NO340695B1 - HCV NS-3 Serine Protease Inhibitors - Google Patents

Description

This invention relates to new compounds which are easier for the NS3-serine protease of the flavivirus HCV, which compounds have the structure and composition as stated in claim 1. The compounds are useful in the treatment or prevention of HCV.

The background of the invention

The NS3 serine protease of HCV is a multifunctional protein containing a serine protease domain and an RNA helicase domain. The protease cofactor NS4A, which is a relatively small protein, is absolutely necessary for enhanced serine protease activity. The NS3 serine protease is essential in the virus life cycle. From analysis of the substrate's binding site demonstrated by X-ray crystal structure, it has been shown that the binding site for the NS3 protease is remarkably surface- and solvent-exposed, making the design of small-molecule inhibitors a challenge.

It is believed that two HCV protease inhibitors have reached clinical trials, namely BILN-2061 described in WO 0059929 and VX-950 described in WO 0387092. A number of similar peptidomimetic HCV protease inhibitors have also been proposed in the academic and patent literature. Common to the vast majority of such previously known peptidomimetics is the presence of an L-proline derivative in the P2 position of the inhibitor and interacting with the S2 subunit of the HCV protease enzyme. Regarding BILN-2061, the L-proline is 4-substituted with a quinoline ether, while VX-950 has a carbocyclic ring fused to the L-proline ring. Most peptidomimetics additionally comprise additional L-amino acid derivatives peptide-linked at the P3 position, with many proposed inhibitors also comprising additional L-amino acid derivatives extending into P4, P5 and P6.

It has already been revealed that the delayed administration of BILN-2061 or VX-950 selects HCV mutants that are resistant to the respective drug, so-called drug escape mutants. These drug escape mutants have characteristic mutations in the HCV protease genome, particularly D168V, D168Y and/or A165S. Treatment paradigms for HCV will thus have to resemble HIV treatment, where drug-escape mutations also occur with ease. Consequently, additional drugs with different resistance patterns will consistently be needed to provide treatment options to unsuccessful patients, and multidrug combination therapy is likely to be the rule in the future, even for first-line therapy.

Experience with HIV drugs, and HlV protease inhibitors in particular, has further emphasized that sub-optimal pharmacokinetics and complex dosing regimens quickly result in unintended compliance failure. This in turn means that a 24 hour vessel concentration (minimum plasma concentration) for the respective drugs in an HIV regimen often falls below the IC90 or ED90 threshold for large parts of the day. It is considered that a 24 hour vessel occurrence of at least the IC50 value and, more realistically, the IC90 or ED90 value is essential to slow the development of drug escape mutants, and to achieve the necessary pharmacokinetics and drug metabolism to enable such vessel occurrences, constitute a major challenge for medicine development. The highly peptidomimetic nature of previously known HCV protease inhibitors, with multiple peptide bonds in naturally occurring configurations, pose pharmacokinetic obstacles to effective dosing systems.

Brief description of the invention

In accordance with a first aspect of the invention there are provided compounds of formula I:

in which

A is C(=0)OR<1>or C(=0)NHS02R<2>wherein;

R<1> is hydrogen, C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl;

R 2 is C 1 -C 6 alkyl, C 0 -C 3 alkylcarbocyclyl, C 0 -C 3 alkylheterocyclyl;

wherein R 2 is optionally substituted with 1 to 3 substituents independently selected from the group consisting of halo, oxo, nitrile, azido, nitro, Ci-

C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl, NH2C(=0)-, Y-NRaRb, Y-0-Rb, Y-C(=0)Rb, Y-(C=0)NRaRb, Y-NRaC(=0) Rb, Y-NHSOpRb, Y-S(=0)pRb, Y-S(=0)pNRaRb, Y-C(=0)ORbog Y-NRaC(=0)ORb;

Y is independently a bond or C 1 -C 3 alkylene;

R a is independently H or C 1 -C 3 alkyl;

R b is independently H, C 1 -C 6 alkyl, C 0 -C 3 alkylcarbocyclyl or C 0 -C 3 alkylheterocyclyl;

p is independently 1 or 2;

M is CR7R7' or NRu;

R<7> is C1-C6alkyl, C0-C3alkylC3-C7cycloalkyl or C2-C6alkenyl, each of which is optionally substituted with 1-3 halogen atoms or an amino-, -SH- or C0-C3alkyl-cycloalkyl group; or R<7>is J;

R7' is H or taken together with R7 forms a C3-C6 cycloalkyl ring optionally substituted with R<7a> wherein;

R<7a> is C 1 -C 6 alkyl, C 3 -C 5 cycloalkyl, C 2 -C 6 alkenyl, each of which may be optionally substituted with halo; or R<7a>can be J;

q is 0 to 3, and k is 0 to 3; where q+k > 1;

W is -CH2-, -O-, -OC(=0)H-, -OC(=0)-, -S-, -NH-, -NRa, -NHS02-, -NHC(=0)NH- or -NHC(=O)-, -NHC(=S)NH- or a bond;

R<8>is a ring system containing 1 or 2 saturated, partially unsaturated or unsaturated rings each having 4-7 ring atoms, and each having 0 to 4 heteroatoms independently selected from S, O and N, wherein the ring system is optionally spaced from W of a C 1 -C 3 alkylene group; or R<8> is C1-C6 alkyl; wherein any R<8> group may be optionally mono-, di- or tri-substituted with R<9>, wherein

R<9> is independently selected from the group consisting of halo, oxo, nitrile, azido, nitro, C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl, NH2C(=0)-, Y-NRaRb, Y-O-Rb, Y-C( =0)Rb, Y-(C=0)NRaRb, Y-NRaC(=0)Rb, Y-NHSOpRb, Y-S(=0)pRb, Y-S(=0)pNRaRb, Y-C(=0)ORb and Y- NRaC(=O)ORb; wherein said carbocyclyl or heterocyclyl group is optionally substituted with R<10>;

in which

R<10> is C 1 -C 6 alkyl, C 3 -C 7 cycloalkyl, C 1 -C 6 alkoxy, amino, sulfonyl, (C 1 -C 3 alkyl)sulfonyl, NO 2 , OH, SH, halo, haloalkyl, carboxyl, amido;

E is -C(=0)-;

X is -NR x - where R x is H, C 1 -C 5 alkyl or J; or in the case where E is -C(=0), X may also be -O- or -NRjNRj-;

wherein one of R 1 is H, and the other is H, C 1 -C 5 alkyl or J;

R<11> is H, C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl, each of which may be substituted with halo, oxo, nitrile, azido, nitro, C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl, NH2CO -, Y-NRaRb, Y-O-Rb, Y-C(=0)Rb, Y-(C=0)NRaRb, Y-NRaC(=0)Rb, Y-NHSOpRb, Y-S(=0)pRb, Y-S(=0 )pNRaRb, Y-C(=0)ORb, Y-NRaC(=0)ORb; or R<11>is J;

J, if present, is a single 3- to 10-membered saturated or partially unsaturated alkylene chain extending from the R7/R7" cycloalkyl or from the carbon atom to which R<7> is attached to one of Rj, Rx, Ry or R<11>to form a macrocycle, which chain is optionally interrupted by one to three heteroatoms independently selected from: -O-, -S- or -NR<12->, and wherein 0 to 3 carbon atoms in the chain are optionally substituted with R<14>;

in which;

R<12> is H, C1-C6 alkyl, C3-C6 cycloalkyl or C(=O)R<13>;

R<13> is C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl;

R<14> is independently selected from the group consisting of H, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkoxy, hydroxy, halo, amino, oxo, thio and C 1 -C 6 thioalkyl;

Ru is independently H or C 1 -C 3 alkyl;

m is 0 or 1; n is 0 or 1;

U is =0 or is absent;

R<15> is H, C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylfiheterocyclyl, each of which may be substituted with halo, oxo, nitrile, azido, nitro, C1-C6 alkyl, C0-C3alkylheterocyclyl, C0-C3alkylcarbocyclyl, NH2CO -, Y-NRaRb, Y-O-Rb, Y-C(=0)Rb, Y-(C=0)NRaRb, Y-NRaC(=0)Rb, Y-NHS(=0)pRb, Y-S(=0)pRb , Y-S(=0)pNRaRb, Y-C(=0)ORb, Y-NRaC(=0)ORb;

G is -O-, -NRy-, -NRjNRj-: where one Rj is H, and the other is H, C 1 -C 5 alkyl or J;

R y is H, C 1 -C 3 alkyl; or Ry is J;

R<16> is H; or C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl, each of which may be substituted with halo, oxo, nitrile, azido, nitro, C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl, NH2CO-, Y-NRaRb , Y-O-Rb, Y-C(=0)Rb, Y-(C=0)NRaRb, Y-NRaC(=0)Rb, Y-NHSOpRb, Y-S(=0)pRb, Y-S(=0)pNRaRb, Y-C( =O)ORb, Y-NRaC(=0)ORb;

with the proviso that when m=n=0 and G is O, then R<16> is not tert-butyl or phenyl;

wherein any C atom in C 1 -C 6 alkyl and C 1 -C 3 alkyl may optionally, unless otherwise indicated, be substituted with one two or, where the valence permits, three halogens;

each aryl and cycloalkyl unit in C0-C3alkylaryl and C0-C3alkylC3C7cycloalkyl is, unless otherwise indicated, substituted with 1-3 substituents selected from halogen, hydroxyl, nitro, cyano, carboxyl, C1-C6alkyl, C1-C6 alkoxy, C 1 -C 6 alkoxyC 1 -C 6 alkyl, C 1 -C 6 alkanoyl, amino, azido, mercapto and C 0 -C 3 alkylheterocyclyl;

each carbocyclyl and heterocyclyl unit in C0-C3alkylcarbocyclyl and C0-C3alkylheterocyclyl is, unless otherwise indicated, optionally substituted with 1 - 3 substituents selected from halogen, hydroxyl, nitro, cyano, carboxyl, C1-C6alkyl, C1-C6alkoxy , C 1 -C 6 alkoxyC 1 -C 6 alkyl, C 1 -C 6 alkanoyl, amino, azido, oxo, mercapto, C 0 -C 3 alkylcarbocyclyl and C 0 -C 3 alkylheterocyclyl;

each amino group is selected from NH 2 , NHC 1 -C 6 alkyl and N(C 1 -C 6 alkyl) 2 ; and

each amido group is selected from C(=O)NH 2 , C(=O)NHC 1 -C 6 alkyl, C(=O)N(C 1 -C 6 alkyl) 2 and -NH(C=O)C 1 -C 6 alkyl;

or a pharmaceutically acceptable salt or prodrug thereof.

Without in any way wishing to be bound by theory or to ascribe tentative binding modes to specific variables, the imaginary concepts P1, P2, P3, and P4 used herein are provided only for convenience and have essentially their conventional meanings, as illustrated by Schechter & Berger, (1976) Biochem Biophys Res Comm 27 157-162, and they denote the parts of the inhibitor which are assumed to fill the S1, S2, S3 and S4 subsites respectively in the enzymes, where S1 is adjacent to the cleavage site, and S4 is distant from the cleavage site. Regardless of binding mode, the components defined by formula I are intended to be within the scope of the invention. For example, cap group R<16->G is expected to interact with the S3 and S4 subsets especially when m and/or n is 0.

Various embodiments of the present invention can theoretically be represented as R<16->G-P4-P3-link-P2-P1, in which P3 and/or P4 may be absent, and P1, P3 and P4 each represent a building block consisting of a derivative of a natural or unnatural amino acid, P2 is a heterocyclic residue, and G -R<16>is a capping group. The link is a carbonyl or another function as defined for E. The P1 and P2 building blocks and the P3 and P4 building blocks are thus usually linked together by amide bonds, while the P2 and P3 building blocks are linked via the link described above. The amide bonds are thereby normally reversed in relation to each other on each side of the link in the compounds according to the invention.

Further aspects of the invention comprise a pharmaceutical composition comprising a compound according to the invention as defined above and a pharmaceutically acceptable carrier or diluent therefor.

The compounds and compositions according to the invention have applicability for the medical treatment or prevention of HCV infections in humans. Accordingly, a further aspect of the invention is the use of a compound as defined above in the preparation of a medicament for the prevention or treatment of flavivirus infections in humans or animals. Examples of flaviviruses include BVDV, dengue and especially HCV.

The compounds according to the invention have a non-peptidic bond at the bond between the P2 and P3 building blocks, which results in the orientation of the P3 and P4 residues being reversed in relation to a natural substrate. This non-peptidic link is also normally longer than the corresponding peptide bond would have been and means that the P3 and/or P4 groups (including the R16 cap to the extent that this interacts with S3 or S4) are displaced outwards in relation to a natural peptide substrate. Flipping and shifting in this way would be expected to support the non-natural D-stereochemistries of the pocket-filling groups (eg side chains) in P3 and/or P4 and/or R16. In reality, such compounds are normally extremely active and within the scope of the invention. However, it has surprisingly been found that even compounds of the invention bearing the L-amino acid side chains in P3 and/or P4 exert good activity, despite the fact that the respective side chain unit must approach the S3 or S4 pocket from a different angle in relative to a natural peptide substrate. Accordingly, L stereochemistry at R<11> and/or R<15> and/or the corresponding configuration at R<16>, to mimic L stereochemistry, represents a preferred aspect of the invention.

The different angle of approach to the S3 and/or S4 pockets also has implications for the ability of the compounds of the invention to avoid resistance patterns exhibited by previously known HCV protease inhibitors which until now have all had a conventional peptide backbone of natural and non-natural L -amino acid residues. As with the reverse transcriptase in HIV, which is notorious for rapidly generating drug-escape mutants under the selective pressure of antiviral therapy, the RNA-dependent RNA polymerase NS5A in HCV has a very poor proofreading ability. This in turn means that the HCV polymerase is highly prone to error, and it is likely that characteristic resistance patterns will emerge when HCV antivirals are administered over long periods. Even before launch, it is apparent that BILN 2061 with a predominantly peptidic backbone (albeit macrocyclized) and Vertex's NS3 protease inhibitor VX-950 with a linear peptide backbone in P3 and P4 rapidly cause characteristic resistance mutations at positions 155, 156 or 168 of NS3 the protease (Lin et al J Biol Chem 2004 279(17): 17808-17).

A preferred group of compounds according to the invention comprises those compounds in which P1 represents a hydrazine derivative, that is, M is Nru, where Ru is normally H or C1-C3 alkyl. Compounds wherein M is CR<7>R<7>' constitute a further preferred aspect of the invention.

Preferred embodiments, wherein M is CR7R7' in formulas I, include formulas IA:

Preferred values for q and k in formula I include 2:1, 2:2, 2:3, 3:2, 3:3, more preferably 1:2 and 1:0; and most preferably 1:1, in which case preferred compounds have the partial structure:

It is currently preferred that E is -C(=0)-.

Compounds according to the invention can include both a P3 and a P4 function, i.e. m and n are each 1. Preferred designs within formula I include formula Ida-Idd below:

Alternative designs include the structures corresponding to Ida, Idb and Idd where M is NRu.

Alternative configurations of the compounds according to the invention include a P3 but no P4 function, ie m is 1 and n is zero. Preferred designs within formula I include the formulas Iea-Iee below:

Alternative designs include the structures corresponding to lea, Ieb, led and lee in which M is NRu.

Even further alternative configurations of the compounds according to the invention include those compounds where m and n are zero, and thus the groups R<16->G around P2, but as mentioned above, the cap groups R16-G can interact favorably with S3 and/or S4 .

Preferred designs within formula I include the formulas Ifa-Ife below:

R<16> in Figure Ifb and elsewhere is normally H, C1-C3alkyl, C5-C6alkyl, C0-C3alkylheterocyclyl, C1-C3alkylcarbocyclyl or C3-C7cycloalkyl, each of which is optionally substituted, as described above. For example, R<16> can be phenyl substituted as described above.

Alternative designs include the structures corresponding to Ifa, Ifb, Ife and Ife where M is NRu.

The compounds according to the invention may comprise linear molecules, as depicted above. Alternatively, in designs in which R7 and R<7>' together constitute a spiro-cycloalkyl group such as spiro-cyclopropyl, the compounds according to the invention can be configured as macrocycles, in which a link group J extends between one of Rj, Rx, Ry or R<11> in formula I. Alternatively, the macrocycle J may extend from the carbon atom adjacent to R<7> to one of Rj, Rx, Ry or R<11>.

Preferred designs of such macrocyclic structures within formula I, in which m is 0, and n is 1, include the structures with Formulas Iga-Igd below:

The corresponding structures in which the J chain is bound to the carbon atom adjacent to R<7> are also preferred.

Further preferred designs of such macrocyclic structures within formula I, wherein m is 0, and n is 1, include the designs with Formulas Ige-Igf below:

The corresponding structures in which the J chain is bound to the carbon atom adjacent to R<7> are also preferred.

Preferred macrocyclic structures within formula I, comprising both a P3 and P4 function, ie wherein m and n are each 1, include the structures of formulas Iha-Ihd below.

At 4

The corresponding structures in which the J chain is bound to the carbon atom adjacent to R<7> are also preferred.

Preferred macrocyclic structures within formula I in which both the P3 and P4 functions are absent, i.e. in which m and n are each 0, include the structures of formulas Ihe-Ihh below, especially lhe and lhf.

The corresponding structures in which the J chain is bound to the carbon atom adjacent to R<7> are also preferred, especially formulas lhe and lhf: In general, in the optionally macrocyclic structures, such as those illustrated above, linker J is a 3 to 10 atom chain , preferably the 5 to 8 atom chain, such as the 6 or 7 atom chain, saturated alkylene chain or a partially unsaturated alkylene chain, i.e. an alkylene chain bearing 1 to 3 unsaturated bonds between adjacent carbon atoms, normally one unsaturation. The length of the chain will naturally depend on whether J extends from Rd, Rj, Rx, Ry, R<11> or the carbon atom adjacent to R<7>. Suitable chains are described in detail in WO 00/59929. Normally, J will be sized to provide a macrocycle of 13 to 16 ring atoms (including the atoms of the P1, P2 and, if present, P3 groups contributing to the ring). Conveniently, J is sized to provide a macrocycle with 14 or 15 ring atoms.

Conveniently, the J chain contains one or two heteroatoms selected from: O, S, NH, NC 1 -C 6 alkyl or N-C(=O)C 1 -C 6 alkyl. More preferably, the J-chain optionally contains one heteroatom selected from: NH or N-C(=O)C1-C6alkyl, most preferably N(Ac). It is most preferred that the chain containing a nitrogen atom is saturated. In an alternative embodiment, J contains one heteroatom selected from O or S. The chain may be substituted with R<14> such as H or methyl.

Normally, the J-linker structure is saturated. Alternatively, J contains 1 to 3, preferably 1 double bond, normally spaced one carbon atom from the cycloalkyl R<7> function, if present. The double bond can be cis or trans.

Representative examples of J thus include pentylene, hexylene, heptylene, each of which may be substituted with C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkoxy, hydroxyl, halo, amino, oxo, thio or C 1 -C 6 thioalkyl; penten-3-yl, hexen-4-yl, hepten-5-yl, where 3, 4 or 5 refers to a double bond between carbon atoms 3 and 4, 4 and 5 etc.

Conveniently, the R<7> and R<7>' groups include those compounds in which R<7>' is H, and R<7> is n-ethyl, n-propyl, cyclopropylmethyl, cyclopropyl, cyclobutylmethyl, cyclobutyl, 2,2-difluoroethyl or mercaptomethyl. Preferred embodiments include those compounds in which R<7> is n-propyl or 2,2-difluoroethyl.

Alternative preferred configurations for R<7> and R<7>' include those compounds in which R7' is H, and R<7> is C3-C7cycloalkyl or C1-C3alkylC3-C7cycloalkyl.

Still further preferred configurations for R<7> and R<7>' include those configurations in which R7' is H, and R<7> is J.

Alternatively, R<7> and R7' together constitute a spiro-cycloalkyl function, such as a spiro-cyclobutyl ring and more preferably a spiro-cyclopropyl ring. "Spiro" in this context simply means that the cycloalkyl ring shares a single carbon atom with the peptidic backbone of the compound. The ring is substituted or unsubstituted. Preferred substituents include mono- or di-substitutions with R<7a>wherein R<7>,<a>is C1-C6alkyl, C3-C5cycloalkyl or C2-C6alkenyl, each of which is optionally substituted with halo. Alternatively, the substituent can be a J linker as described above. Currently preferred stereochemistries for a spiro-cyclopropyl ring are defined below.

Particularly preferred substituents include R<7a> as ethyl, vinyl, cyclopropyl (ie a spiro-cyclopropyl substituent of the "spiro"-cycloalkyl ring of R7/R7'), 1- or 2-bromomethyl, 1- or 2-fluoroethyl, 2 -bromovinyl or 2-fluoroethyl.

In one embodiment of the invention, A is equal to -CR4R4' as illustrated in detail in PCT/EP03/10595.

Suitable R<4>' groups thus include C1-C6 alkyl, such as methyl, ethyl, propyl, ethenyl and -CHCHCH3. Alternative preferred R<4>' groups include aryl or heteroaryl such as optionally substituted phenyl, pyridyl, thiazolyl or benzimidazolyl or C1-C3alkylaryl or C1-C3alkylheteroaryl, where the alkyl part is methyl, ethyl, propyl, ethenyl and -CH=CHCH3. Preferred aryl groups include optionally substituted: phenyl, benzothiazole and benzimidazole.

Preferred R<4> groups include -NH2, fluorine or chlorine. Alternative preferred R<4->groups include -OH and especially =0.

An alternative design for A is C(=O)NHR<3>, where R 3 is optionally substituted C 0 -C 3 alkylaryl, C 0 -C 3 alkylheteroaryl, OC 0 -C 3 alkylaryl or OC 0 -C 3 alkylheteroaryl. Suitable substituents are found in the definition sections below.

A currently preferred configuration for A is C(=O)OR<1>, especially where R<1> is C1-C6 alkyl, such as methyl, ethyl or tert-butyl and most preferably hydrogen.

A particularly preferred configuration for A is C(=0)NHS02R<2>, especially where R<2> is optionally substituted C1-C6alkyl, preferably methyl or optionally substituted C3-C7cycloalkyl, preferably cyclopropyl or optionally substituted C0-C6alkylaryl, preferably optionally substituted phenyl. Suitable substituents are found in the definition sections below.

Substituent -W-R8 on the cyclic P2 group can use any of the proline substituents described in detail in WO 00/59929, WO 00/09543, WO 00/09558, WO 99/07734, WO 99/07733, WO 02/60926 , WO03/35060, WO 03/53349, WO03/064416, W=03/66103, WO03/064455, WO03/064456, WO03/62265, WO03/062228, WO03/87092, WO 03/99274, WO03/99316, WO03 /99274, WO04/03670, , WO04/032827, WO04/037855, WO04/43339, WO04/92161, WO04/72243, 5WO04/93798. WO04/93915, WO04/94452, WO04/101505, WO04/101602, WO04/103996, WO04113365.

Preferred W functions include W such as -OC(=0)NH-, -OC(=0)-,

-NH-, -NR<8>'-, -NHS(0)2-or -NHC(=0)-, especially -OC(=0)NH- or -NH-. Preferred R < 8 -> groups for such W functions include optionally substituted C 0 -C 3 alkylcarbocyclyl or C 0 -C 3 alkyl heterocyclyl, including those described in WO0009543, WO0009558 and WO 00/174768. For example ester substituents, -W-R<8>, on the cyclic P2 group, include those described in WO 01/74768 such as C1-C6alkanoyloxy, C0-C3alkylaryloyloxy, especially (optionally substituted) benzoyloxy or C0-C3alkylheterocycloyloxy, especially

This publication also describes alternative -W-R<8> possibilities for example C1-C6alkyl, such as ethyl, isopropyl, C0-C3alkylcarbocyclyl such as cyclohexyl, 2,2-difluoroethyl, -C(=0)NRc, where Rc is Ci- C 6 alkyl, C 0 -C 3 alkylcyclopropyl, C 0 -C 3 alkylaryl or C 0 -C 3 alkylheterocyclyl.

Currently preferred W functions include -S- and especially -0-. Suitable values for R<8> in such embodiments include C0-C3 alkylaryl or C0-C3 alkylheteroaryl, each of which is optionally mono-, di- or tri-substituted with R<9>, wherein;

R<9> is C 1 -C 6 alkyl, C 1 -C 6 alkoxy, NO 2 , OH, halo, trifluoromethyl, amino or amido (eg amido or amino optionally mono- or di-substituted by C 1 -C 6 alkyl), C 0 -C 3 alkylaryl, C 0 -C 3 alkylheteroaryl or carboxyl, wherein the aryl group or the heteroaryl group is optionally substituted with R<10>; in which

R<10> is C 1 -C 6 alkyl, C 3 -C 7 cycloalkyl, C 1 -C 6 alkoxy, amino, amido, sulfonyl C 1 -C 3 alkyl, NO 2 , OH, halo, trifluoromethyl, carboxyl or heteroaryl.

Normally the C0-C3alkyl component of R<8>, such as C0-C3alkylaryl or C0-C3alkylheteroaryl, is methyl, and in particular it is absent, i.e. C0. The aryl or heteroaryl component is illustrated in detail in the definition section below. Preferably, R<9 > comprises C 1 -C 6 alkyl, C 1 -C 6 alkoxy, amino, (such as di-C 1 -C 3 alkylamino), amido (such as -NHC(O)C 1 -C 6 alkyl or C(=O)NHC 1 -C 6 alkyl) , aryl or fieteroaryl, and the aryl and fieteroaryl are optionally substituted with R<10>; in which

R<10> is C 1 -C 6 alkyl, C 3 -C 7 cycloalkyl, C 1 -C 6 alkoxy, amino, (such as mono- or di-C 1 -C 3 alkylamino), amido (such as -NHC(0)C 1 -C 3 alkyl or C(= 0)NHC 1 -C 3 alkyl), halo, trifluoromethyl or heteroaryl.

Preferably, R<10> comprises C1 -C6 alkyl, C1 -C6 alkoxy, amino, amido (such as - NHC(O)C1 -C6 alkyl or C(=O)NHCi -C6 alkyl), halo or heteroaryl.

Particularly preferred R<10> includes methyl, ethyl, isopropyl, tert-butyl, methoxy, chlorine, amino, amido (such as -NHC(O)C1-C6alkyl, for example -NC(=O)CHC(CH3)3 or C(=O)NHd-C3alkyl) or C1-C3alkylthiazole.

Preferred embodiments of R<8> include 1-naphthylmethyl, 2-naphthylmethyl, benzyl, 1-naphthyl, 2-naphthyl or quinolinyl, each of which is unsubstituted, mono- or disubstituted with R<9> as defined, especially 1-naphthylmethyl or quinolinyl unsubstituted, mono- or disubstituted with R<9> as defined.

A currently preferred R<8> is:

wherein R<9a> is C1-C6 alkyl; C 1 -C 6 alkoxy; thioC 1 -C 3 alkyl; amino optionally substituted with C 1 -C 6 alkyl; C 0 -C 3 alkylaryl; or C0-C3alkylheteroaryl, C0-C3alkylheterocyclyl, wherein said aryl, heteroaryl or heterocycle is optionally substituted with R<10>wherein

R<10> is C 1 -C 6 alkyl, C 3 -C 7 cycloalkyl, C 1 -C 6 alkoxy, amino, amido, heteroaryl or heterocyclyl; and

R<9b>is C 1 -C 6 alkyl, C 1 -C 6 alkoxy, amino, amido, NO 2 , OH, halo, trifluoromethyl, carboxyl.

Suitable R<9a> includes aryl or heteroaryl, all optionally substituted with R<10> as defined, especially where R<9a> is selected from the group consisting of:

wherein R<10> is H, C1-C6alkyl or Co-C3alkyl-C3-C6cycloalkyl, amino optionally mono- or di-substituted with C1-C6alkyl, amido (such as -NHC(0)Ci-C6alkyl or C(=0 )NHC 1 -C 6 alkyl), heteroaryl or heterocyclyl.

R<9a> is conveniently phenyl, and thus R<8> is:

wherein R<10a> is H, C1-C6 alkyl; Q-Cealkoxy; or halo; and R<9b>is C 1 -C 6 alkyl, C 1 -C 6 alkoxy, amino such as di(C 1 -C 3 alkyl)amine, amido (such as -NHC(O)C 1 -C 3 alkyl or C(=O)NHC 1 -C 3 alkyl), NO2, OH, halo, trifluoromethyl, carboxyl.

An alternative preferred R<8> is:

wherein R<10a> is H, C1-C6alkyl or C0-C3alkyl-C3-C6cycloalkyl, amine (such as amine mono- or di-substituted with C1-C6alkyl), amido (such as -NHC(0)C1-C6alkyl or C(=O)NHC 1 -C 6 alkyl), heteroaryl or heterocyclyl; and R<9b>is C 1 -C 6 alkyl, C 1 -C 6 alkoxy, amino (such as di(C 1 -C 3 alkyl)amino), amido (such as -NHC(O)C 1 -C 3 alkyl or C(=O)NHC! -C 3 alkyl), NO 2 , OH, halo, trifluoromethyl or carboxyl.

In the embodiments described immediately above, R<9b> is conveniently C1-C6 alkoxy, preferably methoxy.

A further suitable R<8>, for example when W is an ether, has the formula

where W is N or CH, r is 0 or 1, Ra' is H, C1-C6alkyl, C0-C3alkylcycloalkyl, C1-C6alkyloxy, hydroxy or amine, and Rb' is H, halo, C1-C6alkyl, C0-C3alkylcycloalkyl, C1-Qalkyloxy, C1-Qthioalkyl, cycloalkylC0-C3alkyloxy, C1-CjalkyloxyC1-Cjalkyl, C0-C3alkylaryl or C0-C3alkylheterocyclyl. A particularly preferred ether substituent is 7-methoxy-2-phenyl-quinolin-4-yl-oxy.

When W is a bond, then R<8> is preferably a substituted or unsubstituted heterocyclic ring system as described in WO2004/072243 or WO2004/113665.

Representative examples of R<8>, when W is a bond, include the following aromatic compounds which may be optionally substituted: 1W-pyrrole, 1W-imidazole, 1W-pyrazole, furan, thiophene, oxazole, thiazole, isoxazole, isothiazole, pyridine , pyndazine, pyrimidine, pyrazine, phthalazine, quinoxaline, quinazoline, quinoline, cinnoline, lW-pyrrolo[2,3]-/j]pyridine, lAY-indole, lAY-benzoimidazole, lAY-indazole, 7AY-purine, benzothiazole, benzooxazole , 1H-imidazo[4, 5-c]pyridine, 1W-imidazo[4,5-/>]pyridine, 1,3-dihydro-benzoimidazol-2-one, 1, 3-dihydro-benzoimidazol-2-thione, 2,3-dihydro-1N-indole, 1,3-dihydro-indol-2-one, 1N-indole-2,3-dione, 1, 3-dihydro-benzoimidazol-2-one, 1H, 1N-pyrrolo[ 2, 3-c]pyridine, benzofuran, benzo[^]thiophene, benzo[c/]isoxazole, benzo[c/]isothiazole, lH-quinothin-2-one, lW-quinolin-4-one, lW-quinazolin- 4-one, 9W-carbazole, 1W-quinazolin-2-one.

Further representative examples of R<8>, when W is a bond, include the following non-aromatic compounds, which may be optionally substituted: aziridine, azetidine, pyrrolidine, 4,5-dihydro-1N-pyrazole, pyrazolidine, imidazolidin-2- one, imidazolidin-2-thione, pyrrolidin-2-one, pyrrolidin-2,5-dione, piperidin-2,6-dione, piperidin-2-one, piperazin-2,6-dione, piperazin-2-one, piperazine, morpholine, thiomorpholine-1,1-dioxide, pyrazolidin-3-one, imidazolidin-2,4-dione, piperidine, tetrahydrofuran, tetrahydropyran, [1,4]dioxane, 1,2,3,6-tetrahydropyridine.

Preferred alternatives for R<8> when W is a bond include tetrazole and derivatives thereof. The tetrazole group is attached to the cyclic P2 structure and optionally substituted as shown below:

wherein Q<*>is selected from the group consisting of:

absent, -CH2-, -O-, -NH-, -NCR<1*>), -S-, -S(=O)2- and -(C=0)-; Q<*>is selected from the group consisting of: absent, -CH2- and -NH; Y<*> is selected from the group consisting of: H, C 1 -C 6 alkyl, C 0 -C 3 aryl, C 0 -C 3 heterocyclyl; R<1*>is selected from the group consisting of: H, C1-C6alkyl, carbocyclyl, C0-C3aryl, C0-C3heterocyclyl,

Representative examples of substituted tetrazoles are as described in Table 1 of WO2004/072243 and the structures immediately following or WO2004/113665.

Further preferred alternatives for R<8>, when W is a bond, include triazole and derivatives thereof. The triazole group is attached to the cyclic P2 structure and optionally substituted as shown below:

wherein X<*> and Y<*> are independently selected from the group consisting of: H, halogen, C 1 -C 6 alkyl, C 0 -C 3 carbocyclyl, -CH 2 -amino, -CH 2 -arylamino, -CH 2 -diarylamino, -(C=0 )-amino, -(C=0)-arylamino, -(C=0)-diarylamino, C0-C3aryl, C0-C3heterocyclyl or alternatively, X<*>and Y<*> taken together with the carbon atoms to which they are attached , forms a cyclic group selected from the group consisting of aryl and heteroaryl.

Representative examples of substituted triazoles are as described in Table 2 of WO2004/072243 and the structures immediately following and in the tables of WO2004/113365.

Further preferred alternatives for R<8> when W is a bond include pyridazinone and derivatives thereof. The pyridazinone group is attached to the cyclic P2 structure and optionally substituted as shown below:

wherein X<*>, Y<*> and Z<*> are independently selected from the group consisting of: H, N3, halogen, C1-C6alkyl, carbocyclyl, amino, C0-C3aryl, -S-aryl, -O- aryl, -NH-aryl, diarylamino, diheteroarylamino, C0-C3heterocyclyl, -S-heteroaryl, -O-heteroaryl, NH-heteroaryl, or alternatively X and Y or Y and Z taken together with the carbon atoms to which they are attached form a aryl or heteroaryl cyclic group.

Representative examples of substituted pyridazinones are as described in Table 3 of WO2004/072243 and the structures immediately following and in the tables of WO2004/113365.

Preferred P3 groups, i.e. when m is 1, resemble natural and unnatural amino acids, especially aliphatic amino acids, such as L-valyl, L-leucyl, L-isoleucyl or L-t-leucyl. Further preferred P3 groups, as shown in WO 02/01898, comprise C0-C3alkyl-cycloalkylalanine, especially cyclohexylalanine, optionally substituted with C02Rg, where Rg is H, is C1-C6alkyl, C0-C3alkylaryl, C0-C3alkylheterocyclyl, C0-C3alkylcycloalkyl or amine; or N-acetylpiperidine or tetrahydropyran. Preferably, the R<11> groups thus comprise C1-C6alkyl, C0-C3a I alkyl carbocyclyl, for example C0-C3alkylC3-C7cycloalkylyl, C0-C3alkylaryl or C0-C3alkylheteroaryl, each of which is optionally substituted with hydroxy, halo, amino, Ci- C 6 alkoxy, C 1 -C 6 thioalkyl, C(=0)OR<14>, carboxyl, (C 1 -C 6 alkoxy)carbonyl, aryl, heteroaryl or heterocyclyl, especially where the substituent is hydroxy or C(=0)OR<14>.

Particularly preferred R<11> includes tert-butyl, iso-butyl, cyclohexyl, phenylethyl, 2,2-dimethyl-propyl, cyclohexylmethyl, phenylmethyl, 2-pyridylmethyl, 4-hydroxy-phenylmethyl or carboxyl propyl. The most preferred R<n>" values are currently tert-butyl, isobutyl or cyclohexyl.

One embodiment of the invention includes compounds in which P4 is absent (ie n is 0), and in which the P3 function lacks a carbonyl, ie U is absent. Representative substructures include the structures of formula li below:

in which

Rx and Ry are as defined above, preferably H,

R<11>' is C1-C6 alkyl, preferably branched C3-C5 alkyl such as the side chains in L-valyl, L-leucyl, L-isoleucyl, L-t-leucyl; or C 0 -C 2 alkylC 3 -C 7 cycloalkyl such as cyclohexyl or cyclohexylmethyl;

R 16<a>is -Rba, -S(=O)pRba, -C(=O)Rba;

Rba is C 1 -C 6 alkyl, C 0 -C 3 alkylheterocyclyl, C 0 -C 3a 1 alkyl carbocyclyl.

Alternatively, compounds with the partial structure li may be macrocyclized between an appropriate value of R<7> and one of Rx, Ry or R<11>'.

Representative designs of the P3 groups lacking a carboxy function (ie the variable U is absent) include those of formula Iia-Iid below:

where Ar is carbocyclyl or heterocyclyl, especially aryl or heteroaryl, each of which is optionally substituted with R<9>. Although the partial structures with Formulas lia - lid have been illustrated in the context of a compound within formula I, it will be obvious that such configurations with Formula li also apply to other values of q and k. In the same way, it applies that even if the partial structures of the formulas lic and lid is an R<11->group corresponding to leucine, it will be obvious that these configurations will be applicable to other R<n->groups, especially those resembling the side chains of natural and unnatural L-amino acids, for example t-butyl-alanine/t-leucine.

R<15> in the compounds according to the invention in which n is 1 is preferably optionally substituted C1-C6alkyl or C0-C3alkylcarbocyclyl, for example C0-C3alkylC3-C7cycloalkyl, each of which can be optionally substituted. Preferred P4 groups are normally analogues of natural and unnatural amino acids, especially aliphatic amino acids such as L-valyl, L-leucyl, L-isoleucyl, L-t-leucyl or L-cyclohexylalanine and thus preferred R<15->groups include cyclohexyl, cyclohexylmethyl , tert-butyl, iso-propyl or iso-butyl.

Preferred G values include -NRy-, especially where Ry is methyl or preferably H or hydrazine.

A further preferred G value is O which thereby forms an ester with the carbonyl

in P4 (if present) or the carbonyl in P3 (if present) or an ether in the case of variants in which group U is absent. Conventional pharmaceutical

acceptable ethers or esters capping groups for R<16> include C1-C6alkyl (especially methyl or t-butyl), C0-C3alkylheterocyclyl (especially pyridyl, benzimidazolyl, piperidyl, morpholinyl, piperazinyl) or C0-C3alkylcarbocyclyl (especially phenyl, benzyl, indanyl) each optionally substituted with hydroxy, halo, amino or C 1 -C 6 alkoxy.

It will be obvious that for compounds of formula I, when m=n=0, then R<16>G- is not a BOC or CBz protecting group, but this restriction does not apply to other permutations of m and n. The Boe- or CBz-protected 4-substituted synthetic proline intermediates described for example in WO 0059929 are thus outside the scope of the invention.

Preferred compounds according to the invention may comprise a hydrazine functionality, for example where X is -NHNH- and m is 1; where n is zero or 1. Alternatively, especially where m is zero, G can be -NRjNRj- such as -NHNH-. The compounds will generally not include a hydrazine at both G and X. Typical hydrazines within formula I, wherein m and n are zero, include compounds with the partial structures Ija-Ijb below:

R<16>' in the formulas Ija and Ijb can be considered an alkyl (or C1-C3 alkylheterocyclyl or C1-C3 alkylcarbocyclyl) in which the first alkyl carbon is substituted with an oxo group and constitutes the keto function, and R<16>' is the remainder of the alkyl -, alkylheterocyclyl or alkylcarbocyclyl moiety. Formula Ijb depicts a variant where R<16> is a methylene group, the carbon of which is substituted with an oxo substituent and also -ORb, where Rb is as defined above, normally C1-C6alkyl, such as t-butyl, C0-C3alkylheterocyclyl such as pyridyl or C0-C3a I alkyl carbocyclyl, such as benzyl or phenyl, each of which is optionally substituted as defined above. Compounds with the partial structures Ija and Ijb may be linear molecules as shown (both Rj are H) or preferably one of the depicted Rj groups may be macrocyclized via J to a suitable R<7> group.

Alternative hydrazines of Formula I, where m is 1, include those compounds with the partial structures Ijc and Ijd below:

where R<1>6, G, R11, R<15>, Rj and Ru are as defined for formula I above. Compounds with the partial structures Ijc and Ijd may be linear molecules as shown (both Rj are H) or preferably one of the depicted Rj groups or the R<n->group may be macrocyclized via J to a suitable R<7->group .

Although formulas Ija-Ijd are depicted with a proline analog as P2, it will be apparent that this aspect of the invention is equally adaptable to other configurations of q and k.

Alternate hydrazine-like configuration occurs when G is amino, and m and n are 0, and R<16>is an N-linked unsaturated heterocycle as defined below, such as pyridyl or pyrimidyl or a saturated heterocycle as defined below, such as piperazinyl , piperidinyl and especially morpholinyl. Examples of such designs include the structures with the formulas Ije:

Compounds with the partial structures Ije can be linear molecules as shown,

or preferably Rx can be macrocyclized via J to a suitable R<7> group. Although these partial structures are depicted with a five-membered ring for P2, it will be readily apparent that this configuration extends to other values of q and k. On

likewise, these configurations will be applicable to other N-bonded heterocycles such as R<16>.

Back to Formulas I in general: Preferred R<16->groups for the compounds according to the invention include 2-indanol, indanyl, 2-hydroxy-1-phenyl-ethyl, 2-thiophenemethyl, cyclohexylmethyl, 2,3-methylenedioxybenzyl, cyclohexyl, phenyl, benzyl, 2-pyridylmethyl, cyclobutyl, iso-butyl, n-propyl, methyl or 4-methoxyphenylethyl.

Currently preferred R<16> groups include 2-indanol, indane, 2-hydroxy-1-phenyl-ethyl, 2-thiophenemethyl, 2,3-methylenedioxybenzyl or cyclohexylmethyl.

Unnatural amino acids include L-amino acids in which the side chain is not one of the 20 naturally occurring amino acids. Examples of non-natural amino acids include L-beta-methylsulfonylmethylalanine, L-cyclohexylalanine, L-tertiary-leucine, L-norleucine, L-norvaline, L-ornithine, L-sarcosine, L-citurline, L-homophenylalanine, L-homoserine , L-beta-(l-naphthyl)alanine, L-beta-(2-naphthyl)alanine etc. Non-natural amino acids also include the D-amino acids corresponding to the 20 natural amino acids and D-amino acids bearing other side chains, such as the as indicated above.

'Ci-Cealkyl' (also abbreviated as Ci-Qalk or used in terms such as Q.-C6alkyloxy etc) as used herein is intended to include straight and branched aliphatic carbon chains such as methyl, ethyl, n-propyl, isopropyl , n-butyl, isobutyl, t-butyl, pentyl, isopentyl, hexyl, heptyl and any single isomer thereof. The alkyl group may have an unsaturated bond. In addition, any C atom in C 1 -C 6 alkyl may optionally be substituted with one, two or, where the valence permits, three halogen atoms and/or substituted, or the alkyl chain may be interrupted by a heteroatom S, O, NH. If the heteroatom is located at a chain terminus, then it is suitably substituted with one or 2 hydrogen atoms. Ci-C6alkyl and Ci-C6alkyl have the corresponding meaning as Ci-C6alkyl adapted for the number of carbon atoms as necessary.

'C 1 -C 8 alkyl' as used herein includes methyl, ethyl, propyl, isopropyl, cyclopropyl, each of which may be optionally substituted or heteroatom interrupted, as described in the paragraph above, or in the case of C 2 or C 3 , may carry an unsaturated bond such as CH2=CH.

"C 1 -C 6 alkylene" as used herein describes a divalent C 1 -C 6 alkyldiyl group, including propylene, ethylene and especially methylene. The normally longer alkylene chains for

J may comprise 1 to 3 unsaturations and/or interruptions with heteroatoms as defined above.

'Amino' includes NH 2 , NHC 1 -C 3 alkyl or N 1 -C 8 alkyl) (especially C 1 -C 3 alkyl variants)

'Amido' includes C(=0)NH2 and alkylamido, such as C(=0)NHCi-C6alkyl, C(=0)N(Ci-C6alkyl)2especially C(=0)NHC1-C3alkyl, C(=0)N (C 1 -C 3 alkyl) 2 or -NH(C=O)C 1 -C 6 alkyl, for example -NHC(=O)CHC(CH 3 ) 3 , including -NH(C=O)C 1 -C 3 alkyl.

'Halo' or halogen as used herein is intended to include F, Cl, Br, I, especially chlorine and preferably fluorine.

'C0-C3alkylaryl as used herein is intended to include an aryl group such as a phenyl, naphthyl or phenyl fused to a C3-C7cycloalkyl (eg indanyl), which aryl is directly attached (ie C0) or via an intermediate methyl- , ethyl or propyl group as defined for the C 1 -C 3 alkylene above. If nothing else is stated, the aryl and/or its fused cycloalkyl group is optionally substituted with 1-3 substituents selected from halo, hydroxy, nitro, cyano, carboxy, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkyl, C 1 -C 6 alkyl, C 1 - C6alkanoyl, amino, azido, oxo, mercapto, nitro C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl. "Aryl" has the corresponding meaning, ie where the C0-C3 alkyl bond is absent

'C0-C3alkylC3C7cycloalkyl' as used herein is meant to include a C3-C7cycloalkyl group such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl, which cycloalkyl is directly attached (ie C0alkyl) or via an intermediate methyl, ethyl- , propyl or isopropyl group as defined for the C 1 -C 3 alkylene above. The cycloalkyl group may contain an unsaturated bond. If nothing else is stated, the cycloalkyl group is optionally substituted with 1-3 substituents selected from halo, hydroxy, nitro, cyano, carboxy, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkoxyC 1 -C 6 alkyl, C 1 -C 6 alkanoyl, amino, azido, oxo , mercapto, nitro C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl.

'C0-C3alkylcarbocyclyl' as used herein is intended to include C0-C3alkylaryl and C0-C3alkylC3-C7cycloalkyl. If nothing else is stated, the aryl or cycloalkyl group is optionally substituted with 1-3 substituents selected from halo, hydroxy, nitro, cyano, carboxy, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkoxyC 1 -C 6 alkyl, C 1 -C 6 alkanoyl, amino, azido, oxo, mercapto, nitro, C0-C3a I alkyl carbocyclyl and/or C0-C3 alkylheterocyclyl.

"Carbocyclyl" has the corresponding meaning, ie where the C0-C3 alkyl bond is absent

'C0-C3 alkylheterocyclyl as used herein is intended to include a monocyclic, saturated or unsaturated, heteroatom-containing ring such as piperidinyl, morpholinyl, piperazinyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazinolyl, isothiazinolyl, thiazolyl, oxadiazolyl, 1,2, 3-triazolyl, 1,2,4-triazolyl, tetrazolyl, furanyl, thienyl, pyridyl, pyrimidyl, pyridazinyl, pyrazolyl or any such group fused to a phenyl ring, such as quinolinyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazinolyl, benzisothiazinolyl, benzothiazolyl, benzoxadiazolyl, benzo-l,2,3-triazolyl, benzo-l,2,4-triazolyl, benzotetrazolyl, benzofuranyl, benzothienyl, benzopyridyl, benzopyrimidyl, benzopyridazinyl, benzopyrazolyl etc, which ring is attached directly i.e. (C0) or via an intermediate methyl, ethyl, propyl or

isopropyl group as defined for the C1-C3 alkylene above. Any such unsaturated ring having aromatic character may be referred to herein as heteroaryl. If nothing else is stated, the heteroring and/or its fused phenyl group is optionally substituted with 1-3 substituents selected from halo, hydroxy, nitro, cyano, carboxy, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkoxyC 1 -C 6 alkyl, C 1 -C 6 C6alkanoyl, amino, azido, oxo, mercapto, nitro, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl. "Heterocyclyl" and "Heteroaryl" have the corresponding meaning, ie where the C0-C3 alkyl bond is absent.

Normally heterocyclyl and carbocyclyl groups within the scope of the above definitions are thus a monocyclic ring with 5 or especially 6 ring atoms or a bicyclic ring structure comprising a 6-membered ring condensed to a 4-, 5- or 6-membered ring.

Typical such groups include C3-C8 cycloalkyl, phenyl, benzyl, tetrahydronaphthyl, indenyl, indanyl, heterocyclyl such as from azepanyl, azocanyl, pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, indolinyl, pyranyl, tetrahydropyranyl, tetrahydrothiopyranyl, thiopyranyl, furanyl, tetrahydrofuranyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, imidazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, tetrazolyl, pyrazolyl, indolyl, benzofuranyl, benzothienyl, benzimidazolyl, benzthiazolyl, benzoxazolyl, benzisoxazolyl, quinolinyl, tetrahydro-quinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, quinazolinyl, tetrahydroquinazolinyl and quinoxalinyl, each of which may be optionally substituted as defined herein.

The saturated heterocycle group thus includes radicals such as pyrrolinyl, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, pyranyl, thiopyranyl, piperazinyl, indolinyl, azetidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydrofuranyl, hexahydropyrimidinyl, hexahydropyridazinyl, 1,4,5,6- tetrahydropyrimidinylamine, dihydro-oxazolyl, 1,2-thiazinanyl-1,1-dioxide, 1,2,6-thiadiazinanyl-1,1-dioxide, isothiazolidinyl-1,1-dioxide and imidazolidinyl-2,4-dione, while the unsaturated heterocycles include radicals with an aromatic character such as furanyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, tetrazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, indolyl, isoindolyl. In each case, the heterocycle may be fused with a phenyl ring to form a bicyclic ring system.

Synthesis

Synthesis of the compounds according to the present invention can be carried out using different chemical strategies in solution or solid phase or a combination of both. The suitably protected individual building blocks can first be manufactured and then connected together ie P2+P1-» P2-P1. Alternatively, precursors of the building blocks can be linked together and modified at a later stage of the synthesis of the inhibitor sequence. Further construction items, precursors of construction items or prefabricated larger fragments of the desired structure can then be connected to the growing chain, e.g. R<16->G-P3+ E-P2-Pl-> R<16->G-P3-P2-P1 or R<16->G-P4-P3+E-P2-P1-» R<16- >G-P4-P3-E-P2-P1.

Coupling between two amino acids, an amino acid and a peptide or two peptide fragments can be performed using standardized coupling procedures such as the azide method, the mixed carbon-carboxylic anhydride (isobutyl chloroformate) method, the carbodiimide method (dicyclohexylcarbodiimide, diisopropylcarbodiimide or water-soluble carbodiimide), the active ester method (pnitrophenyl ester, N-hydroxysuccinimido ester), The method with Woodward's reagent K, the carbonyldiimidazole method, the phosphorus reagent or oxidation-reduction methods. Some of these methods (especially the carbodiimide method) can be enhanced by adding 1-hydroxybenzotriazole or 4-DMAP. These coupling reactions can be carried out in either solution (liquid phase) or solid phase.

Specifically, the coupling step involves dehydrative coupling of a free carboxyl in one reactant with the free amino group in the other reactant in the presence of a coupling agent to form a connecting amide bond. Descriptions of such coupling agents appear in general textbooks of peptide chemistry, for example, M.

Bodanszky, "Peptide Chemistry", 2nd rev ed., Springer-Verlag, Berlin, Germany,

(1993) hereafter simply referred to as Bodanszky. Examples of suitable coupling agents are N,N'-dicyclohexylcarbodiimide, 1-hydroxybenzotriazole in the presence of N,N'-dicyclohexylcarbodiimide or N-ethyl-N'-[(3-dimethylamino)propyl]-carbodiimide. A practical and useful coupling agent is the commercially available (benzotriazol-1-yloxy)tris-(dimethylamino)phosphonium hexafluorophosphate, either alone or in the presence of 1-hydroxybenzotriazole or 4-DMAP. Another convenient and useful coupling agent is commercially available 2-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate. Yet another convenient and useful coupling agent is commercially available O-(7-azabenzotrizol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate.

The coupling reaction is carried out in an inert solvent, e.g. dichloromethane, acetonitrile or dimethylformamide. An excess of a tertiary amine, e.g. Diisopropylethylamine, N-methylmorpholine, N-methylpyrrolidine or 4-DMAP are added to keep the reaction mixture at a pH of about 8. The reaction temperature usually varies between 0°C and 50°C, and the reaction time usually varies between 15 min and 24 h.

The functional groups in the existing amino acids must generally be protected during the coupling reactions to avoid the formation of unwanted bonds. The protective groups which may be used are listed in Greene, "Protective Groups in Organic Chemistry", John Wiley & Sons, New York (1981) and "The Peptides: Analysis, Synthesis, Biology", Vol. 3, Academic Press, New York (1981), hereafter referred to simply as Greene.

The <x-Carboxyl group in the C-terminal residue is usually protected as an ester which can be cleaved to give the carboxylic acid. Protecting groups that can be used include 1) alkyl esters such as methyl, trimethylsilyl and t-butyl, 2) aralkyl esters such as benzyl and substituted benzyl or 3) esters that can be cleaved by a mild base or a mild reducing agent such as trichloroethyl and phenacyl esters .

The cc-Amino group in each amino acid to be linked is usually protected. Any protecting group known in the art can be used. Examples of such groups include: 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 (Boe), ethoxycarbonyl, diisopropyl methoxycarbonyl and allyloxycarbonyl; 4) cyclic alkyl carbamate groups such as cyclopentyloxycarbonyl and adamantyloxycarbonyl; 5) alkyl groups such as triphenylmethyl and benzyl; 6) trialkylsilyl such as trimethylsilyl; and 7) thiol-containing groups such as phenylthiocarbonyl and dithiasuccinoyl. The preferred α-amino protecting group is either Boe or Fmoc. Many amino acid derivatives suitably protected for peptide synthesis are commercially available.

The cc-amino protecting group is cleaved before the next coupling step. When the Boc group is used, the methods of choice are trifluoroacetic acid, undiluted or in dichloromethane or HCl in dioxane or in ethyl acetate. The resulting ammonium salt is then neutralized either before the 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. Deprotection is carried out at a temperature between 0°C and room temperature, usually 20-22°C.

Any of the natural or non-natural amino acids having side chain functionalities will normally be protected during the preparation of the peptide by using any of the groups described above. Those skilled in the art will appreciate that the selection and use of appropriate protecting groups for these side chain functionalities depends on the amino acid and the presence of other protecting groups in the peptide. In the selection of such protective groups, it is desirable that the group is not removed during the deprotection and coupling of the α-amino group

For example, when Boe is used as the α-amino protecting group, the following side chain protecting groups are suitable: p-toluenesulfonyl (tosyl) groups can be used to protect the amino side chain of amino acids such as Lys and Arg; acetamidomethyl, benzyl (Bn) or tert-butylsulfonyl groups can be used to protect the sulfide-containing side chain of cysteine; benzyl ethers (Bn) can be used to protect the hydroxy-containing side chains of serine, threonine or hydroxyproline; and benzyl esters can be used to protect the carboxy-containing side chains of aspartic acid and glutamic acid.

When Fmoc is chosen for the α-amine protection, usually tert. butyl-based protecting groups acceptable. For example, Boe can be used for lysine and arginine, tert-butyl ether for serine, threonine and hydroxyproline and tert-butyl ester for aspartic acid and glutamic acid. The triphenylmethyl group (Trityl) can be used to protect the sulphide-containing side chain of cysteine.

Once the inhibitor sequence is complete, any protecting groups are removed in the manner dictated by the choice of protecting groups. These procedures are well known to those skilled in the art.

In compounds of formula I, the P2 unit comprises a nitrogenous ring residue which is substituted with the W and R8 groups.

Synthesis of heterocyclic P2 bvaae subjects

The R<8> group can be attached to the P2 structure at any appropriate stage of the synthesis of compounds of the present invention. One solution is to first connect the R<8->group to the P2 structure and then add the other desired building blocks, i.e. Pl and optionally P3 and P4. Another solution is to connect P1 and, if present, P3 and P4, using an unsubstituted P2 structure and adding the R<8> group afterwards.

Compounds wherein W is O, and R<8> is alkyl, C0-C3 alkylcarbocyclyl, C0-C3 alkylheterocyclyl, can be prepared according to the procedure described by E. M. Smith et al. (J. Med. Chem. (1988), 31, 875-885), as depicted in Scheme 1, which illustrates the technique in a group in which q and k are 1.

Commercially available Boc-4-(R)-hydroxyproline or any suitable hydroxy-substituted proline analog such as a hydroxypiperidine acid is treated with a base such as sodium hydride or potassium t-butoxide in a solvent such as dimethylformamide, and the resulting alkoxide is reacted with a alkylating agent, R<8->X, wherein X is a suitable leaving group such as a halide, mesylate, triflate or tosylate or the like, giving the desired substituted proline derivative. Alternatively, when W is O, or S and R<8> is carbocyclyl such as phenyl or heterocyclylyl such as heteroaryl, the P2 building blocks can also be prepared via 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) as shown in Scheme 2, which illustrates the technique in a group in which q and k are 1.

Treatment of the appropriate hydroxy-substituted proline analog, such as a hydroxypiperidic acid, shown here as commercially available Boc-4-hydroxyproline methyl ester, with the desired alcohol or thiol (R<8->WH) in the presence of triphenylphosphine and an activating agent such as diethyl azodicarboxylate ( DEAD), d i isopropyl azod i carboxylate (DIAD) or the like, provides the ester compound (2b). Hydrolyzing the ester to the acid by standard procedures affords the P2 building block (2c).

Alcohol (2a) can alternatively be treated with phosgene which thus gives the corresponding chloroformate which after reaction with an amine, R<8>NH2, in the presence of a base such as sodium bicarbonate or triethylamine, provides carbamates, i.e. W is -OC(=0 )NH-, while reaction of alcohol (2a) with an acylating agent, R8-CO-X, such as an acid anhydride or acid halide such as the acid chloride, to provide esters, i.e. W is -OC(=0)-.

Various alcohols R<8->OH and alkylating agents R<8->X are described in WO 00/09543 and WO00/59929. An example of the synthesis in which R<8> is a substituted quinoline derivative is shown in Scheme 3.

Friedel-Craft acylation of an appropriately substituted aniline (3a), available either commercially or in the literature, using an acylating agent such as acetyl chloride or the like in the presence of boron trichloride and aluminum trichloride in a solvent such as dichloromethane affords (3b). Coupling of (3b) to a heterocyclic carboxylic acid (3c) under basic conditions, such as in pyridine, in the presence of an activating agent for the carboxylate group, for example POCl3, followed by ring closure and dehydration under basic conditions such as potassium tert-butoxide in tert -butanol provides quinoline derivative (3e). Quinoline derivative (3e) can be coupled in a Mitsunobu reaction to an alcohol as described above, or the hydroxy group can be replaced by a suitable leaving group such as a halide such as chloride, bromide or iodide, by treating quinoline (3e) with a suitable halogenating agent for example phosphoryl chloride or the like.

Many different carboxylic acids with the general structure (3c) can be used in Scheme 3. These acids are available either commercially or in the literature. An example of the preparation of 2-(substituted)-amino-carboxy-aminothiazole derivatives, according to the procedure described by Berdikhina et al. Chem. Heterocycl. Compd. (Engl. Transl.) (1991), 427-433, is shown below.

Thioureas (4c) with different alkyl substituents R' can be formed by reacting the appropriate amine (4a) with tert-butyl isothiocyanate 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 (4c) with 3-bromopyruvic acid affords the acid (4d).

P2 building blocks in which the R<8->substituent is attached via an amine, amide, urea or sulfonamide can be prepared from aminoproline analogues obtained either from a suitable commercially available aminoproline, etc derivative or by converting the hydroxy group in the corresponding hydroxy derivative to an azide group for for example by converting the hydroxy group into a suitable leaving group such as a mesylate or halogen such as chloride, followed by substitution of the leaving group with azide or by using an azide transfer agent such as diphenyl phosphoryl azide (DPPA). Reduction of the azide by catalytic hydrogenation or any other suitable reduction method provides the amine. The amino derivative can be reacted in a substitution reaction with an alkylating agent of the general formula R<8>X wherein R<8>and X are as described for Scheme 1, to form P2 building blocks for use in the preparation of compounds of the general formula I , wherein W is -NH-. Reaction of the aminoproline analog with an acid of the general formula R<8->COOH under normal amide coupling conditions affords compounds in which the R8 substituent is attached via an amide bond, while reaction of the aminoproline analog with an appropriate derivative of sulfonic acid, R<8->S(0 )2-X where X is a leaving group for example chloride, in the presence of a base, provides sulfonamides. Compounds in which the bond between the cyclic structure and the R8 substituent consists of a urea group can be obtained, for example, by treating the aminoproline analogue with phosgene to obtain the corresponding chlorocarbamate followed by reaction with the desired amine. Alternatively, the aminoproline analogue can be reacted with the carbamoyl chloride or the isocyanate of the desired R8 substituent to form the urea bond. It will be obvious that similar reactions will be available for the P2 groups with other ring sizes and substitution patterns.

4-Substituted heterocyclyl derivatives such as 4-substituted proline for use as P2 building blocks where W is -CH2- can be prepared as shown in Scheme 5, which illustrates the technique on a group where q and k are 1, according to the procedures described by J. Ezquerra et al., Tetrahedron, 1993, 38, 8665-8678 and C. Pedregal et al. Tetrahedron Lett., 1994, 35, 2053-2056.

Treatment of suitable acid-protected pyrrolidone or piperidinone such as commercially available Boc-pyroglutamic acid (5a) with a strong base such as lithium diisopropylamide in a solvent such as tetrahydrofuran followed by addition of an alkylating agent R<8->CH2-X where X is a suitable leaving group such as a halide such as chloride or bromide, followed by reduction of the amide and deprotection of the ester gives the desired compound (5d).

Compounds according to the present invention in which a heterocyclic R<8> group is attached directly to the cyclic P2 structure, i.e. W is a bond in the general formula I, can be prepared for example by using a substitution reaction in which a suitable leaving group on P2 -structure is replaced with the desired R<8-> group such as a heterocyclic group.

Alternatively, the R8 group can be introduced by means of a Mitsunobu reaction in which the hydroxy group in the P2 structure is reacted with a nitrogen atom in the heterocyclic R<8> group.

Compounds in which a tetrazole derivative is attached via a carbon atom in the heterocyclic ring are easily prepared by building up the tetrazole group directly on the P2 precursor. This can be achieved, for example, by converting the hydroxy group in the P2 precursor to a cyano group followed by reaction with an azide reagent such as sodium azide. Triazole derivatives can also be built up directly on the P2 precursor, for example by converting the hydroxy group in the P2 precursor to an azide group followed by a 3+2 cycloaddition reaction between the obtained azide and a suitable alkyne derivative.

Structurally different tetrazoles for use in the above-described substitution or Mitsunobu reaction can be prepared by reacting commercially available nitrile compounds with sodium azide. Triazole derivatives can be prepared by reacting an alkyne compound and trimethylsilyl azide. Useful alkyne compounds are available either commercially or they can be prepared for example according to the Sonogashira reaction, i.e. reaction of a primary alkyne, an aryl halide and triethylamine in the presence of PdCl2(PPh)3 and Cul as described for example in A. Elangovan, Y. -H. Wang, T.-I. Ho, Org. Lett., 2003, 5, 1841-1844. The heterocyclic substituent can also be modified when it is attached to the P2 building block, either before or after linking the P2 building block to the other building blocks.

These methods and further alternatives for the preparation of compounds, in which W is a bond, and R 8 is an optionally substituted heterocycle, are described in detail in WO2004/072243.

Compounds with alternative ring size and/or position of the W-R8 substituent in the proline derivatives in schemes 1, 2 and 5 can also be used in the preparation of compounds according to the present invention. For example, alkylation of commercially available 3-hydroxyproline provides compounds of the general formula (I), wherein k is 0, and q is 2. Similarly, alkylation of 5-hydroxyproline, prepared for example as described by Hallberg et al., provides J. Med. Chem. (1999), 4524-4537, compounds of the general formula (I), wherein k is 2, and q is 0.

Various methods for the preparation of hydroxylated 2-piperidine carboxylic acids are described in the literature, see for example Celestini et al., Org. Lett., (2002), 1367-1370, Hoarau et al., Tetrahedron: Asymmetry, (1996), 2585-2594, Zhu et al., Tetrahedron Lett., 41, (2000), 7033-7036. For example, the corresponding pyridine carboxylic acids can be reduced to provide hydroxylated 2-piperidine carboxylic acids. Enzymatic methods can also be used for the production of hydroxylated proline analogues. For example, a 3-hydroxy substituent can be introduced onto commercially available 4-, 5- and 6-membered heterocyclic acids using proline 3-hydroxylase as described by Ozaki et al., Tet. Letters, 40, (1999), 5227-5230.

Synthesis and introduction of Pl-byqqe topics

The amino acids used in the preparation of P1 fragments are available either commercially or in the literature, see for example WO 00/09543 and WO00/59929 from Boehringer-Ingelfieim or US2004/0048802 from BMS.

Scheme 6 shows an example of the preparation of a sulfonamide derivative to be used as a P1 fragment, and the subsequent coupling to a Boc-protected P2 building block.

The sulfonamide group can be introduced onto a suitably protected amino acid (6a) by treating the amino acid with a coupling agent, for example N,N'-carbonyldiimidazole (CDI) or the like, in a solvent such as THF, followed by reaction with the desired sulfonamide (6b) in the presence of a strong base such as 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). Alternatively, the amino acid can be treated with the desired sulfonamide (6b) in the presence of a base such as diisopropylethylamine followed by treatment with a coupling agent such as PyBOP® to effect the introduction of the sulfonamide group. Removal of the amino-protecting group by normal methods and subsequent coupling to a P2 building block, prepared as described above, using standard methods for amide bond formation, such as with a coupling agent such as 0-(7-azabenzotriazol-1-yl) )-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) in the presence of a base such as diisopropylamine in a solvent such as dimethylformamide affords Boc-protected P2-P1 compound (6e). Alternatively, the sulfonamide group can be introduced at a later stage of the synthesis, for example as the last step. In this case, an amino acid with the opposite protection pattern, ie having an unprotected amino function and a protected acid function, is coupled to the acid function of the P2 building block using normal peptide coupling conditions, for example as described above. Removal of the acid protecting group, using the appropriate conditions for the protecting group employed, followed by coupling of the sulfonamide as described above, affords compound 6e

The P1 building block for the preparation of compounds according to the general formula I, in which A is an ester or an amide, can be prepared by reacting amino acid (6a) with the appropriate amine or alcohol, respectively, under standard conditions for amide or ester formation. Compounds of the general formula I, wherein A is CR<4>R<4>', can be prepared by coupling the appropriate P1 building block to the P2 building block as described in Oscarsson et al Bioorg Med Chem 2003 11(13) 2955-2963 and PCT/EP03/10595 filed 23.09.2003.

Compounds comprising an azapeptide P1 residue, i.e. Q is NRu in the general formula I, can be prepared by using an appropriate P1-aza-aminoacyl group in the linkage to the P2 fragment. The preparation of aza-aminoacyl groups is described by M. D. Bailey et al. in J. Med. Chem., 47, (2004), 3788-3799 and an example is shown in Scheme 6A.

Incorporation of the appropriate N-linked side chain, Ru, onto commercially available tert-butylhydrazine can be carried out for example by a reductive amination reaction with the appropriate aldehyde or ketone as described in Scheme 19 below which gives the N-alkyl terecarbazate (6Aa). Condensation of 6Aa with a desired chloroformate in the presence of a base such as triethylamine or diisopropylethylamine in a solvent such as THF affords 6Ab. The R1' group can then optionally be removed by applying the appropriate conditions depending on the specific RI', such as catalytic hydrogenation for RI' when it is benzyl, giving the corresponding acids. Subsequent reaction of the obtained acid with a desired sulfonamide derivative as described in scheme 6 gives sulfonamide-capped building blocks. Alternatively, reaction of carbazate 6Aa with an isocyanate, R3-N=C=0, provides building blocks for the preparation of compounds of general formula I, wherein M is Nru, and A is CONHR<3>.

The P2 and P3 groups can be tied together before or after the introduction of the Pl building block.

Synthesis of capped P3- and P3- P4- bvaae subjects

The building blocks R<16->G-P3 and R<16->G-P4-P3 can be manufactured as generally depicted in scheme 7.

An appropriate N-protected amino acid (7a) can be coupled with an amino cap group (R<16->NHRy) using normal peptide coupling conditions such as with coupling agents such as HATU, DCC, HOBt or the like in the presence of a base such as DIEA or DMAP in a solvent such as dichloromethane, chloroform or dimethylformamide or a mixture thereof and conditions for ester formation such as providing amides, ie G is NHRy (7b). Alternatively, reaction of amino acid (7a) with a compound of the general formula R<16->X, where R<16> is as defined above, and X is a leaving group such as a halide, in the presence of a base such as cesium carbonate or silver(II) oxide, esters, i.e. G is O (7b). On the other hand, amino acid (7a) can be coupled to a second, suitably O-protected, amino acid (7d) using normal peptide coupling conditions as described above, yielding (7e). Replacement of the ester group with a suitable capping group (7b) provides fragment (7f) useful for the preparation of compounds of the present invention, wherein m and n are 1.

When G is N-Ry, the cut P3 or P2 building block can also be produced on a fixed support as exemplified in Form 8.

A suitable N-protected, for example Boc-protected, amino acid (8a) can be immobilized on a solid support, exemplified here by Agronaut resin PS-TFP, by reacting the amino acid with the desired solid support in the presence of coupling reagents such as N,N '-diisopropylcarbodiimide and a base such as DMAP in a solvent such as dichloromethane and dimethylformamide. The immobilized amino acid can then be cleaved from the support with a suitable capping group (8c) and thus yield fragments useful for the preparation of compounds of the present invention, wherein m or n is 1. Optionally, the amino-protecting group can be removed followed by coupling of a appropriate amino acid using standard methods, thus providing fragments useful for the preparation of compounds of the present invention, wherein m and n are 1.

Linking a cover group or a cover building subject to the P2-Pl construct

The R<16->G, R<16->G-P3 or R<16->G-P4-P3 building block linked via a urea functionality to the P2-P1 construct can be introduced as depicted in Scheme 9, which illustrates the technique with a variant where the P2 structure is a 5-membered ring.

A chlorocarba mat group can be formed on the ring amide of the P2-P1 construct (9a) by removal of the amine protecting group using standard procedures, such as acidic treatment with, for example, TFA in dichloromethane or the like when the Boc group is used, followed by reaction of the free amine with phosgene in toluene in the presence of a base such as sodium bicarbonate or triethylamine in a solvent such as tetrahydrofuran. Subsequent reaction of the formed electrophilic center with the amino group in an R<16->NH-2, R<16->NH-NH-2, R<16->G-P3- or R<16->G-P4- P3 building block (9c) in a solvent such as dichloromethane in the presence of a base such as sodium bicarbonate affords (9d). Compounds of the general formula (I) in which E is C=S, S(=0) or S(=0)2 can be prepared according to the above procedure, but using reagents such as thiocarbonyldiimidazole, thionyl chloride or sulphoryl chloride instead of phosgene.

Compounds containing a hydrazine group attached to the P2 unit, ie X is -NRjNRj- in the general formula I, or when the P3 and P4 units are absent, and G is NRjNRj, can be prepared as depicted below. Scheme 10 shows the introduction of a hydrazine derivative into a 5-membered P2 building block.

Reaction of tert-butyl carbazate (10a), optionally alkyl-substituted on one or both nitogens, with p-nitrophenyl chloroformate in the presence of a base such as sodium bicarbonate followed by addition of the P2 building block (10b) affords the urea derivative 10c. The phosgene method described in scheme 9 can alternatively be used to effect binding of the fragments 10a and 10b. Optional removal of the Boc group by standard procedures such as acid treatment with, for example, TFA in a suitable solvent such as dichloromethane affords the hydrazine-containing derivative (10d). Alternatively, any suitable hydrazine derivative such as morpholin-1-ylamine, piperidin-1-ylamine or the like may be attached to 9Ab in place of the tert-butylcarbazate derivative.

The obtained compound can then be further extended by coupling a P3 or P4-P3 building block to the primary amine in compound 9Ad, for example as shown in scheme 11.

Treatment of the α-amino compound (11a) with sodium nitrite, potassium bromide and sulfuric acid (Yang et al. J. Org. Chem. (2001), 66, 7303-7312) provides the corresponding α-bromo compound (11b) which after reaction with the above described derivative (10d) provides the hydrazine-containing derivative (lic).

The bond between the P2 and P3 building blocks can also consist of a carbamate group and a general route to such compounds is depicted in Scheme 12, which illustrates the technique with a variant in which P2 is a proline derivative.

The desired, optionally protected, amino cap group (12a) is coupled to a hydroxy acid (10b) using normal peptide coupling techniques followed by reaction with the electrophilic P2 building block (12d) described above, and optional deprotection provides construct (12e).

Compounds lacking a carboxy group in the P3 unit can be prepared as illustrated in Scheme 13, which illustrates the technique applied to a compound of formula I

Chlorocarbamoyl derivative (13a) can be reacted in a substitution reaction with an azide derivative (13b), prepared by methods known from the literature, in the presence of a base such as sodium bicarbonate to give (13c). X is as described for the general formula (I). Reduction of the azide function for example by polymer-bound triphenylphosphine in a solvent such as methanol or any other suitable reduction method provides intermediate (13d) which can then be reacted with an acid under peptide coupling conditions or with an amine in a reductive amination reaction, giving amides and secondary amines respectively.

Scheme 14 shows an alternative route towards compounds lacking a carboxy group in the P3 unit.

Instead of using the azide derivative (13b) in scheme 13, the corresponding, optionally protected, hydroxy derivative (14b) can be used in the replacement reaction with the chlorocarbamate (14a) and thus introduce a primary alcohol. The alcohol (14c) can then, after optional deprotection, be oxidized with a suitable oxidizing agent such as, for example, Dess-Martin periodinan to form the corresponding aldehyde. Reaction of the aldehyde with a desired amine in a reductive amination reaction using a reagent such as, for example, polystyrene-bound cyanoborohydride in a solvent such as THF affords the amine derivatives (14e).

Alternatively, alcohol (14c) can be reacted with a suitable acylating or alkylating agent under suitable conditions to provide ester or ether compounds, ie G is O in the general formula (I).

Subsequent reaction of the formed alcohol with a suitable acylating or alkylating agent using suitable conditions provides the ester or ether compounds, ie G is O in the general formula (I).

Alternatively, the bond between the P2 and P3 building blocks can be via a guanidine group and a general route to such compounds is depicted in Scheme 15. Treatment of the P2 building block (15a) with thiocarbonyldiimidazole or the like in a solvent such as dimethylformamide followed by condensation with sodium -cyanamide in a solvent such as ethanol gives the thiolate intermediate (15b). Reaction of intermediate (15b) with the desired building block, shown here as a capped P3 building block (12c) affords the cyanoguanidine derivative (15d). Other building blocks, R<16->G or R<16->G-P4-P3, can alternatively be connected to the intermediate product (15b). Hydrolysis of the cyano group by treatment of (15d) with dilute hydrochloric acid gives the guanylurea derivative (15e).

When R7, R7' and A' contain functional groups, these are suitably protected by methods known to those skilled in the art, see for example Bodanzky or Greene indicated above.

Formation of macrocyclic compounds

Compounds according to the present invention in which an alkylene chain which extends from the R7/R7 -cycloalkyl to Rx or R<11> and thus forms a macrocycle, can be prepared as described below. Appropriate P1, P2 and P3 building blocks or precursors thereof are coupled using the strategies described above, followed by a ring closure reaction (macrocyclization). The substituent W-R<8> in the P2 building block can be incorporated via a Mitsunobu reaction as described above, before or after formation of the macrocycle, or the desired building blocks can be linked together using the appropriately substituted P2 building block. For macrocyclic structures extending from the R<7>/R<7>cycloalkyl to R<11>, P3 amino acids containing the appropriate side chain can be prepared as described in WO00/59929.

A typical route to macrocyclic compounds is shown in Scheme 18 which illustrates the technique applied to a compound having a 5-membered P2 structure and a spiro-cyclopropyl group in the P1 unit, where the macrocycle extends from the P3 side chain.

Coupling of proline derivative (16a) with the appropriate acid-protected amino acid (16b) using e.g. the phosgene conditions described above provide (16c). Formation of the macrocycle can then be carried out via an olefin metathesis reaction using a Ru-based catalyst such as that reported by Miller, S.J., Blackwell, H.E.; Grubbs, R.H. J. Am. Chem. Soc. 118, (1996), 9606-9614, Kingsbury, J.S., Harrity, J.P.A., Bonitatebus, P.J., Hoveyda, A.H., J. Am. Chem. Soc. 121, (1999), 791-799 and Huang et al., J. Am. Chem. Soc. 121, (1999), 2674-2678. It will also be known that catalysts containing other transition metals such as Mo can be used for this reaction. Optionally, the double bond is reduced and/or the ethyl ester is hydrolysed using respectively normal hydrogenation and/or hydrolysis methods, which is well known in the art. Alternatively, the methyl ester can be selectively hydrolyzed followed by coupling of an R<16->G-P4 building block under normal peptide coupling conditions. The macro-cyclization step described in Scheme 16 can also be applied to the corresponding carbocyclic analogues described above. When the linker contains a nitrogen atom, the ring closure can be carried out by reductive amination as described in WO00/59929.

Macrocyclic compounds without the cyclopropyl group in the P1 moiety, i.e. the macrocyclic ring extends directly from the peptidic backbone at the carbon atom adjacent to R<7>, can be prepared using the methods described herein. An example in which a proline derivative is used as the cyclic P2 structure is shown in Scheme 17.

Coupling of an appropriate allylglycine derivative (17a) to the acid function of the P2 building block (17b) using normal peptide coupling conditions gives the amide derivative (17c). Removal of the Boc protecting group by acid treatment followed by formation of a chlorocarbamate by treatment with phosgene in the presence of sodium bicarbonate and subsequent reaction with the olefin-substituted amino acid (17d) affords the urea compound (17e). A ring-closing metathesis reaction is then carried out by using, for example, Hoveyda-Grubb's catalyst which gives the macrocyclic compound (17f).

Although Scheme 17 shows the synthetic sequence using a P2 building block in which the R8 substituent is attached to the backbone, it will be apparent that an unsubstituted P2 backbone could be used and the R8 group introduced at any appropriate stage of the synthesis, by using any of the methods described herein. Building blocks that can be used in the preparation of compounds in which the macrocycle extends from the nitrogen in the bond between the P2 and P3 fragments, i.e. X is NRx in the general formula I or in the preparation of compounds in which the P3 and P4 fragments are absent, i.e. m and n is 0, and G is NRj in the general formula I, can normally be prepared as indicated in scheme 18B.

Carbamate 18a, which is commercially available, or can be easily prepared,

for example, by reacting the desired alkylamine with di-tert-butyldicarbonate, can be reacted with an appropriate co-unsaturated alcohol under Mitsunobu conditions to provide the alkylated carbamate (18b). Exposure of 18b to acidic conditions such as, for example, treatment with trifluoroacetic acid in a solvent such as dichloromethane gives the free amine (18c) which can be bound to a P2 fragment using any of the previously described strategies.

Macrocyclic structures containing a hydrazine group, ie X is NRjNRj or m, and n is 0, and G is NRjNRj, in the general formula I, can be prepared by attaching an appropriate N-alkylated carbazate derivative to the P2 fragment. Alkylated carbazate derivatives can be prepared, for example, as described in Scheme 19.

Oxidation of the appropriate alcohol (19a) by a suitable oxidation method such as, for example, with N-methylmorpholine oxide and tetrapropylammonium perruthenate in a solvent such as dichloromethane provides the aldehyde (19b). Reductive alkylation of tert-butylcarbazate with the resulting aldehyde gives the desired N-alkylated building block (19c). Alternatively, any desired hydrazine derivative such as morpholin-1-ylamine, piperidin-1-ylamine or the like can be used instead of tert-butylcarbazate in the reaction with aldehyde 19b.

Scheme 20 illustrates synthetic sequences of building blocks suitable for the preparation of compounds in which the "outer" nitrogen of the hydrazine group is alkylated, either with a co-unsaturated alkyl chain suitable for subsequent macro-

cyclization or with any other suitable alkyl group.

Reaction of a suitable protected hydrazine derivative, for example (1,3-dioxo-1,3-dihydro-isonidol-2-yl)-carbamic acid tert-butyl ester (20a), which can be easily prepared by one skilled in the art, with a desired alcohol , R-OH, under Mitsunobu conditions, provides the N-alkylated hydrazine compound (20b). Removal of the phthalimido group by treatment with hydrazine or a derivative thereof such as hydrazine hydrate or hydrazine acetate affords the carbazate (20c). The primary amine obtained can then either be coupled to any desired P2 fragment using any of the methods previously described to give the urea derivative (20d), or alternatively it can be further alkylated using, for example, the reductive amination method described in scheme 19 followed by coupling to a P2 fragment as previously described to give 20e .

Scheme 21 exemplifies the coupling of a hydrazine-containing P3 building block to a cyclopentane structure followed by macrocyclization.

Coupling of the carbazate derivative (21b) to the P2-P1 building block (21a) using normal peptide coupling conditions affords intermediate (21c). Ring closure of (21c) by an olefin metathesis reaction as described in scheme 18 gives the macrocyclic compound (21d).

The term "N-protecting group" or "N-protected" as used herein refers to those groups intended to protect the N-terminus of an amino acid or peptide or to protect an amino group from unwanted reactions during synthetic procedures. Commonly used N-protecting groups are described in Greene, "Protective Groups in Organic Synthesis" (John Wiley & Sons, New York, 1981). N-protecting groups include acyl groups such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, a-chlorobutyryl, benzoyl, 4-chlorobenzoyl , 4-bromobenzoyl, 4-nitrobenzoyl and the like; sulfonyl groups such as benzenesulfonyl, p-toluenesulfonyl and the like, carbamate-forming groups such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2 -nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, l-(p-biphenylyl)-l-methylethoxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl, t-butoxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl , ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl and the like; alkyl groups such as benzyl, triphenylmethyl, benzyloxymethyl and the like; and silyl groups such as trimethylsilyl and the like. Preferred N-protecting groups include formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, phenylsulfonyl, benzyl, t-butoxycarbonyl (BOC) and benzyloxycarbonyl (Cbz).

Hydroxy protecting group as used herein refers to a substituent that protects hydroxyl groups from unwanted reactions during synthetic procedures such as the O-protecting groups described in Greene, "Protective Groups in Organic Synthesis," (John Wiley & Sons, New York (1981)). Hydroxy protecting groups include substituted methyl ethers, for example, methoxymethyl, benzyloxymethyl, 2-methoxyethoxymethyl, 2-(trimethylsilyl)ethoxymethyl, t-butyl and other lower alkyl ethers, such as isopropyl, ethyl and especially methyl, benzyl and triphenylmethyl; tetrahydropyranyl ethers; substituted ethyl ethers, for example, 2,2,2-trichloroethyl; silyl ethers, for example, trimethylsilyl, t-butyldimethylsilyl and t-butyldiphenylsilyl; and esters prepared by reacting the hydroxyl group with a carboxylic acid, for example acetate, propionate, benzoate and the like.

In the treatment of conditions caused by flaviviruses such as HCV, the compounds of formula I are normally administered in an amount to achieve a plasma level of about 100 to 5000 nM, such as 300 to 2000 nM. This corresponds to a dosage rate, depending on the bioavailability of the formulation, in the order of 0.01 to 10 mg/kg/day, preferably 0.1 to 2 mg/kg/day. A typical dosage rate for a normal adult person will be about 0.05 to 5 g per day, preferably 0.1 to 2 g such as 500-750 mg, in one to four dosage units per day. day. As with all pharmaceuticals, dosage rates will vary with the size and metabolic state of the patient as well as the severity of the infection and may need to be adjusted for concomitant medications.

According to good prescribing practice in antiviral therapy, the compounds of formula I are normally co-administered with other HCV therapies to avoid the generation of drug escape mutants. Examples of such additional HCV antiviral therapies include ribavirin, interferons, including pegylated interferons. In addition, a number of nucleoside analogues and protease inhibitors are in clinical or preclinical development and will be available for co-administration with the compounds of the invention.

Accordingly, a further aspect of the invention provides a composition comprising a compound of formula I and at least one further HCV antiviral agent in a common dosage unit, such as any of the dosage forms described below, but particularly an orally administered tablet or capsule or a liquid suspension or solution for oral application or injection application. Supply of the compounds or compositions according to the invention can be carried out by sequential or simultaneous administration of a compound of formula I and at least one further HCV antiviral agent. A related aspect of the invention provides a patient package comprising a first pharmaceutical composition, preferably in unit dosage form, of the compound of formula I and a second pharmaceutical composition, normally also in unit dosage form and generally in a separate container within the patient package, of another HCV antiviral agent. A patient package will conveniently also be provided with instructions printed on the package or a container therein or on a package insert, for simultaneous or sequential administration of the respective pharmaceutical compositions.

Many HCV patients are co-infected, or have a tendency for superinfection, with other infectious diseases. The compound according to the invention and at least one further anti-infective agent can be administered simultaneously or sequentially, normally in doses corresponding to the monotherapy dose for the agent in question. However, certain anti-infectives can induce a synergistic response, which makes it possible to administer one or both active ingredients at a lower dose than the corresponding monotherapy. For example, in drugs that tend to be rapidly metabolized by Cyp3A4, co-dosing with the HIV protease inhibitor ritonavir may allow lower dosage regimens to be administered.

Typical coinfections or superinfections with HCV include hepatitis B virus or HIV. Accordingly, the compound of the invention is advantageously co-administered (either in the same dosage unit, co-packaged or in a separately prescribed dosage unit) with at least one HIV antiviral agent and/or at least one HBV antiviral agent.

Representative HIV antiviral agents include NRTIs such as alovudine (FLT), zudovudine (AZT, ZDV), stavudine (d4T, Zerit), zalcitabine (ddC), didanosine (ddl, Videx), abacavir, (ABC, Ziagen), lamivudine (3TC , Epivir), emtricitabine (FTC, Emtriva), racevir (racemic FTC), adefovir (ADV), entacavir (BMS 200475), alovudin (FLT), tenofovir disoproxil fumarate (TNF, Viread), amdoxavir (DAPD), D- d4FC (DPC-817), -dOTC (Shire SPD754), elvucitabine (Achillion ACH-126443), BCH 10681 (Shire) SPD-756, racivir, D-FDOC, GS7340, INK-20 (thioetherphospholipid AZT, Kucera), 2 '3'-dideoxy-3'-fluoroguanosine (FLG) & its prodrugs such as MIV-210, reversed (RVT, D-D4FC, Pharmasset DPC-817) .

Representative NNRTIs include delavirdine (Rescriptor), efavirenz (DMP-266, Sustiva), nevirapine (BIRG-587, Viramune), (+)calanolide A and B (Advanced Life Sciences), capravirine (AG1549f S-1153; Pfizer), GW -695634 (GW-8248; GSK), MIV-150 (Medivir), MV026048 (R-1495; Medivir AB/Roche), NV-05 2 2 (Idenix Pharm.), R-278474 (Johnson & Johnson), RS -1588 (Idenix Pharm.), TMC-120/125 (Johnson & Johnson), TMC-125 (R-165335; Johnson & Johnson), UC-781 (Biosyn Inc.) and YM215389 (Yamanoushi).

Representative HIV protease inhibitors include PA-457 (Panacos), KPC-2 (Kucera Pharm.), 5 HGTV-43 (Enzo Biochem), amprenavir (VX-478, Agenerase), atazanavir (Reyataz), indinavir sulfate (MK- 639, Crixivan), Lexiva (fosamprenavir calcium, GW -433908 or 908, VX-175), ritonavir (Norvir), lopinavir + ritonavir (ABT-378, Kaletra), tipranavir, nelfinavir mesylate (Viracept), saquinavir (Invirase , Fortovase), AG1776 (JE-2147, KNI-764; Nippon Mining Holdings), AG-1859 (Pfizer), DPC-681/684 (BMS), GS224338; Gilead Sciences), KNI-272 (Nippon Mining Holdings), Nar-DG-35 (Narhex), P(PL)-100 (P-1946; Procyon Biopharma), P-1946 (Procyon Biopharma), R-944 (Hoffmann -LaRoche), RO-0334649 (Hoffmann-LaRoche), TMC-114 (Johnson & Johnson), VX-385 (GW640385; GSK/Vertex), VX-478 (Vertex/GSK).

Other HIV antiviral agents include entry inhibitors, including fusion inhibitors, inhibitors of the CD4 receptor, inhibitors of the CCR5 co-receptor and inhibitors of the CXCR4 co-receptor or a pharmaceutically acceptable salt or prodrug thereof. Examples of entry inhibitors are AMD-070 (AMD11070; AnorMed), BlockAide/CR (ADVENTRX Pharm.), BMS 806 (BMS-378806; BMS), Enfurvirtide (T-20, R698, Fuzeon), KRH1636 (Kureha Pharmaceuticals), ONO -4128 (GW-873140, AK-602, E-913; ONO Pharmaceuticals), Pro-140 (Progenics Pharm), PR0542 (Progenics Pharm.), SCH-D (SCH-417690; Schering-Plough), T-1249 (R724; Roche/Trimeris), TAK-220 (Takeda Chem. Ind.), TNX-355 (Tanox) and UK-427,857 (Pfizer). Examples of integrase inhibitors are L-870810 (Merck & Co.), c-2507 (Merck & Co.) and S(RSC)-1838 (shionogi/GSK).

Examples of HBV antivirals include adefovir dipivoxil (Hepsera) and especially lamivudine and 2'3'-dideoxy-3'-fluoroguanosine (FLG) & its prodrugs such as MIV-210, the 5'-0-valyl-L-lactyl prodrug of FLG. These latter HBV antiviral agents are particularly suitable as they are also active against HIV.

While it is possible to administer the active agent alone, it is preferable to prepare it as part of a pharmaceutical formulation. Such a formulation will comprise the active agent defined above together with one or more acceptable carriers or excipients and optionally other therapeutic ingredients. The carrier(s) must be acceptable in the sense that it is compatible with the other ingredients in the formulation and not harmful to the recipient.

The formulations include those compounds suitable for rectal, nasal, topical (including buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration, but preferably the formulation is an orally administered formulation. The formulations can conveniently be prepared in unit dose form, e.g. sustained release tablets and capsules, and they may be prepared by any method well known in the art of pharmacy.

Such methods include the step of bringing the above-defined active agent into contact with the carrier. In general, the formulations are prepared by uniformly and thoroughly contacting the active agent with liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product. Thus, a pharmaceutical composition can be prepared by bringing a compound of formula I or its pharmaceutically acceptable salt into combination or contact with a pharmaceutically acceptable carrier or vehicle. If the preparation of pharmaceutical formulations involves thorough mixing of pharmaceutical excipients and the active ingredient in salt form, then it is often preferred to use excipients which are non-basic in nature, i.e. either acidic or neutral.

Formulations for oral administration may be prepared as separate units such as capsules, powder capsules or tablets each containing a predetermined amount of the active agent; as a powder or granules; as a solution or a suspension of the active agent in an aqueous liquid or a non-aqueous liquid; or as a liquid oil in water emulsion or a liquid water in oil emulsion and as a bolus etc.

With respect to compositions for oral administration (e.g. tablets and capsules) the term includes suitable carrier vehicles such as common excipients e.g. binding agents, such as syrup, acacia, gelatin, sorbitol, tragacanth, polyvinylpyrrolidone (Povidone), methylcellulose, ethylcellulose, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, sucrose and starch; fillers and carriers, for example, corn starch, gelatin, lactose, sucrose, microcrystalline cellulose, kaolin, mannitol, dicalcium phosphate, sodium chloride and alginic acid; and lubricants such as magnesium stearate, sodium stearate and other metallic stearates, stearic acid, glycerol stearate, silicone fluid, talc wax, oils and colloidal silicic acid. Aromatic substances such as peppermint, wintergreen oil, cherry aroma or the like can also be used. It may be desirable to add a dye to make the dosage form easy to identify. Tablets can also be coated using methods well known in the art.

A tablet can be prepared by compression or molding, optionally with one or more additional ingredients. Compressed tablets can be prepared by pressing in a suitable machine the active agent in free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surfactant or dispersant. Molded tablets may be prepared by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated to provide slow or controlled release of the active agent.

Other formulations suitable for oral administration include lozenges comprising the active agent in an aromatic base, usually sucrose and acacia or tragacanth; lozenges comprising the active agent in an inert base such as gelatin and glycerin or sucrose and acacia; and mouthwash comprising the active agent in a suitable liquid carrier.

The compounds of formula I can form salts, which constitutes a further aspect of the invention. Suitable pharmaceutically acceptable salts of the compounds of formula I include salts of organic acids, especially carboxylic acids, including acetate, trifluoroacetate, lactate, gluconate, citrate, tartrate, maleate, malate, pantothenate, isethionate, adipate, alginate, aspartate, benzoate, butyrate, digluconate , cyclopentanate, glucoheptanoate, glycerophosphate, oxalate, heptanoate, hexanoate, fumarate, nicotinate, palmoate, pectinate, 3-phenylpropionate, picrate, pivalate, proprionate, tartrate, lactobionate, pivolate, camphorate, undecanoate and succinate, organic sulfonic acids such as methanesulfonate, ethanesulfonate , 2-hydroxyethanesulfonate, camphorsulfonate, 2-naphthalenesulfonate, benzenesulfonate, p-chlorobenzenesulfonate and p-toluenesulfonate; and inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, bisulfate, hemisulfate, thiocyanate, persulfate, phosphoric acid and sulfonic acid.

The compounds according to the invention have different steric centers, and the invention includes racemates and enantiomers that each of these steric centers.

Normally, the stereochemistry of the groups corresponding to the P3 and P4 side chains (ie R<15> and/or R<11>) will correspond to an L-amino acid configuration, although the invention also encompasses D-isomers at one or both of these centers. It is worth noting that the L configuration is active despite the fact that the nature of the E group means that P3 and P4 are normally rearranged one atom relative to a conventional polypeptide and the fact that the inversion of a peptide residue, as planned for P3 and P4 then throws the amino acid side chain to the opposite side compared to a conventional peptide substrate.

The stereochemistry of the backbone component of the cyclic P2 group (ie spanning the carbonyl of the P1 amide bond and the carbonyl or E extending out of P3) will normally correspond to L-proline. The stereochemistry of the P2 ring atom to which W is attached is normally as shown:

In compounds according to the invention in which R7 and R<7>' together form a spiroalkyl group, such a spiro-cycloalkyl will normally comprise an R7a substituent on the spiro-cyclopropyl ring which is oriented syn in relation to A: or anti in relation to A:

Conveniently, the spiro carbon in such a spiro-cyclopropyl ring has the R configuration:

Conveniently, an R7a substituent on a spiro-cyclopropyl ring adjacent to A in syn orientation is in the following absolute configuration:

Particularly preferred variants are when R<7a>comprises ethyl, consequently they have asymmetric carbon atoms in positions 1 and 2 R,R configuration. Alternatively preferably, R<7a> comprises vinyl, consequently the asymmetric carbon atoms in positions 1 and 2 have R,S configuration.

Where the compound according to the invention is a macrocycle comprising a J group, J is preferably a diastereomer represented by the partial structures (i) or (ii):

Detailed description of the designs

Various designs of the invention will now be described only by way of illustration with reference to the following examples.

Example 1

7-Methoxy-2-phenyl-quinolin-4-ol (1)

To a stirred round bottom flask with toluene (100 mL) was added ethyl benzoyl acetate (18.7 g, 97 mmol) and m-anisidine (12 g, 97 mmol). 4 M HCl in dioxane (0.5 mL) was added and the reaction mixture was refluxed for 6 h (140°C). The mixture was co-evaporated with toluene. To the crude mixture was added diphenyl ether (50 mL) and the mixture was heated to 280°C for 2 h. When the theoretical amount of ethanol (6 mL) was collected in a Dean Stark trap, heating was terminated and the mixture was cooled to rt . The crude mixture was dissolved in CH 2 Cl 2 (100 ml) and stirred for 30 min. The precipitate formed was filtered off and dried to give 1 (4.12 g, 16.4 mmol, 17%): pale yellow powder.

<*>H (300 MHz, DMSO-D6): δ 3.8 (s, 3H), 6.24 (s, 1H), 6.88-6.96 (dd, 1H, J = 9.07 Hz , J = 2.47 Hz), 7.19 (d, 1H, J=2.19 Hz), 7.56 (t, 3H, J = 2.19 Hz), 7.8 (dd, 2H, J = 7.14 Hz, J = 2.19 Hz), 8.0 (d, 1H, J = 9.06Hz); 13C (75.5 MHz, DMSO-D6): δ 55.3, 99.6, 106.9, 113.1, 119.1, 126.4, 127.5, 128.8, 130.2, 134.1, 142.2, 149.4, 161.8, 176.4.

Example 2

Boc-L-tert-leucine-OH (2)

Triethylamine (890 µl, 6.40 mmol) was added dropwise to a stirred solution of L-tert-leucine (300 mg, 2.29 mmol) and di-tert-butyl dicarbonate (599 mg, 2.74 mmol) in dioxane/ water 1:1 (8 mL), and the solution was stirred overnight. The mixture was extracted with petroleum ether (2x), and the aqueous phase was cooled to 0°C and gently acidified to pH 3 by slow addition of 4M NaHSO 4 H 2 O. The acidified aqueous phase was extracted with EtOAc (3x) and the combined organic phases were washed with brine (2x) and then dried, filtered and concentrated to give the title compound (522 mg, 99%) as a colorless powder. No further purification was required.

<1>H-NMR (300 MHz, CD3OD) 8 0.99 (s, 9H), 1.44 (s, 9H), 3.96 (s, 1H);<13>C-NMR (75.5 MHz, CD 3 OD) 8 27.1, 28.7, 34.9, 68.0, 80.5, 157.8, 174.7.

Example 3

ffS)- Cyclohexyl- methylcarbamol- methylyl)- carbamide- tert- butyl ester ( 3)

Boc-Chg-OH (387 mg, 1.50 mmol) was coupled to methylamine hydrochloride (111 mg, 1.65 mmol) using the same HATU coupling conditions as in the synthesis of compound 7. The crude product was extracted with EtOAc, washed with brine and concentrated. Purification by flash column chromatography (EtOAc) afforded the title compound (307 mg, 76%) as a colorless solid.

<1>H-NMR (300 MHz, CDCl3) 8 0.91-1.13 (m, 2H), 1.14-1.31 (m, 3H), 1.44 (s, 9H), 1, 61-1.80 (m, 6H), 2.80 (d, J = 4.7 Hz, 3H), 3.91 (dd, J = 7.1, 9.1 Hz, 1H), 5.23

(b, 1H), 6.52 (bs, 1H); 29.6, 40.5, 59.5, 79.7, 155.9, 172.4.

Example 4

• ffS)- l- rffS)- Cvchlorieksvl- metvlcarbamovl- metvn- carbamovl- 2, 2- dimetvl- propvl>-carbamic acid- ethert- butvlester ( 4)

To a solution of compound 3 (98 mg, 0.362 mmol) in methylene chloride (3 mL) was added triethylsilane (115 mL, 0.742 mmol) and TFA (3 mL). The mixture was stirred for 2 h at room temperature and was then evaporated and co-evaporated with toluene. The deprotected amine was dissolved in DMF (5 mL) and coupled to compound 2 (84 mg, 0.363 mmol) using the same HATU coupling conditions as in the synthesis of 7. The crude product was extracted with EtOAc, washed with brine, dried, filtered and concentrated. Purification by flash column chromatography (toluene/EtOAc 1:1) afforded the title compound (128 mg, 92%) as a colorless solid.

<1>H-NMR (300 MHz, CDCl3) 8 0.99 (s, 9H), 1.02-1.30 (m, 5H), 1.44 (s, 9H), 1.58-1, 77 (m, 4H), 1.78-1.89 (m, 2H), 2.79 (d, J = 4.7 Hz, 3H), 4.11 (d, J = 9.3 Hz, 1H ), 4.33 (app, t, J = 8.5 Hz, 1H), 5.65 (b, 1H), 7.25 (b, 1H), 7.39 (b, 1H);<13> C-NMR (75.5 MHz, CDCI3) 8 25.9, 25.9, 26.0, 26.2, 26.8, 28.4, 29.0, 29.7, 34.5, 39, 7, 58.4, 62.4, 79.4, 156.0, 171.4, 171.8.

Example 5

Hept-6- enal ( 5)

Ground molecular sieve (3 .5 g, 4 Å).

The mixture was stirred for 10 min at room temperature under a nitrogen atmosphere before tetrapropylammonium perruthenate (TPAP) (131 mg, 0.37 mmol) was added. After

stirring for a further 2.5 h, the solution was filtered through celite. The solvent was then carefully evaporated and the remaining solution was purified by flash column chromatography (DCM) to give the volatile aldehyde 5 (620 mg, 74%) as an oil.

Example 6

/ V'- Hept- 6- en-( E)- vlidene- hvdrazincarboxylsvre- tert- butyl ester ( 6)

To a solution of 5 (68 mg, 0.610 mmol) and tert-butylcarbazate (81 mg, 0.613 mmol) in MeOH (5 mL) was added ground molecular sieve (115 mg, 3Å). The mixture was stirred for 3 h after which it was filtered through celite and evaporated. The residue was dissolved in dry TH F (3 ml), and AcOH (3 ml). NaBH 3 CN (95 mg, 1.51 mmol) was added and the solution was stirred overnight. The reaction mixture was diluted with saturated NaHCO 3 solution (6 mL) and EtOAc (6 mL). The organic phase was washed with brine, saturated NaHCO 3 , brine, dried over MgSO 4 and evaporated. The cyanoborane adduct was hydrolyzed by treatment with MeOH (3 mL) and 2 M NaOH (1.9 mL). The mixture was stirred for 2 h, and the MeOH was evaporated. H 2 O (5 mL) and DCM (5 mL) were added and the aqueous phase was extracted three times with DCM. The combined organic phases were dried and evaporated. Purification by flash column chromatography (toluene/ethyl acetate 9:1 with 1% triethylamine and toluene/ethyl acetate 6:1 with 1% triethylamine) afforded the title compound (85 mg, 61%) as an oil.

Example 7 ffS)-1-Cyclopentylcarbamoyl-2, 2-dimethyl- propion- carbamides- tert-butyl ester ( 7 )

To a cold solution of 2 (133 mg, 0.575 mmol), cyclopentylamine (64 mL, 0.648 mmol) and DIEA (301 mL, 1.73 mmol) in DMF (3 mL) was added the coupling reagent HATU (240 mg, 0.631 mmol) . The mixture was stirred for half an hour and for another two hours at room temperature. The solvent was removed by heating the reaction flask in a water bath under reduced pressure, and the residue was dissolved in ethyl acetate, after which the organic phase was washed three times with brine, dried, filtered and evaporated. Purification by flash column chromatography (toluene/ethyl acetate 4:1) afforded the title compound (140 mg, 82%) as colorless crystals.

<1>H-NMR (300 MHz, CDCl3): δ 0.95 (s, 9H), 1.28-1.48 (m, overlapped, 2H), 1.40 (s, 9H), 1.49 -1.71 (m, 4H), 1.86-2.01 (m, 2H), 3.76 (b, 1H), 4.09-4.23 (m, 1H), 5.32 (b , 1H), 5.91 (b, 1H); ,1, 34.5, 51.0, 62.2, 79.4, 155.9, 170.3.

Example 8

( S)- tert- Butoxxcarbonvlamino- cvclohexvl- acetic acid- ester ( 8)

To a solution of Boc-Chg-OH (53 mg, 0.206 mmol) in acetone (3 mL) was added methyl iodide (195 mL, 3.1 mmol) and silver(II) oxide (53 mg, 0.229 mmol). The mixture was allowed to stir overnight in a reaction flask which was covered with aluminum foil. The solution was then filtered through celite and evaporated. Purification by flash column chromatography (toluene/ethyl acetate 15:1) afforded methyl ester 8 (56 mg, 100%) as a colorless oil.

<1>H-NMR (300 MHz, CDCl3): 8 1.00-1.34 (m, 5H), 1.44 (s, 9H), 1.54-1.82 (m, 6H), 3 .73 (s, 3H), 4.20 (dd, J = 2.8, 5.0 Hz, 1H), 5.05 (bs, 1H);<13>C-NMR (75.5 MHz, CDCl3 ): 5 26.0, 28.2, 28.3, 29.5, 41.1, 52.0, 58.3, 79.7, 155.6, 172.9.

Example 9

fS)- ffS)- 2- Benzyloxycarbonylamino- 3- methyl- butylamino)- chlorohexyl-acetic acid- methylester ( 9)

Compound 8 (93 mg, 0.343 mmol) was deprotected and coupled to Z-Val-OH (95 mg, 0.378 mmol) according to the method for the preparation of 39. Flash column chromatography (toluene/ethyl acetate 4:1) gave the title compound (131) mg, 94%) as a colorless solid.

<1>H-NMR (300 MHz, CDCl3): δ 0.92-1.30 (m, 11H), 1.54-1.88 (m, 6H), 2.02-2.18 (m, 1H), 3.72 (s, 3H), 4.05-4.18 (m, 1H), 4.52 (dd, J = 3.0, 5.5 Hz, 1H), 5.12 (s , 2H), 5.49 (bs, 1H), 6.52 (bs, 1H), 7.34 (s, 5H);<13>C-NMR (75.5 MHz, CDCl 3 ): δ 17.8 , 19.0, 25.8, 28.2, 29.3, 31.2, 40.5, 51.9, 56.8, 60.0, 66.8, 127.7, 127.9, 128 ,1, 128.3, 136.2, 156.3, 171.3, 172.2.

Example 10

N- Boc- 4R-( 2- phenyl- 7- methoxyquinoline- 4- oxo) proline f 10)

To a stirred solution of N-Boc-trans-4-hydroxy-L-proline (3.9 g, 16.9 mmol) in DMSO (90 ml) was added potassium tert.butoxide (4.5 g, 40.1 mmol). After 1 h, 4-chloro-2-phenyl-7-methoxyquinoline (4.5 g, 16.7 mmol) was added and stirred at RT for 12 h. The mixture was diluted with water (180 mL), washed with ethyl acetate (1 x 30 mL) and neutralized with 1N HCl. The solid was filtered, washed with water and dried to give (4.65g, 10mol) of product. >95% purity according to HPLC. M+H<+>464.2.

Example 11

2- f l- Ethoxycarbonvl- 2- vinvl- cvclopropylcarbamovn- 4- f7- methoxysv- 2- phenvl- quinoline-4- vloxvl)- pyrrolidine- l- carboxyvl- tert. butflester file)

To a solution of l-amino-2-vinyl-cyclopropanecarboxylic acid ethyl ester (41 mg, 0.26 mmol), 10 (11 mg, 0.22 mmol), HATU (204 mg, 0.54 mmol) in DMF (4 ml) was added diisopropylethylamine (187 µl, 1.08 mmol). After stirring at RT for 1 hour, dichloromethane (4 mL) was added. The solution was washed with aqueous NaHCO 3 (saturated) and with two portions of water. The organic layer was dried and concentrated. The product was pure enough (>95% by HPLC) to be used in the next step. M+H<+>602.2.

Example 12

1- ir4- f7- Methoxy- 2- phenyl- quinoline- 4- cyclohex)- pyrrolidin- 2- carbonyl- amino>- 2- vinylcyclopropanecarboxylic acid ester ( 12)

Compound 11 was kept in TFA-DCM 1:2 (3 mL) at RT for 60 min. Toluene (3 mL) was added. The sample was co-evaporated to dryness. Purity according to HPLC >95%. M+H<+>502.4.

Example 13

l- free-ri- f2- Hydroxy- indan- l- vlcarbamovl)- 2, 2- dimethyl- propvlcarbamovl- 4-( 7- methoxy- 2- phenyl- quinolin- 4- yloxy)- pyrrolidin- 2- carbonyl1- amino >- 2- vinyl-cyclopropanecarboxylic acid ester ( 13 ) To a solution of compound 12 (0.13 mmol) in THF (2 mL), was added a large excess of NaHCO 3 (s) and a solution of phosgene in toluene (1 .6 M, 600 µl). After 10 min of shaking, the slurry was filtered and concentrated to dryness. The solid was redissolved in dichloromethane, and a large excess of NaHCO 3 (s) and 2-amino-[v-(2-hydroxy-indan-1-yl)-3,3-dimethyl-butyramide (0.65 mmol) was added. The slurry was shaken for 24-40 hours at RT. The slurry was filtered, concentrated and subjected to silica column chromatography (gradient elution from 100% DCM to MeOH/DCM 2:98) to give the title compound (89.6 mg, 0.11 mmol). Purity according to HPLC >95%. M+H<+>790.3. Example 14

l- ri- ri- f2- Hydroxv- indan- l- vlcarbamovl)- 2, 2- dimethyl- propyll- 4- f6- methoxyl- 3-phenvl- naphthalene- l- vloxv)- pvrrolidine- 2- vll- 2- vinyl-cyclopropanecarboxylic acid ( 14)

To a solution of 13 (76.7 mg, 0.097 mmol) in THF-MeOH 2:3 (2 ml) was added IM LiOH 5 equiv. The solution was kept at 60°C for 60 min. After cooling to RT, HOAc 15-30 equiv. added followed by toluene (2 mL) and then concentrated to dryness. The residue was taken up in DCM and washed with water. The organic layer was dried and concentrated to give the title compound (72 mg, 0.094 mmol). Purity >95% according to HPLC M+H<+>762.2.

Example 15

/ V- f2- Hydroxv- indan- l- vl)- 2- r4- f6- methoxyl- 3- phenyl- naphthalene- l- loxv)- 2- f 1-phenvlmethanesulfonvlaminocarbonvl- 2- vinvl- cyclopropvl)- pyrrolidin- l - vIII- 3, 3- dimethylbutyramide ( 15)

To a solution of 14 (25 mg, 0.033 mmol) in chloroform (1 mL) was added benzenesulfonamide (10.5 mg, 0.066 mmol) followed by diisopropylethylamine (34 µL, 0.197 mmol). The solution was stirred at RT for 10 min and then at -20 °C for 30 min. PyBOP (76 mg, 0.13 mmol) was then added as a solid. The solution was kept at -20 °C for 48 hours. The solution was then poured into aqueous NaHCO 3 (saturated) and washed with water. The organic layer was dried, concentrated and subjected to purification by HPLC to give the title compound as a white solid.

Example 16

Resin-bound 2-tier. butoxxcarbonvlamino- 3, 3- dimethyl butyric acid ( 16)

To Argonaut resin PS-TFP (1.38 mmol/g, 10 g) and 2-tert-butoxycarbonylamino-3,3-dimethylbutyric acid (4.5 g, 20.7 mmol) was added dichloromethane (40 mL) and DMF (10ml). To this mixture was added DMAP (1 g, 8.28 mmol) followed by DIC (9.5 mL, 60.7 mmol). After 3 h of shaking at RT, the resin was filtered and washed successively with DMF, THF, DCM, THF, DCM and ether and then dried in vacuo.

Example 17

ri-(2-Hydroxy-indan-1-vlcarbamovyl)- 2, 2- dimethyl-propvyl-carbamide acid-tertbutyl ester (17)

To a portion of 16 (200 mg) in DCM was added aminoindanol (0.14 mmol). The mixture was shaken for 2 hours. The liquid was filtered off and the resin was washed with 2xDCM. The liquids were combined and concentrated to dryness to afford the title compound (20.5 mg, 0.055 mmol) Purity >95% by HPLC. M+H<+>363.15.

<13>C NMR8C(100 MHz; CDCl3; Me4Si) 27.0, 28.5, 34.2, 39, 8, 50.8, 57.9, 68.2, 73.7, 124.8, 125 .6, 127.4, 128.5, 140.4, 171.6,<*>H NMR 8H (400 MHz; CDCl 3 ; Me 4 Si) 1.07

(9H, s, CCH3), 1.44 (9H, s, OCCH3), 2.93 (1H, dd, Jgem16.4 Hz, J3, 22.3 Hz, CH2), 3.15 (1H, dd, Jgem16.4Hz, J3.25.2Hz, CH2),

Example 18

2- Amino-/V-(2-hydroxvHndan-l- vl)- 3, 3- dimethylbutyramide ( 18)

Compound 17 was kept in DCM-TFA 2:1 (2 mL) for 60 min at RT. The solution was co-evaporated with toluene to dryness.

Example 19

( 2-tert--Butoxycarbonylamino- 3, 3-dimethyl- butylamino)-cyclohexyl-acetic acid ester (19)

To a solution of 2-tert.butoxycarbonylamino-3,3-dimethyl-butyric acid (500 mg, 2.16 mmol), amino-cyclohexyl-acetic acid methyl ester (444 mg, 2.59 mmol) and HATU (2 g, 5 .40 mmol) in DMF (20 mL) was added diisopropylethylamine (1.88 mL, 10.8 mmol). The solution was stirred for 1 hour at r.t. and diluted with dichloromethane (40 mL). This solution was washed with aqueous NaHCO 3 (saturated) and water (x2), dried and concentrated. The product was >95% pure. M+H<+>385.4.

Example 20

• fl- rfCvclohexvl- metvlcarbamovl- metvl)- carbamovl- 2, 2- dimetvl- propvl>-carbamidevre- etrt- butvle ester ( 20

A large excess of methylamine (30% in water) was added to compound 19 in EtOH-THF 1:2, and the solution was allowed to stand at RT for 2 weeks. The solution was concentrated to dryness and the residue was applied to a short silica gel column, eluted with 2% MeOH in dichloromethane to give a pure (>95%) product M+H<+>384.5.

Example 21

2- Amino-/ V-( cyclohexyl- methylcarbamoyl- methyl)- 3, 3- dimethyl- butyramide ( 71)

Compound 20 was kept in dichloromethane-trifluoroacetic acid 2:1 for 1 h at RT and concentrated to dryness. The residue was dried in vacuo for 16 hours. Reverse phase C18-HPLC showed >95% purity M+H<+>283.1.

Example 22

flR, 2SM- frf2S, 4R)- l- fflS, 2R)- 2- Hydroxv- indan- l- vlcarbamovn- 4- f7- methoxyv-2- phenvl- quinoline- 4- vloxv)- pvrrolidine- 2- carvvll- amino >- 2- vinyl- cyclopropanecarboxylic acid ( 22)

Compound 12 was treated as described for the preparation of 13, but using (1S,2R)-cis-1-amino-2-indanol instead of 2-amino-N-(2-hydroxyindan-1-yl)-3,3 -dimethylbutyramide followed by ester hydrolysis as described for the preparation of compound 14, yielding the title compound. Purity according to HPLC >95%. M+H<+>649.1.

Example 23

flR, 2SVl-- frf2S, 4R)- l- rflS)- l- fCvclohexvlmetvl- carbamovn- 2- metvl-propvlcarbamovvl- 4- r7- methoxyv- 2- phenvl- quinoline- 4- vloxv)- pvrrolidine- 2- carbovl- amino>- 2-vinyl-cyclopropanecarboxylic acid (23) N-(tert-butoxycarbonyl)-L-valine was attached to the resin as described for the preparation of compound 16 followed by reaction with cyclohexylamine as described for the preparation of 17 and removal of the Boc group as described for 18. The compound obtained was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13 to give the title compound. Purity according to HPLC >95%. M+H<+>712.3.

Example 24

riR. 2S)- l- frr2S. 4R)- 1- rriR)- 2- Hydroxyl- 1- phenyl- ethylcarbamovn- 4- r7- methoxy- 2- phenyl- quinoline- 4- hydroxy)- pyrrolidin- 2- carboxylic acid- amino>- 2- vinyl- cyclopropanecarboxylic acid ( 24)

Compound 12 was treated as described for the preparation of 13, but using (R)-2-phenylglycinol instead of 2-amino-N-(2-hydroxyindan-1-yl)-3,3-dimethylbutyramide instead of 2-amino-/ v-(2-Hydroxy-indan-1-yl)-3,3-dimethyl-butyramide followed by ester hydrolysis as described for the preparation of compound 14, yielding the title compound. Purity according to HPLC >95%. M+H<+>637.1.

Example 25

flR. 2SM- fr( 2S. 4R)- 1-{ TflS)- Cyclohexyl- fccyclohexylmethyl- carbamovn- metvll-carbamovl>- 4- f7- methoxysv- 2- phenyl- quinoline- 4- cycloxv)- pyrrolidine- 2- carbonvll- amino >-2-vinyl- cyclopropanecarboxylic acid ( 25)

N-(tert-butoxycarbonyl)-L-cyclohexylglycine was attached to the resin as described for the preparation of compound 16 followed by reaction with cyclohexanemethylamine as described for the preparation of 17 and removal of the Boc group as described for 18. The obtained compound was then reacted with kl orka rba mat obtained from 12 as described for the preparation of 13, which gave the title compound. Purity according to HPLC >95%. M+H<+>752.4.

Example 26 flR, 2SVl-^ rf2S, 4RVl- rflSV2- Cyclohexyl- 1- fccyclohexylmetvl- carbamovn-etvlcarbamovvll- 4-( 7- methoxysv- 2- phenvl- quinolin- 4- loxv)- pvrrolidine- 2- carbonyl-amino>- 2-vinyl-cyclopropanecarboxylic acid ( 26)

N-(tert-butoxycarbonyl)-L-cyclohexylalanine was attached to the resin as described for the preparation of compound 16 followed by reaction with

cyclohexanemethylamine as described for the preparation of 17 and removal of the Boc group as described for 18. The resulting compound was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13 to give the title compound. Purity according to HPLC >95%. M+H<+>766.4.

Example 27

flR. 2S)- l- frf2S. 4R)- 1- rflS)- 1- fCvclohexylmethyl- carbamovn- 2. 2- dimethyl-propvlcarbamovn- 4- f7- methoxy- 2- phenyl- quinoline- 4- cyclox)- pyrrolidin- 2- carbonyl-amino>- 2- vinyl-cyclopropanecarboxylic acid (27) N-(tert-butoxycarbonyl)-L-tert-butylglycine was attached to the resin as described for the preparation of compound 16 followed by reaction with cyclohexanemethylamine as described for the preparation of 17 and removal of the Boc group as described for 18. The compound obtained was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13 to give the title compound. Purity according to HPLC >95%. M+H<+>726.3. Example 28

flR, 2SM- frf2S, 4R)- l- rflSM- fCvclohexvlmetvl- carbamovn- 2- phenvl-etvlcarbamovvl- 4- f7- methoxyv- 2- phenvl- quinoline- 4- vloxv)- pyrrolidine- 2- carbonvl-amino>- 2 - vinyl-cyclopropanecarboxylic acid ( 28)

N-(tert-butoxycarbonyl)-L-phenylalanine was attached to the resin as described for the preparation of compound 16 followed by reaction with cyclohexanemethylamine as described for the preparation of 17 and removal of the Boc group as described for 18. The obtained compound was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13, giving the title compound. Purity according to HPLC >95%. M+H<+>760.4.

Example 29 flR, 2SM- frf2S, 4R)- l- rflSM- fflS, 2RV2- Hydroxv- indan- l- vlcarbamovn- 3-phenvl- propvlcarbamovvl- 4-( 7- methoxysv- 2- phenvl- quinoline- 4- vloxav) - pyrrolidin- 2-carbonyl- amino>- 2- vinyl- cyclopropanecarboxylic acid ( 29)

N-(tert.butoxycarbonyl)-L-phenethylglycine was attached to the resin as described for the preparation of compound 16 followed by reaction with (1S,2R)-cis-1-amino-2-indanol as described for the preparation of 17 and removal of The Boc group as described for 18. The resulting compound was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13 to give the title compound. Purity according to HPLC >95%. M+H<+>810.4.

Example 30

flR, 2S)- l- frf2S, 4R)- l- fflS)- l- Benzvlcarbamovl- 2- methyl- propvlcarbamovn- 4- f7-methoxysv- 2- phenvl- quinoline- 4- vloxv)- pyrrolidine- 2- carbonvl- amino>- 2-vinyl-cyclopropanecarboxylic acid (30) N-(tert-butoxycarbonyl)-L-valine was attached to the resin as described for the preparation of compound 16 followed by reaction with benzylamine as described for the preparation of 17 and removal of the Boc group as described for 18. The resulting compound was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13 to give the title compound. Purity according to HPLC >95%. M+H<+>706.2. Example 31

flR, 2SM- frf2S, 4R)- l- rflSM- fflR)- 2- Hydroxv- l- phenvl- etvlcarbamovn- 2, 2-dimetvl- propvvlcarbamovn- 4- f7- methoxysv- 2- phenvl- quinoline- 4- vloxv) - pyrrolidine- 2-carbonyl- amino>- 2- vinyl- cyclopropanecarboxylic acid ( 31)

N-(tert-butoxycarbonyl)-L-tert-butylglycine was attached to the resin as described for the preparation of compound 16 followed by reaction with (R)-2-phenylglycinol as described for the preparation of 17 and removal of the Boc group as described for 18. The compound obtained was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13 to give the title compound. Purity according to HPLC >95%. M+H<+>750.3.

Example 32

f IR. 2S)- 1-( T( 2S. 4R)- l- rflS)- l- fflR)- Indan- l- vlkarbamovn- 2- metvl-propvlcarbamovvl- 4- f7- methoxysv- 2- phenvl- quinoline- 4- vloxv )- pyrrolidin- 2- carbonyl-amino>- 2- vinyl- cyclopropanecarboxylic acid ( 32)

(2S)-tert-butoxycarbonylamino-3-methylbutyric acid was attached to the resin as described for the preparation of compound 16 followed by reaction with (1R)-1-

aminoindane as described for the preparation of 17 and removal of the Boc group as described for 18. The obtained compound was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13, which after purification by HPLC gave the title compound (12.5 mg , 28% yield), purity according to HPLC >90%. M+H<+>732.2.

Example 33

f IR, 2S)- 1-( T( 2S, 4R)- l- rflS)- l- fflS)- Indan- l- vlcarbamovl)- 2- metvl-propylcarbamoyl1- 4-( 7- methoxy- 2- phenyl- quinolin- 4- yloxy)- pyrrolidin- 2- carbonyl-1-amino>- 2- vinyl- cyclopropanecarboxylic acid ( 33)

(2S)-tert-Butoxycarbonylamino-3-methylbutyric acid was attached to the resin as described for the preparation of compound 16 followed by reaction with (1S)-1-aminoindane as described for the preparation of 17 and removal of the Boc group as described for 18. The compound obtained was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13, which after purification by HPLC afforded the title compound (22 mg, 49% yield), HPLC purity >90% M+H<+>732, 2.

Example 34

f IR, 2S)- 1-( T( 2S, 4RVl- rflS)- 1- f2- hvdroxvevlcarbamovn- 2- metvl-propvlcarbamovl- 4- r7- methoxysv- 2- phenvl- quinoline- 4- vloxv)- pvrrolidine- 2 - carbonyl-amino>- 2- vinyl- cyclopropanecarboxylic acid ( 34)

(2S)-tert-Butoxycarbonylamino-3-methylbutyric acid was attached to the resin as described for the preparation of compound 16 followed by reaction with 2-aminoethanol as described for the preparation of 17 and removal of the Boc group as described for 18. The compound obtained was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13, which after purification by HPLC afforded the title compound (3 mg, 8% yield), HPLC purity >90% M+H<+>660.2.

Example 35

( IR, 2S)- 1-( T( 2S, 4R)- l- r( lS)- l-(( lS, 2R)- 2- Hydroxv- indan- l- vlkarbamovn- 2-methyl- propvlcarbamovn- 4- (7-Methoxy-2-phenyl-quinolin-4-cyclohex)-pyrrolidine-2-carbonyl-amino>-2-vinyl-cyclopropanecarboxylic acid (35) (2S)-tert-Butoxycarbonylamino-3-methylbutyric acid was attached to the resin as described for the preparation of compound 16 followed by reaction with (1S,2R)-1-amino-2-indanol as described for the preparation of 17 and removal of the Boc group as described for 18. The obtained compound was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13, which after purification by HPLC afforded the title compound (10 mg, 22% yield), HPLC purity >90% M+H<+>748.2 Example 36

( IR, 2S)- 1-( T( 2S, 4R)- l- r( lS)- l-(( lR, 2S)- 2- Hydroxv- indan- l- vlcarbamovn- 2-metvl- propvlcarbamovn- 4- f7- methoxy- 2- phenyl- quinoline- 4- cyclooxy)- pyrrolidine- 2-carbonyl- amino>- 2- vinyl- cyclopropanecarboxylic acid ( 36)

(2S)-tert-Butoxycarbonylamino-3-methylbutyric acid was attached to the resin as described for the preparation of compound 16 followed by reaction with (1R,2S)-1-amino-2-indanol as described for the preparation of 17 and removal of Boc- group as described for 18. The obtained compound was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13, which after purification by HPLC afforded the title compound (11 mg, 24% yield), HPLC purity >75% M + H<+>748.

Example 37

( IR, 2S)- 1-( T( 2S, 4R)- 1- frCvclohexvKS)- f( 1S, 2R)- 2- hydroxy- indan- 1-vlcarbamovl)- metvll- carbamovl>- 4-( 7- methoxysv - 2-phenyl-quinoline-4-cyclohexyl)-pyrrrolidine-2- carbonyl-amino>-2-vinyl-cyclopropanecarboxylic acid (37) (2S)-tert.Butoxycarbonylamino-cyclohexylacetic acid was attached to the resin as described for the preparation of compound 16 followed by of reaction with (1S,2R)-1-amino-2-indanol as described for the preparation of 17 and removal of the Boc group as described for 18. The obtained compound was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13 , which after purification by HPLC afforded the title compound (7.5 mg, 16% yield), HPLC purity >95% M+H<+>788.3.

Example 38

( IR, 2SM-( T( 2S, 4RM- r( 1S)- 1-(( 1S, 2R)- 2- Hydroxiv- indan- 1- vlcarbamovn- 2. 2-dimethyl- propvvlcarbamovn- 4-( 7- methoxysv - 2- phenyl- quinoline- 4- oxalic)- pyrrolidine- 2-carboxylic acid- amino>- 2- vinyl- cyclopropanecarboxylic acid ( 38)

(2S)-tert-Butoxycarbonylamino-3,3-dimethylbutyric acid was attached to the resin as described for the preparation of compound 16 followed by reaction with (1S,2R)-1-amino-2-indanol as described for the preparation of 17 and removal of Boc group as described for 18. The compound obtained was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13, which after purification by HPLC afforded the title compound (12 mg, 26% yield), HPLC purity >95% M+H<+>762.3.

Example 39

( IR, 2SM-( T( 2S, 4RM- r( 1S)- 1-(( 1S, 2R)- 2- Hydroxiv- indan- 1- vlcarbamovn- 3, 3-dimethyl- butvlcarbamovn- 4-( 7- methoxysv - 2- phenyl- quinoline- 4- hydroxy)- pyrrolidin- 2-carboxylic acid- amino>- 2- vinyl- cyclopropanecarboxylic acid ( 39)

(2S)-tert-Butoxycarbonylamino-4,4-dimethylpentanoic acid was attached to the resin as described for the preparation of compound 16 followed by reaction with (1S,2R)-1-amino-2-indanol as described for the preparation of 17 and removal of Boc group as described for 18. The compound obtained was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13, which after purification by HPLC afforded the title compound (14.2 mg, 30% yield), purity according to HPLC > 95% M+H<+>776.3.

Example 40

( IR, 2SM-( T( 2S, 4RM- r( 1S)- 1-(( 1S, 2R)- 2- Hydroxv- indan- 1- vlcarbamovn- 2- phenyl- etvlcarbamovn- 4-( 7- methoxysv- 2 - phenyl- quinolin- 4- hydroxy)- pyrrolidine- 2-carbonyl- amino>- 2- vinyl- cyclopropanecarboxylic acid ( 40) (2S)-tert-Butoxycarbonylamino-3-phenylpropanoic acid was attached to the resin as described for the preparation of compound 16 followed by of reaction with (1S,2R)-1-amino-2-indanol as described for the preparation of 17 and removal of the Boc group as described for 18. The resulting compound was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13 , which after purification by HPLC afforded the title compound (2.4 mg, 5% yield), HPLC purity >95% M+H<+>796.2 Example 41

( IR, 2S)- 1-( T( 2S, 4R)- 1- r( 1S)- 2- Cyclohexyl- 1-(( 1S, 2RV 2- hydroxy- indan- 1-vlcarbamovl)- etvlcarbamovl- 4-( 7 - methoxy- 2- phenyl- quinoline- 4- cyclohex)- pyrrolidin- 2-carbonyl- amino>- 2- vinyl- cyclopropanecarboxylic acid ( 41)

(2S)-tert-Butoxycarbonylamino-3-cyclohexylpropanoic acid was attached to the resin as described for the preparation of compound 16 followed by reaction with (1S,2R)-1-amino-2-indanol as described for the preparation of 17 and removal of Boc- group as described for 18. The compound obtained was then reacted with the chlorocarbamate obtained from 12 as described for the preparation of 13, which after purification by HPLC afforded the title compound (12.3 mg, 25% yield), HPLC purity >95% M+H<+>802.3.

Example 42

( IR, 2S)- 1-( T( 2S, 4R)- l- f( lS)- l- r( S)-( Cyclohexyl- metvlcarbamovl- metvl)-carbamovl- 2, 2- dimethyl- propvlcarbamovl>- 4 -( 7- methoxy- 2- phenyl- quinoline- 4- cyclohexyl)-pyrrolidin- 2- carbonyl- amino>- 2- vinyl- cyclopropanecarboxylic acid ( 42)

Compound 12 was treated as described for the preparation of 13, but using 21 instead of 2-amino-[v-(2-hydroxy-indan-1-yl)-3,3-dimethyl-butyramide followed by ester hydrolysis as described for the preparation of compound 14, which after purification by HPLC gave the title compound (8.6 mg, 18% yield). Purity according to HPLC >95%. M+H<+>783.3.

Example 43

l-(2-Amino-4- methoxyphenyl)ethanone ( 43 )

m-Anisidine (10.0 g, 82 mmol) was dissolved in CH 2 Cl 2 (50 mL), and the solution was cooled to -50°C. BCl 3 (1 M in CH 2 Cl 2 , 82 mL, 82 mmol) was added slowly over 20 min, after which the mixture was stirred at -50 °C for 30 min, followed by sequential addition of AcCl (6.0 mL, 84 mmol) and AlCl 3 (11 g, 82 mmol). The mixture was stirred at -50°C for 1 h and then allowed to reach rt. After stirring at RT overnight, the solution was heated at 40 °C for 4 h, after which the mixture was poured over ice. The aqueous mixture was made alkaline with 10% NaOH (w/v) and extracted with EtOAc (4 x 200 mL). The combined organic phases were washed with brine, dried (MgSO 4 ) and evaporated to give a black solid, which was purified by flash column chromatography (ether/CH 2 Cl 2 2 O:80). The resulting solid was recrystallized from ether/hexane to give compound 93 as shiny beige small leaves (5.6 g, 42%).

Example 44

/v-(tert-Butvl)-/V'- isopropylthiourea ( 44)

To a solution of tert-butyl isothiocyanate (5.0 mL, 39 mmol) in CH 2 Cl 2 (200 mL) was added isopropylamine (4.0 mL, 47 mmol) and diisopropylethylamine (DIEA) (6.8 mL, 39 mmol), and the mixture was stirred at RT for 2 h. The reaction mixture was diluted with EtOAc, washed with 10% citric acid (2x), saturated NaHCO 3 (2x), H 2 O (2x) and brine (1x). The organic layer was dried (MgSO 4 ) and evaporated to give compound 94 (3.3 g, 52%) as a white solid which was used without further purification.

Example 45

/ V- Isopropvlthiourea ( 45)

Compound 44 (3.3 g, 20 mmol) was dissolved in conc. HCl (45 mL), and the solution was refluxed for 40 min. The mixture was allowed to cool to rt and then allowed to cool in an ice bath and acidified to pH 9.5 with solid saturated NaHCO 3 , after which the product was extracted into EtOAc (3x). The combined organic phases were washed with H 2 O (2x) and brine (1x), dried (MgSO 4 ) and evaporated to give crude compound 95 (2.1 g, 90%) which was used without further purification.

Example 46

2-(Isopropylamino)-1,3-thiazole-4-carboxylic acid hydrobromide (46)

A suspension of compound 45 (2.1 g, 18 mmol) and 3-bromopyruvic acid (3.0 g, 18 mmol) in dioxane (180 mL) was heated to 80 °C. When it reached 80°C, the mixture became clear and immediately thereafter the product began to precipitate as a white solid. After 2 h of heating, the reaction mixture was cooled to rt, and the precipitate was filtered off and collected. This gave the pure compound 46 (4.4 g, 94%).

Example 47

/V-(2-Acetyl-5-methoxyphenyl)-2-(isopropylamino)-1,3-thiazole-4-carboxamide (47)

A mixture of compound 46 (4.4 g, 16.5 mmol) and the aniline derivative 93 (2.75 g, 16.5 mmol) in pyridine (140 mL) was cooled to -30 °C (after cooling, the clear solution partly a suspension). POCl 3 (3.3 mL, 35 mmol) was added slowly over a 5 minute period. The mixture was stirred at -30°C for 1 h and then allowed to reach rt. After stirring at RT for 1.5 h, the reaction mixture was poured over ice and the pH was adjusted to about 9-10 using solid saturated NaHCO 3 . The crude product was extracted into CH 2 Cl 2 (3x), and the combined organic phases were dried (MgSO 4 ) and evaporated. The crude dark beige solid was purified by flash column chromatography (hexane/EtOAc 55:45) to give compound 47 (5.6 g, 76%) as a pale yellow solid.

Example 48

2- r2-(Isopropvlamino)-1,3-thiazol-4-vll-7- methoxyquinolin-4-ol ( 48)

A solution of t.BuOK (2.42 g, 21 mmol) in anhydrous t.BuOH (40 mL) was heated to reflux. Compound 47 (1.8 g, 5.4 mmol) was added portionwise over a 5 min period and the dark red solution that formed was stirred at reflux for an additional 20 min. The mixture was cooled to rt and HCl (4 M in dioxane, 8.0 mL, 32 mmol) was added, after which the reaction mixture was concentrated in vacuo. To ensure that all the HCl and dioxane were removed, the crude product was redissolved in CH 2 Cl 2 twice and thoroughly evaporated to afford the slightly impure HCl salt of compound 98 (1.62 g) as a brown solid. The product was dissolved in CH 2 Cl 2 and washed with saturated NaHCO 3 , after which the aqueous phase was extracted several times with CH 2 Cl 2 . The combined organic phases were dried (MgSO 4 ) and evaporated to give the title compound (1.38 g, 81%) as a light brown solid (>95% pure according to HPLC tests).<1>H-NMR (MeOH -c/4, 400 MHz): 5 1.30 (d, J = 6.0 Hz, 6H), 3.93 (s, 3H), 3.95-4.07 (m, 1H), 6, 73 (s, 1H), 6.99 (dd, J = 2.4, 9.2 Hz, 1H), 7.26 (d, J = 2.4 Hz, 1H), 7.37 (s, 1H ), 8.10 (d, J = 9.2 Hz, 1H).

Example 49

(!S)- l-{Tf2S, 4R)- 2- fl- Methoxxcarbonyl- butvlcarbamovn- 4- f7- methoxyl- 2- phenvlquinoline- 4- loxxv) l- pyrrolidineT- carboxyvl- tert- butylester ( 49)

Reaction of 10 with Nva-OMe hydrochloride according to the method described in Example 11 gave the title compound. Purity >95% according to HPLC, M+H<+>578.24.

Example 50

flS)- 1-{ Tf2S, 4R)- 2- r4- f7- Methoxy- 2- phenyl- quinoline- 4- cyclohex)- pyrrolidin- 2- carbonyl-aminoT- pentanesic acid- methyl ester ( 50)

Compound 49 was kept in TFA-DCM 1:2 (3 mL) at RT for 60 min. Toluene (3 mL) was added. The sample was co-evaporated to dryness. Purity according to HPLC >95%. M+H<+>478.21.

Example 51

flS)- 2-{ Tf2S, 4RM- rflS)- 1- fCvclohexylmethyl- carbamovn- 2- methyl-propvlcarbamovyl- 4-( 7- methoxy- 2- phenyl- quinoline- 4- cyclox)- pyrrolidine- 2- carbonyl-aminoT - pentanoic acid methyl ester ( 51)

To a solution of 50 (0.1 mmol) in THF (4 mL), cooled to 0 °C, was added a large excess of NaHCO 3 (s) and a solution of phosgene in toluene (0.2 mmol, 21^. 1). After 10 min of shaking, the slurry was filtered and concentrated to dryness. The solid was redissolved in dichloromethane, and a large excess of NaHCO 3 (s) and 2-amino-N-cyclohexylmethyl-3-methyl-butyramide, described in Example 23, (0.15 mmol) was added. The slurry was shaken for 30 h at RT. The slurry was filtered, concentrated and subjected to silica column chromatography (gradient elution from 100% DCM to MeOH/DCM 2:98) to give the title compound (30 mg, 0.042 mmol). Purity according to HPLC >95%. M+H<+>716.40.

Example 52 flSV2-- frf2S, 4R)- 1- rflS)- 1- fCvclohexvlmetvl- carbamovn- 2- metvl-propvlcarbamovvl- 4- r7- methoxysv- 2- phenvl- quinoline- 4- vloxv)- pvrrolidine- 2- carbovl- amino>- pentanoic acid ( 52)

To a solution of 51 (26 mg, 0.036 mmol) in THF-MeOH 2:3 (2 mL) was added IM LiOH 1.5 equiv. The solution was kept at 60°C for 60 min. After cooling to RT, HOAc was added followed by toluene (2 mL) and then concentrated to dryness to give the title compound (25 mg, 0.035 mmol). Purity >95% according to HPLC M+H<+>702.34.

Example 53

flR, 2S)- l- frf2S, 4R)- l- r2- f2- Methoxv- phenoxyl)- etvlcarbamovll- 4- f7- methoxyl- 2-phenvl- quinoline- 4- vloxv)- pvrrolidine- 2- carbonvn- amino> - 2-vinyl-cyclopropanecarboxylic acid ester (53)

To a solution of 12 (0.06 mmol) in THF (2 mL) was added a large excess of NaHCO 3 (s) and a solution of phosgene in toluene (0.078 mmol). After 10 min of shaking, the slurry was filtered and concentrated to dryness. The solid was redissolved in dichloromethane and a large excess of NaHCO 3 (s) and 2-(2-methoxy-phenoxy)-ethylamine (15 mg, 0.09 mmol) were added. The slurry was shaken for 30 h at RT. The slurry was filtered, concentrated to dryness, redissolved in MeOH and subjected to HPLC purification to give the title compound (10.6 mg, 0.015 mmol). Purity according to HPLC >95%. M+H<+>695.17.

Example 54

flR, 2S)- 1- frf2S, 4R)- 1- r2- f2- Methoxys- phenoxy)- ethvlcarbamovvll- 4- f7- methoxysv- 2-phenvl- quinolin- 4- yloxy)- pyrrolidin- 2- carbonyl- amino> - 2-vinyl-cyclopropanecarboxylic acid ( 54)

To a solution of 53 (10.6 mg, 0.0153 mmol) in THF-MeOH 2:3 (2 mL) was added IM LiOH 10 equiv. The solution was kept at 50°C for 60 min. After cooling to RT, HOAc 25 equiv. Added followed by toluene (2 mL) and then concentrated to dryness. The residue was taken up in ethyl acetate, filtered and concentrated to dryness to give the title compound (9.4 mg, 0.014 mmol). Purity >95% according to HPLC M+H<+>667.14.

Example 55

flR, 2S)- l- frf2S, 4R)- l- fflS, 2R)- 5- Hydroxv- 4, 5, 6, 7- tetrahydro- benzorb1thiophene- 4-vl- carbamovl))- 4- f7- methoxysv- 2 - phenyl- quinoline- 4- hydroxy)- pyrrolidine- 2- carboxylic acid- amino>- 2- vinyl- cyclopropanecarboxylic acid ( 55)

The procedure described in Example 53 was followed, but using 2-amino-4,5,6,7-tetrahydro-benzo[b]thiophen-5-ol instead of 2-(2-methoxy-phenoxy)-ethylamine, followed by hydrolysis of the ethyl ester as described in Example 54, yielding the title compound (7.5 mg, 0.011 mmol). Purity >95% according to HPLC M+H<+>669.

Example 56

flR, 2SM- frf2S, 4R)- l- rf3RV3- Hydroxv- pvrrolidine- l- carbonvni- 4- r7- methoxyv- 2-phenyl- quinoline- 4- vloxv)- pvrrolidine- 2- carbonvn- amino>- 2- vinvl - cyclopropanecarboxylic acid ( 56)

The procedure described in Example 53 was followed, but using (R)-3-pyrrolidinol instead of 2-(2-methoxy-phenoxy)-ethylamine, followed by hydrolysis of the ethyl ester as described in Example 54, yielding the title compound (4 mg, 0.007 mmol). Purity >95% according to HPLC M+H<+>587.1.

Example 57 flR, 2SM- frf2S, 4R)- 4- f7- Methoxv- 2- phenyl- quinoline- 4- loxvM- rthiophene- 2- vl-metvl)- carbamovl- pyrrolidin- 2- carbovl>- amino)- 2- vinylcyclopropanecarboxylic acid (57)

The procedure described in Example 53 was followed, but using thiophene-2-methylamine instead of 2-(2-methoxy-phenoxy)-ethylamine, followed by hydrolysis of the ethyl ester as described in Example 54, giving the title compound (8 mg, 0.013 mmol ). Purity >95% according to HPLC M+H<+>613.08.

Example 58

flR, 2S)- l- frf2S, 4R)- irfl, l- Dioxo- tetrahydro- l- A6- thiophen- 3- vl - carbamovl)- 4-( 7-methoxy- 2- phenyl- quinolin- 4- yloxy) - pyrrolidine- 2- carbonyl-amino>- 2- vinyl-cyclopropanecarboxylic acid ( 58)

The procedure described in Example 53 was followed, but using 3-aminotetrahydro-1H-1A<6->thiophene-1,1-dione instead of 2-(2-methoxy-phenoxy)-ethylamine, followed by hydrolysis of the ethyl ester as described in Example 54, giving the title compound (13 mg, 0.02 mmol). Purity >95% according to HPLC M+H<+>635.05.

Example 59

2- Amino- 3, 3- dimethyl- N- thiophene- 2- yl- methyl- butyramide ( 59)

The title compound was prepared as described in Example 17, but using thiophene-2-methylamine instead of aminoindanol followed by removal of the Boc group as described in Example 18.

Example 60

2- Amino- N-( 6- hydroxy- 4, 5, 6, 7- tetrahydro- benzolthiophene- 5- vl)- 3, 3- dimethylbutyramide ( 60)

The title compound was prepared as described in Example 17, but using 2-amino-4,5,6,7-tetrahydro-benzo[b]thiophen-5-ol instead of aminoindanol followed by removal of the Boc group as described in Example 18 .

Example 61

2- Amino- N-(2- diethylamino-ethyl)- 3, 3- dimethyl-butyramide (61)

The title compound was prepared as described in Example 17, but using N,N-diethylethylenediamine instead of aminoindanol followed by removal of the Boc group as described in Example 18.

Example 62

2- Amino-N-r2-(2-methoxy-phenoxy)-ethyl-3,3-dimethyl-butyramide (62)

The title compound was prepared as described in Example 17, but using 2-methoxyphenoxyethylamine instead of aminoindanol followed by removal of the Boc group as described in Example 18.

Example 63

2- Amino-l-(3-hydroxv- pyrrolidin-l-vl)- 3, 3- dimethyl- butan-l-one ( 63)

The title compound was prepared as described in Example 17, but using (R)-3-pyrrolidinone instead of aminoindanol followed by removal of the Boc group as described in Example 18.

Example 64

2- Amino- N-(1,1- dioxo-tetrahydro-1-D6-thiophene-3-v1)-3,3- dimethylbutyramide (64)

The title compound was prepared as described in Example 17, but using 2-methoxyphenoxyethylamine instead of aminoindanol followed by removal of the Boc group as described in Example 18.

Example 65

flR. 2SVl- frf2S. 4RVl- rflSVl- f2. 2- Dimethyl-1- rthiophene- 2- methyl- methvn- carbamol- propylcarbamol>- 4- f7- methoxy- 2- phenvl- quinoline- 4- cyclox)- pyrrolidin- 2- carbonvl-amino>- 2- vinvl- cyclopropanecarboxylic acid - etflester ( 65)

To a solution of 12 (0.06 mmol) in THF (2 mL), was added a large excess of NaHCO 3 (s) and a solution of phosgene in toluene (0.078 mmol). After 10 min of shaking, the slurry was filtered and concentrated to dryness. The solid was redissolved in dichloromethane, and a large excess of NaHCO 3 (s) and 59 (0.09 mmol) was added. The slurry was shaken for 30 h at RT. The slurry was filtered, concentrated to dryness, redissolved in MeOH and subjected to HPLC purification to give the title compound (15.5 mg, 0.02 mmol). Purity according to HPLC >95%. M+H<+>754.2.

Example 66 flR, 2SVl-^ rf2S, 4RVl- rflSVl- f2, 2- Dimetvl- l- rftiophen- 2- vlmetvn- carbamovl1-propvlcarbamovl>- 4-( 7- methoxysv- 2- phenvl- quinoline- 4- vloxv)- Pyrrolidine-2-carbonyl-amino>-2-vinyl-cyclopropanecarboxylic acid (66)

To a solution of 65 (14 mg, 0.017 mmol) in THF-MeOH 2:3 (2 mL) was added IM LiOH 10 equiv. The solution was kept at 50°C for 60 min. After cooling to RT, HOAc 20 equiv. added followed by toluene (2 mL) and then concentrated to dryness. The residue was taken up in ethyl acetate, filtered and concentrated to dryness to give the title compound (13 mg, 0.017 mmol). Purity >95% according to HPLC M+H<+>748.13.

Example 67

flR, 2S)- l- frf2S, 4R)- flS)- l- rriS, 2R)- l- ri- f5- Hvdroksv- 4, 5, 6, 7- tetrahydro-benzorbltiofen- 4- vl- carbamovl)- 2 .

The procedure described in Example 65 was followed, but using 60 instead of 59, followed by hydrolysis of the ethyl ester as described in Example 66, yielding the title compound (4 mg, 0.005 mmol). Purity >95% according to HPLC M+H<+>782.16.

Example 68

riR. 2SVl- frr2S. 4RVl- rriSVl- r2- Diethylamino- etvlcarbamovn- 2. 2- dimethyl-propvlcarbamovn- 4- f7- methoxysv- 2- phenyl- quinoline- 4- cyclosv)- pyrrolidin- 2- carbonyl-amino>- 2- vinvl- cyclopropanecarboxylic acid ( 68)

The procedure described in Example 65 was followed, but using 61 in place of 59, followed by hydrolysis of the ethyl ester as described in Example 66, yielding the title compound (6 mg, 0.008 mmol). Purity >95% according to HPLC M+H<+>729.24.

Example 69

iriR, 2S)- l-{ Tf2S, 4R)- l- rflS)- l- r2- f2- Methoxy- phenoxy)- ethylcarbamoyl- 2, 2-dimethyl- propylcarbamoyl>- 4-( 7- methoxy- 2- phenyl - quinoline- 4- hydroxy)-pyrrolidine- 2-carboxylic acid- amino>- 2- vinyl-cyclopropanecarboxylic acid ( 69)

The procedure described in Example 65 was followed, but using 62 instead of 59, followed by hydrolysis of the ethyl ester as described in Example 66, yielding the title compound (3 mg, 0.004 mmol). Purity >95% according to HPLC M+H<+>780.19.

Example 70

riR. 2SVl- frr2S. 4RVriSVl- rr3RVl- r3- Hydroxv- Pvrrolidine- 1- carbonvn- 2. 2-dimethyl- propvlcarbamovvll- 4- f7- methoxyv- 2- phenvl- quinoline- 4- vloxv)- pvrrolidine- 2-carbonvll- amino>- 2- vinyl-cyclopropanecarboxylic acid (70)

The procedure described in Example 65 was followed, but using 63 instead of 59, followed by hydrolysis of the ethyl ester as described in Example 66, yielding the title compound (12.4 mg, 0.02 mmol). Purity >95% according to HPLC M+H<+>700.16.

Example 71

(lR, 2S)- l- fr( 2S, 4R)- l- r( lS)- l-( l, l- Dioxo- tetrahydro- l- D6- thiophene - 3- vlcarbamovn- 2, 2- dimethyl-propvlcarbamovl - 4-( 7- methoxy- 2- phenyl- quinoline- 4- chloro)-pyrrrolidine- 2- carbonyl- amino>- 2- vinyl- cyclopropanecarboxylic acid ( 71)

The procedure described in Example 65 was followed, but using 64 instead of 59, followed by hydrolysis of the ethyl ester as described in Example 66, yielding the title compound (13 mg, 0.014 mmol). Purity >95% according to HPLC M+H<+>748.13.

Example 72

f4/ ?)- l- phetrf-- butoxxcarbonvn- 4- rf7- methoxysv- 2- phenvlquinoline- 4- venoxvl- L- prolvl-/V1-( phenvlsulfonvl)- L- norvalinamide ( 72)

To a solution of 10 (60 mg, 0.13 mmol) in DMF was added HATU (124 mg, 0.325 mmol), diisopropylethylamine (114 µl, 0.65 mmol) and shaken for 30 min at RT. A solution of 75 (0.157 mmol) in DMF was added. The slurry was shaken for 16 h at RT followed by concentration to dryness. The residue was taken up in DCM and washed with NaHCO 3 (saturated) and water. The organic layer was dried, concentrated and subjected to silica column chromatography (gradient elution from 100% DCM to 2% MeOH/DCM) to give the title compound (61 mg, 0.087 mmol). Purity >90% according to HPLC. M+H<+>703.23.

Example 73 f4/ ?)- 4- rf7- methoxy- 2- phenylquinoline- 4- vl) oxvl- L- prolvl-/ Vl- phenvl sulfonvl)- L-norvalinamide ( 73)

Compound 72 was kept in DCM-TFA 2:1 (2 mL) for 2.5 h at RT. The solution was co-evaporated with toluene to dryness. Yield 100%. M+H 603.12

Example 74

Urea acid, r( lS)- l- rr( phenylsulfonyl) aminolcarbonylbutyl- phenylmethyl ester ( 74)

To a stirred solution of Z-Nva-OH (150 mg, 0.59 mmol) in THF (6 mL) was added CDI (400 mg, 2.4 mmol). The slurry was shaken for 30 min at RT followed by the addition of DBU (200 µl, 1.3 mmol) and a solution of benzenesulfonamide (250 mg, 1.59 mmol) in THF (2 mL). The mixture was stirred at 60°C for 48 hours followed by concentration to dryness. The residue was dissolved in MeOH and subjected to HPLC purification to give the title compound (118.5 mg, 0.304 mmol). Purity >95% according to HPLC. M-H+ 389.0, +Na 412.96.

Example 75

( 2S )- 2- Amino- N -(phenylsulfonyl) pentanamide ( 75 )

Compound 74 was dissolved in MeOH (5 mL) followed by addition of Pd/C and subjected to hydrogenation for 2 h. The slurry was filtered through celite, washed with MeOH and concentrated to dryness to give the title compound. Yield 100%. M+H<+>257.3.

Example 76

4-( 7- Methoxy- 2- phenyl- quinolin- 4- cyclohexyl)- pyrrolidin- 1, 2- dicarboxylic acid 1-(- fl-rfcvchlorohexyl- methylcarbamoyl- methyl)- carbamol- 2, 2- dimethyl- propyl>- amide )- 2-Tf 1- phenylmethanesulfonylaminocarbonyl- 2- vinyl- cyclopropionamidl ( 76)

To a solution of 42 (8.7 mg, 0.011 mmol) in chloroform (1 mL) was added α-toluenesulfonamide (7 mg, 0.04 mmol) followed by diisopropylethylamine (21 µL, 0.12 mmol). The solution was stirred at RT for 10 min and then at -20°C for 30 min. PyBOP (46.5 mg, 0.08 mmol) was then added as a solid. The solution was kept at -20°C for 48 hours. The solution was then poured into aqueous NaHCO 3 (saturated) and washed with water. The organic layer was dried, concentrated and subjected to purification by HPLC, to give the title compound as a white solid (2.8 mg, 0.0049 mmol), HPLC purity >95%, M+H<+> 936.26.

Example 77

N- f2- Hydroxv- indan- l- vn- 2- r4- f6- methoxysv- 3- phenyl- naphthalene- l- loxv)- 2- f 1-methanesulfonvlaminocarbonvl- 2- vinvl- cyclopropvl)- pyrrolidin- l- vll - 3, 3- dimethylbutyramide (77)

The title compound was prepared as described in Example 76, using 14 as carboxylic acid starting material and methanesulfonamide instead of α-toluenesulfonamide. Yield 13%, purity according to HPLC >95%, M+H<+>839.16.

Example 78

4- f7- Methoxv- 2- phenyl- quinoline- 4- cyclox)- pyrrolidine- 1, 2- dicarboxylic acid 1-- T Tl-( cyclohexylmethyl- carbamovl)- 2- methyl- propyl- amide> 2 -\( 1- Phenylmethanesulfonylaminocarbonyl-2-vinyl-cyclopropyl)-amidl (78)

The title compound was prepared as described in Example 76, using 23 as the carboxylic acid starting material. Dividend 2%. Purity >95% according to HPLC. M+H<+>865.28.

Example 79

4- f7- Methoxy- 2- phenyl- quinoline- 4- cyclox)- pyrrolidine- 1, 2- dicarboxylic acid 1-- T Tl-( cyclohexylmethyl- carbamovl)- 2- methyl- propyl- amide> 2 -\( l - phenylmethanesulfonylaminocarbonyl- butvD- amidl ( 79)

The title compound was prepared as described in Example 76, using 52 as the carboxylic acid starting material. Dividend 8%. Purity >95% according to HPLC. M+H<+>855.28.

Example 80

4-( 7- Methoxy- 2- phenyl- quinolin- 4- cyclohex)- pyrrolidin- 1, 2- dicarboxylic acid 2- T( 1-benzenesulfonylaminocarbonyl- butyl)- amide 1-( Tl-( cyclohexylmethyl- carbamoyl)- 2- methyl propyl amide> ( 80)

The title compound was prepared as described in Example 76, using 52 as carboxylic acid starting material and benzenesulfonamide instead of α-toluenesulfonamide. Dividend 21.5%. Purity >95% according to HPLC. M+H<+>841.28.

Example 81

4- f7- Methoxv- 2- phenyl- quinoline- 4- cyclox)- pyrrolidine- 1, 2- dicarboxylic acid 2-\( 1-benzenesulfonvlaminocarbonvl- 2- vinvl- cvclopropvl)- amidl l-(- d- r( cvclohexvl- methylcarbamoyl- metyl)-carbamoyl- 2, 2- dimethyl- propyl>- amide) ( 81)

The title compound was prepared as described in Example 76, using benzenesulfonamide instead of α-toluenesulfonamide. Dividend 26%. Purity according to HPLC

>95%, M+H<+>922.23.

Example 82

4-( 7- Methoxy- 2- phenyl- quinolin- 4- lox)- pyrrolidin- 1, 2- dicarboxylic acid- 2- r( 1- benzenesulfonvlaminocarbonyl- butvD- amidl l- d" l-( 2- hydroxy- indan- 1-ylcarbamoyl)-2-methyl- propyl-amide> (82)

To a solution of 73 (24.1 mg, 0.04 mmol) in DCM (2 mL) was added a large excess of NaHCO 3 (s) and a solution of phosgene in toluene (50 µL, 0.096 mmol). After 10 min of shaking, the slurry was filtered and concentrated to dryness. The solid was redissolved in DCM, and a large excess of NaHCO 3 (s) and 2-amino-[N-(2-hydroxy-indan-1-yl)-3-methyl-butyramide, described in Example 35, (0.1 mmol) was added. The slurry was shaken for 40 h at RT. The slurry was filtered, concentrated and subjected to HPLC purification to give the title compound (1.6 mg, 0.0018 mmol). Purity >95% according to HPLC. M+H<+>877.21.

Example 83

4-( 7-Methoxv- 2- phenyl- quinoline- 4- cycloxv)- pyrrolidin- 1, 2- dicarboxylic acid 2- Tf 1-benzenesulfonvlaminocarbonvl- butvD- amidl l- f- fl- r( cvclohexvl- methylcarbamovl- metvl)- carbamol- 2, 2- dimethyl- propyl>- amide) ( 83)

The title compound was prepared as described in Example 82, but using 21 instead of 2-amino- N -(2-hydroxy-indan-1-yl)-3-methyl-butyramide. Dividend 2%. Purity >95% according to HPLC. M+H<+>912.25.

Example 84

flR, 2S)- l- frf4R, 2S) l- fl- flS)- Hydroxvmetvl- 2, 2- dimethyl- propvlcarbamovn- 4- f7-methoxysv- 2- phenvl- quinoline- 4- vloxv)- pvrrolidine- 2- carbovl - amino>- 2- vinylcyclopropanecarboxylic acid ester ( 84)

Treatment of compound 12 as described for the preparation of 13, but using (S)-tert-leucinol instead of 2-amino-N-(2-hydroxy-indan-1-yl)-3,3-dimethyl-butyramide gave title I the product. M+H<+>645.2.

Example 85

riR. 2SVl- frr4R. 2Sn- ri- riSVFormvl- 2. 2- dimethyl- propvlcarbamovn- 4- r7- methoxysv- 2- phenyl- quinoline- 4- cycloxv)- pyrrolidin- 2- carbovl- amino>- 2- vinvlcvclopropanecarboxylic acid- ethyl ester ( 85)

To a stirred solution of compound 84 (64 mg) in dichloromethane was added Dess-Martin periodinane (80 mg) at ambient temperature. After 4 hours, the slurry was filtered through basic alumina and concentrated to dryness. M+H<+>643.2.

Example 86

( lR. 2S)- l- fr( 4R. 2S) l- fl- r(( lS. 2R)- 2- Hvdroksv- indan- l- vlamino)- metvll- 2. 2-dimetvl- propvlcarbamovl>- 4- ( 7- methoxy- 2- phenyl- quinoline- 4- cyclohexyl)- pyrrolidin- 2-carbonyl- amino>- 2- vinyl- cyclopropanecarboxylic acid ester ( 86)

To a solution of compound 85 in THF (2 mL) and HOAc (0.5 mL) was added polystyrene-bound cyanoborohydride (2.36 mmol/g, 100 mg) and (1S,2R)-1-aminoindan-2-ol (18 mg) added and shaken for 4 hours. The mixture was filtered, concentrated and purified on prep.

HPLC. Purity according to HPLC >90%. M+H<+>776.5

Example 87

flR. 2SVl- frf4R. 2SU- fl- rfflS. 2RV2- Hydroxiv- indan- 1- vlaminoVmetvll- 2. 2-dimethyl- propvlcarbamovl>- 4- f7- methoxyv- 2- phenvl- quinoline- 4- vloxv)- pyrrolidine- 2-carbonvl- amino>- 2- vinvl- cyclopropanecarboxylic acid (87)

To a solution of compound 86 in THF (2 mL) and MeOH (1 mL) 1N LiOH (0.2 mL) was added, and the solution was kept at 60 °C for 1.5 h. The slurry was neutralized with 1N HCl to pH 7, concentrated and purified on prep. HPLC, which gave a pure product according to HPLC >95%. M+H<+>748.4.

Example 88

flR, 2SVl-^ rf4R, 2SU- fl-^ rflSVfCvclohexvl- metvlcarbamovl- metvn- aminol-metvl>- 2, 2- dimetvl- propvlcarbamovn- 4-( 7- methoxyv- 2- phenvl- quinoline- 4- vloxv)-pvrrolidine - 2-carbonyl-amino>- 2-vinyl-cyclopropanecarboxylic acid ( 88) Treatment of compound 85 as described for the preparation of 86, but using 2-amino-2-cyclohexyl-N-methyl-acetamide (17 mg) instead of ( 1S,2R)-1-aminoindan-2-ol followed by hydrolysis of the ethyl ester as described in Example 87 gave the product. Purity according to HPLC >95%. M+H<+>769.5 Example 89

Acetic acid ( 1S, 2R)- 1-(( 2S)- 2- amino- 3, 3- dimethyl- butvnflamino)- indan- 2- ester ( 89)

A solution of compound 17 (4g) was kept in pyridine-acetic anhydride 2:1 for 30 min. DCM was added and the solution was washed with citric acid (aq) and NaHCO 3 (aq). The organic layer was concentrated to dryness, giving the acetylated product >90% pure according to HPLC. The obtained compound was then kept in a solution of 30% TFA in DCM for 1.5 hours and then concentrated to dryness. Co-evaporation twice from toluene gave the title product >90% pure by HPLC.

Example 90

( 2S, 4R)- 2-(( 1S, 2R) 1- Ethoxycarbonyl- 2- vinyl- cyclopropylcarbamol)- 4- hydroxy-pyrrolidine- 1- carboxyl ester. buttlester ( 90)

A solution of HATU (6 g), diisopropylethylamine (6.8 ml), (1R,2S)-1-amino-2-vinyl-cyclopropanecarboxylic acid ethyl ester (1.5 g) and BOC-L-hydroxyproline (1.6 g) in dichloromethane was stirred for 1 hour. The mixture was extracted with DCM-NaHCO 3 (aq), dried and concentrated. HPLC purity approx. 90% M+H<+>369.1.

Example 91

( 1S, 2R)- 1- r( 2S, 4R)-( 4- Hydroxy- pyrrolidine- 2- carboxylic acid)- aminol- 2- vinylcyclopropanecarboxylic acid ester ( 91).

Compound 90 was kept in 30% trifluoroacetic acid in dichloromethane and 1% MeOH for 2 h before being concentrated to dryness. The residue was redissolved in dichloromethane, and with stirring 1N NaOH was added to pH 10-11. The organic layer was separated and concentrated, yielding 1.6 g of title I product. HPLC purity approx. 90% M+H<+>269.1.

Example 92 flR, 2SM- r- ff2S, 4R)- l- rflSM- fflS, 2RV2- Acetoksv- indan- l- vlcarbamovn- 2, 2-dimethyl- propvlcarbamovnll- 4- rivdroxv- pvrrolidine- 2- carbonvl>- amino) - 2- vinylcyclopropanecarboxylic acid ester ( 92).

To a stirred solution of compound 89 (1.81 g) in acetonitrile at 0 °C was added solid NaHCO 3 (800 mg) and p-nitrophenyl chlorocarbonate (1.2 g). The slurry was brought to ambient temperature and stirred for a further 30 min. To this slurry was added a solution of compound 91 (1.6 g) in acetonitrile (5 mL) and diisopropylethylamine (1 mL). After 10 min, the resulting mixture was concentrated, redissolved in ethyl acetate and washed with K 2 CO 3 (aq) and then with 0.5 N HCl. Dried and concentrated, which gave a >80% pure product according to HPLC M+H<+>599.6

Example 93

flR, 2SM- r- ff2S, 4R)- l- rflM- fflS, 2R)- 2- Acetoxv- indan- l- vlcarbamovn- 2, 2-dimethyl- propvlcarbamovvll- 4- phenvlcarbamovvloxv- pyrrolidine- 2- carbonvl>- amino )- 2-vinyl- cyclopropanecarboxylic acid- ethyl ester ( 93)

To a stirred solution of compound 92 (20mg) in DCM and solid K 2 CO 3 (200 mg) was added 20% phosgene in toluene (1 ml). After 6 hours, the slurry was filtered and concentrated to dryness. To this residue was added a mixture of aniline (30 mg) DCM (3 mL) and solid NaHCO 3 (50 mg) and shaken for 10 h. The mixture was filtered, concentrated and purified on prep. HPLC, which gave a look at the product, >95% pure M+H<+>718.6.

Example 94

flR, 2SM- r- ff2S, 4R)- l- ri- fflS, 2R)- 2- Hydroxv- indan- l- vlcarbamovn- 2, 2- dimethyl-propvlcarbamovvll- 4- phenvlcarbamovvlxv- pyrrolidine- 2- carbonvl>- amino )- 2- vinylcyclopropanecarboxylic acid ( 94

To a solution of compound 93 in THF-MeOH 2:1 (3 mL) was added 1N LiOH (0.2 mL). The solution was heated to 60°C for 2 hours. After cooling to ambient temperature, acetic acid (0.5 mL) was added and the solution was concentrated to dryness. The remaining residue was purified on prep. HPLC, which gave the title product >95% pure M+H+ 648.5.

Example 95

( 5S, 3R)- 3, 4- Dihydro- 1H- isoquinoline- 2- carboxylic acid 5- f( 1R, 2S)- 1- carboxylic- 2- vinvl-cyclopropvlcarbamovl)- 1- ri- fflS, 2R)- 2- Hydroxy- indan-1- vylcarbamovyl)- 2, 2-dimethyl- propylcarbamovyl- pyrrolidin- 3-yl ester ( 95)

Treatment of compound 92 as described for the preparation of 93, but using 1,2,3,4-tetrahydro-isoquinoline instead of aniline followed by hydrolysis of the ethyl ester as described in Example 94 gave the title compound. Purity >90%. M+H<+>688.6.

Example 96

f5S, 3R)- 3, 4- Dihydro- 2H- quinoline- l- carboxylic acid- 5- fflR, 2S)- l- carboxyl- 2- vinvl-cvclopropvlcarbamovn- l- ri-( riS, 2R)- 2- hydroxy- indan-l- vlcarbamovyl)- 2, 2-dimethyl-propvlcarbamovyl- pyrrolidin- 3- ester ( 96)

Treatment of compound 92 as described for the preparation of 93, but using 1,2,3,4-tetrahydroquinoline instead of aniline followed by hydrolysis of the ethyl ester as described in Example 94 gave the title compound. Purity >90%. M+H<+>688.6.

Example 97 flR, 2SM- frf2S, 4R)- l- rflSM- fflS, 2RV2- Hydroxv- indan- l- vlcarbamovn- 2, 2-dimethyl-propvlcarbamovll- 4-( pvridin- 3- vlmetvlcarbamovlxv)- pvrrolidine- 2-carbovll - amino>- 2- vinyl-cyclopropanecarboxylic acid ( 97)

Treatment of compound 92 as described for the preparation of 93, but using 2-pyridin-3-yl-ethylamine instead of aniline followed by hydrolysis of the ethyl ester as described in Example 94 gave the title compound. Purity >90%. M+H<+>663.5.

Example 98

flR, 2S)- l- frf2S, 4R)- l- rflS)- l- fflS, 2R)- 2- Hvdroksv- indan- l- vlkarbamovn- 2, 2-dimetvl- propvlcarbamovvll- 4- fmetvl- fenetvl- carbamovlovskv) - pyrrolidine- 2- carbonyl-amino>- 2- vinyl- cyclopropanecarboxylic acid ( 98)

Treatment of compound 92 as described for the preparation of 93, but using N-methylphenethylamine instead of aniline followed by hydrolysis of the ethyl ester as described in Example 94 gave the title compound. Purity >90%. M+H<+>690.6.

Example 99 flR, 2SM- r- ff2S, 4R)- 4- Benzvlkarbamovloxv- l- rriS)- l- fflS, 2R)- 2- hvdroksv-indan- l- vlkarbamovn- 2, 2- dimetvl- propvlkarbamovl- pvrrolidine- 2 - carboxyl>-amino)- 2- vinyl- cyclopropanecarboxylic acid ( 99)

Treatment of compound 92 as described for the preparation of 93, but using benzylamine instead of aniline followed by hydrolysis of the ethyl ester as described in Example 94 gave the title compound. Purity >90%. M+H<+>662.4.

Example 100

riR. 2S)- l- r- fr2S. 4R)- l- rriS)- l- rriS. 2R)- 2- Hydroxiv- indan- 1- vlcarbamovn- 2. 2-dimethyl- propvvlcarbamovn- 4- phenethvlcarbamovvlxv- pyrrolidin- 2- carbovl>- amino)-2- vinvl- cyclopropanecarboxylic acid ( 100 )

Treatment of compound 92 as described for the preparation of 93, but using phenethylamine instead of aniline followed by hydrolysis of the ethyl ester as described in Example 94 gave the title compound. Purity >90%. M+H<+>676.5.

Example 101 fl/ ?, 2S)-l- r- ff4RVl- fr2- tert- butoxxcarbonvnrivdrazinolcarbonvl>- 4- rf7-methoxysv- 2- phenylquinoline- 4- vl)oxvl- L- prolvl>amino)- 2-vinvlcvclopropanecarboxylic acid ester ( 101 )

To a solution of tert-butylcarbazate (0.3 mmol) and p-nitro-phenylchloroformate (0.3 mmol) in acetonitrile (6 ml) was added sodium bicarbonate (0.48 mmol) as a solid. The solution was stirred at RT for 5 h and then allowed to cool to 0°C. Compound 62 (0.3 mmol) dissolved in acetonitrile (10 mL) was mixed with diisopropylethylamine (0.75 mmol) at 0°C and then added to the previous solution. The mixture was stirred at RT overnight and then concentrated to dryness. The residue was dissolved in DCM and then washed with citric acid pH 4, followed by NaHCO 3 (aq) and water, dried over anhydrous sodium sulfate, filtered and concentrated to dryness. The crude product was dissolved in DCM and purified by column chromatography eluting with 0.1 to 0.2% MeOH/DCM to give the title compound (101 mg). Purity >95% according to HPLC, M+H<+>660.1.

Example 102

fl/ ?, 2S)- l- r- ff4RVl- fr2- fetrt- butoxxcarbonvnhvdrazinolcarbonvl>- 4- rf7-methoxysv- 2- phenylquinoline- 4- vl) oxvl- L- prolvl>amino)- 2-vinvlcvclopropanecarboxylic acid ( 159)

Method A: To a solution of compound 101 (0.0115 mmol) in THF-MeOH 2:3 (2 mL) was added 1M LiOH (10 eq). The solution was kept at 50°C for 60 min. After cooling to RT, HOAc (20 eq) was added followed by toluene (2 mL) and then concentrated to dryness. The residue was taken up in MeOH and then purified by prep. LCMS, which gave the title compound (0.7 mg). Purity >95% according to HPLC M+H<+>732.2.

Method B: To a solution of tert-butylcarbazate (0.07 mmol) and p-nitrophenyl chloroformate (0.07 mmol) in acetonitrile (3 ml) was added sodium bicarbonate (0.112 mmol) as a solid. The solution was stirred at RT for 2.5 hours and then allowed to cool to 0°C. Compound 103 (described below) (0.07 mmol) dissolved in acetonitrile (10 mL) was mixed with diisopropylethylamine (0.175 mmol) at 0°C and then added to the previous solution. The mixture was stirred at RT overnight and then concentrated to dryness. The crude material was dissolved in MeOH and purified by prep. LCMS, which gave the title compound (4.8 mg). Purity >95% according to HPLC M+H<+>632.2

Example 103

flR. 2S)- l- frf2S. 4R)- 4- f7- Methoxv- 2- phenyl- quinoline- 4- cycloxv)- Pyrrolidine- 2-carbonvl- amino>- 2- vinvl- cyclopropanecarboxylic acid f 103)

To a solution of compound 12 (0.067 mmol) in THF-MeOH 2:3 (2 ml) was added IM LiOH 10 equiv. The solution was kept at 50°C for 2.5 hours. After cooling to RT, HOAc 20 equiv. added followed by toluene (2 mL) and then concentrated to dryness. The residue was taken up in DCM and filtered from the salts to give the title compound (0.07 mmol). Purity >95% according to HPLC M+H<+>474.

Example 104

( l/ ?, 2S)- l- fif4R)- l- fhvdrazinocarbonvl)- 4- rf7- niethoxv- 2- phenylquinoline- 4- vl) oxvl-L- prolvl>amino)- 2- vinvlcvclopropanecarboxylic acid ( 104)

Compound (102) was kept in TFA-DCM 1:2 (3 mL) at RT for 60 min. Toluene (1 mL) was added. The sample was co-evaporated to dryness to give the title compound (10.5 mg) as the trifluoroacetic acid salt. Purity according to HPLC >95%. M+H<+>532.

Example 105

(1/?,2S)-1-(-{'(4R)-1-(hydrazinocarbonyl)-4-[(7-methoxy-2-phenylquinolin-4-yl)oxy1-L-prolvl>amino)-2 - vinylcyclopropanecarboxylic acid ester f 105)

Compound 101 (50 mg) was kept in TFA-DCM 1:2 (3 mL) at RT for 60 min. Toluene (1 mL) was added. The sample was co-evaporated to dryness then taken up in DCM and washed with K 2 CO 3 , dried over anhydrous sodium sulfate and concentrated to dryness to give the title compound (41.8 mg). Purity according to HPLC >95%. M+H<+>560.

Example 106 fl/ ?, 2S)-1-fif4R)-1-rf2-Benzylhvdrazino)carbonyl-4-rf7- methoxy-2- phenylquinoline- 4-vinoxyl- L-prolvl>amino)- 2- vinylcyclopropanecarboxylic acid ester ( 106 )

To a solution of compound 105 (0.037 mmol) in MeOH:THF (4:1) was added benzaldehyde (0.0448 mmol). The solution was stirred at RT for 18 h. Borane-pyridine complex (0.37 mmol) was added followed by HCl (37%, 400 µl). The solution was stirred for 1.5 hours and then filtered and concentrated to dryness. The crude material was dissolved in MeOH and purified by prep. LCMS, which gave the title compound (0.01 mmol). Purity according to HPLC >95%. M+H<+>650.

Example 107

( l/ ?, 2S)- l- f-{'( 4R)- l- r( 2- benzvlhvdrazino) carbovll- 4- r( 7- methoxysv- 2- phenvlquinoline- 4-vl) oxvl- L- prolvl> amino)- 2-vinylcyclopropanecarboxylic acid f 164107

To a solution of compound 106 (0.0101 mmol) in THF-MeOH 2:3 (3 ml) was added IM LiOH 10 equiv. The solution was kept at 50 °C for 18 hours. After cooling to RT, the sample was neutralized with HCl and concentrated to dryness. The crude material was dissolved in DCM (2 mL) and a solution of TFA: TES 1:1 (1 mL) was added. The mixture was stirred for 3 h at RT and then concentrated to dryness. The crude material was dissolved in MeOH and purified by prep. LCMS, which gave the title compound (0.6 mg). Purity according to HPLC >95%. M+H<+>622.

Example 108

f IR, 2SM-( T( 2S, 4RVl- rflS)- 1- Azidometvl- 3- metvl- butvlcarbamovn- 4- f7- methoxysv- 2- phenvl- quinolin- 4- vloxv)- pvrrolidine- 2- carvvlvl- amino> - 2- vinylcyclopropanecarboxylic acid ester ( 108)

i) (2S)-Methanesulfonic acid 2-tert-butoxycarbonylamino-4-methyl-pentyl ester

Diisopropylethylamine ( 35.7 g, 276 mmol) and methanesulfonyl chloride (15.81 g, 138 mmol). The resulting solution was stirred overnight during which time the mixture was allowed to gradually warm to ambient temperature. The mixture was washed successively with water, 10% citric acid (aq), water and saturated NaHCO 3 (aq), then dried with Na 2 SO 4 and concentrated to a brown solid (32.6 g, 96%) which was used in the next reaction without further purification.

ii) ((1S)-1-Azidomethyl-3-methyl-butyl)-carbamic acid tert-butyl ester

The mesylate from step i (32.6 g, 110 mmol) was treated with sodium azide (21.45 g, 330 mmol) in DMF at 80°C for 24 h. The solvent was evaporated, the residue was taken up in DCM, filtered and washed with saturated NaHCO 3 (aq). The solution was dried with Na 2 SO 4 and concentrated to a brown oil which was purified by flash chromatography using a gradient of ethyl acetate and hexane to afford the title compound as a white solid (19.55 g, 73%).

iii) (1S)-1-Azidomethyl-3-methyl-butylamine

((1S)-1-Azidomethyl-3-methyl-butyl)-carbamic acid tert-butyl ester (9.64 g, 39.78 mmol) was treated with TFA (30 mL) in DCM (150 mL) for 3 h, the mixture was evaporated under reduced pressure and the residue was dissolved in ethyl acetate and washed with aqueous 1 M K 2 CO 3 , dried with Na 2 SO 4 and concentrated to a yellow liquid (4.55 g, 80%).

Compound 12 was treated with phosgene as described in Example 13, which gave the corresponding chlorocarbamate compound. The obtained chlorocarbamate (568 mg, 1.13 mmol) was dissolved in a solution of DCM-THF (1:1, 10 mL) and (1S)-1-azidomethyl-3-methyl-butylamine (401 mg, 2.82 mmol), and a large excess of NaHCO 3 (s) was added. The resulting mixture was stirred for 18 hours, filtered and washed with dilute citric acid (aqueous, pH 5). The organic layer was dried with Na 2 SO 4 and evaporated to give the desired product as a light yellow oil (837 mg, 99%) sufficiently pure to be used in the next step. M+H<+>670.1.

Example 109

f IR, 2SM-( T( 2S, 4RVl- rflS)- 1- Aminometvl- 3- metvl- butvlcarbamovn- 4- f7- methoxysv- 2- phenvl- quinolin- 4- vloxv)- pvrrolidine- 2- carbonvlvl- amino> - 2- vinylcyclopropanecarboxylic acid ester ( 109)

A solution of 108 (717 mg, 1.07 mmol) in THF (25 mL) was shaken with PS triphenylphosphine resin (diphenylphosphinopolystyrene) (3.24 g, 1.65 mmol PPh3/g) and methanol (2, 5 ml) for 78 hours. The mixture was filtered and the polymer was washed repeatedly with DCM and methanol. The combined filtrates were evaporated to give the title compound as a light brown solid foam (685 mg, 99%) with greater than 95% purity as determined by reverse phase HPLC. M+H<+>644.1.

General procedure IA for the preparation of compounds 110-116

To a solution of the acyl chloride (0.075 mmol) in DCM (0.5 mL) was added NaHCO 3 (s)

(60 mg, 07 mmol) and a solution of amine 109 (25 mg, 0.037 mmol) in THF (1 mL). The resulting mixture was stirred at room temperature overnight, filtered and then shaken in the presence of PS-trisamine resin (tris-(2-aminoethyl)aminomethylpolystyrene) (3.91 mmol/g, 50 mg, 0.2 mmol) for 5 h. The mixture was filtered and evaporated. The resulting solid residue was dissolved in MeOH-THF (2:1, 1.5 mL) and treated with 1 M LiOH (aq) (170 µl) at 50°C between 2 and 16 h. The reaction was monitored by HPLC-MS. The mixture was acidified with acetic acid and evaporated to dryness. The residue was dissolved in methanol and purified by reverse-phase HPLC.

General procedure IB for the preparation of compounds 110-116

To the acid (0.039 mmol) was successively added a solution of HATU (14.7 mg,

0.039 mmol) in DMF (0.5 mL), a solution of amine 109 (20 mg, 0.031 mmol) in DMF (0.5 mL) and diisopropylethylamine (30 µL, 0.155 mmol). The resulting mixture was stirred for 16 h, then the solvent was evaporated, and the residue was dissolved in DCM and washed with water and aqueous saturated NaHCO 3 . The solvent was

evaporated, and the residue was dissolved in methanol-THF (2:1, 1.5 mL). To this was added 1 M LiOH (aqueous) (155 µl), and the mixture was stirred at 60°C for 3-5 hours. Glacial acetic acid (50 µl) was added and the mixture was concentrated, dissolved in methanol and purified by reverse phase HPLC.

Example 110

f IR. 2S)- l-(Tf2S. 4RVl- fflS)- l- rr3- Fluoro- benzovlamino)- metvll- 3-metvlbutvlcarbamovl>- f7- methoxysv- 2- phenvl- quinoline- 4- vloxv)- pyrrolidine- 2- carbonvol- amino>- 2- vinvol- cyclopropanecarboxylic acid fl 10)

General procedure IA was followed using 3-fluorobenzoyl chloride (12 mg) as the acyl chloride to give the title compound as a solid (13.6 mg, 50%). M+H<+>738,l.

Example 111

f IR, 2S)- l-{ Tf2S, 4R)- 4- f7- Methoxv- 2- phenvl- quinoline- 4- vloxv)- l- fflS)- 3- metvl- l-• rr( pyridine- 3- carbonyl )- amino1- methyl>- butylcarbamoyl)- pyrrolidine- 2- carbonyl1-amino>- 2- vinyl- cyclopropanecarboxylic acid fill)

General procedure IA was followed using nicotinoyl chloride (10.5 mg) as the acyl chloride to give the title compound as a solid (10 mg, 37%). M+H<+>721,l.

Example 112

f IR, 2S)- 1-( T( 2S, 4RV4- r7- Methoxv- 2- phenvl- quinolin- 4- vloxvM- fflS)- 3- metvl- l-• rrfPvrazin- 2- carbonvn- amino1- metvl>- butylcarbamoyl)- pyrrolidine- 2- carbonyl-amino>- 2- vinyl- cyclopropanecarboxylic acid ( 112)

General procedure IB was followed using pyrazine-2-carboxylic acid (5 mg) as acid to give the title compound as a solid (5.7 mg, 25%). M+H<+>722,l.

Example 113

f IR, 2S)- 1-( T( 2S, 4R)- 4- f7- Methoxv- 2- phenvl- quinoline- 4- vloxv)- l- fflS)- 3- metvl- l-• rr( thiophene- 3 - carbonyl)- amino1- methyl>- butylcarbamoyl)- pyrrolidine- 2- carbonyl1-amino>- 2- vinyl- cyclopropanecarboxylic acid f 113)

General procedure IA was followed using thiophene-3-carbonyl chloride (11 mg) to give the title compound as a solid (4.3 mg, 16%). M+H<+>726,l.

Example 114

f IR, 2S)- 1-( T( 2S, 4R)- 4- f7- Methoxv- 2- phenvl- quinoline- 4- vloxv)- l- fflS)- 3- metvl- l-•} Tftetrarivdro- furan- 2-carbonyl-aminol-methyl>-butylcarbamoven- pyrrolidin- 2-carbonyl- amino>- 2-vinyl-cyclopropanecarboxylic acid ( 114 )

General procedure IB was followed using tetrahydrofuran-2-carboxylic acid (4.5 mg) as acid to give the title compound as a solid (7.9 mg, 36%). M+H<+>714,l.

Example 115

f IR, 2SM-( T( 2S, 4RV4- r7- Methoxv- 2- phenvl- quinolin- 4- vloxvM- fflS)- 3- metvl- l-• fr( 5- phenvl- oxazole- 4- carbovl)- aminol - methyl>- butylcarbamoven- pyrrolidin- 2-carbonyl- amino>- 2- vinyl- cyclopropanecarboxylic acid ( 115)

General procedure IB was followed using 5-phenyl-oxazole-4-carboxylic acid (7.5 mg) as acid to give the title compound as a solid (7.5 mg, 31%). M+H<+>787.1.

Example 116

f IR, 2SM-( T( 2S, 4RVl- rflS)- 1-{ TfBenzofuran- 2- carbovn- aminol- metvl>- 3- mevl- butvlcarbamovn- 4-( 7- methoxysv- 2- phenvl- quinoline- 4- (cyclopropanecarboxylic acid)

General procedure IB was followed using benzofuran-2-carboxylic acid (6.5 mg) as acid to give the title compound as a solid (5.4 mg, 23%). M+H<+>760,l.

General procedure 2 for the preparation of compounds 117-119

To a solution of the sulfonyl chloride (0.075 mmol) in DCM (0.5 mL) was added NaHCO 3 (s) (60 mg) and a solution of amine 109 (25 mg, 0.037 mmol) in THF (1 mL). The resulting mixture was stirred at room temperature for 18 hours, filtered and then shaken with PS-trisamine (tris-(2-aminoethyl)aminomethylpolystyrene, 3.91 mmol/g, ~50 mg) for 5 hours.

The mixture was filtered and the polymer was washed successively with DCM, THF and methanol. The solid residue resulting from evaporation of the combined filtrates was dissolved in MeOH-THF (2:1, 1.5 mL) and treated with 1 M LiOH (aq) (170 µl) at 50 °C for reaction times varying from 18 hours to a week depending on the actual structure. The reaction was monitored by HPLC-MS. The mixture was acidified with acetic acid and evaporated to dryness. The residue was dissolved in methanol and purified by reverse-phase HPLC.

Example 117

f IR, 2S)- l-(- f( 2S, 4RV4- r7- Methoxv- 2- phenyl- quinoline- 4- loxvM- rflS)- 3- methylvl- 1- phenvlmethanesulfonvlamino- metvl)- butvlcarbamovl- pyrrolidine- 2- carboxylic acid (amino)-2-vinyl- cyclopropanecarboxylic acid ( 117 )

General procedure 2 was followed using α-toluenesulfonyl chloride (14 mg) as the sulfonyl chloride to give the title compound as a white solid (4.9 mg, 17%). M+H<+>770.1.

Example 118

f IR, 2SM- r(( 2S, 4R)- 4- f7- Methoxv- 2- phenyl- quinoline- 4- cycloxvM- fflS)- 3- methylvl- 1-rf5- methylvl- isoxazole- 4- sulfonvlamino)- metvl - butylcarbamoyl>- pyrrolidine- 2-carbonyl)- aminol- 2- vinyl- cyclopropanecarboxylic acid f 118)

General procedure 2 was followed using 5-methyl-isoxazole-4-sulfonyl chloride (14 mg) as the sulfonyl chloride to give the title compound as a white solid (1.6 mg, 6%). M+H<+>761.0.

Example 119

f IR, 2S)- 1-( T( 2S, 4R)- l- fflS)- l- rf5- Isoxazole- 3- vl- thiophene- 2- sulfonvlamino)-methyl- 3- methyl- butvlcarbamovl>- 4- f7 - methoxy- 2- phenyl- quinoline- 4- cyclooxy)- pyrrolidine-2- carbonyl- amino>- 2- vinyl- cyclopropanecarboxylic acid ( 119)

General procedure 2 was followed using 5-isoxazol-3-yl-thiophene-2-sulfonyl chloride (19 mg) as the sulfonyl chloride to give the title compound as a white solid (3.0 mg, 10%). M+H<+>828.98.

Example 120

1- free-(N,- tert. Butoxycarbonyl- N- hept- 6-enyl- hvdrazinocarbonyl)- 4-( 7- methoxy- 2-phenyl- quinolin- 4- cyclox)- pyrrolidin- 2- carbonvn- amino>- 2-vinyl-cyclopropanecarboxylic acid ester (120)

Compound 12 (200 mg, 0.4 mmol) was dissolved in tetrahydrofuran (10 mL). A teaspoon of sodium bicarbonate was added, followed by phosgene (1.8 µl, 1.9 M in toluene). The reaction mixture was stirred for 30 min and filtered. The solvent was evaporated and the crude chloride redissolved in dichloromethane (10 mL). Sodium bicarbonate (1 teaspoon) and N'-hept-6-enyl-hydrazinecarboxylic acid tert-butyl ester (182 mg, 0.8 mmol) were added. The reaction mixture was stirred at room temperature for 40 h and then filtered and purified by silica chromatography (1% methanol in ether -> 2% methanol in ether) to give pure title product (240 mg, 79%).

Example 121

14- tert. Butoxxcarbonvlamino- 18- f7- methoxyx- 2- phenvl- quinoline- 4- xoxx)- 2, 15-dioxo- 3, 14, 16- triaza- trichloro 14. 3. 0. 0* 4, 6* 1 nonadec- 7 - en- 4- carboxylic acid ester ( 121 )

Compound 120 (200 mg, 0.26 mmol) was dissolved in degassed dichloromethane (30 mL). Hoveyda-Grubbs catalyst II generation (16 mg, 0.026 mmol) was then added and the mixture was refluxed under argon overnight. The solvent was then evaporated, and the crude product was purified by silica chromatography (1% methanol in ether) to give 39 mg (20%) of the title product. MS (M+H<+>) 728.2

Example 122

14- tert. Butoxxcarbonvlamino- 18- f7- methoxyx- 2- phenvl- quinoline- 4- xoxx)- 2, 15-dioxo- 3, 14, 16- triaza- trichloro 14. 3. 0. 0* 4, 6* 1 nonadec- 7 - en- 4- carboxylic acid ( 122)

Compound 121 (39 mg, 0.054 mmol) was dissolved in tetrahydrofuran (3.5 mL), water (1.75 mL) and methanol (1.75 mL). Lithium hydroxide (430 µl, 1 M in water) was then added and the reaction was stirred at room temperature for 24 h. The volume was reduced to half and water (10 mL) was added. Acidification (pH=5) followed by extraction with chloroform gave 34 mg (90%) of the pure acid 179. MS (M+H<+>) 700.2

Example 123

l- rri- fN,- tert. Butoxxcarbonyl- N- hex- 5-enyl- hydrazinocarbyl)- 4- f7- methoxy- 2-phenyl- quinoline- 4- loxyl)- pyrrolidin- 2- carbonyl- amino>- 2- vinyl- cyclopropanecarboxylic acid ester ( 123)

The title compound was prepared from compound 12 (800 mg, 1.6 mmol) and γ-hex-5-enyl-hydrazinecarboxylic acid tert-butyl ester (620 mg, 2.9 mmol) according to the procedure described in Example 120, giving 1 g (85%). MS (M+H<+>) 742.37

Example 124

13- tert. Butoxxcarbonvlamino- 17- f7- methoxyx- 2- phenvl- quinoline- 4- xoxxv)- 2, 14-dioxo- 3, 13, 15- triaza- trichloro 13. 3. 0. 0* 4, 6* loctadec- 7- en- 4- carboxylic acid ethyl ester ( 124)

Treatment of compound 123 (400 mg, 0.54 mmol) according to the procedure described in Example 121 gave a crude product. Purification by silica gel chromatography (1% methanol in ether) afforded the product (67 mg, 17%). MS (M+H<+>) 714.29

Example 125

13- tert. Butoxxcarbonvlamino- 17-( 7- methoxyx- 2- phenyl- quinolin- 4-xxx)- 2, 14-dioxo- 3, 13, 15- triaza- trichloro 13. 3. 0. 0* 4, 6* loctadec- 7 - en- 4- carboxylic acid ( 125)

The title compound was prepared from compound 124 (67 mg, 0.09 mmol) by the same procedure as described in 122 to give 46 mg (71%) of the pure acid. Chloroform was replaced with 1,2-dichloroethane in the extraction step for the preparation of this compound. MS (M+H<+>) 686.33

Example 126

13- tert. Amino- 17- f7- methoxy- 2- phenyl- quinoline- 4- cyclox)- 2, 14- dioxo- 3, 13, 15- triaza- trichloro3. 3. 0. 0* 4, 6* loctadec- 7- en- 4- carboxylic acid f 126)

Compound 125 (10 mg) was dissolved in dichloromethane (4 mL). Trifluoromethanesulfonic acid (4 mL) was added and the mixture was allowed to stand at 50°C for 6 hours. The solvent was removed and the residue was washed with acetonitrile to give 3 mg of the pure title product (35%). MS (M+H<+>) 586.25

Example 127

1- free-(1- Methoxycarbonyl- oct- 7-enylcarbamoyl)- 4-( 7- methoxy- 2- phenyl- quinoline- 4-cyclohexyl)- pyrrolidin- 2- carbonyl- amino>- 2- vinyl- cyclopropanecarboxylic acid ester a.m

The title compound was prepared from compound 12 (380 mg, 0.758 mmol) and 2-aminononan-8-enyl-carboxylic acid methyl ester (250 mg, 1.89 mmol) using the conditions described in Example 120, giving the pure product (405 mg, 75%).

Example 128

19-( 7- Methoxy- 2- phenyl- quinolin- 4- yloxy)- 2, 16- dioxo- 3, 15, 17- triaza- trichloro5. 3. 0. 0* 4, 6* licos- 7- en- 4, 14- dicarboxvlsvre- 4- etvlester- 14- metvlester

(128)

Compound 127 (170 mg, 0.2385 mmol) was dissolved in dichloromethane (40 mL) and degassed by bubbling through nitrogen for 20 min. Hoveyda-Grubbs catalyst II generation (10 mg, 0.016 mmol, 6.7 mol%) was then added and the mixture was refluxed under nitrogen atmosphere overnight. The solvent was then evaporated, catalyst and salts were removed by flash chromatography (5% methanol in chloroform) and the crude product (120 mg, 73% yield, 85-90% purity) was used in the next step MS (M+H<+>) 685

Example 129

19-( 7-Methoxy- 2- phenyl- quinolin- 4- chloro)- 2, 16- dioxo- 3, 15, 17- triaza- trichloro5. 3. 0. 0* 4, 6* licos- 7- en- 3, 14- dicarboxvlsvre- 3- etvlester ( 129)

Compound 128 (120 mg, 0.175 mmol) was dissolved in dioxane (9 mL) and water (6 mL). Lithium hydroxide (12 mg, 0.526 mmol) was added and the reaction mixture was stirred at room temperature for 3.5 h. The mixture was acidified with acetic acid to pH=5 and co-evaporated with toluene. The crude product was used in the next step. MS (M+H<+>) 671

Example 130

14- rfCvclohexvl- metvlcarbamovl- metvl)- f7- methoxysv- 2- phenvl- quinoline- 4- vloxv)-2, 16- dioxo- 3, 15, 17- triaza- tricvchlor 15. 3. 0. 0* 4 , 6* 1 icos- 7- en- 4- carboxylic acid 3-ethyl ester ( 130)

Compound 129 (crude, 100 mg), indanolamine (33 mg, 0.209 mmol) and Hunig's base (DIEA) (0.2 mL) were dissolved in DMF (14 mL). After stirring at 0°C for 10 min, HATU was added. The reaction was monitored by LC-MS. After 5h the conversion was 100%. DMF and DIEA were removed in vacuo. The residue was partitioned between ethyl acetate and water. The organic layer was washed with brine, dried and concentrated in vacuo. The crude product yield was 120 mg, purification by prep. HPLC gave 21 mg (25%) of the title product. MS (M+H<+>) 802

Example 131

14- rfCvclohexvl- metvlcarbamovl- metvl)- f7- methoxysv- 2- phenvl- quinoline- 4- vloxv)-2, 16- dioxo- 3, 15, 17- triaza- tricvchlor 15. 3. 0. 0* 4 , 6* 1 icos- 7- en- 4- carboxylic acid ( 131)

To a solution of the ester 130 (19 mg, 0.024 mmol) in the mixture of THF (0.2 mL) and methanol (0.3 mL) was added a solution of LiOH (6 mg, 0.237 mmol) in 0.15 mL of water. The resulting mixture was stirred at 60°C for 3.5 h. After cooling to room temperature, acetic acid was added (30 eq). The mixture was co-evaporated with toluene. The residue was partitioned between chloroform and the aqueous phase, the aqueous phase was extracted with chloroform and ethyl acetate, the organic phases were combined, dried over sodium sulfate, evaporated to give 15 mg of pure product. MS (M+H<+>) 774

Example 132 ri4- Cyclopropanesulfonvlaminocarbonvl- 17- f7- methoxy- 2- phenyl- quinolin- 4- cyclox)-2, 14- dioxo- 3. 13. 15- triaza- trichloro 13. 3. 0. 0* 4. 6* loctadec- 7- en- 13- vll-urea tert. buttlester ( 132)

To the acid 125 (19 mg, 0.028 mmol) in 0.5 mL of DMF was added 5.5 mg (0.044 mmol) of DMAP and 10.7 mg (0.056 mmol) of EDC. After 6.5 hours of stirring, 20 mg of cyclopropyl sulfonamide and 0.04 ml of DBU were added. The mixture was stirred overnight, acidified with 5% citric acid (in water) and extracted with ethyl acetate. Dried, evaporated, purified by 5% to 10% methanol in chloroform (or prep LC-MS), yielding 8 mg of the title compound (37%). MS (M+H<+>) 783

Example 133

4- r2- f2- Isopropvlamino- thiazole- 4- vl)- 7- methoxy- quinoline- 4- vloxvl- pyrrolidin- 1, 2-dicarboxylic acid 1- tert. buttlester ( 133)

To a stirred solution of N-Boc-trans-4-hydroxy-L-proline (221 mg, 0.96 mmol) in DMSO was added potassium tert-butoxide (320 mg, 2.9 mmol). After 1h, 2-[2-isopropylamino)-1,3-thiazol-4-yl]-7-methoxyquinolin-4-ol (319 mg, 0.96 mmol) was added and the mixture was stirred at 70°C for 72 hours. The mixture was diluted with water and extracted with ethyl acetate. The product was used without further purification. Yield 429 mg, 85%.

Example 134

2- f l- Ethoxysvl- 2- vinvl- cvcloproplvlcarbamovn- 4- r2- f2- isopropylamino- thiazole-4- vn- 7- methoxysv- quinoline- 4- vloxvl- pyrrolidin- l- carboxyvl- tert. buttlester ( 134 )

Compound 133 (300 mg, 0.56 mmol) was reacted with 1-amino-2-vinyl-cyclopropanecarboxylic acid ethyl ester (130 mg, 0.84 mmol) as described in Example 11 to give the title compound (302 mg, 80%) .

Example 135

l- f-{' 4- r2- f2- Isopropvlamino- thiazole- 4- vl)- 7- methoxy- quinoline- 4- vloxvl- pyrrolidin- 2-carbovl>- amino)- 2- vinvl- cyclopropanecarboxylic acid vl- ethyl ester ( 135 )

Compound 134 (302 mg, 0.45 mmol) was treated as described in Example 12 to give the title compound (195 mg, 76%).

Example 136

l- f-{' l- ri- f2- Hydroxv- indan- l- vlcarbamovn- 2, 2- dimethyl- propvlcarbamovl- 4- r2-f2- isopropvlamino- thiazole- 4- vl)- 7- methoxysv- quinoline- 4 - cyclopropane- pyrrolidin- 2- carboxyl>-amino- 2- vinyl- cyclopropanecarboxylic acid ester ( 136)

Compound 135 (80 mg, 0.14 mmol) was treated as described in Example 13 to give the title product (87 mg, 72%).

Example 137

l-(- fl- ri-( 2- Hydroxy- indan- 1- vlcarbamovyl)- 2, 2- dimethyl- propvlcarbamovyl- 4- r2-( 2- isopropylamino- thiazol- 4- yl)- 7- methoxy- quinolin- 4-yloxy1-pyrrolidine-2-carbonyl>-amino- 2-vinyl- cyclopropanecarboxylic acid ( 137)

The ethyl ester of compound 136 (80 mg, 0.09 mmol) was hydrolyzed according to the procedure described in Example 14 to give the title product. Yield by preparative LC-MS (7.5 mg, 10%).

Example 138

l- rri- Ethylcarbamovl- 4- f7- methoxy- 2- phenyl- quinoline- 4- cyclox)- pyrrolidin- 2-carbovllamino>- 2- vinyl- cyclopropanecarboxylic acid- ethyl ester ( 138)

Reaction of compound 12 (330 mg, 0.66 mmol), phosgene (1.6 mL, 1.9 M in toluene, 3 mmol) and hex-5-enylamine hydrochloride (500 mg, 3.68 mmol) according to the procedure described in Example 120 gave the pure title product (328 mg, 80%), MS (M+H<+>) 627.

Example 139

17- f7- Methoxv- 2- phenvl- quinoline- 4- vloxv)- 2, 14- dioxo- 3, 13, 15- triaza- trichloro 13. 3. 0. 0* 4, 6* loctadec- 7- en- 4- carboxylic acid ethyl ester f 139)

Compound 138 (200 mg, mol) was dissolved in degassed dry dichloromethane (200 mL), bubbled through with nitrogen. Then, Hoveyda-Grubbs catalyst (second generation) (5 mg, 2 mol%) was added, and the reaction mixture was refluxed for 20 h under nitrogen. The resulting mixture was cooled to room temperature and concentrated by rotary evaporation. The resulting oil was purified by column chromatography on YMC silica (ethyl acetate - toluene 1:1 to 9:1) to give 55 mg of the title compound as a beige solid. Dividend 29%. MS (M+H<+>) 599.

Example 140

17-( 7- Methoxy- 2- phenyl- quinoline- 4- cyclooxy)- 2, 14- dioxo- 3, 13, 15- triaza- trichloro3. 3. 0. 0* 4, 6* loctadec- 7- en- 4- carboxylic acid f 140)

Compound 139 (55 mg, mol) was dissolved in 2 mL of methanol and mixed with 3 equiv. of aqueous NaOH and heated for 2 h at 60°C in a closed bottle. The reaction mixture was then extracted into ethyl acetate. The aqueous solution was collected and acidified with IN HCl solution to pH 2. The resulting solution was concentrated by rotary evaporation, dissolved in methanol and purified by preparative HPLC (acetonitrile-water) to give 34 mg of the title product. Yield 65%. MS (M+H<+>) 571.

Example 141 1- iri- yl- rfCcyclohexyl- methoxycarbonyl- methvn- carbamol- 2, 2- dimethyl-propylcarbamol>- 4-( 7- methoxy- 2- phenyl- quinoline- 4- cyclohexyl)- pyrrolidin- 2- carbonyl- amino >- 2- vinyl-cyclopropanecarboxylic acid ( 141).

Compound 103 was dissolved in dichloromethane (3 mL), and solid sodium bicarbonate (100 mg) and phosgene 20% in toluene (0.1 mL) were added. After 30 min at room temperature, the mixture was concentrated to dryness. (S)-(2S-2-Amino-3,3-dimethyl-butyrylamino)-cyclohexyl-acetic acid methyl ester (12 mg in dichloromethane 2 ml) was added. After 3 days of shaking at room temperature, the reaction mixture was filtered, concentrated to dryness and purified on preparative HPLC-MS to give the title product (4.4 mg). M+H<+>784.7.

Example 142

1-- TT l-( l- Aminomethyl- 2, 2- dimethyl- propylcarbamoyl- 4- f7- methoxy- 2- phenyl- quinoline-4- cyclohexyl)- pyrrolidin- 2- carbonyl- amino>- 2- vinyl- cyclopropanecarboxylic acid -etlester f 142)

The title compound was prepared from compound 12 (1.22 g, 2.43 mmol) by following

the procedure described for the preparation of compound 108, but using methanesulfonic acid 2-tert.butoxycarbonylamino-3,3-dimethyl-butyl ester instead of methanesulfonic acid 2-tert.butoxycarbonylamino-4-methylpentyl ester, in Example 165 step i). Reduction of the azide as described in Example 109 gave the title compound

(1.49 g, 95%). Purity according to HPLC >95%, M+H<+>644.2.

Example 143

l- Cri- f2, 2- Dimethyl- l-{'rthiophene- 3- carbonvn- aminol- metvl>- propvlcarbamovl)- f7- methoxysv- 2- phenvl- quinoline- 4- cycloxv)- pyrrolidine- 2- carbonvl - amino>- 2-vinylcyclopropanecarboxylic acid ( 143)

Compound 142 (100 mg, 0.155 mmol) was reacted according to the general procedure IA for the preparation of compounds 110-116, using thiophene-3-carbonyl chloride (28.5 mg, 0.194 mmol) as the acyl chloride, giving the title compound as a white solid (45 mg, 40%). Purity according to HPLC

>95%, M+H<+>726.

Example 144

l- Cn-{' l- r( 5- Isoxazole- 3- vl- thiophene- 2- sulfonvlamino)- methylvl- 2, 2- dimethyl-propvlcarbamovl>- f7- methoxysv- 2- phenyl- quinoline- 4- cyclopropane carboxylic acid f 144)

Compound 142 (25 mg, 0.039 mmol) was reacted according to the general procedure IA for the preparation of compounds 110-116, using 5-isoxazol-3-yl-thiophen-2-sulfonyl chloride (14.5 mg, 0.058 mmol ) as the acyl chloride, affording the title compound as a white solid (1.8 mg, 6%). Purity according to HPLC was >94%, M+H<+>829.

Example 145

l- rri- f3- Fluoro- benzovlamino)- metvll- propvlcarbamovl>- 4- f7- methoxyvl>- 2- phenvlquinoline- 4- vloxvl)- pyrrolidin- 2- carvvll- amino>- 2- vinvl- cyclopropanecarboxylic acid

(145)

Compound 142 (25 mg, 0.039 mmol) was reacted according to the general procedure IA for the preparation of compounds 110-116, using 3-fluorobenzoyl chloride (12.3 mg, 0.078 mmol) as the acyl chloride, affording the title compound as a white solid (4.1 mg, 14%). Purity according to HPLC was >94%, M+H<+>738.

Example 146

1- ri- f lirf- Furan- 3bcarbonvl)- aminol- metvll- 2, 2- dimethyl- propvlcarbamovl>- 4- f7- methoxysv- 2- phenvl- quinoline- 4- cycloxv)- pvrrolidine- 2- carbonvl- amino> - 2- vinylcyclopropanecarboxylic acid ( 146)

Compound 142 (25 mg, 0.039 mmol) was reacted according to the general procedure IB for the preparation of compounds 110-116, using 3-furanic acid (5.5 mg, 0.049 mmol) as the acyl chloride, affording the title compound as a white solid (4.1 mg, 14%). Purity according to HPLC was >99%, M+H<+>710.

Example 147

4- f7- Methoxv- 2- phenyl- quinoline- 4- cyclox)- pyrrolidin- 1, 2- dicarboxylic acid 2-\( 1-cyclopropanesulfonvlaminocarbonvl- 2- vinvl- cyclopropyl)- amidl l- r( 2, 2- dimethyl- l-• f r( thiophene- 3- carbonvl) aminol- metvl>- propyl)- amide f 147)

To a solution of compound 143 (42.2mg, 0.058mmol) in chloroform (3ml) was added cyclopropylsulfonamide (14mg, 0.116mmol) followed by diisopropylethylamine (60.5µl, 0.17mmol). The solution was stirred at RT for 10 min and then at -20°C for 30 min. PyBOP (121 mg, 0.116 mmol) was then added as a solid. The solution was kept at -20°C for 10 days. The solution was then poured into aqueous NaHCO 3 (saturated) and washed with water. The organic layer was dried, concentrated and subjected to purification by HPLC to give the title compound as a white solid (2.3 mg, 0.0028 mmol), HPLC purity>95%, M+H<+>830 .

Example 148

Fmoc- 4-amino- 2-(1-ethoxycarbonyl-2-vinyl-cyclopropylcarbamoven- pyrrolidine-1-carbocyclovolvyl tert. butyl ester ( 148 )

(2S,4R) Fmoc-4-amino-1-Boc-pyrrolidine-2-carboxylic acid (5.3 g, 11.8 mmol) was dissolved in DCM (100 mL), HATU (4.94 g, 12.99 mmol), DIEA (4.63 mL, 26.57 mmol) and vinylcyclopropyl glycine ethyl ester (2.26 g, 11.81 mmol) were added one after the other. The mixture was stirred for 16 h at room temperature and was then diluted with

DCM (50 mL), washed with citric acid (10% aqueous), water, NaHCO 3 (saturated aqueous) and water. The organic phase was dried over Na 2 SO 4 and concentrated to give a beige solid foam (8.11 g) which was subjected to silica gel column chromatography to give the title compound (7.14 g, 12.11 mmol).

Example 149

l- r( Fmoc- 4- amino- pyrrolidine- 2- carbonyl)- aminol- 2- vinylcyclopropanecarboxylic acid ester ( 149)

Compound 148 (3.65 g, 6.04 mmol) was treated with a solution of TFA/DCM (10 ml TFA, 50 ml DCM) for 2.5 h and then concentrated to give the title compound (2.99 g, 6.12 mmol) ).

Example 150

l- f-{' Fmoc- 4- amino- l- ri- f2- rivdroxv- indan- l- vlcarbamovl- 2, 2- dimethyl-propvlcarbamovl- pvrrolidine- 2- carbovl>- amino)- 2- vinvl- cyclopropanecarboxvlvre- etvlester ( 150)

The aminoproline derivative 149 (2.96 g, 6.04 mmol) was stirred with phosgene (1.93 M in toluene, 4 mL, 7.55 mmol) for 10 min. The solvents and excess phosgene were evaporated. The residue was dissolved in DCM (30 mL), and t-Bug-aminoindanol (1.9 g, 7.24 mmol) was added as a solution in DCM (30 mL), followed by NaHCO 3 (2 g). The mixture was stirred for 48 h, then diluted with DCM, washed with water, 10% citric acid and NaHCO 3 (saturated, aqueous), dried over Na 2 SO 4 and evaporated to dryness. The residue was subjected to column chromatography purification, EtOAc-hexane 0-30% to obtain the title compound (1g, 1.3 mmol).

Example 151

l-(- f4- Amino- l- ri-( 2- hydroxy- indan- l- vylcarbamovn- 2, 2- dimethyl-propvlcarbamovl- pyrrolidin- 2- carbovl>- amino)- 2- vinvl- cyclopropanecarboxylic acid ester ( 151 )

Compound 150 (595 mg, 0.765 mmol) was dissolved in DMF (20 mL) and treated with Si-piperazine (0.08 mmol/g, 4.78 g, 3.82 mmol) for 48 h. The silica was filtered and washed once with DMF and then with several portions of DCM. The solvents were evaporated and the residue subjected to column chromatography to obtain the title compound (170 mg, 0.3 mmol).

Example 152

l- f-{' l- ri- f2- Hydroxv- indan- l- vlcarbamovl)- 2, 2- dimethyl- propvlcarbamovl- 4-rfpvridine- 3- carbonvl)- aminol- pyrrolidine- 2- carbonvl>- amino)- 2- vinylcyclopropanecarboxylic acid ( 152)

To a stirred solution of compound 151 (35 mg, 0.064 mmol) in DCM (1 mL) was added DIEA (0.12 mmol, 19 µl) and nicotinoyl chloride hydrochloride (0.12 mmol, 17 mg). The solution was stirred at RT for 18h, PS-trisamine was added, then stirred at RT for 4h. After filtration, the solution was washed with citric acid (10% aqueous) and NaHCO 3 (saturated, aqueous), and the organic phase was dried over Na 2 SO 4 and concentrated. The residue was dissolved in THF:MeOH (2:1, 1.5 mL). LiOH (1N aqueous, 3.2 mmol, 320 µl) was added. The solution was stirred at 60°C for 24h. Acetic acid was added and then concentrated. The residue was dissolved in MeOH and subjected to purification by HPLC to give the title compound (19.5 mg, 0.03 mmol). Purity according to HPLC>98%, M+H<+>633.1.

Example 153

l- fil- ri- f2- Hydroxv- indan- l- vlcarbamovn- 2, 2- dimethyl- propylcarbamovvl- 4-phenvlacetamino- pvrrolidine- 2- carbovl>- amino)- 2- vinvl- cyclopropanecarboxylic acid £1531

The procedure described in Example 152, but using phenylacetyl chloride instead of nicotinoyl chloride hydrochloride, was followed to give the title compound (12.7 mg, 0.019 mmol). Purity according to HPLC>90%, M+H<+>646.1.

Example 154

l- f-{' l- ri- f2- Hydroxv- indan- l- vlcarbamovl)- 2, 2- dimethyl- propvlcarbamovl- 4- rf5-metvl- 3- phenvl- isoxazole- 4- carbovl)- aminol- pvrrolidine- 2- carboxyl>- amino)- 2- vinylcyclopropanecarboxylic acid ( 154)

The procedure described in Example 152, but using 5-methyl-3-phenyl-isoxazole-4-carbonyl chloride instead of nicotinoyl chloride hydrochloride, was followed to give the title compound (3.6 mg, 0.0055 mmol). Purity according to HPLC>98%, M+H<+>713.1.

Example 155

l- iri- ri- f2- Hydroxv- indan- l- vlcarbamovn- 2, 2- dimethyl- propylcarbamovn- 4- f3-phenyl- ureido)- pyrrolidin- 2- carbonvn- amino>- 2- vinvl- cyclopropanecarboxylic acid £1551

To a stirred solution of compound 151 (30 mg, 0.054 mmol) in acetonitrile:dichloromethane (2:1, 3 mL) was added triethylamine (0.0648 mmol, 9 µL) and phenyl isocyanate (0.0648 mmol, 7 µL). The solution was stirred at room temperature for 3h, methanol was added (1ml) and then it was concentrated. The residue was dissolved in methanol and subjected to purification by HPLC, which gave the ester compound as a white solid (32.7mg, 0.047 mmol), HPLC purity>95%, M+H<+>675.31. LiOH IN aqueous (0.47 mmol, 475 µl) was added to the ester dissolved in THF:MeOH (2:1). The reaction mixture was stirred at 50°C for 15 min and then at 8°C for 12 h followed by the addition of acetic acid (0.98 mmol, 53 µl) before concentration. The residue was dissolved in MeOH and subjected to purification by HPLC to give the title compound as a white solid (3.8 mg, 0.006 mmol), HPLC purity>98%, M+H<+>675.31.

Example 156

l- f-{' 4- Benzenesulfonvlamino- l- ri- f2- hydroxy- indan- l- vlcarbamovl)- 2, 2- dimethyl-propvlcarbamovl- pyrrolidin- 2- carbovl>- amino)- 2- vinvl- cyclopropanecarboxylic acid

(156)

To a stirred solution of compound 151 (30 mg, 0.054 mmol) in DCM (2 mL), DIEA (0.0648 mmol, 11.5 µl) and phenylsulfonyl chloride (0.0648 mmol, 11.5 µl) were successively added . The solution was stirred at RT for 3h, and then it was concentrated. The residue was dissolved in MeOH and subjected to purification by HPLC to give the ester compound as a white solid (17.9 mg, 0.0257 mmol), HPLC purity>95%, M+H<+>696.24 . LiOH IN aqueous, (0.25 mmol, 257 µl) was added to the ester dissolved in THF:MeOH (2:1). The reaction mixture was stirred at 50°C for 1.5 h before the addition of acetic acid (0.98 mmol, 53 µl). The solution was concentrated. The residue was dissolved in DCM and washed with water; the aqueous phase was acidified to pH 5 and then extracted with dichloromethane and ethyl acetate. The combined organic phases were dried over Na 2 SO 4 and concentrated to give the title compound as a white solid (7.1 mg, 0.01 mmol), purity by HPLC>98%, M+H<+>668.19.

Example 157

l- rri- ri- f2- Hydroxv- indan- l- vlcarbamovl)- 2, 2- dimethyl- propylcarbamovl- 4- f3-phenvl- thioureido)- pvrrolidine- 2- carbovl>- amino)- 2- vinvl- cyclopropanecarboxylic acid £ 15Z1

To a stirred solution of compound 151 (30 mg, 0.054 mmol) in acetonitrile (3 mL) TEA (0.0648 mmol, 9 µl) and phenylthioisocyanate (0.0648 mmol, 7.8 µl) were successively added. The solution was stirred at RT for 16h, and then it was concentrated. The residue was dissolved in MeOH and subjected to purification by HPLC to give the ester compound as a white solid (22.7mg, 0.0328mmol), HPLC purity>95%, M+H<+>691.2. LiOH IN aqueous, (0.33 mmol, 328 µl) was added to the ester dissolved in THF:MeOH (2:1). The reaction mixture was stirred at 50°C for 2.5 h before the addition of acetic acid (0.98 mmol, 53 µl). The solution was concentrated. The residue was dissolved in dichloromethane and washed with water, the aqueous phase was extracted with EtOAc. The combined organic phases were dried over Na 2 SO 4 and concentrated to give the title compound as a white solid (7.2 mg, 0.01 mmol), purity by HPLC>98%, M+H<+>663.26.

Analyze (assavs)

The compounds according to the invention are readily analyzed for activity against the NS3 protease of flaviviruses such as HCV using conventional assays or cell culture assays in vitro (enzyme).

A useful assay is Bartenshlager's replicon assay described in EP 1043399. An alternative replicon assay is described in WO 03064416.

A suitable enzyme assay involving inhibition of full-length hepatitis C NS3 is essentially as described in Poliakov, 2002 Prot Expression & purification 25 363 371. Briefly, the hydrolysis of a depsipeptide substrate, Ac-DED(Edans)EEAbu \j/[COO]ASK(Dabcyl)-NH2 (AnaSpec, San José, USA), is measured spectrofluorometrically in the presence of a peptide cofactor, KKGSVVIVGRIVLSGK, as described by Landro, 1997 Biochem 36 9340-9348. The enzyme (1 nM) is incubated in a buffer such as 50 mM HEPES, pH 7.5, 10 mM DTT, 40% glycerol, 0.1% n-octyl-β-D-glucoside, with 25 nM cofactor and inhibitor at about 30° C for 10 min, after which the reaction is initiated by adding substrate, normally 0.5 nM substrate. The inhibitors are normally dissolved in DMSO, sonicated for 30 s and vortexed. The solutions are generally stored at -20°C between measurements.

An alternative enzyme assay is described in WO 0399316 and uses an HCV NS3/4A protease complex FRET peptide assay. The purpose of this in vitro assay is to measure the inhibition of HCV NS3 protease complexes, derived from BMS, H77Cell or J416S strains, as described below, by compounds according to the present invention. This assay provides an indication of how effective compounds of the present invention would be in inhibiting HCV proteolytic activity.

Serum is taken from an HCV-infected patient. An engineered full-length cDNA template of the HCV genome (BMS strain) was constructed from DNA fragments obtained by reverse transcription-PCR (RT-PCR) of serum RNA and by using primers selected on the basis of homology between other genotype la - tribes. From the determination of the genome sequence, a genotype Ia was assigned to the HCV isolate according to the classification of Simmonds et al. (See P Simmonds, KA Rose, S Graham, SW Chan, F McOmish, BC Dow, EA Follett, PL Yap and H Marsden, J.CIin. Microbiol.,

31(6), 1493-1503 (1993)). The amino acid sequence of the non-structural region, NS2-5B, was found to be >97% identical to HCV genotype la (H77C) and 87% identical to genotype Ib (J4L6S). The infectious clones, H77C (Ia genotype) and J4L6S

(Ib genotype) can be obtained from R. Purcell (NIH) and the sequences are published in Genbank (AAB67036, see Yanagi, M, Purcell, RH, Emerson, SU and Bukh. Proe. Nati. Acad. Sei. USA 94 (16 ) 8738-8743 (1997); AF054247, see Yanagi, M, St Claire, M, Shapiro, M, Emerson, SU, Purcell, RH and Bukhj, Virology 244 (1), 161 (1998)).

The BMS, H77C and J4L6S strains are conventional for the production of recombinant NS3/4A protease complexes. DNA encoding the recombinant HCV NS3/4A protease complex (amino acids 1027 to 1711) for these strains was manipulated as described by P. Gallinari et al. (see Gallinari P, Paolini C, Brennan D, Nardi C, Steinkuhler C, De Francesco R. Biochemistry. 38(17):562032, (1999)). Briefly, a three-lysine solubilizing tail was added at the 3' end of the NS4A coding region. The cysteine in the P1 position of the NS4A-NS4B cleavage site (amino acid 1711) was changed to a glycine to avoid proteolytic cleavage of the lysine tag. Also, a cysteine to serine mutation can be introduced by PCR at amino acid position 1454 to prevent autolytic cleavage in the NS3 helicase domain. The aberrant DNA fragment can be cloned into the pET21b bacterial expression vector (Novagen), and the NS3/4A complex can be expressed in the Escherichia coli strain BL21 (DE3) (Invitrogen) according to the protocol described by P. Gallinari et al. (see Gallinari P, Brennan D, Nardi C, Brunetti M, Tomei I, Steinkuhler C, De Francesco R., J Virol. 72(8):6758-69 (1998)) with modifications. Briefly, NS3/4A expression can be induced with 0.5mM Isopropyl beta-D-thiogalactopyranoside (IPTG) for 22 hours at 20°C. A typical fermentation (101) yields approximately 80g of wet cell paste. The cells are resuspended in lysis buffer (10 ml/g) consisting of 25 mM N-(2hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid)

(HEPES), pH7.5, 20% glycerol, 500mM sodium chloride (NaCl), 0.5% Triton-X100, 1 ug/ml lysozyme, 5mM magnesium chloride (MgCl2), 1 ug/ml Dnasel, 5mM beta-mercaptoethanol (BME ), Protease inhibitor - ethylenediaminetetraacetic acid (EDTA) free (Roche), homogenized and incubated for 20 min at VC. The homogenate is sonicated and made clear by ultra-centrifugation at 235,000 g for 1 hour at 4°C.

Imidazole is added to the supernatant to a final concentration of 15mM and the pH value adjusted to 8. The untreated protein extract is applied to a nickel-nitrilotriacetic acid (Ni-NTA) column, previously equilibrated with buffer B (25n-tM 2 0 HEPES, pH8 20% glycerol, 500mM NaCl, 0.5% Triton-XIOO, 15mM imidazole, 5mM BME). The sample is placed at a flow rate of Imi/min. The column is washed with 15 column volumes of buffer C (same as buffer B except with 0.2% Triton-XIOO). The protein is eluted with 5 column volumes of buffer D (same as buffer C except with 200 mM imidazole).

NS3/4A protease complex-containing fractions are pooled and applied to a Superdex-S200 desalting column pre-equilibrated with buffer D (25 mM HEPES, pH7.5, 20% glycerol, 300 mM NaCl, 0.2% Triton-XIOO, 10 mM BME). The sample is placed at a flow rate of ImUmin. NS3/4A protease complex 30-containing fractions are pooled and concentrated to approximately 0.5 mg/ml. The purity of the NS3/4A protease complexes, derived from the BMS, H77C and J4L6S strains, is generally considered to be greater than 90% by SDS-PAGE and

mass spectrometry analyses.

The enzyme is normally stored at -80°C, thawed on ice and diluted before use in assay buffer. The substrate used for the NS3/4A protease assay is conveniently RET S 1 (Resonance Energy Transfer Depsipeptide Substrate; AnaSpec, Inc. eat # 22991)(FRET peptide), described by Taliani et al. in Anal. Biochem. 240(2):6067

(1996). The sequence of this peptide is loosely based on the natural NS4A/NS4B cleavage site except that there is an ester bond and not an amide bond at the cleavage site. The peptide substrate is incubated with one of the three recombinant NS3/4A complexes, in the absence or presence of a compound of the present invention, and the formation of fluorescent reaction product is monitored continuously using a Cytofluor Series 4000. Useful reagents are as follows: HEPES and Glycerol (Ultrapure) can be obtained from GIBCO-BRL. Dimethyl sulfoxide (DMSO) is from Sigma. Beta-Mercaptoethanol is from Bio Rad.

Assay buffer: 50mM HEPES, pH7.5; 0.15 M NaCl; 0.1% Triton; 15% Glycerol; lOmM BME. Substrate: 2 µM final concentration (from a 2 mM stock solution in DMSO stored at -20°C). HCV NS3/4A type la (Ib), 2-3 nM final concentration (from a 5 µM stock solution in 25 mM HEPES, pH7.5, 20% glycerol, 300 mM NaCl, 0.2% Triton-XIOO, 10 mM BME). For compounds with potency approaching the assay limit, the assay can be made more sensitive by adding 50 µg/ml BSA to the assay buffer and/or reducing the final protease concentration to 300 µM.

The analysis is conveniently carried out in a 96-well black polystyrene plate from Falcon. Each well contains 25ul NS3/4A protease complex in assay buffer, 50ul of a compound according to the present invention in 10% DMSO/assay buffer and 25ul substrate in assay buffer. A control (no compound) is also prepared on the same assay plate. The enzyme complex is mixed with compound or control solution, normally for 1 min before initiation of the enzymatic reaction by addition of substrate. The assay plate is normally read immediately using a spectrophotometer such as a Cytofluor Series 4000 (Perspective Biosystems). The instrument is conveniently set to read emission at 340nm and excitation at 490nm at 25°C. The reactions are generally followed for approximately 15 minutes.

The percentage inhibition can be calculated with the following equation. where dF is the change in fluorescence over the linear range of the curve. A non-linear curve fit is applied to the inhibition concentration data and the 50% effective concentration (IC50) is calculated using software such as "Excel XI-fit" software by applying the equation: Enzyme assays conveniently utilize a principle of fluorescence resonance energy transfer (FRET) to generate a spectroscopic response to an HCV NS3 serine protease-catalyzed NS4A/4B cleavage event. The activity is normally measured in a continuous fluorimetric assay using an excitation wavelength of 355 nm and an emission wavelength of 500 nm. The initial rate can be determined from 10 minutes of continuous reading of increased fluorescence intensities as a result of the NS3 protease-catalyzed cleavage event.

An alternative enzyme assay can be performed as follows:

Materials

Recombinant full-length HCV NS3 enzyme can be prepared as shown in Poliakov et al Protein Expression & purification 25 (2002) 363-371.

The NS4A cofactor advantageously has an amino acid sequence of KKGSVVIVGRIVLSGK (commercially available), generally prepared as a 10 mM stock solution in DMSO.

The FRET substrate (Ac-Asp-Glu-Asp(EDANS)-Glu-Glu-Abu-i(j-[COO)Ala-Ser-Lys(DABCYL)-NH2, MW1548.60 can be purchased from AnaSpec RET Sl, CA . USA) and is normally prepared as a 1.61 mM stock solution in DMSO. Aliquots (50pl/tube) should be wrapped in aluminum foil to protect from direct light and stored at -20°C. Reference compound-1, N-1725 with a sequence of AcAsp-D-Gla-Leu-Ile-Cha-Cys, MW 830.95 can be purchased from BACHEM, Switzerland and is generally prepared as a 2 mM stock solution in DMSO and stored in aliquots at -20 °C.

IM HEPES buffer can be purchased from Invitrogen Corporation, store at 20°C.

Glycerol can be purchased from Sigma, 99% purity.

CHAPS, 3-[(3-cholamidopropyl)dimethylammonio]-l-propanesulfonate: can be purchased from Research Organics, Cleveland, OH44125, USA. MW614.90 DTT, DL-Dithiothreitol (Cleland Reagent: DL-DTT) 99% purity, MW. 154.2 Storage: +4°C.

DMSO can be purchased from SDS, 13124 Peypin, France. 99.5% purity.

TRIS, ultrapure (TRIS-(hydroxymethylaminomethane), can be purchased from ICN Biomedicals Inc.

N-dodecyl-p-D-maltoside, minimum 98%, can be purchased from Sigma, storage -20°C.

Equipment

Microtiter plates ("white cliniplate", ThermoLab Systems catalog number 9502890)

Eppendorf pipettes.

Biohit pipette, multidosing.

Ascent fluorimeter, filter pair ex 355nm, em500 nm.

Method

Experimental Procedure:

10 mM stock solutions of the compounds are prepared in DMSO. The stock solutions are stored at room temperature during testing and placed at -20°C for long-term storage.

Assay Buffer A:

50 mM HEPES buffer, pH=7.5, 40% Glycerol, 0.1% CHAPS

Storage: room temperature

10 mM DTT (stored in aliquots at -20°C and added fresh at each experiment)

Assay Buffer B:

25 mM TRIS pH7.5, 0.15 M NaCl, 10% glycerol, 0.05% n-dodecyl-p-D-maltoside 5mM DTT (stored in aliquots at -20°C and added fresh at each experiment)

Experiment questions:

Preparation of reaction buffer (for one plate, 100 reactionsHbuffer A)

1. Prepare 9500 µl assay buffer (HEPES, pH=7.5, 40% glycerol and 0.1% CHAPS in deionized water. Add DTT, giving a final concentration of 100 mM (freshly prepared for each run).

2. Rapidly thaw the NS3 protease

3. Add 13.6 µl NS3 protease and 13.6 µl NS4A peptide and mix thoroughly. Let the mixture stand for 15 minutes at room temperature. 4. Place the enzyme stock solution back into liquid nitrogen or -80°C as soon as possible.

Preparation of reaction buffer (for one plate, 100 reactions Hbuffer B)

5. Prepare 9500 µl assay buffer (TRIS, pH=7.5, 0.15 M NaCl, 0.5 mM EDTA, 10% glycerol and 0.05% n-dodecyl-β-D-maltoside in deionized water. Add DTT , giving a final concentration of 5mM (freshly prepared for each run).

6. Thaw the NS3 protease rapidly.

7. Add 27.2 µl NS3 protease and 13.6 µl NS4A peptide and mix thoroughly. Let the mixture stand for 15 minutes at room temperature. 8. Place the enzyme stock solution back into liquid nitrogen or -80°C as soon as possible.

Preparation of inhibitor/reference compound

Prepare a dilution series of the inhibitors in DMSO to 100x final concentrations of 10, 1, 0.1, 0.01 and 0.001 nM. The final DMSO concentration in lOOul total reaction volume is 1%.

Prepare a dilution series of the reference compound, N-1725 in DMSO to 100x final concentrations of 120, 60, 30, 15, 7.5 and 3.75 nM.

Eight enzyme control wells are needed for each run.

Black wells contain 95 µl buffer (without NS3 PR), lul DMSO and 5 µl substrate.

Preparation of FRET substrate

Dilute the substrate stock solution (1.61 mM) with assay buffer to 40 µM working solution. Do not expose to light.

Assav- see quens

Use the 96-well "cliniplate", the total analysis volume per well is 100 pl.

1. Add 95 µl assay buffer to each well

2. Add lpl inhibitor/reference compound

3. Pre-incubate for 30 minutes at room temperature

4. Start the reaction by adding 5 µl of 40 pM substrate solution (final concentration 2 pM) 5. Read continuously for 20 minutes at ex=355nm and em=500nm, and monitor the increased fluorescence per minute. 6. Prepare the progression curve (within the linear range, 8-10 time points) and determine the direction coefficient as an initial rate with respect to each individual inhibitor concentration.

7. Calculate % inhibition with respect to the enzyme control.

Processing of the results

The result is expressed as %inhibition at a certain concentration (shielding) or as a Ki value in nM or pM.

Calculation of % inhibition

The initial rate is determined from 10 minutes of continuous reading of increased fluorescence intensities as a result of the NS3 protease-catalyzed cleavage event. The change in the direction coefficient for the inhibitor compared to the enzyme control gives % inhibition at a given concentration.

Calculation of Ki

All inhibitors are treated as if they follow the rules for competitive inhibition.

The IC50 value is calculated from the inhibition values for a series of inhibitor concentrations. The calculated value is used in the following equation:

The production of the curve is carried out using two calculation programs: Grafit and Graphpad

Various compounds according to the invention exemplified above exhibited IC50 values in the range lnM to 6.9 micromolar and ED50 values in the sub-micromolar to micromolar range.

Pattern and rate of drug escape resistance

Replicon cultures in microtiter plates can be used to determine rates of resistance development and to select for drug escape mutants. The compounds being tested are added at concentrations around their ED50 value using approximately 8 duplicates per concentration. After the appropriate replicon incubation period, the protease activity in the supernatant or the lysed cells is measured.

The following procedure is followed for successive passages of the cultures. Virus produced at the concentration of test compound that showed >50% of the protease activity of untreated infected cells (SIC, Starting Inhibitory Concentration) is passed on to healthy replicon cultures. An aliquot of approximately 15 1 of supernatant from each of the eight duplicates is transferred to replicon cells without the test compound (control) and to cells with the test compound at the same concentration and also at respective fivefold higher concentrations. (See table below)

When the viral component of replicon propagation (eg measured by HCV protease activity) is allowed at the highest non-toxic concentration (5 - 40 nM), 2-4 parallel wells are collected and diluted to provide material for sequence analysis and cross-resistance.

Key:

Virus growth allowed

Virus production inhibited

Alternative methods for determining activity against

drug escape mutants involves the preparation of mutant enzyme carrying the characteristic mutation for use in standardized Ki determinations as shown above. For example, WO 04/039970 describes constructs that allow access to HCV proteases carrying the 155, 156 and/or 168 drug efflux mutants arising from the selective pressure of BILN-2061 and VX-950. Such constructs can then be made in replicon vectors instead of the wild-type protease and thereby enable easy determination in a cell assay of whether a given compound is active against a given cell escape mutant.

P450 metabolism

The metabolism of compounds according to the invention via the major isoforms of the human cytochrome P450 system is conveniently determined in baculovirus-infected insect cells transfected with human cytochrome P450 cDNA (supersome) Gentest Corp. Woburn USA.

The test compounds at concentrations of 0.5, 5 and 50 nM are incubated in duplicate in the presence of supersomes overexpressing different cytochrome P450 isoforms, including CYP1A2 + P450 reductase, CYP2A6 + P450 reductase, CYP2C9-Arg 144 + P450 reductase, CYP2C19 + P450 reductase, CYP2D6-Val 374 + P450 reductase and CYP3A4 + P 450 reductase. The incubations contain a fixed concentration of cytochrome P450 (e.g. 50 pmol) and are carried out for 1 hour. The involvement of a given isoform in the metabolism of the test compound is determined by chromatographic UV HPLC measurement of the disappearance of the parent compound.

Claims (64)

1. Compound of formula I: in which A is C(=0)OR<1>or C(=0)NHS02R<2>wherein; R<1> is hydrogen, C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl; R<2> is C 1 -C 6 alkyl, C 0 -C 3 alkyl carbocyclyl, C 0 -C 3 alkylheterocyclyl; in which R<2> is optionally substituted with 1 to 3 substituents independently selected from the group consisting of halo, oxo, nitrile, azido, nitro, C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl, NH2CO-, Y-NRaRb, Y- 0-Rb, Y-C(=0)Rb, Y-(C=0)NRaRb, Y-NRaC(=0)Rb, Y-NHSOpRb, Y-S(=0)pRbog Y-S(=0)pNRaRb, Y-C(=0 )ORb, Y-NRaC(=0)ORb; Y is independently a bond or C 1 -C 3 alkylene; R a is independently H or C 1 -C 3 alkyl; R b is independently H, C 1 -C 6 alkyl, C 0 -C 3 alkylcarbocyclyl or C 0 -C 3 alkylheterocyclyl; p is independently 1 or 2; M is CR7R<7>' or NRu; R<7> is C 1 -C 6 alkyl, C 1 -C 3 alkylC 3 -C 7 cycloalkyl or C 2 -C 6 alkenyl, each of which is optionally substituted with 1-3 halogen atoms or an amino, -SH or C 0 -C 3 alkylcycloalkyl group; or R 7 is J; R7' is H or taken together with R<7> forms a C3-C6 cycloalkyl ring optionally substituted with R<7a> wherein; R<7a> is C 1 -C 6 alkyl, C 3 -C 5 cycloalkyl, C 2 -C 6 alkenyl, each of which may be optionally substituted with halo; or R<7a>can be J; q is 0 to 3 and k is 0 to 3; where q+k > 1; W is -CH2-, -O-, -OC(=0)H-, -OC(=0)-, -S-, -NH-, -NRa, -NHS02-, -NHC(=0)NH- or -NHC(=O)-, -NHC(=S)NH- or a bond; R<8>is a ring system containing 1 or 2 saturated, partially saturated or unsaturated rings each having 4-7 ring atoms, and each having 0 to 4 heteroatoms independently selected from S, O and N, wherein the ring system is optionally spaced from W of a C 1 -C 3 alkylene group; or R<8> is C1-C6 alkyl; where any of the R<8> groups may be optionally mono-, di- or tri-substituted with R<9>, wherein R<9> is independently selected from the group consisting of halo, oxo, nitrile, azido, nitro, C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl, NH2C(=0)-, Y-N Ra'Rb, Y-O-Rb, Y-C (=0)Rb, Y-(C=0)NRa'Rb, Y-NRaC(=0)Rb, Y-NHSOpRb, Y-S(=0)pRb, Y-S(=0)pNRaRb, Y-C(=0)ORb , Y-NRaC(=0)ORb; wherein Ra' is Ra with the proviso that when W is -S- or -O-, R<8> is C0-C3alkylaryl or C0-C3alkylheteroaryl, Y is a bond and Rb is H or C1-C6alkyl, then Ra' R a or C 1 -C 6 alkyl; and where said carbocyclyl or heterocyclyl is optionally substituted with R<10>; where R<10> is C 1 -C 6 alkyl, C 3 -C 7 cycloalkyl, C 1 -C 6 alkoxy, amino, amido, sulfonyl, (C 1 -C 1 alkyl)sulfonyl, NO 2 , OH, SH, halo, haloalkyl, carboxyl; E is -C(=0)-; X is -NR x - where R x is H, C 1 -C 5 alkyl or J; or in the case where E is -C(=0), X may also be -O- or -NRjNRj-; wherein one of R 1 is H, and the other is H, C 1 -C 8 alkyl or J; R<11> is H, C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl, each of which may be substituted with halo, oxo, nitrile, azido, nitro, C1-Qalkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl, NH2C(= 0)-, Y-NRaRb, Y-O-Rb, Y-C(=0)Rb, Y-(C=0)NRaRb, Y-NRaC(=0)Rb, Y-NHSOpRb, Y-S(=0)pRb, Y-S( =O)pNRaRb, Y-C(=0)ORb, Y-NRaC(=0)ORb; or R<11>is J; J, if present, is a single 3- to 10-membered saturated or partially unsaturated alkylene chain extending from the R7/R7" cycloalkyl or from the carbon atom to which R<7> is attached, to one of Rj, Rx, Ry or R<11>to form a macrocycle, which chain is optionally interrupted by one to three heteroatoms independently selected from: -O-, -S- or -NR<12->, and wherein 0 to 3 carbon atoms in the chain are optionally substituted with R<14>; wherein; R<12> is H, C1-C6 alkyl, C3-C6 cycloalkyl or C(=O)R<13>; R<13> is C1-C6 alkyl, C0-C3 alkylcarbocyclyl, C0-C3 alkylheterocyclyl; R<14> is independently selected from the group consisting of H, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkoxy, hydroxy, halo, amino, oxo, thio and C 1 -C 6 thioalkyl; Ru is independently H or C 1 -C 3 alkyl; m is 0 or 1; n is 0 or 1; U is =0 or is absent; R<15> is H, C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl, each of which may be substituted with halo, oxo, nitrile, azido, nitro, C1-C6alkyl, C0-C3alkylheterocyclyl, C0-C3a I alkylcarbocyclyl , NH2CO-, Y-NRaRb, Y-O-Rb, Y-C(=0)Rb, Y-(C=0)NRaRb, Y-NRaC(=0)Rb, Y-NHSOpRb, Y-S(=0)pRb, Y-S( =O)pNRaRb, Y-C(=0)ORb, Y-NRaC(=0)ORb; G is -O-, -NRy-, -NRjNRj-: where one Rj is H, and the other Rj is H, C 1 -C 5 alkyl or J; R y is H, C 1 -C 3 alkyl; Ry is J; R<16> is H; or C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl, each of which may be substituted with halo, oxo, nitrile, azido, nitro, C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylheterocyclyl, NH2CO-, Y-NRaRb, Y-O -Rb, Y-C(=0)Rb, Y-(C=0)NRaRb, Y-NRaC(=0)Rb, Y-NHSOpRb, Y-S(=0)pRb, Y-S(=0)pNRaRb, Y-C(=0 )ORb, Y-NRaC(=0)ORb; with the proviso that when m=n=0, and G is O, then R<16> is not tert-butyl or phenyl; wherein any C atom in C 1 -C 6 alkyl and C 1 -C 3 alkyl may optionally, unless otherwise indicated, be substituted with one two or, where the valence permits, three halogens; each aryl and cycloalkyl unit in C0-C3alkylaryl and C0-C3alkylC3C7cycloalkyl is, unless otherwise indicated, substituted with 1-3 substituents selected from halogen, hydroxyl, nitro, cyano, carboxyl, C1-C6alkyl, C1-C6 alkoxy, C 1 -C 6 alkoxyC 1 -C 6 alkyl, C 1 -C 6 alkanoyl, amino, azido, mercapto and C 0 -C 3 alkylheterocyclyl; each carbocyclyl and heterocyclyl unit in C0-C3alkylcarbocyclyl and C0-C3alkylheterocyclyl is, unless otherwise indicated, optionally substituted with 1-3 substituents selected from halogen, hydroxyl, nitro, cyano, carboxyl, C1-C6alkyl, C! -C 6 alkoxy, C 1 -C 6 alkoxyC 1 -C 6 alkyl, C 1 -C 6 alkanoyl, amino, azido, oxo, mercapto, C 0 -C 3 alkylcarbocyclyl and C 0 -C 3 alkylheterocyclyl; each amino group is selected from NH 2 , NHC 1 -C 6 alkyl and N(C 1 -C 6 alkyl) 2 ; and each amido group is selected from C(=O)NH 2 , C(=O)NHC 1 -C 6 alkyl, C(=O)N(C 1 -C 6 alkyl) 2 and -NH(C=O)C 1 -C 6 alkyl; or a pharmaceutically acceptable salt thereof. 2. Compound according to claim 1, in which R7' is H and R7 is n-ethyl, cyclopropylmethyl, cyclopropyl, cyclobutylmethyl, cyclobutyl or mercaptomethyl, preferably n-propyl or 2,2-difluoroethyl. 3. Compound according to claim 1, wherein R7 and R<7>' together form a spiro-cyclopropyl or spiro-cyclobutyl ring, both optionally mono- or di-substituted with R<7>,<a>wherein; R<7a> is C 1 -C 6 alkyl, C 3 -C 5 cycloalkyl or C 2 -C 6 alkenyl, each of which is optionally substituted with halogen; or R<7a>is J. 4. Compound according to claim 3, wherein the ring is a spiro-cyclopropyl ring substituted with R<7>,<a>wherein; R<7a> is ethyl, vinyl, cyclopropyl, 1- or 2-bromomethyl, 1- or 2-fluoroethyl, 2-bromovinyl or 2-fluoroethyl. 5. Compound according to claim 1, wherein R<7> is J, and R<7>' is H. 6. Compound according to any one of claims 1-5, with the partial structure Ia: 7. A compound according to claim 1, wherein m is 0 and n is 0. 8. Compound according to claim 7, wherein G is -NRy- or -NRjNRj-. 9. Compound according to claim 8, where Ry or one of the Rj groups is J, and thereby constitutes a macrocyclic compound. 10. A compound according to claim 9, wherein R<16> is H, C3-C6 alkyl or C3-C6 cycloalkyl. 11. Compound according to claim 1, wherein m is 1. 12. A compound according to claim 11, wherein X is -NRx-. 13. A compound according to claim 11, wherein U is O. 14. A compound according to claim 11, wherein R<11> is C1-C6alkyl, C0-C3alkylcarbocyclyl, C0-C3alkylaryl, or C0-C3alkylheteroaryl, each of which is optionally substituted with halo, amino, C1-C6alkoxy, C1-C6thioalkyl, carboxyl, ( C 1 -C 6 alkoxy)carbonyl, aryl, heteroaryl or heterocyclyl, and especially wherein the substituent is hydroxy or C(=O)OR<14>. 15. A compound according to claim 14, wherein R<11> is phenylethyl, 2,2-dimethyl-propyl, cyclohexylmethyl, phenylmethyl, 2-pyridylmethyl, 4-hydroxy-phenylmethyl or carboxylpropyl; or especially tert-butyl, iso-butyl or cyclohexyl. 16. Compound according to claim 11, in which one of Rx or R<11> is J, thereby forming a macrocyclic compound. 17. Compound according to claim 11, wherein n is 1. 18. A compound according to claim 17, wherein R<15> is C 1 -C 6 alkyl or C 0 -C 3 alkylcarbocyclyl, each of which is optionally substituted. 19. Compound according to claim 18, wherein R<15> is cyclohexyl, cyclohexylmethyl, tert-butyl, iso-propyl or iso-butyl. 20. A compound according to claim 11, wherein G is NRy or -NRjNRj-, where Ry or one Rj is H or methyl, and the other Rj is H. 21. Compound according to claim 20, wherein R<16> is H, C1-C6 alkyl or a 5- or 6-membered heterocycle, especially morpholine, piperidine or piperazine. 22. A compound according to claim 11, wherein R<16> is C 1 -C 6 alkyl, C 0 -C 3 alkylheterocyclyl, C 0 -C 3 a 1 alkyl carbocyclyl, each of which is optionally substituted with hydroxy, halo, amino or C 1 -C 6 alkoxy. 23. Compound according to claim 22, wherein R<16> is 2-indanol, indanyl, 2-hydroxy-1-phenyl-ethyl, 2-thiophenemethyl, cyclohexylmethyl, 2,3-methylenedioxybenzyl, cyclohexyl, benzyl, 2-pyridylmethyl, cyclobutyl , iso-butyl, n-propyl or 4-methoxyphenylethyl. 24. A compound according to claim 1, wherein W is -OC(=O)-, -NRa-, -NHS(O)2-or -NHC(=O)-; or especially -OC(=0)NH- or -NH. 25. Compound according to claim 1, wherein W is -S-, a bond or especially -0-. 26. A compound according to claim 24 or 25, wherein R<8> is optionally substituted C0-C3a I alkyl carbocyclyl or optionally substituted C0-C3 alkylheterocyclyl. 27. Compound according to claim 26, in which the C0-C3 alkyl group is methylene or preferably a bond. 28. A compound according to claim 27, wherein R<8> is C0-C3 alkylaryl or C0-C3 alkylheteroaryl, each of which is optionally mono-, di- or tri-substituted with R<9>, wherein; R<9> is C1-C6alkyl, C1-C6alkyl, NO2, OH, halo, trifluoromethyl, amino or amido optionally mono- or di-substituted with C1-C6alkyl, C0-C3alkylaryl, C0-C3-alkylheteroaryl, carboxyl, aryl or heteroaryl optionally substituted with R<10>; in which R<10> is C 1 -C 6 alkyl, C 3 -C 7 cycloalkyl, C 1 -C 6 alkoxy, amino optionally mono- or di-substituted with C 1 -C 6 alkyl, amido, sulfonyl C 1 -C 3 alkyl, NO 2 , OH, halo, trifluoromethyl, carboxyl or heteroaryl. 29. A compound according to claim 28, wherein R<9> is C1-C6alkyl, C1-C6alkyl, amino, di-(C1-C3alkyl)amino, C1-C6alkylamide, aryl or heteroaryl, and the aryl and heteroaryl are optionally substituted with R< 10>; in which R<10> is C 1 -C 6 alkyl, C 3 -C 7 cycloalkyl, C 1 -C 6 alkoxy, amino, mono- or di-C 1 -C 3 alkylamino, amido, halo, trifluoromethyl or heteroaryl. 30. Compound according to claim 29, wherein R<10> is C1-C6 alkyl, C1-C6 alkoxy, amino optionally mono- or di-substituted with C1-C3 alkyl, amido, C1-C3 alkylamide, halo or heteroaryl. 31. Compound according to claim 30, wherein R<10> is methyl, ethyl, isopropyl, tert-butyl, methoxy, chlorine, amino optionally mono- or di-substituted with C1-C3 alkyl, amido, or C1-C3 alkyl thiazolyl. 32. A compound according to claim 31, wherein R<8> is 1-naphthylmethyl, 2-naphthylmethyl, benzyl, 1-naphthyl, 2-naphthyl or quinolinyl each of which is unsubstituted, mono or disubstituted with R<9> as defined. 33. A compound according to claim 32, wherein R<8> is 1-naphthylmethyl or quinolinyl each of which is unsubstituted, mono or disubstituted with R<9> as defined. 34. Compound according to claim 33, wherein R<8> is: wherein R<9a> is C1-C6 alkyl; C 1 -C 6 alkoxy; thioC 1 -C 3 alkyl; amino optionally substituted with C 1 -C 6 alkyl; C 0 -C 3 alkylaryl; or C0-C3alkylheteroaryl, C0-C3alkylheterocyclyl, wherein said aryl, heteroaryl or heterocycle is optionally substituted with R<10>wherein R<10> is C 1 -C 6 alkyl, C 0 -C 3 alkylC 3 -C 7 cycloalkyl, C 1 -C 6 alkoxy, amino optionally mono- or di-substituted with C 1 -C 6 alkyl, amido, C 1 -C 3 alkylamide; and R<9b>is C 1 -C 6 alkyl, C 1 -C 6 alkoxy, amino, di(C 1 -C 3 alkyl)amino, (C 1 -C 3 alkyl)amide, NO 2 , OH, halo, trifluoromethyl, carboxyl. 35. A compound according to claim 34, wherein R<9a> is aryl or heteroaryl, each of which is optionally substituted with R<10> as defined. 36. A compound according to 35, wherein R<9a>is selected from the group consisting of: wherein R<10> is H, C1-C6alkyl or C0-C3alkylcycloalkyl, amino optionally mono- or di-substituted with C1-C6alkyl, amido, (C1-C3alkyl)amide. 37. A compound according to claim 35, wherein R<9a> is optionally substituted phenyl, preferably phenyl substituted with C 1 -C 6 alkyl; C 1 -C 6 alkoxy; or hello. 38. Compound according to claim 34, wherein R<8> is: wherein R<10a> is H, C1-C6alkyl or C0-C3alkylcarbocyclyl, amino optionally mono- or di-substituted with C1-C6alkyl, amido, heteroaryl or heterocyclyl; and R<9b>is C 1 -C 6 alkyl, C 1 -C 6 alkoxy, amino, di(C 1 -C 3 alkyl)amino, amido, NO 2 , OH, halo, trifluoromethyl or carboxyl. 39. A compound according to claim 34, wherein R<9b>is C1-C6 alkoxy, preferably methoxy. 40. A compound according to claim 1, wherein A is C(=O)NHS02R<2>. 41. A compound according to claim 42, wherein R 2 is optionally substituted C 1 -C 6 alkyl, preferably methyl. 42. Compound according to claim 40, where R 2 is optionally substituted C 3 -C 7 cycloalkyl, preferably cyclopropyl. 43. A compound according to claim 40, wherein R<2> is optionally substituted C0-C3 alkylaryl, preferably substituted phenyl. 44. A compound according to claim 1, wherein A is C(=0)OR<1>. 45. Compound according to claim 44, wherein R<1> is H or C1-C6 alkyl, preferably hydrogen, methyl, ethyl or tert-butyl. 46. A compound according to claim 1, wherein J is a 3- to 8-membered saturated or unsaturated alkylene chain optionally containing one to two heteroatoms independently selected from: -O-, -S- or -NR<12->, wherein R<12 >is H, C 1 -C 6 alkyl, such as methyl or -C(=O)C 1 -C 6 alkyl, such as acetyl. 47. A compound according to claim 46, wherein J is a 4- to 7-membered saturated or unsaturated, fully carboalkylene chain. 48. A compound according to claim 46, wherein J is saturated or mono-unsaturated. 49. A compound according to claim 46, wherein J is sized to provide a macrocycle with 14 or 15 ring atoms. 50. Compound according to claim 1 with formula lhe 51. Compound according to claim 50, where J is a simple 6- or 7-membered atomically saturated or partially unsaturated alkylene chain. 52. A compound according to any one of claims 50-51, wherein J has an unsaturation. 53. A compound according to any one of claims 50-52, wherein W is O and R<8> is aryl or heteroaryl, each of which is optionally mono-, di- or tri-substituted by R<9>, wherein R<9> is C1-C6alkyl, C1-C6alkyl, NO2, OH, halogen, trifluoromethyl, amino, amido, C0-C3alkylaryl, C0-C3alkylheteroaryl or carboxyl, where the aryl or heteroaryl unit is optionally substituted with R<10> ; in which R<10> is C 1 -C 6 alkyl, C 3 -C 7 cycloalkyl, C 1 -C 6 alkoxy, amino, amido, sulfonyl C 1 -C 6 alkyl, NO 2 , OH, halogen, trifluoromethyl, carboxyl or heteroaryl. 54. A compound according to claim 53, wherein R<9> is selected from C1-C6 alkyl, C1-C6 alkoxy, amino, amido, aryl or heteroaryl, where aryl or heteroaryl is optionally substituted with R<10>; in which R<10> is C 1 -C 6 alkyl, C 3 -C 7 cycloalkyl, C 1 -C 6 alkoxy, amino, amido, halogen, trifluoromethyl or heteroaryl. 55. A compound according to any one of claims 50-54, wherein R<8> is where R<9a> is C0-C3alkylaryl, C0-C3alkylheteroaryl or C0-C3alkylheterocyclyl, where said aryl, heteroraryl or heterocyclyl is optionally substituted with R<10>, where R<10> is C 1 -C 6 alkyl, C 3 -C 7 cycloalkyl, C 1 -C 6 alkoxy, amino, amido, heteroaryl or heterocyclyl; and R<9b> is C 1 -C 6 alkyl, C 1 -C 6 alkoxy, amino, amido, NO 2 , OH, halogen, trifluoromethyl or carboxyl. 56. A compound according to claim 55, wherein R<9a> is selected from the group consisting of where R<10> is H, C1-C6alkyl, C3-C6cycloalkyl, amino which is optionally mono- or di-substituted with C1-C3alkyl, amido, heteroaryl or heterocyclyl. 57. A compound according to any one of claims 51-56, wherein A is C(=O)NHS02R<2>. 58. Compound according to claim 57, where R 2 is methyl, cyclopropyl or optionally substituted phenyl. 59. A pharmaceutical composition comprising a compound as defined in any of claims 1-58 and a pharmaceutically acceptable carrier therefor. 60. The pharmaceutical composition of claim 59, further comprising an additional HCV antiviral agent selected from nucleoside analog polymerase inhibitors, protease inhibitors, ribavirin and interferon. 61. A compound according to any one of claims 1-58 for use in therapy. 62. Use of a compound according to any one of claims 1-58 for the preparation of a medicament for the prophylaxis or treatment of flavivirus infections including HCV. 63. A compound according to any one of claims 1-58 for use in the prophylaxis or treatment of flavivirus infections including HCV. 64. Compound according to claim 63 for use in the treatment of HCV infection.