile (13 mL, 246 mmol) was added at 5°C. After 0.5 h at 5°C, the solution was transferred into a dropping funnel and slowly added at 5°C to AICI slurry.₃ (26.7 g, 200 mmol) in CH₂IC₂ (150 mi). After 45 min at 5°C, the reaction mixture was warmed to

70°C under a stream of nitrogen. After CH evaporation₂IC₂, the temperature of the reaction mixture reached 65°C. After 12h at 65°C, the reaction mixture was cooled to 0°C, poured onto ice (300g) and slowly refluxed for 7h. After 2 days at room temperature, 6N NaOH (50 mL) was added. The pH of the resulting solution was 2-3. The xylene layer was decanted. The organic layer was extracted with CH₂IC₂. The xylene and CH layers₂IC₂ were combined, subsequently washed with water, 1N NaOH and brine, dried (Na₂SW₄) and evaporated. The residue was triturated from diisopropyl ether at 0^D.C, filtered and washed with diisopropylether to give 13.6 g (40%) of the title product as a yellowish solid: m/z = 180 (M+H)⁺.

Step 4: Synthesis of 2'-[[(4-isopropylthiazol-2-yl)(oxo)methyl]amino]-4'-methoxy-3'-methylacetophenone (35)

<img file="ECSP088150A_D0100.tif" />

⁰

<img file="ECSP088150A_D0101.tif" />

A solution of (2-amino-4-methoxy¡-3-methylphenyl)(methyl)ketone (18.6 g, 104 mmol) in dioxane (50 mL) was added under nitrogen to a suspension of 4-isopropylthiazole-2-carbonyl chloride in dioxane (250 ml). After 2h at room temperature, the reaction mixture was concentrated to dryness. The residue was then partitioned between aqueous NaHCOg solution and AcOEt, the organic layer washed with brine, dried (Na₂SW₄) and evaporated. The residue was triturated in diisopropyl ether, filtered and washed with diisopropyl ether to give 30.8 g (90%) of the title product 35.

Step 5: Synthesis of 4-hydroxy-2-(4-¡sopropylthiazol-2-yl)-7-methoxy¡-8-methylquinol¡na (36)

<img file="ECSP088150A_D0102.tif" />

Potassium tert-butoxide (21.8 g, 195 mmol) was added to a suspension of 2'-[[(4-isopropylthiazol-2¡l)(oxo)methyl]am¡no]-4'-methoxy¡- 3'-Methylacetophenone (35, 30.8 g, 92.7 mmol) in ferr-butanol. The resulting reaction mixtures were heated at 100°C overnight. The reaction mixture was then cooled to room temperature and diluted with ether (100 ml). The precipitate was filtered and washed with Et₂Or to give a powder (fraction A). The mother liquor was concentrated in vacuo, triturated in ether, filtered and washed with ether to give a powder (fraction 2). Fractions 1 and 2 were mixed and poured into water (250 ml). The pH of the resulting solution was adjusted to 6-7 (control with pH paper) with 1N HCl. The precipitate was filtered off, washed with water and dried. The solid was then triturated in diisopropyl ether, filtered and dried to give 26 g (88%) of the title product 36 as a brownish solid: m/z =315 (M+H)⁺.

Synthesis of íhex-5-enil)(met¡l)am¡na (38)

<img file="ECSP088150A_D0103.tif" />

Yo

NH

Step A:

Sodium hydride (1.05 eq) was added slowly at 0^andC to a solution of /V-methyltrifluoro-acetamide (25 g) in DMF (140 ml). The mixture was stirred for 1h at room temperature under nitrogen. Then, a solution of bromohexene (32.1 g) in DMF (25 ml) was added dropwise and the mixture was heated at 70°C for 12 hours. The reaction mixture was poured into water (200 ml) and extracted with ether (4 x 50 ml), dried (MgSO₄), filtered and evaporated to give 35 g of target product 37 as a yellow oil which was used without further purification in the next step.

Step B:

A solution of potassium hydroxide (187.7 g) in water (130 ml) was added dropwise to a solution of 37 (35 g) in methanol (200 ml). The mixture was stirred at room temperature for 12 hours. The reaction mixture was then poured into water (100 ml) and extracted with ether (4 x 50 ml), dried (MgSO₄), was filtered and the ether was distilled at atmospheric pressure. The resulting oil was purified by vacuum distillation (13 mm Hg pressure, 50 °C) to give 7.4 g (34%) of the title product 38 as a colorless oil:¹H-NMR (CDCI₃): δ 5.8 (m, 1H), 5 (ddd, J= 17.2 Hz, 3.5 Hz, 1.8 Hz, 1H), 4.95 (m, 1H), 2.5 (t , J= 7.0 Hz, 2H), 2.43 (s, 3H), 2.08 (q, J = 7.0 Hz, 2H), 1.4 (m, 4H), 1.3 (br s, 1H).

Preparation of acid 17-[2-(4-¡soprop¡lt¡azol-2-¡l)-7-methox¡-8-met¡lquinol¡n-4-¡lox¡]-13-methyl- 2₁14-d¡oxo-3,13d¡azatr¡c¡clo[13.3.0.0^4,61octadec-7-en-4-carboxylic (46)

step A

3-Oxo-2-oxa-bicyclo[2,2,1]heptane-5-carboxylic acid 39 (500 mg, 3.2 mmol) in 4 mL DMF was added to 0^yesC to HATU (1.34 g, 3.52 mmol) and /V-methylhex-5-enylamine (435 mg, 3.84 mmol) in DMF (3 mL), followed by DIPEA. After stirring for 40 min at 0^toC, the mixture was stirred at room temperature for 5 h. Then the solvent was evaporated, the residue was dissolved in EtOAc (70 ml) and washed with NaHCO₃saturated (10 ml). The aqueous layer was extracted with EtOAc (2 x 25 mL). The organic phases were combined, washed with saturated NaCI (20 mL), dried (Na₂SW₄) and evaporated. Purification by flash chromatography (EtOAc/petroleum ether, 2:1) gave 550 mg (68%) of the target product as a colorless oil: m/z = 252 (M+H)⁺.

step B

ooh

<img file="ECSP088150A_D0104.tif" />

A solution of LiOH (105 mg in 4 mL of water) was added at 0^andC to the lactone amide 40. After 1h, the conversion was complete (HPLC). The mixture was acidified to pH 2-3 with 1N HCI, extracted with AcOEt, dried (MgSO₄), evaporated, co-evaporated with toluene several times and dried in high vacuum overnight to give 520 mg (88%) of the target product 41: m/z = 270 (M+H)⁺.

<img file="ECSP088150A_D0105.tif" />

.HCI .COOEt

<img file="ECSP088150A_D0106.tif" />

1 -(Amino)-2-(vinyl)cyclopropane-carboxylic acid ethyl ester hydrochloride 42 (4.92 g, 31.7 mmol) and HATU (12.6 g, 33.2 mmol) were added to 41 (8.14g, 30.2mmol). The mixture was cooled in an ice bath under argon and then DMF (100 mL) and DIPEA (12.5 mL, 11.5 mmol) were added subsequently. After 30 min at 0 °C, the solution was stirred at room temperature for an additional 3 h. The reaction mixture was then partitioned between EtOAc and water, post-washed with 0.5 N HCI (20 mL) and saturated NaCI (2 x 20 mL) and dried (Na₂SW₄). Purification by flash chromatography (AcOEt/CH₂IC₂/petroleum ether, 1:1:1) gave 7.41 g (60%) of the desired product 43 as a colorless oil: m/z=407 (M+H)⁺.

Step D

<img file="ECSP088150A_D0107.tif" />

DIAD (1.02 mL, 5.17 mmol) was added at -15°C under a nitrogen atmosphere to a solution of 43 (1.5 g, 3.69 mmol), quinoline 36 (1.39 g, 4, 43 mmol) and triphenylphosphine (1.26 g, 4.80 mmol) in dry THF (40 mL). After 4.5 h, at -15 °C, the reaction mixture was partitioned between ice water and AcOEt, dried (Na₂SW₄) and evaporated. The crude material was purified by flash column chromatography (AcOEt/CH oil gradient₂IC₂0.1:9 to 2:8) to give 1.45 g (56%) of the desired product 44: m/z=703 (M+H)⁺.

<img file="ECSP088150A_D0108.tif" />

<img file="ECSP088150A_D0109.tif" />

A solution of 44 (1.07 g, 1.524 mmol) and HoveydaGrubbs 1st generation catalyst (33 mg, 0.03 eq) in dry, degassed 1,2-dichloroethane (900 mL) was heated at 75°C under nitrogen for 12 noon Then the solvent was evaporated and the residue purified by silica gel chromatography (25% EtOAc in CH₂IC₂). 620 mg (60%) of pure macrocycle 45 were obtained. m/z = 674 (M+H)⁺. ¹H NMR (CDCI₃): 1.18-1.39 (m, 12H), 1.59 (m, 1H), 1.70-2.08 (m, 5H), 2.28 (m, 1H), 2.38 ( m, 1H), 2.62 (m, 2H), 2.68 (s, 3H), 2.83 (m, 1H), 3.06 (s, 3H), 3.19 (sep, J= 6 0.7 Hz, 1H), 3.36 (m, 1H), 3.83 (m, 1H), 3.97 (s, 3H), 4.09 (m, 2H), 4.65 (td, J = 4 Hz, 14 Hz, 1H), 5.19 (dd, J= 4 Hz, 10 Hz, 1H), 5.31 (m, 1H), 5.65 (td, J= 4 Hz, 8 Hz, 1H), 7.00 (s, 1H), 7.18 (s, 1H), 7.46 (d, J= 9 Hz, 1H), 7.48 (s, 1H), 8.03 (d, 9hz, 1H).

Step F

Z°vkA

<img file="ECSP088150A_D0110.tif" />

A solution of lithium hydroxide (1.65 g, 38.53 mmol) in water (15 mL) was added to a stirred solution of ester 45 (620 mg, 0.920 mmol) in THF (30 mL) and MeOH (20 mL ). After 16 h at room temperature, the reaction mixture was quenched with NH₄Cl sat., concentrated under reduced pressure, acidified to pH 3 with 1N HCI and extracted with CH₂IC₂, dried (MgSO₄) and evaporated to give 560 mg (88%) of carboxylic acid 46, m/z=647 (M+H)⁺. ¹H NMR (CDCI₃): 1.11-1.40 (m, 8H), 1.42-1.57 (m, 2H), 1.74 (m, 2H), 1.88-2.00 (m, 2H), 2.13 (m, 1H), 2.28 (m, 1H), 2.40 (m, 1H), 2.59 (m, 2H), 2.67 (S, 3H), 2.81 (m , 1H), 2.97 (s, 3H), 3.19 (m, 1H), 3.31 (m, 1H), 3.71 (m, 1H), 3.96 (s, 3H), 4 .56 (sd, J = 4 Hz, 12 Hz, 1H), 5.23 (m, 2H), 5.66 (m, 1H), 7.01 (s, 1H), 7.10 (s, 1H ), 7.22 (d, J= 10 Hz, 1H), 7.45 (s, 1H), 8.00 (d, 10 Hz, 1H).

<img file="ECSP088150A_D0111.tif" />

<img file="ECSP088150A_D0112.tif" />

A solution of 17-[2-(4-¡soprop¡lt¡zol-2-¡l)-7-methoxy-8-methylquinol¡n-4-¡lox¡]-13- acid was stirred methyl2,14-dioxo-3,13-diazatricyclo[13.3.0.04,6]octadec-7-ene-4-carboxylic acid 46 (138.3 mg, 0.214 mmol) prepared according to the procedure described above and carbonyldiimidazole ( 96.9 mg, 0.598 mmol) in dry THF (5 mL) at reflux under nitrogen for 2h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was partitioned between EtOAc and 11N HC, the organic layer washed with brine, dried (Na2SO₄) and evaporated. The solid was then triturated in i-Pr ether to obtain 46' as a white powder: m/z = 629 (M+H)⁺. ¹H NMR (CDCI₃): 0.99-1.00 (m, 1H), 1,201.35 (m, 2H), 1.39 (d, J = 6.9 Hz, 6H), 1.55-1.7 (m, 1H), 1.9-2 (m, 2H), 2.15-2.25 (m, 2H), 2.3-2.60 (m, 4H), 2.68 (s, 3H), 2 .71-2.82 (m, 1H), 2.82-2.9 (m, 1H), 3.08 (s, 3H), 3.1-3.2 (m, 1H), 3.4 -3.5 (m, 1H), 3.65-3.71 (m, 1H), 3.91 (s, 3H), 4.28-4.4 (m, 1H), 5.32-5 .46 (m, 2H), 5.85-5.95 (m, 1H), 7.00 (s, 1H), 7.22 (d, J = 9.2 Hz, 1H), 7.45 ( S, 1H), 8.09 (d, J= 9.2 Hz, 1H).

Example 5: Preparation of A/-[17-[2-(4-isoprooylthiazol-2-yl)-7-methoxy¡-8-methyllauinol¡n-4-yloxyl-13-methyl-2.14d¡ oxo-3,13-diazatricyclo[13.3.0.0⁴⁶loctadec-7-en-4-carbonyl-(cyclopropyl)sulfonamide (47)

<img file="ECSP088150A_D0113.tif" />

<img file="ECSP088150A_D0114.tif" />

A solution of 17-[2-(4-isopropylthazol-2-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl2,14-dioxo-3,13-diazatricyclo acid was stirred [13.3.0.04,6]octadec-7-ene-4-carboxylic acid 46 (560mg, 0.867mmol) prepared according to Example 4 and carbonyldiimidazole (308mg, 1.90mmol) in dry THF (10ml) at reflux in nitrogen for 2h. The reaction mixture was cooled to room temperature and cyclopropylsulfonamide (400 mg, 3.301 mmol) and DBU (286 mg, 1.881 mmol) were added. This solution was heated at 50 °C for 15 h. The reaction mixture was then cooled to room temperature and concentrated under reduced pressure. The residue was partitioned between CH₂IC₂ and 1N HCI, the organic layer was washed with brine, dried (MgSO₄) and evaporated. Purification by flash chromatography (gradient EtOAc (0 to 25%) in CH₂IC₂) gave 314 mg of an off-white solid which was further washed with water, then isopropyl ether and dried in the vacuum oven to give 282 mg (40%) of the pure product title 47 as a white powder: m/z = 750 (M+H)⁺. ¹H NMR (CDCI₃): 0.99-1.52 (m, 14H), 1.64-2.05 (m, 4H), 2.77 (m, 1H), 2.41 (m, 2H), 2.59 ( m, 2H), 2.69 (s, 3H), 2.92 (m, 2H), 3.04 (s, 3H), 3.19 (m, 1H), 3.40 (m, 2H), 3.98 (s, 3H), 4.60 (t, J= 13 Hz, 1H), 5.04 (t, J = 11 Hz, 1H), 5.37 (m, 1H), 5.66 ( m, 1H), 6.21 (s, 1H), 7.02 (s, 1H), 7.22 (d, J= 10 Hz, 1H), 7.45 (s, 1H), 7.99 ( d, J= 10 Hz, 1H), 10.82 (broad s, 1H).

Example 6: Preparation of Λ/-[17-[2-(4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14dioxo-3,13 -diazatricyclo[13.3.0.0^{4 6}]octadec-7-en-4-carbonyl](1-methylcyclopropyl)sulfonamide (48)

<img file="ECSP088150A_D0115.tif" />

A solution of carboxylic acid 46 (240 mg, 0.38 mmol) and carbonyldiimidazole (2 eq) in dry THF (5 mL) was stirred under reflux under nitrogen for 2h. The reaction mixture was cooled to room temperature and 1-methylcyclopropylsulfonamide (2 eq) and DBU (2 eq) were added. This solution was heated at 50 °C for 15h. The reaction mixture was then cooled to room temperature and concentrated under reduced pressure. The residue was partitioned between CH₂IC₂ and 1N HCI, the organic layer was washed with brine, dried (MgSO₄) and evaporated. Purification by flash chromatography (gradient EtOAc (0 to 25%) in CH₂IC₂) gave 170 mg (58%) of the title compound 48 as an off-white solid which was further washed with water, then isopropyl ether and dried in a vacuum oven: m/z = 764 (M+H)⁺. ¹H NMR (Ci6-acetone): 0.86 (m, 2H), 1.15-1.78 (m, 19H), 1.87 (m, 2H), 2.13-2.54 (m, 3H ), 2.57-2.71 (m, 4H), 2.96-3.25 (m, 4H), 3.54 (m, 2H), 4.02 (s, 3H), 4.58 ( t, J= 13 Hz, 1H), 5.04 (m, 1H), 5.46 (m, 1H), 5.62 (m, 1H), 7.31 (S, 1H), 7.43 ( d, J = 9 Hz, 1H), 7.58 (s, 1H), 8.07 (d, J = 13 Hz, 1H), 8.19 (broad S, 1H), 11.44 (broad s, 1 HOUR).

Example 7: Preparation of 17-[8-chloro-2-(4-isopropylthiazol-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3. 0.0^{4 yes}]octadec-7-en-4-carboxylic acid (25)

Step A: Synthesis of (2-amino-3-chloro-4-methoxyphenyl)(methyl)ketone (50)

<img file="ECSP088150A_D0116.tif" />

Cl

<img file="ECSP088150A_D0117.tif" />

EITHER

Added a BCI fix₃ (1.0 M, 138 mL, 138 mmol) in CH₂IC₂ slowly under nitrogen to a solution of 2-chloro-3-methoxyaniline 49 (20.6 g, 131 mmol) in xylene (225 mL). The temperature was monitored during the addition and was kept below 10°C. The reaction mixture was stirred at 5°C for 0.5h. Then, dry acetonitrile (9.0 mL, 170 mmol) was added at 5°C. After 0.5 h at 5°C, the solution was transferred into a dropping funnel and slowly added at 5°C to AICI slurry.₃(18.4 g, 138 mmol) in CH₂IC₂ (80 mi). After 45 min at 5°C, the reaction mixture was heated to 70°C under a stream of nitrogen. After CH evaporation₂IC₂, the temperature of the reaction mixture reached 65°C. After 12h at 65°C, the reaction mixture was cooled to Q°C, poured onto ice (200g) and slowly refluxed for 7h. after 2 days at room temperature, 6N NaOH (25 mL) and CH₂IC₂ (100 mi). The mixture was filtered, the filtrate was washed with CH₂IC₂. The organic layer was decanted and subsequently washed with water, 1N NaOH and brine, dried (Na₂SW₄) and evaporated. The residue was triturated from diisopropyl ether at 0^yesC, filtered and washed with diisopropylether to give 19.0 g (73%) of the title product 50 as a white solid: m/z = 200 (M+H)⁺.

Step B: Synthesis of 2'-[[(4-¡sopropilt¡azol-2-¡l)(oxo)methyl]am¡no]-3'-chloro-4'-methoxy¡acetophenone (51)

or or

The title product 51 (79%) was prepared from (2-amino-3-chloro-4-methoxyphenyl)-(methyl)ketone (50) following the procedure reported for 2'-[[( 4-¡soprop¡lt¡azol-2-yl)(oxo)methyl]am¡no]-4'-methox¡-3'methylacetophenone (35): m/z= 353 (M+H)⁺.

Step C: Synthesis of 8-chloro-4-h¡drox¡-2-(4-¡soprop¡lt¡azol-2-yl)-7-methox¡-quinol¡na (52) or

ooh

Title product 52 was prepared (58%) from 2'-[[(4-¡soprop¡ltiazol-2-¡l)(oxo)-methyl]am¡no]-3'-chloro4'-methoxyacetophenone (51) following the procedure reported for 4-hydroxy-2-(4-isopropylthiazol-2-yl)-7methoxy-8-methylquinoline (36): m/z= 335 (M+H)⁺.

<img file="ECSP088150A_D0118.tif" />

Compound 53 was prepared from alcohol 43 and 8-chloro-4-hydroxy-2-(4-isopropylthazol-2-yl)-7-methoxy-quinoline (52) following the procedure described for 44: m/z = 723 (M+H)⁺.

Step E: Preparation of compound 54

<img file="ECSP088150A_D0119.tif" />

<img file="ECSP088150A_D0120.tif" />

Compound 54 was prepared from 53 following the procedure described for 45: m/z = 695 (M+H)⁺.

Step F: Preparation of compound 55

<img file="ECSP088150A_D0121.tif" />

<img file="ECSP088150A_D0122.tif" />

A solution of lithium hydroxide (3.85 g, 90.1 mmol) in water (30 mL) was added to a stirred solution of ester 54 (1.64 g, 2.36 mmol) in THF (55 mL) and MeOH (40 mi). After 16h at room temperature, more LiOH (1.0g) was added. After 20 h at room temperature, the reaction mixture was quenched with saturated NH solution.₄CI, concentrated under reduced pressure, acidified to pH 5 with 1N HCl, extracted with EtOAc, dried (MgSO₄) and evaporated to give 1.37 g (87%) of carboxylic acid 55, m/z=667 (M+H)⁺.

Example 8: Preparation of /V-[17-[8-chloro-2-(4-isopropylthiazol-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14dioxo-3,13-diazatr cycle[13.3.0.0^{4 6}]octadec-7-en-4-carbonyl](cyclopropyl)sulfonamide (56).

<img file="ECSP088150A_D0123.tif" />

<img file="ECSP088150A_D0124.tif" />

A solution of carboxylic acid 55 (1.37 g, 2.52 mmol) and carbonyldiimidazole (2 eq) in dry THF (75 mL) was stirred under reflux under nitrogen for 2h. The reaction mixture was cooled to room temperature and cyclopropylsulfonamide (2 eq) and DBU (2 eq) were added. This solution was heated at 50 °C for 36 h. The reaction mixture was then cooled to room temperature and concentrated under reduced pressure. The residue was partitioned between EtOAc and 1N HCI, the organic layer washed with brine, dried (MgSO₄) and evaporated. Purification by flash chromatography (gradient EtOAc (0 to 25%) in CH₂IC₂) gave 880 mg (55%) of the title compound 56 as an off-white solid: m/z = 770 (M+H)⁺. ¹H NMR (CDCh, main rotamer): 0.93-1.52 (m, 13H), 1.60-2.07 (m, 5H), 2.21-2.64 (m, 5H), 2, 92 (m, 2H), 3.04 (s, 3H), 3.19 (m, 1H), 3.41 (m, 2H), 4.07 (s, 3H), 4.60 (t, J = 13 Hz, 1H), 5.04 (t, J= 11 Hz, 1H), 5.37 (m, 1H), 5.66 (m, 1H), 6.33 (s, 1H), 7, 07 (s, 1H), 7.24 (d, J= 9 Hz, 1H), 7.52 (s, 1H), 8.05 (d, J= 9 Hz, 1H), 10.81 (broad s , 1 HOUR).

Example 9: Preparation of /V-[17-[8-chloro-2-(4-isopropylthiazol-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14dioxo-3,13-diazatricyclo [13.3.0.0^{4 yes}]octadec-7-en-4-carbonyl](1-methylcyclopropyl)sulfonamide (57).

<img file="ECSP088150A_D0125.tif" />

<img file="ECSP088150A_D0126.tif" />

A solution of carboxylic acid 55 (49 mg, 0.073 mmol) and carbonyldiimidazole (2 eq) in dry THF (5 mL) was stirred under reflux under nitrogen for 2h. The reaction mixture was cooled to room temperature and 1-methylcyclopropylsulfonamide (2 eq) and DBU (2 eq) were added. This solution was heated at 50 °C for 15h. The reaction mixture was then cooled to room temperature and concentrated under reduced pressure. The residue was partitioned between EtOAc and 1N HCI, the organic layer washed with brine, dried (MgSO₄) and evaporated. Purification by flash chromatography (gradient EtOAc (0 to 25%) in DCM) gave 10 mg (20%) of the title compound 57: m/z = 784 (M+H)⁺.

Example 10: Preparation of 17-[2-(3-isopropylpyrazol-1-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl2,14-dioxo-3,13-diazatriciclo[ 13.3.0.0⁴⁶]octadec-7-en-4-carboxylic acid (65).

Step 1: Synthesis of ethyl 4-hydroxy-7-methoxy-8-methylquinoline-3-carboxylate (58).

<img file="ECSP088150A_D0127.tif" />

Ό

Diethyl ethoxymethylene malonate (17.2 g, 79.6 mmol) was added to 2-methyl-m-anisidine (8.4 g, 61.2 mmol) (exothermic reaction). Then diethyl ether (100 ml) was added and the mixture was stirred overnight at room temperature. The solvent was evaporated and the residue redissolved in ether (50 ml), filtered, washed with heptane and dried to give 12 g of intermediate. This intermediate was added in portions to diphenyl ether (50 mL) preheated to 230 °C. The reaction mixture was subsequently heated at 250°C for 1.5h, cooled to room temperature and diluted with heptane (200ml). The precipitate was filtered and subsequently washed with heptane and ether to give 9.2 g (57.5%) of the target product 58 as a yellow powder: m/z=262 (M + H)⁺.

Step 2: Synthesis of 4-Hydroxy-7-methoxy-8-methylquinoline (59).

<img file="ECSP088150A_D0128.tif" />

<img file="ECSP088150A_D0129.tif" />

A suspension of ethyl 4-hydroxy-7-methoxy-8-methylquinoline-3-carboxylate (58, 9.2 g,

35.2 mmol) in 5N NaOH (150 mL) for 1.5 h (until a clear solution was obtained). The solution was then cooled to 0 °C and the pH was adjusted to 2-3 with concentrated HCI. The solid was filtered and subsequently washed with water, acetone and ether. This powder was added in small portions to diphenyl ether (40 mL), preheated to 250 °C. The resulting suspension became a solution after 20 min (the formation of CO was observed₂). After 1h at 250°C the brown solution was cooled to room temperature and diluted with heptanes (200ml). The precipitate was filtered and washed with heptanes and ether to give 6.4 g (96%) of the target product 59 as a yellow powder: m/z = 190 (M + H)⁺.

Step 3: Synthesis of 4-Chloro-7-methoxy-8-methylquinolin (60).

<img file="ECSP088150A_D0130.tif" />

OH 60Cl

A solution of 4-hydroxy-7-methoxy-8-methylquinoline (59, 6.4 g, 33.8 mmol) was heated in POCI₃(17.2 g, 111.6 mmol) at reflux for 1 h under nitrogen. The resulting solution was then cooled to room temperature and the excess POCI₃ evaporated under reduced pressure. The residue was partitioned between cold 1N NaOH and AcOEt. The organic layer was dried (Na₂SW₄) and evaporated. The product was purified by silica gel filtration (AcOEt/CH₂IC₂/Heptane, 4:4:2) to give 6.5 g (92.5%) of the desired product 60 as yellow needles: m/z=208 (M+H)⁺.

Step 4: Synthesis of 4-chloro-7-methoxy-8-methylquinolin N-oxide (61).

OR, AND

<img file="ECSP088150A_D0131.tif" />

<img file="ECSP088150A_D0132.tif" />

Cl

Cl 61 Cl

Metachloroperbenzoic acid (90.2 g, 366.0 mmol) was added portionwise over 3 h to a solution of 4-chloro-7-methoxy-8-methylquinolin (60, 15.2 g, 73.2 mmol ) in CHCI₃ (1L). The solution was then partitioned between cold 1N NaOH and CH₂IC₂ (8 successive extractions). The organic layers were combined, dried (Na₂SW₄) and evaporated. The product was purified by column chromatography (AcOEt/CH gradient₂IC₂0.1:2 to 1:0) to give 3.0 g (18.3%) of the title product 61 as a pale yellow powder: m/z = 224 (M + H)⁺.

Step 5: Synthesis of 4-benz¡loxy-7-methoxy¡-8-methylquinoline Μ-oxide (62).

NaH (973 mg, 60% in mineral oil, 24.3 mmol) was added at 0 °C, under an inert atmosphere, to benzyl alcohol (2.96 mL, 28.6 mmol) in DMF (10 mL). After 5 min at 0 °C, the solution was warmed to room temperature. After 10 min at room temperature, 4-chloro-7-methoxy-8-methylquinoline /V-oxide (61, 3.2 g, 14.3 mmol) was added in one part. The resulting black solution was stirred at room temperature under an inert atmosphere for another 30 min, then poured into cold water and extracted 4 times with AcOEt. The combined organic layers were dried (Na₂SW₄) and evaporated. The product was purified by column chromatography (AcOEt/CH gradient₂IC₂1:1 to 1:0, then AcOEt/MeOH 9:1) to give 2.5 g (59%) of the target product 62 as a yellow powder: m/z=296 (M + H)⁺.

Step 6: Synthesis of 4-benzyloxy-2-chloro-7-methoxy-8-methylquinoline (63).

.0.

.0.

Cl

POCI added₃ under an inert atmosphere at -78 °C to A/-4-benzyloxy-7-methoxy-8-methylquinoline oxide (62, 2.5 g, 8.47 mmol). The reaction mixture was then allowed to warm to room temperature, then heated to reflux. After 35 min, the solution was cooled to room temperature and excess POCI₃ evaporated under reduced pressure. The residue was partitioned between cold water and AcOEt, dried (Na₂SW₄) and evaporated. The residue was triturated in ether, then filtered and subsequently washed with small parts of methanol and ether to give 2.4 g (90.4%) of the target product 63 as a white powder: m/z = 314 (M + h)⁺.

Step 7: Synthesis of 4-hydroxy-2-(3-isopropylpyrazol-1-yl)-7-methoxy-8-methylquinoline (64).

<img file="ECSP088150A_D0133.tif" />

A mixture of 4-benzyloxy¡-2-chloro-7-methoxy¡-8-methylquinoline (63.1.00 g, 3.19 mmol) and 3-isopropylpyrazole was heated at 155 °C for 12h. The reaction mixture was then partitioned between AcOEt and water, dried (Na₂SW₄) and evaporated. The product was purified by column chromatography (AcOEt/CH₂IC₂, 1:1) to give 900 mg (95%) of the desired product 64 as a yellowish powder: m/z = 298 (M + H)⁺. Step 8: Synthesis of 17-[2-(3-¡sopropylpyrazol-1-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo acid [13.3.0.0^{4 yes}]octadec-7-en-4-carboxylic acid (65).

<img file="ECSP088150A_D0134.tif" />

either

<img file="ECSP088150A_D0135.tif" />

The title compound was prepared from 4-hydroxy-2-(3-isopropylpyrazol-1-yl)-7-methoxy-8-methylquinoline (64) and intermediate 26 following the procedure (Step DF) reported for the preparation of acid 17 -[7-methoxy-8-methyl-2-(thazol-2-yl)quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13d¡azatr¡ciclo[13.3. 0.0^4,6]octadec-7-en-4-carboxylic (29): m/z= 630 (M+H)⁺.

Example 11: Preparation of /V-[ 17-[2-(3-isopropylpyrazol-1 -yl)-7-methoxy-8-methylqui nolin-4-yloxy]-13-methyl I-2,14dioxo-3 ,13-diazatricyclo[13.3.0.0^4s]octadec-7-en-4-carbonyl](cyclopropyl)sulfonamide (66).

<img file="ECSP088150A_D0136.tif" />

The title compound was prepared from 17-[2-(3-isopropylpyrazol-1-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13 acid -diazatricyclo-[13.3.0.0^4,6]octadec-7-en-4-carboxylic acid (65) and cyclopropylsulfonamide following the procedure reported for the preparation of /V-[17-[8-chloro-2-(4isopropylthiazol-2-yl)-7-methoxyquinolin-4- yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^{4 yes}]-octadec-7-in-

4-carbonyl](cyclopropyl)sulfonamide (56): m/z= 733 (M+H)⁺. ¹H NMR (CDCI₃): 0.80-1.50 (m, 12H), 1.65-1.78 (m, 1H), 1.79-2.05 (m, 4H), 2.15-2.31 (m , 1H), 2.32-2.48 (m, 2H), 2.49-2.63 (m, 5H), 2.84-2.96 (m, 2H), 3.03 (s, 3H ), 3.05-3.14 (m, 1H), 3.33-3.42 (m, 2H), 3.61-3.70 (m, 1H), 3.96 (s, 3H), 4.60 (t, J = 12.3 Hz, 1H), 5.04 (t, J= 10.6 Hz, 1H), 5.26-5.46 (m, 1H), 5.61-5 .69 (m, 1H), 6.32 (d, J = 2.5 Hz, 1H), 6.37 (br s, 1H), 7.13 (d, J = 9.0 Hz, 1H), 7.30 (s, 1H), 7.95 (d, 9.0 Hz, 1H), 8.68 (d, J = 2.5 Hz, 1H), 10.88 (br s,

1 HOUR).

Example 12: Preparation of 17-[8-ethyl-2-(4-isopropylthiazol-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3. 0.0^{4 6}]octadec-7-en-4-carboxylic acid (70).

Step 1: Synthesis of Λ/-[2-(1-hydroxyethyl)-3-methoxyphenyl]pivalo¡lam¡dida (66).

<img file="ECSP088150A_D0137.tif" />

<img file="ECSP088150A_D0138.tif" />

A solution of /V-butyllithium (2.5 M in hexanes, 4.4 mL, 11.1 mmol) was added dropwise at 0 °C under nitrogen to a stirred solution of A/-(3-methoxyphenyl )p¡valoylamide. After 1h at room temperature, the reaction mixture was cooled to -78°C. Then, a solution of acetaldehyde (544 μΙ, 9.64 mmol) in THF (1 mL) was added. After 10 min, the reaction mixture was allowed to warm to room temperature over 30 min. The reaction mixture was then partitioned between AcOEt and water, dried (Na₂SW₄) and evaporated to give 500 mg (45%) of the target product 66 as a yellow solid: m/z = 252 (M+H)⁺.

Step 2: Synthesis of A/-[2-ethyl-3-methoxyphenyl]pivaloylamide (67).

<img file="ECSP088150A_D0139.tif" />

<img file="ECSP088150A_D0140.tif" />

A mixture of /V-[2-(1-hydroxyethyl)-3-methoxyphenyl]pivaloylamide (66.42 g, 167 mmol), Pd/C (10%, 2.00 g) and H was stirred.₂SW₄ (10 ml) in acetic acid (400 ml) at room temperature for 30 minutes. The resulting reaction mixture was then hydrogenated for 4 days, after which the catalyst was removed by filtration over kieselghur. The filtrate was concentrated to 300 mL, then poured into 1.0 L of water. The solid formed was filtered, washed with water to give the desired product 67 as a yellow solid: m/z = 236 (M+H)⁺.

Step 3: Synthesis of 2-ethyl-m-anisidine (68).

<img file="ECSP088150A_D0141.tif" />

A solution of /V-[2-ethyl-3-methoxyphenyl]pivaloylamide (67.167 mmol) and 37% HCl (700 mL) in EtOH (700 mL) was refluxed for 48 h. The reaction mixture was then cooled to room temperature and concentrated under reduced pressure (1/3 volume). This solution was kept at 5 °C for 6 h. The solid that appeared was filtered and washed with diisopropylether to give 22.35 g of the sought product as its HCI salt. The free base was generated by treatment with K₂CO₃ to give 20.85 g (83%) of the desired product 68: m/z =152 (M+H)⁺.

Step 4: Synthesis of 8-eth¡l-4-h¡drox¡-2-(4-¡soprop¡lt¡zol-2-yl)-7-methoxy¡quinol¡na (69).

<img file="ECSP088150A_D0142.tif" />

<img file="ECSP088150A_D0143.tif" />

The title compound was prepared from 2-ethyl-m-anisidine (68) following the procedure (Steps 3-5) reported for the preparation of 4-hydroxy¡-2-(4-isopropylthiazol-2-yl)- 7-methoxy-8-methylquinoline (36): m/z = 329 (M+H)⁺.

Step 5: Synthesis of 17-[8-ethyl-2-(4-isopropylthiazol-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo3,13-diazatricyclo[13.3 acid .0.0⁴⁶]octadec-7-en-4-carboxylic acid (70).

<img file="ECSP088150A_D0144.tif" />

The title compound was prepared from 8-ethyl-4-hydroxy-2-(4-isopropylthazol-2-yl)-7-methoxyquinoline (69) and intermediate 43 following the procedure (Steps DF) reported for the preparation of 17-[2-(4-¡soprop¡lt¡azol-2-¡l)-7-methoxy-8-methylquinol¡n-4-¡lox¡]-13-methyl-2,14 acid -d¡oxo-3,13diazatricyclo[13.3.0.0⁴⁶]octadec-7-en-4-carboxylic acid (46): m/z= 661 (M+H)⁺.

Example 13: Λ/-[17-[8-ethyl-2-(4-isopropylthiazol-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13diazatricyclo[13.3 .0.0^{4 6}]octadec-7-en-4-carbonyl](cyclopropyl)-sulfonamide (71).

<img file="ECSP088150A_D0145.tif" />

The title compound was prepared from 17-[8-ethyl-2-(4-isopropylthiazol-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14 acid -dioxo-3,13-diazatricyclo[13.3.0.0⁴⁶]-octadec-7-en-4-carboxylic acid (70) and cyclopropylsulfonamide following the procedure reported for the preparation of W-[17-[8-chloro-2-(4isopropylthiazol-2-yl)-7-methoxyquinolin- 4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatriciclo-[13.3.0.0^{4 6}]-octadec-7en-4-carbonyl](cyclopropyl)sulfonamide (56): m/z = 764 (M+H)⁺.

Example 14: Preparation of 17-[8-fluoro-2-(4-isopropylthiazol-2-¡l)-7-methoxyquinolin-4-yloxy]-13-methyl2,14-dioxo-3,13-diazatricyclo acid [13.3.0.0^4,6]-octadec-7-en-4-carboxylic acid (73).

Step 1: 8-fluoro-4-hydroxy-2-(4-isopropylthiazol-2-yl)-7-methoxyquinoline (72).

<img file="ECSP088150A_D0146.tif" />

The title compound was prepared from 2-fluoro-3-methoxybenzoic acid following the procedure (steps 1-5) reported for the preparation of 4-hydroxy-2-(4-isoprop¡ltiazol-2-¡l)- 7-methoxy-8-methylquinoline (36): m/z = 319 (M+H)⁺.

Step 2: Synthesis of 17-[8-fluoro-2-(4-isopropylthiazol-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo3,13-diazatric acid [13.3.0.0⁴⁵]octadec-7-en-4-carboxylic (73)

<img file="ECSP088150A_D0147.tif" />

The title compound was prepared from 8-fluoro-4-hydroxy-2-(4-isopropylthiazol-2-yl)-7-methoxyquinoline (72) and alcohol 43 following the procedure (steps DF) reported for the preparation of acid 17 -[2-(4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3. 0.0⁴⁶]octadec-7-en-4-carboxylic acid (46): m/z= 651 (M+H)⁺.

Example 15: /\/-[17-[8-fluoro-2-(4-¡soprop¡lt¡azol-2-¡l)-7-methox¡au¡nol¡n-4-¡lox¡l- 13-methyl-2_he14-d¡oxo-3₁13diazatricyclo[13.3.0.0^{4 6}]octadec-7-en-4-carbonyl](cyclopropyl)sulfonamide (74).

<img file="ECSP088150A_D0148.tif" />

The title compound was prepared from 17-[8-fluoro-2-(4-isoprop¡lt¡zol-2-yl)-7methoxyquinolin-4-yloxy¡]-13-methyl-2,14-dioxo acid -3,13-diazatricyclo[13.3.0.0⁴⁶]-octadec-7-en-4-carboxylic acid (73) and cyclopropylsulfonamide following the procedure reported for the preparation of /V-[17-[8-chloro-2-(4isopropylthiazol-2-yl)-7-methoxyquinolin -4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0⁴ °]-octadec-7-in-

4-carbonyl](cyclopropyl)sulfonamide (56): m/z=754 (M+H)⁺. ¹H NMR (CDCI3):¹H NMR (CDCI3): 0.75-1.52 (m, 15H), 1.64-2.05 (m, 4H), 2.77 (m, 1H), 2.41 (m, 2H), 2.59 (m, 2H), 2.92 (m, 2H), 3.04 (s, 3H), 3.19 (m,

1H), 3.40 (m, 2H), 4.07 (s, 3H), 4.60 (m, 1H), 5.05 (t, J= 10.5 Hz, 1H), 5.37 ( m, 1H), 5.66 (m, 1H), 6.17 (s,

1H), 7.07 (s, 1H), 7.54 (s, 1H), 7.86 (m, 1H), 10.77 (broad s, 1H).

Example 16: 18-[2-(4-¡soproo¡lt¡azol-2-¡l)-7-methox¡-8-met¡lau¡nol¡n-4-¡lox¡1-2.15 acid -dioxo-3,14d¡azatr¡c¡clo[14.3.0.0^4,6lnonadec-7-en-4-carboxylic acid (80).

Step 1. Synthesis of /V-(hept-6-enyl)phthalimide (75).

<img file="ECSP088150A_D0149.tif" />

<img file="ECSP088150A_D0150.tif" />

A solution of potassium phthalamide (627 mg, 3.38 mmol) and 7-bromohept-1-ene in dry DMF (10 mL) was stirred at 100 °C under nitrogen for 1 h. The reaction mixture was then cooled to room temperature, filtered, diluted with ether and filtered again. The filtrate was concentrated under reduced pressure to give the target product75 as an oil, which was used without further purification in the next step: m/z=244 (M+H)⁺.

Step 2. Synthesis of 6-heptenylamine (76).

either

<img file="ECSP088150A_D0151.tif" />

A solution of /V-(hept-6-enyl)phthalimide (75, 66.2 g, 272 mmol) and hydrazine hydrate (19.8 mL, 408 mmol) in MeOH (1.0 I) was stirred at room temperature. atmosphere until the next day. Then the reaction mixture was cooled to room temperature and the solid was removed by filtration. The filtrate was diluted with ether and the solid formed was removed by filtration. The ether was evaporated under reduced pressure. Then, 5N HCl (50 mL) was added and the resulting mixture was stirred at reflux. After 45 min, the reaction mixture was cooled to room temperature and the solid formed was filtered. The pH of the filtrate was regulated to 3 at 0 °C with NaOH. The reaction mixture was then extracted with ether and dried (Na₂SW₄) and evaporated. The crude product was purified by distillation to give 34.57 g of the target product 76 as an oil: m/z=114 (M+H)⁺.

Step 3. Synthesis of the intermediary 77.

<img file="ECSP088150A_D0152.tif" />

The title compound was prepared from 6-heptenylamine (76) and 3-Oxo-2-oxabicyclo[2,2,1]heptane-5-carboxylic acid (22) following the procedure reported for the preparation of intermediate 23: m/z = 252 (M+H)⁺. The title compound was also prepared (82 isolated yield) using other coupling conditions (EDCI.HCI (1.1 eq.), HOAT (1.1 eq.) and diisopropylethylamine in

dry DMF).

Step 4. Synthesis of the intermediary 78.

<img file="ECSP088150A_D0153.tif" />

<img file="ECSP088150A_D0154.tif" />

<img file="ECSP088150A_D0155.tif" />

The title compound was prepared (65%) from intermediate 77 and LiOH following the reported procedure for the preparation of intermediate 24: m/z = 270 (M+H)⁺.

Step 5. Synthesis of the intermediary 79.

ooh

<img file="ECSP088150A_D0156.tif" />

The title compound was prepared (65%) from intermediate 78 and 1-(amino)-2-(vinyl)cyclopropanecarboxylic acid ethyl ester hydrochloride 25 following the procedure reported for the preparation of intermediate 26: m/z = 407 (M+F)⁺.

Step 6. Synthesis of acid 18-[2-(4-¡soprop¡lt¡zol-2-¡l)-7-methox¡-8-methylquinol¡n-4-¡lox¡]-2,15 -d¡oxo-3,14diazatricyclo[14.3.0.0⁴⁶]nonadec-7-en-4-carboxylic acid (80).

<img file="ECSP088150A_D0157.tif" />

The title compound was prepared from intermediate 79 and quinoline 36 following the procedure (Steps DF) reported for the preparation of 17-[2-(4-isopropylthiazol-2-yl)-7-methoxy¡-8-methylquinolin-4-acid. -yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^{4 6}] octadec-7-en-4-carboxylic acid (46): m/z = 647 (M+H)⁺.

Example_________17:_________A/-[18-[2-(4-isopropylthazol-2-yl)-7-methoxy-8-methylquinolin-4-yloxy]-2,15-dioxo-3,14 Diazatricyclo[ 14.3.0.0^{4 6}]n onadec-7-e η-4-carbon yl](cyclopropyl)-su If o namide (81).

<img file="ECSP088150A_D0158.tif" />

<img file="ECSP088150A_D0159.tif" />

The title compound was prepared from 18-[2-(4-isopropylthiazol-2-yl)-7-methoxy¡-8-methylquinolin-4-yloxy]-2,15-dioxo-3,14-diazatric¡ acid. clo[14.3.0.0^4,6]-nonadec-7-en-4-carboxylic acid (80) and cyclopropylsulfonamide following the procedure reported for the preparation of A/-[17-(2-(4isopropylthazol-2-yl)-7-methoxy-8-methylquinolin -4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0⁴ ’⁶]octadec-7-en-4-carbonyl](cyclopropyl)sulfonamide (47): m/z = 750 (M+H)⁺. ¹H NMR (CDCI₃): 0.90-0.96 (m, 1H), 1.1-1.2 (m, 4H), 1.39 (d, J= 6.9 Hz, 6H), 1.4-1, 55 (m, 5H), 1.80-1.92 (m, 5H), 2.15-2.25 (m, 1H), 2,302.40 (m, 1H), 2.45-2.55 ( m, 2H), 2.68 (s, 3H), 2.85-2.92 (m, 1H), 3.15-3.30 (m, 2H), 3.45-3.55 (m, 2H), 3.96 (s, 3H), 4.09 (dd, J = 11.5 Hz, J= 3.8 Hz, 1H), 4.61 (t, J = 7.9 Hz, 1H) , 4.99 (t, J = 9.0 Hz, 1H), 5.51-5.53 (m, 1H), 5.71 (dd, J = 18.6 Hz, J= 8.2 Hz, 1H), 6.86 (s, 1H), 7.03 (s, 1H), 7.20 (d, J= 9.2 Hz, 1H), 7.50 (s, 1H), 7.88 ( d, J= 9.2 Hz, 1H), 9.40 (br s, 1H).

Example_______18:_______A/-íí18-í2-[4-(¡soDrop¡l)t¡azol-2-¡ll-7-methox¡-8-met¡lau¡nolin-4-¡lox¡1-2.15 -d¡oxo-14-(4methox¡benz¡l)-3,14,16-tr¡azatr¡c¡cloí14.3.0.0⁴⁵1nonadec-7-en-4-11carbon11(cyclopropyl)sulfonamide (90).

<img file="ECSP088150A_D0160.tif" />

Step A: Synthesis of the intermediary 82.

<img file="ECSP088150A_D0161.tif" />

<img file="ECSP088150A_D0162.tif" />

<img file="ECSP088150A_D0163.tif" />

Boc-c/s-hydroxy-L-proline methyl ester (500 mg, 2.04 mmol), 4-hydroxy¡-2-[4(¡sopropyl)thiazol-2-yl]-7-methoxy- 8-methylquinoline (36, 769 mg, 2.04 mmol) and 2-diphenylphosphanylpyridine (751 mg,

2.86 mmol) in high vacuum for 1 h. Dry THF was then added under nitrogen and the resulting reaction mixture cooled to -15Ό. Then, DIAD drop exhaust was added. After 1h -5°C the solution was allowed to warm to room temperature. After 16h, the reaction mixture was partitioned between cold water and AcOEt. The organic layer was subsequently washed vigorously with 1M HCI and brine, dried (MgSO₄), was filtered and evaporated. Purification by column chromatography on silica gel (AcOEt/CH gradient₂IC₂, 0:10 to 5:95) gave 940 mg (85%) of the desired product 82 as a colorless oil: m/z=542 (M+H)⁺.

Step B: Synthesis of the intermediary 83.

<img file="ECSP088150A_D0164.tif" />

<img file="ECSP088150A_D0165.tif" />

83

A solution of LiOH (592 mg, 13.8 mmol) in water was added to a solution of intermediate 82 (1.5 g, 2.77 mmol) in 1:1 MeOH/THF. After 16h at room temperature, the reaction mixture was acidified to pH 3-4 with dil. HCI, extracted with AcOEt, washed with brine, dried (MgSO₄) and evaporated. The product was purified by flash chromatography (EtOAc/CH Gradient₂IC₂, 1:9 to 4:6) to give 1.26 g (86%) of the title product 83 as an orange oil: m/z = 528 (M+H)⁺.

Step C: Synthesis of the intermediary 84.

<img file="ECSP088150A_D0166.tif" />

<img file="ECSP088150A_D0167.tif" />

To a stirred solution of carboxylic acid 83 (1.26 g, 2.39 mmol) in dry DMF (20 mL) was added (1R,2S)-1-Amino-2-vinylcyclopropanecarboxylic acid ethyl ester tosylate (860 mg , 2.63 mmol) and diisopropylethylamine (1.04 mL, 5.98 mmol). Then, HATU (999 mg, 2.63 mmol) was added at 0 °C under nitrogen. The resulting solution was stirred at 0°C for 30 minutes, then at room temperature. After 4 h, the reaction mixture was diluted with water and extracted with AcOEt. The organic layers were combined and subsequently washed with saturated NaHCO solution.₃, water and brine, dried (MgSO₄) and evaporated. Purification by column chromatography (AcOEt/CH gradient₂IC₂, 0:1 to 2:8) gave 1.44 g (90%) of the title product 84 as a white solid: m/z = 665 (M+H)⁺.

Step D: Synthesis of the intermediary 85.

<img file="ECSP088150A_D0168.tif" />

<img file="ECSP088150A_D0169.tif" />

To a stirred solution of Boc-protected proline derivative 84 (1.44 g, 2.16 mmol) in CH₂IC₂(20 ml) trifluoroacetic acid (5 ml) was added. After 2h at room temperature, the reaction mixture was concentrated and the residue was partitioned between saturated NaHCO solution.₃ and CH₂IC₂. The organic layer was dried (MgSO₄) was filtered and concentrated to give 1.0 g (81%) of the title product 85 as a colorless oil: m/z=565 (M+H)⁺.

Step E: Synthesis of A/-(hept-6-enyl)-/\/-(4-methoxybenzyl)amine 86.

<img file="ECSP088150A_D0170.tif" />

_XEITHER_/0

A solution of hept-6-enylamine (2.0 g, 13.4 mmol) and anisaldehyde (1.79 mL, 14.7 mmol) in EtOH (50 mL) was stirred at room temperature for 1 h. then NaBH₄ (556 mg, 14.7 mmol) was added at 0 °C under nitrogen. The resulting solution was allowed to warm to room temperature over 4 h. The reaction mixture was then partitioned between ice water and CH₂IC₂, washed with brine, dried (Na₂SW₄) and evaporated. The product was purified by chromatography (AcOEt/CH gradient₂IC₂ 0:1 to 2:8, then CH₂IC₂/MeOH 9:1) to give 1.8 g (34%) of the title product 86 as a colorless oil: m/z =

2. 3. 4 (M+H)⁺.

Step F: Synthesis of the intermediary 87.

<img file="ECSP088150A_D0171.tif" />

To a solution of proline derivative 85 in THF (50 mL) was added NaHCO₃ (1.0g). Then, phosgene (4.7 mL, 20% solution in toluene) was added at 0 °C under nitrogen. After 1.5h the white solid was filtered and washed with THF and CH₂IC₂. The filtrate was then concentrated under reduced pressure and the residue redissolved in dry dichloromethane (50 ml). To this solution, NaHCOg (1.0 g) and protected amine 86 were added subsequently. After 16 h at room temperature, the reaction mixture was filtered. The filtrate was concentrated under reduced pressure, the resulting residue was purified by silica chromatography (AcOEt/CH gradient₂IC₂, 0:1 to 2:8) to give 1.36 g (90%) of the title product 87: m/z = 824 (M+H)⁺.

Step G: Synthesis of the intermediary 88.

<img file="ECSP088150A_D0172.tif" />

Hoveyda-Grubbs 1st generation catalyst (50 mg, 0.082 mmol) was added to a degassed solution of diene 87 (1.36 g, 1.65 mmol) in toluene (170 mL). The resulting solution was heated at 80 °C under nitrogen for 4 h. Then, the reaction mixture was concentrated and purified by flash chromatography (AcOEt/CH gradient₂IC₂, 0:1 to 2:8) to give 900 mg (65%) of the title product 88 as a brownish foam: m/z = 796 (M+H)⁺.

Step H: Synthesis of the intermediary 89.

<img file="ECSP088150A_D0173.tif" />

<img file="ECSP088150A_D0174.tif" />

89

A solution of LiOH (242 mg, 5.65 mmol) in water (20 mL) was added to a solution of ester 88 (900 mg, 1.13 mmol) in 1:1 MeOH/THF. The reaction mixture was stirred at 50 °C for 2 h, then cooled to room temperature, acidified to pH 3-4 with dil. HCI and extracted with AcOEt. The organic layers were subsequently combined, washed with brine, dried (MgSO₄), filtered and evaporated to give 840 mg (97%) of the title product 89 as a slightly yellow solid: m/z = 768 (M+H)⁺.

Step I: Synthesis of Λ/-[[18-[2-[4-(¡sopropyl)thiazol-2-yl]-7-methoxy-8-methylquinol¡n-4-yloxy]-2,15-dioxo- 14-(4methoxybenzyl)-3,14,16-triazatricyclo[14.3.0.0^{4 5}]nonadec-7-en-4-yl]carbonyl](cyclopropyl)sulfonamide (90).

<img file="ECSP088150A_D0175.tif" />

<img file="ECSP088150A_D0176.tif" />

A solution of carboxylic acid 65 (830 mg, 1.03 mmol) and carbonyldiimidazole (333 mg,

2.06 mmol) in dry THF (20 mL) at reflux under nitrogen for 2 h. The reaction mixture was then cooled to room temperature and cyclopropylsulfonamide (249 mg, 2.06 mmol) and DBU (313 mg, 2.06 mmol) were added. The resulting solution was stirred at 50°C for 12h, then cooled to room temperature. The reaction mixture was quenched with water and extracted with CH₂IC₂, washed with dil. HCl, dried (MgSO₄), was filtered and evaporated. The crude material was purified by column chromatography (CH₂IC₂/EtOAc, 80:20) and recrystallized from CH₂IC₂/ether to give 450 mg (50%) of the title product 90 as a white powder: m/z=871 (M+H)⁺; ¹H-NMR (CDCI₃): 1.05-1.61 (m, 18H), 2.00 (m, 1H), 2.12-2.22 (m, 2H), 2.59-2.70 (m, 5H), 2.96 (m, 1H), 3.15-3.20 (m, 3H), 3.63 (s, 3H), 3.71-3.78 (m, 2H), 3.88-3, 94 (m, 4H), 4.54 (d, J = 15 Hz, 1H), 5.08 (t, J = 8.5 Hz, 1H), 5.16 (t, J = 9.4 Hz, 1H), 5.38 (m, 1H), 5.75 (m, 1H), 6.45 (d, J = 8.4 Hz, 2H), 6.65 (d, J = 8.4 Hz, 2H), 7.03 (s, 1H), 7.10 (d, J = 9.1 Hz, 1H), 7.41 (s, 1H), 7.73 (d, J = 9.1 Hz, 1H), 7.76 (br s, 1H), 10.15 (br s, 1H).

Example 19: N-[[18-[2-[4-(isopropyl)thiazol-2-¡l]-7-methox¡-8-methylquinolin-4-¡lox¡]-2,15- dloxo-3,14,16triazatricyclo[14.3.0.0^fl’⁶]nonadec-7-en-4-yl]carbonyl](cyclopropyl)-sulfonamide (91).

EITHER

TFA (10 ml) was added to a solution of /V-[[18-[2-[4-(isopropyl)thazol-2-yl]-7-methoxy-8-methylquinolin-

4-yloxy]-2,15-dioxo-14-(4-methoxybenzyl)-3,14,16-triazatricyclo[14.3.0.0^4,6]nonadec-7-en-4yl]carbonyl](cyclopropyl)sulfonamide (90) in DCM (20 mL). After 30 min at room temperature, water (20 ml) was added to the reaction mixture and the pH was adjusted to 3-4 with NaHCO3. The organic layer was washed with brine, dried (Na2SO4), filtered, and evaporated. The product was purified by column chromatography (MeOH/CH2CI2 gradient, 0:1 to 1:99, then 1:1 EtOAc/CH2CI2) to give 313 mg (73%) of the desired title product 91 as a yellowish solid. : m/z = 751 (M+H)⁺. ¹H-NMR (CDCI3): 0.88-1.64 (m, 16H), 1.96 (m, 2H), 2.52 (m, 1H), 2.68 (ms, 5H), 2.79 -2.92 (m, 3H), 3.18 (m, 1H), 3.63-3.69 (m, 2H), 3.86 (m, 1H), 3.97 (s, 3H), 4.34 (m, 1H), 4.59 (m, 1H), 5.08 (m, 1H), 5.40 (m, 1H), 5.80 (m, 1H),

6.73 (S, 1H), 7.03 (s, 1H), 7.21 (d, J = 8.9 Hz, 1H), 7.26 (br s, 1H), 7.47 (s, 1H), 7.92 (d, J=8.9Hz, 1H),

10.20 (brs, 1H).

Example 20: /V-[[18-[8-chloro-2-[4-(¡soDroD¡l)t¡azol-2-¡l1-7-methox¡au¡nolin-4-¡lox¡1- 2.15-dioxo-3.14.16 triazatricyclo[14.3.0.0^{4 6}]nonadec-7-en-4-yl]carbonyl](cyclopropyl)sulfonamide (94).

Step A: Synthesis of 4,8-dichloro-2-(4-isopropylthiazol-2-yl)-7-methoxyquinoline (92).

<img file="ECSP088150A_D0177.tif" />

<img file="ECSP088150A_D0178.tif" />

A solution of 8-chloro-4-hydroxy-2-(4-isopropylthiazol-2-yl)-7-methoxy-quinoline (2.0 g, 5.97 mmol) was heated in POCI₃ (10 ml) at 85°C for 30 min. Then the reaction mixture was concentrated under reduced pressure. The residue was poured into cold water (20 mL), the pH adjusted to 10 with 50% NaOH, and extracted with CH₂IC₂The organic layer was washed with brine, dried (MgSO₄), filtered and evaporated to give 2.05 g (97%) of the title compound 92 as a yellow solid: m/z = 353 (M+H)⁺.

Step B: Synthesis of the intermediary 93.

<img file="ECSP088150A_D0179.tif" />

<img file="ECSP088150A_D0180.tif" />

<img file="ECSP088150A_D0181.tif" />

NaH (60% in mineral oil, 679 mg, 17.0 mmol) was added under nitrogen to a solution of Boctrans-hydroxy-L-proline-OH (2.0 g, 5.661 mmol) in dry DMF (50 mL). After 30 min at room temperature, a solution of 4,8-dichloro-2-(4-isopropylthiazol-2-yl)-7-methoxyquinoline (92, 1.38 g, 5.94 mmol) was added ) in dry DMF and the resulting solution was stirred overnight at room temperature. The reaction mixture was then quenched with dil. HCl to pH 2, extracted twice with AcOEt, and the combined organic layers washed with brine, dried (MgSO₄) and evaporated. The product was purified by column chromatography (AcOEt/CH gradient₂IC₂, 0:1 to 1:1) to give 2.35 g (75%) of title 93: m/z = 548 (M+H)⁺.

Step C: Synthesis of /V-[[18-[8-chloro-2-[4-(¡soprop¡l)t¡azol-2-yl]-7-methox¡quinol¡n-4-¡ lox¡]-2,15-d¡oxo-3,14,16triazatricyclo[ 14.3.0.0^{4 yes}]nonadec-7-en-4-iIjcarbonyl]-(cyclopropiI)sulfonamide (94).

<img file="ECSP088150A_D0182.tif" />

The title compound was synthesized from intermediate 93 following the procedure (Steps

Cl) reported for M-[[18-[2-[4-(¡sopropyl)thiazol-2-yl]-7-methoxy-8-methylquinol¡n-4-yloxy]-2,15-dioxo-14- (4methoxybenzyl)-3,14,16-triazatricyclo-[14.3.0.0^{4 6}]nonadec-7-en-4-yl]carbonyl](cyclopropyl)sulfonamide (90) and for W-[[18-[2-[4-(isopropyl)thiazol-2-yl]-7-methoxy- 8-methylquinolin-4-yloxy]-2,15-dioxo-3,14,16triazatricyclo[14.3.0.0^4s]nonadec-7-en-4-yl]carbonyl](cyclopropyl)-sulfonamide (91): m/z = 771 (M)⁺; ¹H-NMR (CDCI_g): 0.93 (m, 1H), 1.06-1.63 (m, 15H), 1.92 (m, 3H), 2.50 (m, 1H), 2.64 (m, 2H) , 2.76 (m, 1H), 2.87 (m, 2H), 3.20 (m, J= 6.9 Hz, 1H), 3.70 (m, 1H), 3.77-3, 87 (m, 1H), 4.00 (dd, J= 4.0 Hz, 10.1 Hz, 1H), 4.04 (s, 3H), 4.42 (m, 1H), 4.59 ( t, J= 7.3 Hz, 1H), 5.05 (dd, J = 8.3 Hz, 9.9 Hz, 1H), 5.51 (m, 1H), 5.79 (m, 1H) , 7.03 (m, 1H), 7.08 (s, 1H), 7.22 (d, J= 9.3 Hz, 1H), 7.54 (s, 1H), 7.95 (d, 9.3Hz, 1H).

Example 21: /V-[[18-[8-chloro-2-[4-(isopropyl)t¡azol-2-¡l]-7-methoxyquinolin-4-yloxy]-2,15-d¡oxo- 3,14,16triazatricyclo [ 14.3.0.0^{4 6}] nonadec-7-en-4-i Ijcarbon yl]( 1 -methylcyclopropi I )sulio nam id (95)

<img file="ECSP088150A_D0183.tif" />

The title compound was synthesized from intermediate 93 and 1-Methyl-cyclopropylsulfonamide following the procedure (Steps Cl) reported for A/-[[18-[2-[4-(isopropyl)thiazol-2-yl]-7 -methoxy-8-methylquinolin-4-yloxy]-2,15-dioxo-14-(4-methoxybenzyl)-3,14,16-triazatricyclo[14.3.0.0^{4 6}]nonadec-7-en-4yl]carbonyl]-(c¡cloprop¡l)sulfonam¡de (90) and for Λ/-[[18-[2-[4-(¡soprop¡l)t¡azol- 2-¡l]-7-methoxy-8-methylquinol¡n-4yloxy]-2,15-dioxo-3,14,16-triazatricyclo[14.3.0.0^4s]-nonadec-7-en-4-yl]carbonyl](cyclopropyl)sulfonamide (91): m/z = 785 (M)⁺. ¹H-NMR (CDCI₃): 0.90 (m, 1H), 1.12-1.60 (m, 16H), 1.74 (m, 1H), 1.90-1.99 (m, 4H), 2.51 ( m, 1H), 2.65-2.78 (m, 3H), 2.88 (m, 1H), 3.20 (m, J= 6.7 Hz, 1H), 3.69 (m, 1H ), 3.84 (m, 1H), 3.96-4.00 (m, 1H), 4.01 (s, 3H), 4.46 (m, 1H), 4.63 (t, J= 7.4 Hz, 1H), 5.09 (t, J = 9.1 Hz, 1H), 5.50 (m, 1H), 5.79 (m, 1H), 7.08 (m, 2H) , 7.22 (d, J= 9.2 Hz, 1H), 7.52 (s, 1H), 7.95 (d, J= 9.2 Hz, 1H), 10.08 (br s, 1H ).

Example 22: {17-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13- Diazatricyclo[13.3.0.0^{4 6}Cyclopropanesulfonic acid ]octadec-7-en-4-carbonyl}-amide (103).

<img file="ECSP088150A_D0184.tif" />

Step A: Synthesis of 6-methylpyridine-2-carboxylic acid (6-acetyl-3-methoxy-2-methylphenyl)-amide (96).

<img file="ECSP088150A_D0185.tif" />

6-Methylpicolinic acid (1.12g, 8.167mmol) was dissolved in dry DCM (100ml) and kept in an ice bath in an ice bath. Then, 6-acetyl-3-methoxy-2-methylanyl-line (1.48 g, 8.17 mmol) and pyridine (6.6 mL, 0.082 mol) were added followed by dropwise addition of POCI₃ (1.53 mL, 0.016 mol) for 15 minutes. The resulting solution was stirred at -5 °C for 1h. Then, water (100 mL) was carefully added and after 5 min of stirring, NaOH (40%, 20 mL) was added dropwise consecutively, followed by separation of the organic layer. The water layer was extracted three times with CH₂IC₂ and the combined organic layers were washed with brine, dried (MgSO₄), were filtered and evaporated. The product was purified by column chromatography (Heptane/AcOEt, 3:1) to give the title compound (2.1 g, 86%): m/z = 299 (M+H)⁺.

Step B: Synthesis of 4-hydroxy-2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinoline (97).

<img file="ECSP088150A_D0186.tif" />

<img file="ECSP088150A_D0187.tif" />

To a solution of 6-methylpyridine-2-carboxylic acid (6-acetyl-3-methoxy-2-methylphenyl)-amide (96) in pyridine (15 mL) was added 2.5 equivalents of freshly ground KOH together with water ( 200 μΙ). The mixture was heated by microwave irradiation at 150 °C for 30 min, then 80-85% of the pyridine was evaporated under reduced pressure. The residue was poured on ice and neutralized with acetic acid. The precipitate was filtered, then dried to give the title compound (1.8 g, 95%): m/z=299 (M+H)⁺.

Step C: Synthesis of 2-(1-ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinoline-4- acid tert-butyl ester yloxy]-cyclopentanecarboxylic acid (98).

<img file="ECSP088150A_D0188.tif" />

A solution of 2-(1-ethoxycarbonyl-2-vinylcyclopropylcarbamoyl)-4-hydroxycyclopentanecarboxylic acid tert-butyl ester (500 mg, 1.5 mmol), prepared as described in W02005/073195, 4hydroxy¡-2-(6 -methyl-2-pyridyl)-7-methoxy-8-methylquinoline (97, 504 mg, 1.8 mmol) and triphenylphosphine (990 mg, 3.75 mmol) in dry THF (40 mL) at 0 °C for 10 min. DIAD (0.74 mL, 3.75 mmol) was then added dropwise. The resulting reaction mixture was stirred at a temperature of 0^yesC at 22°C until the next day. The volatiles were then evaporated and the product was purified by column chromatography on silica gel (gradient CH₂IC₂/AcOEt, 1:0 to 95:5) to give 1.1 g (88%) of the title compound 98: m/z=630 (M+H)⁺.

Step D: Synthesis of 2-(1-ethoxycarbonyl-2-vinylcyclopropylcarbamoyl)-4-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxyjcyclopentanecarboxylic acid (99).

<img file="ECSP088150A_D0189.tif" />

TFA (24 mL) was added at room temperature to a solution of 2-(1-ethoxycarbonyl-2-vinylcyclopropylcarbamoyl)-4-[2-(6-methyl-2-pyridyl) acid tert-butyl ester. -7-methoxy-8-methylquinol¡n-4yloxyjcyclopentanecarboxylic acid (98, 1.1 g, 1.75 mmol) and triethylsilane (510 mg, 2.5 eq) in CH₂IC₂ (24 mi). After 2h the reaction mixture was concentrated under reduced pressure and then co-evaporated with toluene. The residue was redissolved in AcOEt and subsequently washed with NaHCO solution.₃ and brine. The organic layer was dried (MgSO₄), filtered and evaporated to give 800 mg (80%) of the title compound 99 (800 mg, 80%): m/z=574 (M+H)⁺.

Step E: Synthesis of 1 -{2-(hex-5-en¡lmethylcarbamoyl)-4-[2-(6-methyl-2-pyridyl)-7-methoxy¡-8-methylquinol¡n-4- acid ethyl ester yloxy]cyclopentanecarbonyl}amino-2-vinylcyclopropanecarboxylic acid (100).

<img file="ECSP088150A_D0190.tif" />

A solution of 2-(1-ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-[2-(6-methyl-2-pyridyl)7-methoxy-8-methylquinolin-4- yloxy]-cyclopentanecarboxylic acid (99, 0.77 g, 1.344 mmol), /V-methylhex-

5-enylamine (221 mg, 1.95 mmol) and diisopropylethylamine (1.17 mL, 6.72 mmol) in DMF (25 mL) at 0 °C under an inert atmosphere. After 30 min, HATU (741 mg, 1.95 mmol) was added and the reaction mixture was allowed to warm to room temperature overnight. The DMF was then evaporated and the residue was partitioned between AcOEt and NaHCO solution.₃. The organic layer was washed successively with water and brine, dried (MgSO₄), was filtered and evaporated. The crude product was purified by silica gel chromatography (Heptane/AcOEt gradient 80:20 to 50:50) to give 735 mg (82%) of the title compound: m/z = 669 (M+H)⁺.

Step F: Synthesis of 17-[2-(6-methylpyridin-2-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-acid ethyl ester

2,14-dioxo-3,13-diaza-tricyclo[13.3.0.0^4,6]octadec-7-en-4-carboxylic acid (101).

<img file="ECSP088150A_D0191.tif" />

1-{2-(Hex-5-enylmethylcarbamoyl)-4-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]cyclopentanecarbonyl} ethyl ester was dissolved amino-2-vinyl-cyclopropanecarboxylic acid (100, 250 mg, 0.37 mmol) in

Dry 1,2-dichloroethane (250 ml). Nitrogen gas was then bubbled through the solution for 30 min before adding 2nd generation Hoveyda-Grubbs (25 mg). The resulting solution was refluxed overnight, then cooled to room temperature and evaporated. The product was purified by silica gel column chromatography (AcOEt/Heptane gradient, 3:7 to 5:5) to give 139 mg (58%) of the title compound 101.

Step G: Synthesis of 17-[2-(6-methyl-2-pyrid¡l)-7-methox¡-8-methylquinolin-4-¡lox¡]-13-methyl-2,14-dioxo acid -3,13diazatricyclo[13.3.0.0⁴⁶]octadec-7-en-4-carboxylic acid (102).

<img file="ECSP088150A_D0192.tif" />

LiOH (0.42 mL, 1M) was added to a solution of 17-[2-(6-methylpyridin-2-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-acid ethyl ester. -methyl-2,14-dioxo-3,13-diaza-tricyclo[13.3.0.0^{4 6}]octadec-7-en-4-carboxylic acid (101.27 mg, 0.042 mmol) in a THF:MeOH:H mixture₂Or, 2:1:1 (6 mi). The resulting solution was stirred at room temperature overnight, then the pH was adjusted to 6 with acetic acid. The reaction mixture was further diluted with water, extracted with CH₂IC₂, dried (MgSO₄), filtered and evaporated to give 17 mg (65%) of the title compound: m/z=613 (M+H)⁺.

Step H: Synthesis of {17-[2-(6-methyl-2-p¡r¡d¡l)-7-methoxy¡-8-methyl-quinolin-4-yloxy¡]-13-methyl-2, 14-dioxo-3,13diaza-tricyclo[13.3.0.0^{4 6}Cyclopropanesulfonic acid ]octadec-7-en-4-carbonyl}-amide (103).

either

103

A mixture of 17-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-acid was heated

2,14-dioxo-3,13-diazatricyclo[13.3.0.0⁴⁶]octadec-7-en-4-carboxylic acid (102.28 mg, 0.046 mmol) and CDI (15 mg, 0.092 mmol) in dry THF (3 mL) at reflux for 2 h under nitrogen. Activation was monitored by LC-MS. The reaction mixture was cooled to room temperature and cyclopropylsulfonamide (17 mg, 0.137 mmol) was added. Then, DBU (16 pL, 0.105 mmol) was added and the reaction was warmed to 55 °C. After 24 h, the pH of the reaction mixture was adjusted to 3 with citric acid (5%). Then the solvent was evaporated and the residue was partitioned between AcOEt and water. The crude material was purified by preparative HPLC to give 17 mg (52%) of the target compound 103: m/z=716 (M+H)⁺. Example 23: {17-í2-(6-¡soprop¡l-2-p¡r¡d¡l)-7-methox¡-8-met¡l-qu¡nol¡n-4-ilox¡l- 13-methyl-2_he14-d¡oxo-3₁13-diazatricyclo[13.3.0.0⁴⁶Cyclopropanesulfonic acid ]octadec-7-en-4-carbonyl}-amide (114).

either

114

Step A: Synthesis of 2-¡sopropylpyridine-/\/-oxide (104).

either

104

A mixture of isorpopylpyridine (2.1 g, 17.75 mmol) and m-CPBA (5.0 g, 1.3 eq.) was stirred in CH₂IC₂overnight at room temperature. Then the reaction mixture was diluted with CH₂IC₂ (twice the volume) and subsequently washed with aqueous sodium carbonate (twice) and brine, dried (Na₂SW₄) and evaporated to give 2.0 g (85%) of the title compound 104.

Step B: Synthesis of 2-cyano-6-isopropylpyridine (105).

A mixture of 2-isopropylpyridine-/V-oxide (104, 1.33 g, 9.7 mmol), cyanotrimethylsilane (TMSCN) (1.42 mL, 1.06 g, 11.0 mmol) was stirred in 1, 2-dichloroethane (40 mL) at room temperature for 5 min. Then, diethylcarbamoyl chloride (Et₂NCOCI, 1.23 mL, 9.7 mmol) and the mixture was stirred at room temperature under an inert atmosphere. After 2 days, an aqueous potassium carbonate solution (10%) was added and stirring was continued for 10 min. The organic layer was separated and the water layer was extracted twice with 1,2-dichloroethane. The combined organic layers were washed with brine, dried (Na₂SW₄) and evaporated. The product was purified by silica gel column chromatography (Hexanes/AcOEt, 3:1) to give 1.06 (74%) of the title compound: m/z = 147 (M+H)⁺.

Step C: Synthesis of 6-¡sopropylpyridine-2-carboxylic acid (106).

A solution of 2-cyano-6-isorpopylpyridine (105, 1.06 g, 7.3 mmol) in 37% HCI-aq. MeOH (1:2) was heated at reflux overnight. Then the solvent was evaporated and the residue was poured into saturated KOH solution. The resulting solution was refluxed overnight. The solution was then further cooled to room temperature and the pH was adjusted to 5 by adding aqueous HCI. The resulting reaction mixture was subsequently extracted with chloroform, washed with brine, dried (Na₂SW₄) and evaporated to give 0.97 g (81%) of the title compound 106: m/z = 166 (M+H)⁺.

Step D: Synthesis of 6-isopropyl-pyridine-2-carboxylic acid (6-acetyl-3-methoxy-2-methylphenyl)amide (107).

either

<img file="ECSP088150A_D0193.tif" />

<img file="ECSP088150A_D0194.tif" />

POCI added₃ (0.88 mL, 9.53 mmol) at -25 °C dropwise over 5 min under nitrogen, to a stirred solution of 6-isopropylpyridine-2-carboxylic acid (106, 1.43 g, 8.66 mmol ) and 6-acetyl-3-methoxy-2-methylaniline (1.55 g, 8.66 mmol) in dry pyridine (70 mL). The resulting solution was stirred at -10 °C for

2.5 hrs. Then the reaction mixture was poured on ice, neutralized with aqueous sodium carbonate and extracted 3 times with AcOEt. The organic layers were combined, washed with brine, dried (Na₂SW₄) and evaporated. The product was purified by column chromatography (hexanes/AcOEt,

3:1) to give 3.54 g (72%) of the title compound 107: m/z = 327 (M+H)⁺.

Step E: Synthesis of 4-hydroxy-2-(6-isopropyl-2-pyridyl)-7-methoxy-8-methylquinoline (108).

<img file="ECSP088150A_D0195.tif" />

108

To a solution of 6-isopropylpyridine-2-carboxylic acid (6-acetyl-3-methoxy-2-methylphenyl)amide (107, 0.70 g, 2.14 mmol) in pyridine (5 mL) is 2.5 equivalents of freshly ground KOH were added together with water (50 μΙ). The mixture was heated by microwave irradiation at 133 °C for 55 min, then 80-85% of the pyridine was evaporated under reduced pressure. The residue was poured on ice and neutralized with acetic acid. The precipitate was filtered, then dried to give 0.62 g (95%) of the title compound 108 (1.8 g, 95%): m/z = 309 (M+H)⁺.

Step F: Synthesis of 2-(1-ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-[2-(6-isopropylpyridin-2-yl)-7-methoxy-8-meth acid tert-butyl ester ¡l-quinolin-4-yloxy]-cyclopentanecarboxylic acid (109).

<img file="ECSP088150A_D0196.tif" />

The title compound was prepared in 62% isolated yield of 4-hydroxy-2-(6-hydroxy-cyclopentanecarboxylic acid) tert-butyl ester. isopropyl-2-pyridyl)7-methoxy-8-methylquinoline (108) following the procedure reported for the preparation of 2-(1-ethoxycarbonyl-2-vinylcyclopropylcarbamoyl)-4-[2-( 6-methyl-2-pyridyl)-7-methoxy-8-methylquinolin-4yloxyjcyclopentanecarboxylic (98): m/z= 658 (M+H)⁺.

Step G: Synthesis of 2-(1-ethoxycarbon¡l-2-vinylcyclopropylcarbamoyl)-4-[2-(6-isopropyl-2-pyridyl)-7-methoxy8-methí Iquino I yn-4-ylox ¡ acid Jcyclope ntancarboxylic (110)

<img file="ECSP088150A_D0197.tif" />

TFA (5 mL) was added at room temperature to a solution of 2-(1-ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-[2-(6-isopropyl-pyridin-2- ¡l)-7-methoxy-8-methyl-quinolin-4-yloxy]cyclopentanecarboxylic acid (109, 590 mg, 0.90 mmol) and triethylsilane (280 mg, 2.5 eq) in CH₂IC₂ (5 mi). After 2h, the reaction mixture was concentrated under reduced pressure to give the desired product 110, which was used in the next step without further purification.

Step H: Synthesis of 1-{2-(hex-5-enylmethylcarbamo¡l)-4-[2-(6-¡soprop¡l-2-p¡r¡d¡l)- 7-methox¡-

8-methylquinoline-4-¡loxyjcyclopentanecarbon¡l}am¡no-2-vinyl-cyclopropanecarboxylic (111)

<img file="ECSP088150A_D0198.tif" />

Title compound 111 was prepared in 70% yield isolated from 2-(1-ethoxycarbonyl-2vin¡lcyclopropylcarbamoyl)-4-[2-(6-¡sopropyl-2-pyridyl)-7-methoxy acid -8-methylquinolin-4-yloxy]cyclopentanecarboxylic acid (110) following the procedure reported for the preparation of 1-{2-(hex-5-enylmethylcarbamoyl)-4-[2-(6-methyl-2- pyrid¡l)-7-methoxy-8-methylquinolin-4-yloxy¡]-cyclopentanecarbonyl}am¡no-2vinylcyclopropanecarboxylic acid (100): m/z = 697 (M+H)⁺.

Step I: Synthesis of 17-[2-(6-isopropyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl- acid ethyl ester

2,14-dioxo-3,13-diazatricyclo[13.3.0.0^4,6]octadec-7-en-4-carboxylic acid (112).

either

112

1-{2-(Hex-5-enylmethylcarbamoyl)-4-[2-(6-isopropyl-2-pyridyl)-7-methoxy¡-8-methylquinol¡n-4-yloxy]cyclopentanecarbonyl}amino acid ethyl ester was dissolved -2-vinyl-cyclopropanecarboxylic acid (111.438 mg, 0.50 mmol) in

dry 1,2-dichloroethane. Nitrogen gas was then bubbled through the solution for 30 min before adding 1st generation Hoveyda-Grubbs (15 mg). The resulting solution was refluxed for 3h, then more catalyst (20mg) was added. After 2 h at reflux, another 10 mg of the catalyst was added. After 12h at reflux, the reaction mixture was cooled to room temperature. Then, MP-TMT scavenging agent (Agronaut Technologies Inc.) (-300 mg) was added and the mixture was stirred at room temperature for 45 min. The catalyst was removed by silica gel filtration (CHClg/MeOH gradient, 1:0 to 98:2) to give 220 mg (66%) of the title compound 112: m/z=669 (M+H)⁺.

Step J: Synthesis of acid 17-[2-(6-¡soprop¡l-2-pyr¡d¡l)-7-methox¡-8-met¡lqu¡nol¡n-4-¡lox¡]- 13-methyl-2,14-dioxo-

3,13-diazatricyclo[13.3.0.0^4S]octadec-7-en-4-carboxylic acid (113).

either

ΌΗ

113

A solution of LiOH (40 mg) in water (1.5 mL) was added to a solution of 17[2-(6-¡sopropyl-2-pyridyl)-7-methoxy-8-acid ethyl ester. -methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13diazatricyclo[13.3.0.0⁴⁶]octadec-7-en-4-carboxylic acid (112, 220 mg, 0.33 mmol) in a mixture of MeOH (3 mL) and THF (1 mL). The resulting solution was subsequently heated at 55°C for 3h, then stirred at room temperature for 5h. Then the pH of the reaction mixture was adjusted to pH 6 with acetic acid and water (3 mL) was added. The resulting solution was extracted with CHCI.₃. Then the organic layer was dried (Na₂SW₄), filtered and evaporated to give 200 mg (95%) of the title compound 113 as a white powder: m/z=641 (M+H)⁺.

Step K: Synthesis of {17-[2-(6-¡sopropyl-2-p¡r¡d¡l)-7-methoxy-8-methyl-quinol¡n-4-yloxy]-13-methyl-2 ,14-dioxo-3,13d¡aza-tric¡clo[13.3.0.0⁴⁶Cyclopropanesulfonic acid ]octadec-7-en-4-carbon¡l}amide (114).

either

114

A solution of 17-[2-(6-isopropyl-2-pindyl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-acid was stirred

2,14-dioxo-3,13-diazatricyclo[13.3.0.0^4,6]octadec-7-en-4-carboxylic acid (113, 200 mg, 0.31 mmol), DMAP (76.5 mg, 0.62 mmol) and EDC (151 mg, 0.78 mmol) in DMF (5 mL ) at room temperature overnight (acid activation was monitored by LC-MS). Then, cyclopropylsulfonamide (191 mg, 1.56 mmol) was added, followed by DBU (228 μι, 1.56 mmol). The resulting solution was stirred overnight at room temperature, then neutralized with acetic acid and evaporated. The residue was redissolved in MeOH and purified by preparative HPLC to give 90 mg (39%) of the title compound 114: m/z = 744 (M+H)⁺.

Example 24: f17-[2-(2-c¡clohex¡lt¡azol-4-¡l)-7-methox¡-8-met¡lau¡nol¡n-4-ylox¡1-13-methyl¡ l-2,14-d¡oxo-3.13-d¡azatricyclo[13.3.0.0⁴⁶(6S)-Cyclopropanesulfonic acid ]octadec-7-en-4-carbonyl}amide (123) and {17-(2-(2-cyclohexylthiazol-4-yl)-7-methoxy-8-methylquinolin-4-yloxy]- 13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^4,6(6R)-cyclopropanesulfonic acid ]octadec-7-en-4-carbon¡l}amide (124).

Step A: Synthesis of cyclohexanecarbothioic acid amide (115).

yes

<img file="ECSP088150A_D0199.tif" />

115

To a suspension of cyclohexanecarboxamide (10 g, 78.6 mmol) in diethyl ether (300 mL) was added phosphorus pentasulfide (9.0 g, 200 mmol) in three parts over 5 h. After stirring overnight the reaction mixture was filtered. The mother liquor was evaporated to give 5.5 g (49%) of the title compound 115.

Step B: Synthesis of 2-cyclohexylthiazole-4-carboxylic acid ethyl ester (116).

<img file="ECSP088150A_D0200.tif" />

116 ⁰

A solution of cyclohexanecarbothioic acid amide (115, 5.5 g, 38.3 mmol) and ethyl 3-bromopyruvate (90%, 8.3 g, 38.3 mmol) in THF (200 mL) was heated to reflux. After 2h, the reaction mixture was cooled to room temperature over 12h. The solvent was then evaporated and the product was purified by column chromatography (heptane/AcOEt gradient, 90:10 to 75:25) to give 6.8 g (74%) of the title compound 116 as a clear liquid.

Step C: Synthesis of 2-cyclohexylthiazole-4-carboxylic acid (117).

<img file="ECSP088150A_D0201.tif" />

117 ⁰

To a solution of 2-cyclohex¡lt¡azole-4-carboxylic acid ethyl ester (116, 6.8g, 28.5mmol) in water was added 1M LiOH (50mL). The solution was kept at room temperature and monitored by LCMS. When the hydrolysis was complete, the reaction mixture was neutralized with myriatic acid and extracted with ethyl acetate and diethyl ether. The organic phase was dried (Na₂SW₄), filtered and concentrated under reduced pressure to give 5.0 g (83%) of the title compound 117: m/z=212 (M+H)⁺.

Step D: Synthesis of 2-cyclohexylthiazole-4-carboxylic acid (6-acetyl-3-methoxy-2-methylphenyl)amide (118).

<img file="ECSP088150A_D0202.tif" />

either

118

POCI added₃ (1.4 mL, 14.9 mmol) dropwise at -35 °C for 5 min, to a stirred solution of 2-cyclohexylthiazole-4-carboxylic acid (117.1.5 g, 7, 1 mmol) and 2-acetyl-5-methoxy-6-methylaniline (1.27 g, 7.1 mmol) in dry pyridine (40 mL). After 1h, the reaction mixture was subsequently warmed to room temperature for 2.5h, evaporated and neutralized with aqueous sodium bicarbonate solution. The precipitate was filtered, washed with water and dried to give 2.6 g (95%) of the title compound 118: m/z =373 (M+H)⁺.

Step E: Synthesis of 2-(2-cyclohexylthiazol-4-yl)-4-hydroxy-7-methoxy-8-methylquinoline (119).

<img file="ECSP088150A_D0203.tif" />

ooh

119

Freshly ground KOH (2 mmol, 112 mg) was added to a 2-cyclohexylthiazol-acid solution.

4-carboxylic (6-acetyl-3-methoxy¡-2-methylphenyl)amide (118, 373 mg, 2 mmol) in pyridine (20 mL). The mixture was divided into several batches and each batch was individually heated by microwave irradiation at 150 °C for 30 min. The different batches were then combined and the pyridine was evaporated. The residue was treated with aqueous citric acid to give a suspension, which was further diluted with a small volume of EtOH, then partitioned between water and CH₂IC₂. The organic layer was dried (Na₂SW₄) and evaporated. The product was purified by column chromatography (CH gradient₂IC₂:MeOH, 1:0 to 93:7) to give 1.8 g (72.5%) of the title compound 119 as a white powder: m/z = 355 (M+H)⁺.

Step F: Synthesis of 1-{[4-[2-(2-cyclohexylthiazol-4-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-2(hex-5-enylmethylcarbamoyl) ethyl ester )cyclopentanecarbonyl]amino}-2-vinylcyclopropanecarboxylic acid (120).

<img file="ECSP088150A_D0204.tif" />

Title compound 120 was prepared in 42% yield from 1-{[4[2-(2-c¡clohex¡lt¡azol-4-¡l)-7-methoxy¡-8-acid ethyl ester. -methyl-quinolin-4-¡lox¡]-2-(hex-5-en¡lmethylcarbamo¡l)cyclopentanecarbonyl]amino}-2-vinylcyclopropane-carboxylic acid (120) following the procedure reported for the Preparation of 1-{2-(hex-5-enylmethylcarbamoyl)-4-[2-(6-isopropyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]cyclopentanecarbonyl}amino-acid ethyl ester 2-vinyl-cyclopropanecarboxylic (111): m/z= 743 (M+H)⁺.

Step G: Synthesis of 17-[2-(2-cyclohexylthiazol-4-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13methyl-2,14-dioxo-3, ethyl ester 13-diazatricyclo[13.3.0.0^{4 6}]octadec-7-en-4-carboxylic acid (121).

<img file="ECSP088150A_D0205.tif" />

Title compound 121 was prepared in 50% yield from 2-(2-cyclohexylthiazol-4-yl)-

4-hydroxy-7-methoxy-8-methylquinoline (119) and 2-(1-ethoxycarbonyl-2vinylcyclopropylcarbamoyl)-4-hydroxycyclopentanecarboxylic acid tert-butyl ester following the procedure reported for the preparation of ethyl ester 17-[2-(6-methylpyridin-2-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diaza-tricyclo[13.3. 0.0⁴⁶]octadec-7-en-4-carboxylic acid (101): m/z = 715 (M+H)⁺.

Step H: Synthesis of 17-[2-(2-cyclohexylthiazol-4-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-acid

3,13-diazatricyclo[13.3.0.0⁴⁶]octadec-7-en-4-carboxylic acid (122).

<img file="ECSP088150A_D0206.tif" />

An aqueous LiOH solution (1M, 5 mL) was added to a solution of 17-(2-(2-cyclohexylthiazol-4-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-acid ethyl ester. -methyl-2,14-dioxo-3,13diazatriciclo[13.3.0.0⁴⁶]octadec-7-en-4-carboxylic acid (121) in MeOH (10 ml), THF (20 ml) and water (5 ml). The resulting solution was stirred at 50 °C for 19 h. Then the pH of the reaction mixture was adjusted to 6 with myriatic acid (3M, 1.7 mL). The resulting solution was evaporated on silica gel and purified by column chromatography (AcOEt/MeOH/AcOH, 74:25:1) to give 273 mg (95%) of the title compound 122 as a white powder: m/z = 687 (M+H)⁺.

Step I: Synthesis of {17-[2-(2-cyclohex¡ltiazol-4-yl)-7-methoxy¡-8-methylquinol¡n-4-¡loxy]-13-methyl-2,14-dioxo- 3,13diaza-tricyclo[13.3.0.0^{4 6}(6S)-cyclopropanesulfonic acid ]octadec-7-en-4-carbonyl}amide (123) and {17-[2(2-cyclohexylthiazol-4-i I)-7-methoxy-8-methylqu i η o I yn-4-yloxy]-13-methyl I-2,14-dioxo-3,13-diazatriciclo[13.3.0.0⁴⁶(6R)-cyclopropanesulfonic acid ]octadec-7-en-4-carbon¡l}amide (124).

<img file="ECSP088150A_D0207.tif" />

<img file="ECSP088150A_D0208.tif" />

123 124

A solution of 17-[2-(2-cyclohex¡lt¡zol-4-yl)-7-methoxy-8-methylquinolin-4-yloxy¡]-13methyl-2,14-dioxo-3 acid was heated ,13-diazatricyclo[13.3.0.0⁴⁶]octadec-7-en-4-carboxylic acid (122, 173 mg, 0.25 mmol) and CDI (81 mg, 0.5 mmol) in THF (7.5 mL) at reflux for 2h (acid activation is monitored by LCMS). The reaction mixture was then cooled to room temperature and cyclopropylsulfonamide (91 mg, 0.75 mmol) and DBU (8 μΙ, 0.575 mmol) were added subsequently. After 12h the reaction mixture was neutralized with acetic acid, evaporated. The residue was redissolved in water and acetonitrile, then purified by preparative HPLC to give 21 mg (11%) of the title compound (123, first isomer): m/z=790 (M+H)⁺ and 35 mg (18%) of the second isomer 124: m/z= 790 (M+H)⁺.

Example 25: Preparation of /V-[ 17-[2-(3-isopropylpyrazol-1 -yl)-7-methoxy-8-methylqu i nolin-4-yloxy]-13-methyl I-2,14dioxo-3 ,13-diazatricyclo[13.3.0.0^{4 6}]octadec-7-en-4-carbonyl][1-(methyl)cyclopropyl]sulfonamide (125).

<img file="ECSP088150A_D0209.tif" />

125

The title compound was prepared from 17-[2-(3-isopropylpyrazol-1-yl)-7-methoxy¡-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13 acid -diazatric¡clo-[13.3.0.0^4,6]octadec-7-en-4-carboxylic acid (65) and 1-methylcyclopropylsulfonamide following the procedure reported for the preparation of /V-[17-[8-chloro-2(4-isopropylthiazol-2-yl)-7-methoxyquinolin- 4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^{4 6}]octadec-7en-4-carbonyl](cyclopropyl)sulfonamide (56): m/z = 747 (M+H)⁺. ¹H NMR (CDCI₃): 0.79-0.92 (m, 2H), 1,202.03 (m, 19H), 2.20-2.32 (m, 1H), 2.35-2.48 (m, 2H), 2.52-2.64 (m, 5H), 2.85-2.93 (m, 1H), 3.04 (s, 3H), 3.05-3.14 (m, 1H), 3, 35-3.46 (m, 2H), 3.97 (s, 3H), 4.60 (td, J = 13.2 Hz, J = 2.2 Hz, 1H), 5.04 (t, J = 10.5 Hz, 1H), 5.30-5.47 (m, 1H), 5.61-5.69 (m, 1H), 6.30 (s, 1H), 6.32 (d, J=2.4Hz, 1H), 7.12 (d, J=9.2Hz, 1H), 7.30 (s, 1H), 7.95 (d, J=9.0Hz, 1H), 8.61 (d, J= 2.5 Hz, 1H), 10.9 (brs, 1H).

Example 26: Preparation of acid 17-[2-(3-tert-butylpyrazol-1 -yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-

2,14-dioxo-3,13-diazatricyclo[13.3.0.0⁴⁶]octadec-7-en-4-carboxylic acid (127).

Step 1: Synthesis of 4-hydroxy-2-(3-tert-but¡lpírazol-1 -yl)-7-methoxy¡-8-methylquinoline (126).

The title compound was prepared from 4-benzyloxy¡-2-chloro-7-methox¡-8-methylquinol¡ne (63) and 3ter-butylpyrazole following the procedure reported for the preparation of 4-hydroxy -2-(3-isopropylpyrazol1 -iI)-7-methoxy-8-meth¡Iquinoline (64): m/z =312 (M+H)⁺.

Step 2: Synthesis of acid 17-[2-(3-ier-but¡lpyrazol-1-yl)-7-methoxy-8-methylquinol¡n-4-yloxy]-13-methyl-2,14- dioxo3,13-diazatricyclo[13.3.0.0^4S]octadec-7-en-4-carboxylic acid (127).

ooh

The title compound was prepared from 4-hydroxy-2-(3-tert-butylpyrazol-1-yl)-7-methoxy-8methylquinoline (126) and intermediate 26 following the procedure (Step DF) reported for the preparation of 17-[7-methoxy-8-methyl-2-(thiazol-2-yl)quinolin-4-ylox¡]-13-methyl-2,14-dioxo-3,13diazatricyclo[13.3. 0.0^{4 6}]octadec-7-en-4-carboxylic acid (29): m/z= 644 (M+H)⁺.

Example 27: Preparation of /V-[17-[2-(3-tert-butylpyrazol-1-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14dioxo-3 ,13-diazatricyclo[13.3.0.0^{4 6}]octadec-7-en-4-carbonyl](cyclopropyl)sulfonamide (128).

The title compound was prepared from 17-[2-(3-tert-butylpyrazol-1-yl)-7-methoxy¡-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3-acid. ,13-diazatricyclo-[13.3.0.0^4,6]octadec-7-en-4-carboxylic acid (127) and cyclopropylsulfonamide following the procedure reported for the preparation of /V-[17-[8-chloro-2-(4isopropylthiazol-2-yl)-7-methoxyquinolin- 4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^{4 yes}]-octadec-7-in-

4-carbonyl](cyclopropyl)sulfonamide (56): m/z=747 (M+H)⁺. ¹H NMR (CDCI₃): 0.95-1.12 (m, 2H), 1.13-1.30 (m, 2H), 1.31-1.55 (m, 11H), 1.63-2.05 (m , 4H), 2.20-2.55 (m, 9H), 2.80-2.98 (m, 1H), 3.03 (s, 3H), 3,363.47 (m, 2H), 3, 61-3.70 (m, 1H), 3.97 (s, 3H), 4.60 (t, J= 12.2 Hz, 1H), 5.04 (t, J= 10.3 Hz, 1H ), 5.26-5.46 (m, 1H), 5.61-5.69 (m, 1H), 6.35 (d, J= 2.5 Hz, 1H), 6.42 (brs, 1H), 7.13 (d, J= 9.1 Hz, 1H), 7.32 (s, 1H), 7.95 (d, J= 9.1 Hz, 1H), 8.67 (d, 2.5 Hz, 1H), 10.9 (br s, 1H).

Example 28: Preparation of 17-[2-(3,5-dimethylpyrazol-1-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-acid

2,14-dioxo-3,13-diazatricyclo[13.3.0.0⁴⁶]octadec-7-en-4-carboxylic acid (130).

Step 1: Synthesis of 4-hydroxy-2-(3,5-dimethylpyrazol-1 -yl)-7-methoxy-8-methylquinoline (129).

The title compound was prepared from 4-benzyloxy¡-2-chloro-7-methoxy¡-8-methylquinol¡ne (63) and

3,5-dimethylpyrazole following the procedure reported for the preparation of 4-hydroxy-2-(3isopropylpyrazol-1 -yl)-7-methoxy-8-methylquinoline (64): m/z= 284 (M+H)⁺.

Step 2: Synthesis of 17-[2-(3,5-dimethylpyrazol-1-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14dioxo-3,13-diazatric acid cycle[13.3.0.0^4S]octadec-7-en-4-carboxylic (130).

<img file="ECSP088150A_D0210.tif" />

The title compound was prepared from 4-hydroxy-2-(3,5-dimethylpyrazol-1-yl)-7-methoxy-8-methylquinoline (129) and intermediate 26 following the procedure (Step DF) reported for the preparation of the 17-[7-methoxy-8-methyl-2-(thiazol-2-yl)quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13diazatricyclo[13.3.0.0 acid⁴⁶]octadec-7-en-4-carboxylic acid (29): m/z = 616 (M+H)⁺.

Example 29: Preparation of W-[ 17-[2-(3,5-dimethylpyrazol-1 -yl)-7-methoxy-8-methylqui nolin-4-yloxy]-13-methyl I-2,14dioxo- 3,13-diazatricyclo[13.3.0.0^{4 6}]octadec-7-en-4-carbonyl](cyclopropyl)sulfonamide (131).

<img file="ECSP088150A_D0211.tif" />

The title compound was prepared from 17-[2-(3,5-dimethylpyrazol-1-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3 acid, 13-diazatricyclo-[13.3.0.0⁴⁶]octadec-7-en-4-carboxylic acid (130) and cyclopropylsulfonamide following the procedure reported for the preparation of /V-[17-[8-chloro-2-(4isopropylthazol-2-yl)-7-methoxyquinolin- 4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^{4 3}]-octadec-7-en4-carbonyl](cyclopropyl)sulfonamide (56): m/z= 719 (M+H)⁺. ¹H NMR (CDCI₃): 0.70-0.96 (m, 1H), 1.1-1.2 (m, 5H), 1.4-1.55 (m, 2H), 1.80-1.93 (m , 4H), 2.15-2.25 (m, 1H), 2.30-2.40 (m, 2H), 3.30 (s, 3H), 2.45-2.55 (m, 2H ), 2.52 (s, 3H), 2.80 (s, 3H), 2.82-2.91 (m, 2H), 3.00 (s, 3H), 3.45-3.55 ( m, 2H), 3.95 (s, 3H), 4.51-4.60 (m, 1H), 4.99-5.1 (m, 1H), 5.21-5.33 (m, 1H), 5.51 (m, 1H), 6.00 (s, 1H), 7.03 (s, 1H), 7.10 (d, 9.1 Hz,

1H), 7.20 (s, 1H), 7.98 (d, J= 9.1 Hz, 1H), 10.80 (br s, 1H).

Example 30

ooh

<img file="ECSP088150A_D0212.tif" />

2-(1-Ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester (132) Boc-protected proline (4 g, 17.3 mmol), HATU ( 6.9 g, 18.2 mmol) and 1-amino-2-vinyl-cyclopropanecarboxylic acid ethyl ester, prepared as described in WO03/099274, (3.5 g, 18.3 mmol) in DMF (60 mL) and cooled down to 0^θ in an ice bath. Diisopropylethyl amine (DIPEA) (6ml) was added. The ice bath was removed and the mixture was allowed to stand at room temperature overnight. Dichloromethane (~80 mL) was then added and the organic phase was washed with aqueous sodium hydrogen carbonate, citric acid, water, brine, and dried over sodium sulfate. Purification by flash chromatography (ether -&gt; 7% methanol in ether) gave pure title compound (6.13 g, 96%).

<img file="ECSP088150A_D0213.tif" />

2-(1-ethoxy¡carbon¡l-2-v¡n¡lc¡cloprop¡lcarbamo¡l)-4-(4-n¡tro-benzoyloxy¡)-pyrrol¡d acid tert-butyl ester n-1carboxylic (133)

Compound 132 (6.13 g, 16.6 mmol), 4-nitrobenzoic acid (4.17 g, 25 mmol) and PPh were dissolved.₃ (6.55 g, 25 mmol) in THF (130 mL). The solution cooled to ~0^θ and diisopropyl azidocarboxylate (5.1 g, 25 mmol) was added slowly. The cooling was then removed and the mixture was left overnight at ambient conditions. Aqueous sodium hydrogen carbonate (60 mL) was added and the mixture was extracted with dichloromethane. Purification by flash chromatography (pentane-ether, 2:1 -»pentane-ether, 1:2 -&gt; 2% methanol in ether) gave pure title compound (6.2 g, 72%).

<img file="ECSP088150A_D0214.tif" />

4-nitro-benzoic acid 5-(1-ethoxy¡carbon¡l-2-v¡n¡lc¡cloprop¡lcarbamo¡l)-p¡rrol¡d¡n-3-¡l ester (134)

Compound 133 (6.2 g, 12 mmol) was dissolved in a cold mixture of trifluoromethanesulfonic acid, 33% CO in dichloromethane. The ice bath was then removed and the mixture was left at room temperature for ~1.5h. The solvent was evaporated and 0.25M sodium carbonate was added and the mixture was extracted with dichloromethane. Evaporation gave the title compound (4.8g, 95%) as a yellowish powder. Example 33

<img file="ECSP088150A_D0215.tif" />

5-(1-ethoxy¡carbon¡l-2-v¡n¡lc¡cloprop¡lcarbamo¡l)-1-[hept-6-en¡l-(4-methoxy¡-benzyl)-carbamo¡l] 4-nitro-benzoic acid -p¡rrol¡d¡n-3-¡l ester (135)

To a solution of compound 134 (4.5 g, 10.8 mmol) in THF (160 mL) was added NaHCO₃ (1 tablespoon) and phosgene in toluene (1.93 M, 11.5 mL, 22 mmol). The mixture was stirred vigorously for 1h at room temperature, then filtered and evaporated. The residue was dissolved in CH₂IC₂ (160 ml) and NaHCO were added₃ (1 tablespoon) and hept-5-enyl-(p-methoxybenzyl)-amine (4.3 g, 18.5 mmol). After stirring overnight at room temperature the reaction mixture was filtered and evaporated to dryness. Flash column chromatography on silica gel (25:75 -&gt; 40:60 EtOAc:toluene) gave the title compound (6.59 g, 90%) as a light brown jelly.

Example 34

ooh

<img file="ECSP088150A_D0216.tif" />

18-H¡drox¡-14-(4-methoxy¡-benz¡l)-2,15-d¡oxo-3,14,16-tñaza-tric¡clo[14.3.0.0* acid ethyl ester 4,61nonadec7-en-4-carboxylic (136)

Compound 135 (1g, 1.48 mmol) was dissolved in 1,2-dichloroethane (2 I). The mixture was degassed for 15 min using a stream of argon. Hoveyda-Grubbs (II) catalyst (50 mg, 5 mol%) was added and the mixture was refluxed for 4h. The solvent was evaporated and the crude ester was dissolved in tetrahydrofuran (100 ml), methanol (50 ml) and water (50 ml). The mixture was cooled 0^B.C in an ice bath. Aqueous lithium hydroxide (20 mL, 1M) was added and the mixture was stirred at 0^SC for 4h. The volume was then doubled with water and the mixture acidified with acetic acid. Extraction (dichloromethane) followed by flash chromatography (1-&gt;5% methanol in ether) gave pure title compound (450 mg, 61%).

MS (M+H)⁺ 500.

<img file="ECSP088150A_D0217.tif" />

18-[2-(4-lsoprop¡lt¡zol-2-¡l)-7-methox¡-8-methyl-quinol¡n-4-¡lox¡l-14- acid ethyl ester (4-methoxy-benz¡l)-2,15d¡oxo-3,14,16-t¡aza-tr¡c¡clo[14.3.0.0*4,6*lnonadec-7-en-4-carboxylic ( 137)

Alcohol 136 (230 mg, 0.460 mmol), quinolinol 36 (218 mg, 0.690 mmol), and tphenylphosphine (182 mg, 0.690 mmol) were dissolved in dry THF and the mixture cooled to 0°C. DIAD (130 μΙ, 0.690 mmol) was added dropwise to the stirred solution at 0°C over 30 minutes after which the solution was allowed to warm to room temperature and subsequently stirred overnight. The solvent was evaporated and the crude material was purified by flash column chromatography (toluene/ethyl acetate 1:1) to give the title compound (366 mg) (M+H)⁺ stall: 796.4; experimental: 796.7.

Example 36

<img file="ECSP088150A_D0218.tif" />

acid_________18-[2-(4-lsoorop¡lt¡azol-2-¡l)-7-methox¡-8-methyl-au¡nol¡n-4-¡lox¡l-14-(4-methox ¡-benz¡l)-2,15-d¡oxo3,14,16-t¡aza-tr¡c¡clo[14.3.0.0*4,6*1nonadec-7-en-4-carboxylic (138)

Ethyl ester 137 (366 mg, 0.460 mmol) was dissolved in THF/MeOH/H₂O 2:1:1 (30 mL) and 1M LiOH (4.6 mL, 4.40 mmol) was added dropwise at room temperature over 5 minutes after which the solution was stirred overnight. The mixture was acidified to pH 3-4 by the addition of solid citric acid and the organic solvents were evaporated. The water phase was diluted with brine (50 mL) and then extracted twice with DCM. The combined organic phase was washed twice with brine and subsequently dried, filtered and concentrated. The crude product was then purified by flash column chromatography (7:1 ethyl acetate/methanol) to give the title compound (212 mg, 60%). (M+H)⁺ stall: 768.3; experimental: 768.7.

<img file="ECSP088150A_D0219.tif" />

[18-í2-(4-¡soprop¡lt¡azol-2-¡l)-7-methox¡-8-methyl-au¡nol¡n-4-¡lox¡1-14-(4- methoxy-benz¡l)-2,15-d¡oxo-3,14,16t¡aza-tr¡c¡clo[14.3.0.0*4,6*lnonadec-7-en-4-carbonyl-am¡de of l-methyl-cyclopropanesulfonic acid (139) To acid 138 (212 mg, 0.276 mmol) dissolved in dichloromethane (7 mL) was added EDC (69 mg, 0.359 mmol) and the reaction mixture was stirred at room temperature. After 7 hours TLC and LC-MS indicated complete conversion of the starting material to the corresponding oxazolidinone. The reaction mixture was diluted with dichloromethane (20ml) and the organic phase washed twice with water after which the organic phase was dried, filtered and concentrated. The residue was dissolved in dichloromethane (5 mL) and cyclopropylmethyl sulfonamide (53 mg, 0.394 mmol) and DBU (78 μΙ, 0.525 mmol) were added and the reaction mixture stirred at room temperature for 20 hours. The mixture was diluted with dichloromethane (30 mL) and the organic phase was washed twice with 10% citric acid and once with brine. The organic phase was dried, filtered and concentrated and the crude product was purified by flash column chromatography (toluene/ethyl acetate 1:1, 1:2, ethyl acetate, ethyl acetate/methanol 9:1) to give the title compound (108 mg, 44%) as a colorless solid. LC-MS purity: &gt;95%. (M+H)⁺ stall: 885.4; experimental: 885.7.

Example 38 or

{18-[2-(4-isopropyl-thazol-2-¡l)-7-methoxy¡-8-methyl-au¡nolin-4-ylox¡l-2,15-dioxo-3, 1-Methyl-cyclopropanesulfonic acid 14,16-triazatricyclo[14.3.0.0*4,6*lnonadec-7-en-4-carbonyl}-amide (140)

To compound 139 (106 mg, 0.120 mmol) dissolved in dichloromethane (18 mL) were added triethylsilane (38 μΙ, 0.240 mmol) and TFA (9 mL) and the reaction mixture was stirred at room temperature for 1 hour. Solvents were evaporated and quickly co-evaporated with toluene. The residue was dissolved in dichloromethane and the organic phase was washed twice with NaHCO solution.₃ crowded. The organic phase was dried, filtered and concentrated and the crude product was purified by flash column chromatography (1:1 toluene/ethyl acetate) to give the title compound (73 mg, 80%) as a slightly yellow solid. . LC-MS purity: &gt;95%. (M+H)⁺ stall: 765.3; experimental: 765.7.

Example 39: Alternative route for the preparation of compound 34

Step A: Synthesis of 4-amino-5-cyano-2-hydroxy¡-3-methylbenzo¡co acid ethyl ester (141)

ΗΟ

<img file="ECSP088150A_D0220.tif" />

EITHER

To a solution of sodium ethoxide (1.3 L) (prepared freshly by adding sodium metal (7.9 g, 0.35 mol) to ethanol (1.3L)) at 0 °C was added sodium acetate. ethylpropionyl (25 g, 0.17 mol) and the solution stirred at rt for 1 h. To the above solution was added ethoxymethylene malononitrile (21 g, 0.17 mol) at rt and the reaction mixture was refluxed at 80 °C for 2h. The reaction mixture was cooled, neutralized to pH=7 by the addition of 11.5 N HC and concentrated in vacuo. The obtained residue was diluted with water (100 ml) and filtered. The solid was washed with water and dried in vacuo at 50 °C to give the crude product (27 g). The crude solid was washed with 5% ethyl acetate in pet. which gave pure title compound (22.5 g, 59%).

TLC: EtOAc/ Pet. ether, 3:7, R_F=0,4

Step B: Synthesis of 4-amino-5-cyano-2-hydroxy-3-methylbenzoic acid (142)

<img file="ECSP088150A_D0221.tif" />

nh₂

CN

To a solution of L¡OHxH₂0 (8.4 g, 0.2 mol) in ethanol/water (1:1, 300 mL) was added compound 74 (22 g, 0.1 mol) at rt and the reaction mixture was refluxed at 80° C for 4h. The reaction mixture was concentrated in vacuo, the residue obtained was diluted with water (100 ml), washed with pet. ether/ethyl acetate (1:1, 2x200 ml). The aqueous layer was separated, acidified to pH=5 using HC11.5N and the solid product obtained was filtered. The aqueous layer was further extracted with ethyl acetate (2x300 mL), dried, and concentrated to give more product. The combined products were washed with 5% ethyl acetate in pet. ether to give the pure title compound (19 g, &gt;95%).

TLC: MeOH/chloroform, 1:4, R_F=0,2

Step C: Synthesis of 2-Amino-4-hydroxy-3-methylbenzonitrile (143)^hovX/^nh₂

CN

A mixture of compound 75 (19 g, 0.1 mol) in quinoline (50 mL) was heated at 170 °C for 2h (until effervescence stopped). The reaction mixture was cooled to RT and aqueous NaOH solution (1M, 500 mL) was added followed by pet. ether (500 ml). The reaction mixture was stirred for 15 min and the aqueous layer was separated. The aqueous layer was further washed with pet. ether (2x300 ml) to completely remove the quinoline. The aqueous layer was acidified with 11.5N HC to pH=5, the solid filtered and dried in vacuo. The obtained solid was further washed with 5% ethyl acetate in pet. ether to give pure title compound (12 g, 82%).

TLC: EtOAc/Pet ether, 3:7, R_F=0,35

Step D: Synthesis of 2-Amino-4-methoxy-3-methylbenzonitrile (144)

<img file="ECSP088150A_D0222.tif" />

A mixture of compound 76 (12 g, 0.08 mol) was stirred, K₂CO₃ (11 g, 0.08 mol) in dry DMF (200 ml) for 15 min at RT. To this, Mel (13.6 g, 0.096 mol) was added and the mixture was stirred for 4h at RT. The reaction mixture was diluted with water (800 ml), extracted with 30% ethyl acetate in pet. ether (3x300 ml). The combined organic layers were washed with water and brine, dried, and concentrated to give a crude product. The crude product was washed with pet. ether to give pure title compound (12 g, 93%).

TLC: Pet. ether/ EtOAc, 7:3, R_F=0,4

Step E: Synthesis of 1-(2-Amino-4-methoxy-3-methyl-phenyl)-ethanone (34)

<img file="ECSP088150A_D0223.tif" />

To a solution of compound 77 (12 g, 0.074 mol) in THF (150 mL) was added MeMgBr in diethyl ether (3M, 100 mL, 0.296 mol) at 0 °C dropwise. The reaction mixture was stirred at rt for 1h and then at 55 °C for 3h. The reaction mixture was cooled to 0 °C, quenched with cold HC11.5N until effervescence stopped (pH=6). The reaction mixture was diluted with water (100ml), extracted with ethyl acetate (2x300ml). The combined organic layers were washed with brine, dried and concentrated to give a brown solid. The crude solid was dissolved in ethyl acetate (150 mL), pet. ether (150 mL) and passed through a pad of silica gel to remove color impurities and concentrated. The obtained solid was washed with 5% ethyl acetate in pet. ether which gave pure title compound (9 g, 68%) as a yellow solid.

TLC: Pet. ether/ EtOAc, 7:3, R_F=0,4,

Example 40:

Synthesis of 3-oxo-2-oxa-bicyclo[2,2,1]heptane-5-carboxylic acid tert-butyl ester (146)

<img file="ECSP088150A_D0224.tif" />

DMAP (14 mg, 0.115 mmol) and Boc were added₂0 (252 mg, 1.44 mmol) to a stirred solution of 145 (180 mg, 1.15 mmol) in 2 mL CH₂IC₂ under an inert argon atmosphere at 0 °C. The reaction was allowed to warm to room temperature and stirred overnight. The reaction mixture was concentrated and the crude product was purified by flash column chromatography (gradient toluene/ethyl acetate 15:1, 9:1, 6:1, 4:1, 2:1) to give the title compound (124 mg, 51%) as white crystals.

¹H-NMR (300 MHz, CD3OD) δ 1.45 (s, 9H), 1.90 (d, J=11.0 Hz, 1H), 2.10-2.19 (m, 3H), 2, 76-2.83 (m, 1H), 3.10 (s, 1H), 4.99 (s, 1H);¹³C-NMR (75.5 MHz, CD3OD) δ 27.1, 33.0, 37.7, 40.8, 46.1, 81.1, 81.6, 172.0,

177,7.

Alternative method for the preparation of compound 146

<img file="ECSP088150A_D0225.tif" />

, BF₃.et₂OR (0.5 eq.)

<img file="ECSP088150A_D0226.tif" />

DCM, -10°C, 70min

<img file="ECSP088150A_D0227.tif" />

Compound 145 (13.9 g, 89 mmol) was dissolved in dichloromethane (200 mL) and then cooled to approximately -10°C, under nitrogen. Isobutylene was then bubbled into the solution until the total volume increased to approximately 250 mL which gave a cloudy solution. BF added_3iet₂O (5.6 mL, 44.5 mmol, 0.5 eq.) and the reaction mixture was kept at ca -10°C under nitrogen. After 10 min, a clear solution was obtained. The reaction was monitored by TLC (3:2 EtOAc-Toluene acidified with a few drops of acetic acid and 4:1 hexane-EtOAc, immunoblotting with basic permanganate solution). At 70 min, only traces of compound 145 remained and NaHCO was added.₃ saturated aqueous solution (200 mL) to the reaction mixture, which was then stirred vigorously for 10 min. The organic layer was washed with NaHCO₃ saturated (3 x 200 ml) and brine (1 x 150 ml), then dried with sodium sulphite, filtered and the residue evaporated to give an oily residue. On adding hexane to the residue, the product precipitated. Addition of more hexane and heating to reflux gave a clear solution from which the product crystallized. The crystals were collected by filtration and washed with hexane (rt), then air-dried for 72 h to give colorless needles (12.45 g, 58.7 mmol, 66%).

Synthesis of tert-butyl ester of the acid (1fí.2fí,4S)-2-((1fí.2S)-1-Ethox¡carbon¡l-2-vinyl-c¡cloDroD¡lcarbamo¡l)-4hydroxy- cycloDentancarboxylic (147)

ooh

<img file="ECSP088150A_D0228.tif" />

Compound 146 (56 mg, 0.264 mmol) was dissolved in 1:1 dioxane/water (5 mL) and the mixture cooled to 0 Ό. 1M Lithium hydroxide (0.52 mL, 0.520 mmol) was added and the mixture was stirred at 0 °C for 45 minutes, after which the mixture was neutralized with 1M hydrochloric acid and evaporated and coevaporated with toluene. The crystalline residue was dissolved in DMF (5 mL) and (1R2S)-1-amino-2-vinylcyclopropane carboxylic acid ethyl ester hydrochloride (60 mg, 0.313 mmol) and diisopropylethylamine (DIEA) (138 pL, 0.792 mmol) and the solution was cooled to 0^B.C. HATU (120 mg, 0.316 mmol) was added and the mixture was stirred for 0.5 h.^yesC and for an additional 2 h at room temperature. The mixture was then evaporated and extracted with EtOAc, washed with brine, dried, filtered, and concentrated. Purification by flash column chromatography (1:1 toluene/EtOAc) gave the title compound (86 mg, 89%) as a colorless oil. The oil crystallized from ethyl acetate-hexane.

Example 41: Activity of the compounds of formula (I)

replicon assay

Compounds of formula (I) were examined for activity in inhibiting HCV RNA replication in a cellular assay. The assay demonstrated that the compounds of formula (I) exhibited activity against functional HCV replicons in cell culture. The cell assay was based on a bicistronic expression construct, as described in the text written by Lohmann et al. (1999) Science vol. 285pp. 110-113 with the modifications described by Krieger et al. (2001) Journal of Virology 75:4614-4624, in a multi-target selection strategy. In essence, the method was as follows.

The assay used the stably transfected cell line Huh-7 luc/neo (hereinafter referred to as Huh-Luc). This cell line harbors a bicistronic expression construct encoding an RNA comprising the wild-type regions of HCV type 1b NS3-NS5B transfected from an encephalomyocarditis virus Internal Ribosome Entry Site (IRES). (EMCV), preceded by a reporter moiety (FfL-luciferase), and a selectable marker moiety (neo^R., neomycin phosphotransferase). The construct is bordered by 5' and 3' NTRs (untranslated regions) from HCV type 1b. Continuous cultivation of replicon cells in the presence of G418 (neo^R.) is dependent on HCV RNA replication. Stably transfected replicon cells expressing autonomously replicating HCV RNA to high levels, encoding inter alia luciferase, are used for evaluation of antiviral compounds.

Replicon cells were plated in 384-well plates in the presence of test and control compounds which are added at various concentrations. After a three-day incubation, HCV replication was measured by luciferase activity assay (using standard luciferase assay substrates and reagents and a Perkin Elmer ViewLux mycoplate imaging device).^tmultraHTS). Replicon cells in control cultures have high luciferase expression in the absence of an inhibitor. The inhibitory activity of the compound on luciferase activity was monitored on Huh-Luc cells, allowing a dose-response curve to be made for each test compound. EC50 values ​​were then calculated, which value represents the amount of compound required to reduce by 50% the level of detected luciferase activity, or more specifically, the replication capacity of genetically linked HCV replicon RNA.

inhibition assay

The objective of this in vitro assay was to measure the inhibition of HCV NS3/4A protease complexes by the compounds of the present invention. This assay provides an indication of the effectiveness of the compounds of the present invention in inhibiting the proteolytic activity of HCV NS3/4A.

Inhibition of the full-length hepatitis C NS3 protease enzyme was measured essentially as described in Poliakov, 2002 Prot Expression &amp; Purification 25 363 371. Briefly, the synthesis of a dipsipeptide substrate, Ac-DED(Edans) EEAbuip[COO]ASK(Dabcyl)-NH₂ (AnaSpec, San José, USA), was measured by spectrofluorometry in the presence of a peptide co-factor, KKGSVVIVGRIVLSGK (Áke Engstróm, Department of Medical Biochemistry and Microbiology, Uppsala University, Sweden). [Landro, 1997 #Biochem 36 9340-9348]. The enzyme (1 nM) was incubated in 50 mM HEPES, pH 7.5, 10 mM DTT, 40% glycerol, 0.1% n-octyl-D-glucoside, with 25 μΜ NS4A cofactor and inhibitor at 30⁵C for 10 min, after which the reaction was started with the addition of 0.5 μΜ substrate. Inhibitors were dissolved in DMSO, sonicated for 30 sec. and shaken with vertex. Solutions were stored at -20 °C between measurements.

The final concentration of DMSO in the test sample was adjusted to 3.3%. The hydrolysis rate was corrected for internal filter effects according to published procedures. [Liu, 1999 Analytical Biochemistry 267 331-335]. K¡ values ​​were estimated by nonlinear regression analysis (GraFit, Erithacus Software, Staines, MX, UK), using a model for competitive inhibition and a fixed value for Km (0.15 μΜ). A minimum of two replications was performed for all measurements.

Table 1 below lists compounds that were prepared according to any of the above examples. The activities of the tested compounds are also shown in Table 1.

<td>Compound No.</td><td>Structure</td><td>ΕΟ_ω(μΜ) test of replicon</td><td>K¡ (μΜ) enzyme assay</td>

<td> 29</td><td>iY φ⁵0 A'9.....i</td><td> 10</td><td></td>

<td> 47</td><td>V γγ 0 λ AA υϋϋ</td><td> 0,00618</td><td> 0,00050</td>

<td> 46</td><td>/0. xx _.χ A9 ' U7 ' 0 γο L1 k™ ¡¡ A° Φ</td><td> 0,91</td><td></td>

<td> 91</td><td>ι 'Ή'. ί 0 OR χ, - τμ......&lt; X™-ν</td><td>8.54 x 10'³</td><td>5.00 x 10'⁵</td>

<td> 55</td><td>. X , i » ji) 'V</td><td></td><td></td>

<td></td><td>/ Χθ: /.....</td><td> 0,36743075</td><td>5.00x10'³</td>

<td> 81</td><td>γ C. twenty</td><td></td><td></td>

<td></td><td>1 x LJ</td><td> 10</td><td> 1</td>

<td> 82</td><td>'a ι A''¹</td><td></td><td></td>

<td></td><td>F either. Q</td><td></td><td></td>

<td></td><td>yeah</td><td></td><td></td>

<td></td><td></td><td> 8,321539</td><td>9.40 x 10'³</td>

<td></td><td>X ΊX /UvA ?</td><td></td><td></td>

<td></td><td>,..Q í X</td><td></td><td></td>

<td> 56</td><td>XXX</td><td>2.93 x 10'³</td><td>1.00 x 10'⁴</td>

<td></td><td>\ - 3X</td><td></td><td></td>

<td></td><td>'X' p</td><td></td><td></td>

<td> 57</td><td>X Χχ·</td><td>1.87 x 10'³</td><td>3.00 x 10'⁴</td>

<td></td><td>VΪr$</td><td></td><td></td>

<td> 94</td><td>° 0 IX. = . XI......L¿</td><td>3.26 x 10'³</td><td>1.00 x 10'⁴</td>

<td></td><td></td><td></td><td></td>

<td></td><td>X⁷ 0</td><td></td><td></td>

<td> 48</td><td> \^XYx</td><td>2.33x10³</td><td>2.50 x 10'⁴</td>

<td></td><td> '&gt; ^ci Ά☆A^x Yo 0</td><td></td><td></td>

<td> 95</td><td>0 Y) rx- %k......&lt;k k&gt;</td><td>4.04 x 10'³</td><td>1.00 x 10'⁴</td>

<td></td><td>,X^k c</td><td></td><td></td>

<td> 75</td><td>Χζ⁰YS 4^z '^Vwhat^TO</td><td>5.75x10'²</td><td></td>

<td></td><td>---k” TO</td><td></td><td></td>

<td> 71</td><td>Χ,ι TΛ^m'rj_ or &lt;-4 { j</td><td> 10</td><td></td>

<td></td><td>AA</td><td></td><td></td>

<td> 103</td><td>°kX)k ,p xxk</td><td>6.30 x 10'³</td><td></td>

<td> 72</td><td>\_ J 0 Λ or o=¥ Ϊ &gt; hn-s·· 00V</td><td>6.60 x 10'²</td><td></td>

<td></td><td>ΧγΑΑ-/Χ 0777</td><td></td><td></td>

<td> 66</td><td>I f) kx^ y</td><td> 0,0036</td><td></td>

Example 42: In Vivo Effects of Ritonavir on the Pharmacokinetics of Compound No. 47 in Rats

The oral pharmacokinetics of compound No. 47 in male and female Sprague-Dawley rats were investigated after a single 10 mg/kg dose, using a formulation in 50% PEG400 in water and the influence of "boosting" with 10 mg/kg of ritonavir.

Four male and female Sprague-Dawley (SD) rats (approximately 200-250g body weight) were randomly divided into 2 groups of 2 males and 2 females each (boosted and non-boosted) based on body weight. The weight of individual animals did not differ too much from the group mean.

Animals were briefly fasted prior to testing. The ingested water remained available ad libitum.

Rats in the non-boosted group received a single oral dose of 10 mg/kg of compound No. 47, formulated as 3 mg/mL in PEG400/50% water pH 8. Rats in the boosted group received a single oral dose of ritonavir, about 30 minutes prior to receiving the single 10 mg/kg oral dosing of compound No. 47. Drug formulations were administered by oral priming.

A 0.5 ml blood sample was collected from each rat at 0.5h, 1h, 2h, 4h and 8h post dosing. Plasma concentrations were determined by using HPLC-MS. The results are shown in Table 2 below, expressed as a factor of change in the pharmacokinetic parameter of the boosted group, compared to the non-boosted group.

Table 2

<td></td><td>Pharmacokinetic parameter</td><td>ritonavir</td>

<td>compound no. 47</td><td>Cmax</td><td> 2,2</td>

<td></td><td>AUC</td><td> 2,5</td>

These results show that ritonavir essentially improves the pharmacokinetics of compound No.

in rats, increasing overall exposures expressed as AUC by more than 2-fold.