HK1100881B - Hcv ns-3 serine protease inhibitors - Google Patents

Description

Hcv NS-3 serine protease inhibitors

Technical Field

The present invention relates to novel inhibitors of the flavivirus HCV NS3 serine protease and methods for their use in the treatment or prevention of HCV.

Background

The HCV NS3 serine protease is a multifunctional protein comprising a serine protease domain and an RNA helicase domain. The protease cofactor NS4A, a relatively small protein, is a protein essential for enhancing serine protease activity. The NS3 serine protease is essential in the viral life cycle. Matrix binding site analysis revealed from the X-ray crystal structure revealed that the binding site for the NS3 protease was significantly shallow and solvent exposed, making small molecule inhibitor design difficult.

Two HCV protease inhibitors are believed to have entered clinical trials, Boehringer Ingelheim's BILN-2061 disclosed in WO0059929 and Vertex' VX-950 disclosed in WO 0387092. A number of similar peptidomimetic HCV protease inhibitors have also been proposed in the academic and patent literature. Most of the above prior art peptidomimetics are usually present as L-proline derivatives at position P2 of the inhibitor and interact with the S2 subsite of the HCV protease. In the case of BILN-2061, the L-proline is 4-substituted with quinolineether, however there is one carbocyclic ring fused to the L-proline ring in VX-950. Most peptidomimetics also include other L-amino acid derivative peptides linked at position P3, and many of the above-mentioned proposed inhibitors also include L-amino acid derivatives that are additionally extended to P4, P5 and P6.

It has become apparent that continuous administration of BILN-2061 or VX-950 selects for HCV mutants which are resistant to the corresponding drug, so-called drug escape mutants. These drug escape mutants have characteristic mutations in the HCV protease genome, particularly D168V, D168Y and/or a 165S. Thus, the therapeutic paradigm for HCV has to resemble HIV therapy, where drug escape mutations are also likely to occur. Accordingly, there is a continuing need for other drugs with different resistance properties in order to provide a treatment regimen for non-effective patients, and combination therapy with multiple drugs is likely to be a normal form in the future, even for the first treatment.

The practice of using HIV drugs, and HIV protease inhibitors in particular, is further emphasized that sub-optimal pharmacokinetics and complex dosage regimes will soon result in compliance being unintentionally compromised. This in turn means that in HIV conditions, the 24 hour trough concentration (minimum plasma concentration) of the corresponding drug tends to decrease in IC for most of the day₉₀Or ED₉₀Below the limit. It is generally accepted that at least IC₅₀And more realistic, IC₉₀Or ED₉₀The 24 hour trough concentration is essential to delay the production of drug escape mutants and to obtain the necessary pharmacokinetics and drug metabolism, which makes the trough concentration a powerful challenge for drug design. The strong peptidomimetic nature of prior art HCV protease inhibitors, as well as the multiple peptide bonds of their own structure, form a pharmacokinetic barrier to effective dosage forms.

Brief description of the invention

According to a first aspect of the present invention there is provided a compound of formula VI, or a pharmaceutically acceptable salt or prodrug thereof.

Wherein:

a is C (═ O) OR¹、C(＝O)NHSO₂R²、C(＝O)NHR³Or CR⁴R⁴', wherein:

R¹is hydrogen, C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃An alkyl heterocyclic group;

R²is C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃An alkyl heterocyclic group;

R³is C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃Alkylheterocyclyl, -OC₁-C₆Alkyl, -OC₀-C₃Alkyl carbocyclyl, -OC₀-C₃An alkyl heterocyclic group;

R⁴is halogen, amino or OH; or R⁴And R⁴Both are ═ O;

R⁴' is C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃An alkyl heterocyclic group;

wherein R is²、R³And R⁴' each optionally substituted with 1 to 3 substituents independently selected from the group consisting of: halogen, oxo, nitrile, azido, nitro, C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃Alkyl heterocyclic group, NH₂C(＝O)-、Y-NRaRb、Y-O-Rb、Y-C(＝O)Rb、Y-(C＝O)NRaRb、Y-NRaC(＝O)Rb、Y-NHSO_pRb、Y-S(＝O)_pRb、Y-S(＝O)_pNRaRb, Y-C (═ O) ORb, and Y-NRaC (═ O) ORb;

y is independently a bond or C₁-C₃An alkylene group;

ra is independently H or C₁-C₃An alkyl group;

rb is independently H, C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl or C₀-C₃An alkyl heterocyclic group;

p is independently 1 or 2;

m is CR⁷R⁷Either of NRu;

ru is H or C₁～C₃An alkyl group;

R⁷is C₁-C₆Alkyl radical, C₀-C₃Alkyl radical C₃-C₇Cycloalkyl or C₂-C₆Alkenyl, each of which is optionally substituted by 1 to 3 halogen atoms or by amino, -SH or C₀-C₃Alkyl cycloalkyl substituted; or R⁷Is J;

R⁷' is H or with R⁷Together form optionally substituted R^7＇aSubstituted C₃-C₆A cycloalkyl ring, wherein;

R⁷' a is C₁-C₆Alkyl radical, C₃-C₅Cycloalkyl radical, C₂-C₆Alkenyl, each of which may be optionally substituted with halogen; or R^7＇aCan be J;

q' is 0 or 1 and k is 0-3;

rz is H, or forms an olefinic bond with the asterisked carbon;

rq is H or C₁-C₆An alkyl group;

w is-CH₂-、-O-、-OC(＝O)NH-、-OC(＝O)-、-S-、-NH-、-NRa、-NHSO₂-, -NHC (═ O) NH — or-NHC (═ O) -, -NHC (═ S) NH — or a bond;

R⁸is a ring system comprising 1 or 2 saturated, partially saturated or unsaturated rings each having 4 to 7 ring atoms and each having 0 to 4 heteroatoms selected from S, O and N, optionally via C₁-C₃Alkyl and W are interrupted; or R⁸Is C₁-C₆An alkyl group; any of the above R⁸All of which may be optionally substituted by R⁹Mono-, di-, or tri-substituted, wherein:

R⁹independently selected from: halogen, oxo, nitrile, azido, nitro, C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃Alkyl heterocyclic group, NH₂CO-、Y-NRaRb、Y-O-Rb、Y-C(＝O)Rb、Y-(C＝O)NRaRb、Y-NRaC(＝O)Rb、Y- NHSO_pRb、Y-S(＝O)_pRb、Y-S(＝O)_pNRaRb, Y-C (═ O) ORb, and Y-NRaC (═ O) ORb; wherein said carbocyclyl or heterocyclyl moiety is optionally substituted with R¹⁰Substituted; wherein

R¹⁰Is C₁-C₆Alkyl radical, C₃-C₇Cycloalkyl radical, C₁-C₆Alkoxy, amino, sulfonyl, (C)₁-C₃Alkyl) sulfonyl, NO₂OH, SH, halogen, haloalkyl, carboxyl, amido;

rx is H or C₁-C₅An alkyl group; or Rx is J;

t is-CHR¹¹-or-NRd-, wherein Rd is H, C₁-C₃An alkyl group; or Rd is J;

R¹¹is H, C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃An alkylheterocyclyl group, each of which may be substituted with: halogen, oxo, nitrile, azido, nitro, C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃Alkyl heterocyclic group, NH₂CO-、Y-NRaRb、Y-O-Rb、Y-C(＝O)Rb、Y-(C＝O)NRaRb、Y-NRaC(＝O)Rb、Y-NHSO_pRb、Y-S(＝O)_pRb、Y-S(＝O)_pNRaRb, Y-C (═ O) ORb, and Y-NRaC (═ O) ORb; or R¹¹Is J;

j, if present, is a single 3-to 10-membered saturated or partially unsaturated alkylene chain derived from R⁷/R⁷"" cycloalkyl "or" from R⁷The carbon atom to which it is attached extends to Rd, Rj, Rx, Ry or R¹¹One thereby forming a macrocycle, said chain being optionally substituted with one to three groups independently selected from-O-, -S-or-NR¹²-is interrupted by a heteroatom, and wherein 0 to 3 carbon atoms in the chain are optionally interrupted by R¹⁴Substitution;

wherein;

R¹²is H, C₁-C₆Alkyl radical, C₃-C₆Cycloalkyl or-C (═ O) R¹³；

R¹³Is C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃An alkyl heterocyclic group;

R¹⁴independently selected from: H. c₁-C₆Alkyl radical, C₁-C₆Haloalkyl, C₁-C₆Alkoxy, hydroxy, halogen, amino, oxo, thio and C₁-C₆A thioalkyl group;

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

u is ═ O or absent;

R¹⁵is H, C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃An alkylheterocyclyl group, each of which may be substituted with: halogen, oxo, nitrile, azido, nitro, C₁ -C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃Alkyl heterocyclic group, NH₂CO-、Y-NRaRb、Y-O-Rb、Y-C(＝O)Rb、Y-(C＝O)NRaRb、Y-NRaC(＝O)Rb、Y-NHS(＝O)_pRb、Y-S(＝O)_pRb、Y-S(＝O)_pNRaRb、 Y-C(＝O)ORb、Y-NRaC(＝O)ORb；

G is-O-, -NRy-, -NRjNRj-;

ry is H, C₁-C₃An alkyl group; or Ry is J;

one Rj is H and the other Rj is H or J;

R¹⁶is H or R¹⁶Is C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃An alkylheterocyclyl group, each of which may be substituted with: halogen, oxo, nitrile, azido, nitro, C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃Alkyl heterocyclic group, NH₂CO-、Y-NRaRb、Y-O-Rb、Y-C(＝O)Rb、Y-(C＝O)NRaRb、Y-NRaC(＝O)Rb、Y-NHSO_pRb、Y-S(＝O)_pRb、Y-S(＝O)_pNRaRb、Y-C(＝O)ORb、Y-NRaC(＝O)ORb。

Without wishing to be bound in any way by a pattern of temporary constraints of theory or particular variables, the ideographic concepts P1, P2, P3 and P4 applied herein are provided merely for convenience and basically have the same meaning as Schechter&Berger, (1976) Biochem Biophys ResComm 27157-162, which refers to those portions of inhibitors identified as filling the S1, S2, S3 and S4 subsites, respectively, with S1 proximal and S4 distal to the cleavage site. Regardless of the manner of attachment, the components defined by formula VI, etc. are intended to be included within the scope of the present invention. For example, it is contemplated that the end-capping group R is especially when m and/or n is 0¹⁶G can interact with S3 and S4 subsites.

Various embodiments of the present invention may be symbolically represented as R¹⁶-G-P4-P3-P2-P1, wherein P3 and/or P4 may be absent. Wherein P1, P3 and P4 each represent a structural unit constituting a natural or unnatural amino acid derivative, P2 is a substituted carbocyclic residue and G-R¹⁶Is a capping group. The structural units are typically linked together by amide bonds. In the compounds of the present invention, the amide linkages are inverted with respect to each other on each side of the P2 building block.

Another aspect of the invention includes a pharmaceutical composition comprising a compound of the invention as defined above together with a pharmaceutically acceptable carrier or diluent therefor.

The compounds and compositions of the present invention may be used in methods of medical treatment or prevention of HCV infection in humans. Accordingly, another aspect of the invention is the use of a compound of the invention as defined above in therapy, such as in the manufacture of a medicament for the prevention or treatment of infection by a flavivirus in a human or animal. Exemplary flaviviruses include BVDV, dengue and in particular HCV.

Amides of P2 and P3 linked together in the compounds of the inventionThe bonds are reversed relative to the amide bond linking P1 and P2, i.e. the amino acid derivatives P1 and P3 on each side of the P2 scaffold are both coupled through their amino functional groups to acid groups on each side of the P2 scaffold. This means that the P3 and P4 side chains (including R)¹⁶Capped to the extent that it interacts with S3 or S4) points in the opposite direction to the propeptide substrate. Another consequence of the inverted orientation of the amino acids P3 and P4 is that the side chains of these amino acids transfer one atom out relative to the propeptide substrate.

It is expected that changes in orientation of the P3 and P4 side chains in this manner will contribute to the formation of P3 and/or P4 and/or R¹⁶Non-natural D-type stereochemistry of pocket filling groups (e.g., side chains). Indeed, the above compounds are generally highly active compounds and are within the scope of the present invention. However, it has now surprisingly been found that even though the compounds according to the invention with L-amino acid side chains at positions P3 and/or P4 show good activity, the corresponding side chain units have to approach the S3 or S4 pocket from different angles with respect to the original peptidic matrix. Accordingly, R¹¹And/or R¹⁵At L-stereochemistry and/or R corresponding to pseudo-L-stereochemistry¹⁶The configurations all represent preferred aspects of the invention.

The different approach angles to the S3 and/or S4 pockets also imply the ability of the compounds of the present invention to avoid the resistance exhibited by prior art HCV protease inhibitors, which to date have a conventional peptide backbone of natural or non-natural L-amino acid residues. RNA-associated HCV RNA polymerase NS5A has very poor read-ability, as well as the well-known HIV reverse transcriptase which rapidly generates drug escape mutants under antiviral therapy selection pressure. This also means that HCV polymerase is very error prone and may develop characteristic resistance when HCV antiviral agents are administered for long periods of time. Even before the input, it is clear that BILN 2061 with a substantially peptide backbone (even if macrocyclization is performed) and the Vertex' NS3 protease inhibitor VX-950 with a linear peptide backbone at P3 and P4 rapidly develop characteristic resistance mutations at positions 155, 156 or 168 of the NS3 protease (Lin et al, J Biol Chem 2004279 (17): 17808-17).

A preferred group of compounds of the invention include those wherein P1 represents a hydrazine derivative, i.e. M is NRu wherein Ru is typically H or C₁-C₃An alkyl group. Wherein M is CR⁷R⁷The compounds of' form a further preferred aspect of the invention.

In formula VI M is CR⁷R⁷Preferred embodiments of' include the following formula VIA:

preferably, the values of q' and k in formula VI include 1: 1, 1: 2, 1: 3, 2: 2, 2: 3, more preferably 0: 2 and 0: 0; and most preferably 0: 1, in which case the preferred compound has one of the following partial structures:

in particular wherein Rz is H or Rq is H or methyl.

The compounds of the invention may contain two functional groups, P3 and P4, i.e. m and n are each 1. Preferred embodiments within formula VI that include P3 and P4 functional groups include the following formulas VIda-VIdb:

additional embodiments include structures corresponding to VIda and VIdb, wherein M is NRu.

Another structure of the compounds of the present invention comprises the P3 functional group, but does not include the P4 functional group, i.e., m is 1 and n is 0. Preferred embodiments within formula VI that include P3, but do not include the P4 functional group include the following formulas VIea-VIeb:

additional embodiments include structures corresponding to VIea and VIeb, where M is NRu.

Other structures of the compounds of the invention include those in which m and n are both 0 and thus the group R¹⁶-G is adjacent to the configuration of P2, but as mentioned above, the end-capping group R¹⁶G may advantageously interact with S3 and/or S4.

Preferred embodiments within formula VI wherein m and n are both 0 include the following formula VIfa:

additional embodiments include structures corresponding to VIfa, wherein M is NRu.

The compounds of the invention may include linear molecules, as described above. In addition, in which R⁷And R⁷In embodiments where' together is defined as spirocycloalkyl (such as spirocyclopropyl), compounds of the invention may be configured as macrocycles, where the linking group J is Rj, Rx, Ry, Rd or R in formula VI¹¹One group extending between them. In addition, the macrocyclic ring J can be substituted with R⁷The contiguous carbon extends to one of Rj, Rx, Ry, Rd, or Ru.

Within compounds of formula VI wherein m is 0 and n is 1, preferred embodiments of the above macrocyclic structures include the following formulas VIga-VIgc:

also preferred is a group wherein the J chain is linked to R⁷The corresponding structure of adjacent carbon atoms.

In compounds of formula VI containing both P3 and P4 functional groups, i.e., in which m and n are both 1, preferred embodiments of the above macrocyclic structures include the following formulas VIha-VIhc:

also preferred is a group wherein the J chain is linked to R⁷The corresponding structure of adjacent carbon atoms.

In compounds of formula VI in which neither the P3 nor P4 functional groups are present, i.e., in which m and n are each 0, preferred macrocyclic structures include the following formulas VIhe-VIhf:

also preferred is a group wherein the J chain is linked to R⁷The corresponding structure of adjacent carbon atoms.

Generally, in optional macrocyclic structures, such as those illustrated above, the linker J is a saturated alkylene or partially unsaturated alkylene chain having from 3 to 10 chain atoms, preferably from 4 to 7 chain atoms, such as 5 or 6 chain atoms, i.e., an alkylene chain with from 1 to 3 unsaturated bonds between adjacent carbon atoms, typically one unsaturation. The length of the chain will naturally depend on whether J is selected from Rd, Rj, Rx, Ry, R¹¹Or from R⁷Adjacent carbon extensions. Suitable chains are described in detail in WO 00/59929. Typically J is of a size that provides a macrocycle of 13 to 16 ring atoms (including those atoms contained within the ring in the P1, P2, and P3 groups if present). Suitably J isFor macrocycles having 14 or 15 ring atoms.

Desirably, the J chain contains one or two heteroatoms selected from: o, S, NH, NC₁ -C₆Alkyl or N-C (═ O) C₁-C₆An alkyl group. More preferably, the J chain optionally comprises one of the following heteroatoms: NH or N-C (═ O) C₁-C₆Alkyl, most preferably n (ac). Most preferably, the chain comprising nitrogen atoms is a saturated chain. In another embodiment, J comprises one heteroatom selected from O or S. The chain may be substituted with R¹⁴Substitution, such as H or methyl.

Typically the J-link is saturated. In addition, J contains 1-3 double bonds, preferably 1 double bond, and is generally bonded to cycloalkyl R⁷The functional groups are separated by one carbon atom, if present. The double bond may be a cis-or trans-double bond.

Thus, representative examples of J include pentene, hexene, heptene, each of which is substituted with: c₁-C₆Alkyl radical, C₁-C₆Haloalkyl, C₁-C₆Alkoxy, hydroxy, halogen, amino, oxo, thio or C₁-C₆A thioalkyl group; penten-3-yl, hexen-4-yl, hepten-5-yl, wherein 3, 4 or 5 refers to the double bond between the carbon atoms in positions 3 and 4, 4 and 5, and the like.

Suitable R⁷And R⁷A group comprising⁷' is H, and R⁷Those which are n-ethyl, n-propyl, cyclopropyl, cyclopropylmethyl, cyclobutyl, cyclobutylmethyl, 2-difluoroethyl or mercaptomethyl. Preferred embodiments include those wherein R⁷Those which are n-propyl or 2, 2-difluoroethyl.

R⁷And R⁷Another preferred structure of' includes wherein R⁷Is H and R⁷Is C₃-C₇Cycloalkyl or C₁-C₃Alkyl radical C₃-C₇Those structures of cycloalkyl。

For R⁷And R⁷Additionally preferred structures include those wherein R⁷Is H and R⁷Those structures that are J.

In addition, R⁷And R⁷Together are defined as a spiro-cycloalkyl functional group, such as a spiro-cyclobutyl ring, and more preferably a spiro-cyclopropyl ring. In the context of the present invention, "spiro" simply means that the cycloalkyl ring shares one carbon atom with the peptide backbone of the compound. The ring is a substituted or unsubstituted ring. Preferred substituents include R^7＇aMono-or di-substituted, wherein R^7＇aIs C₁-C₆Alkyl radical, C₃-C₅Cycloalkyl or C₂-C₆Alkenyl, each optionally substituted with halogen.

Alternatively, the substituent may be a J-linker as described above. For spiro-cyclopropyl rings, the generally preferred stereochemistry is defined as follows.

Particularly preferred substituents include R^7＇aIs ethyl, vinyl, cyclopropyl (i.e. to R)⁷/R⁷The spiro-cyclopropyl substituent of the "spiro" cycloalkyl ring), 1-or 2-bromoethyl, 1-or 2-fluoroethyl, 2-bromovinyl or 2-fluoroethyl.

In one embodiment of the invention, A is-CR as detailed in PCT/EP03/10595⁴R⁴', the contents of which are incorporated herein by reference.

Thus, R is appropriate⁴' the group includes C₁-C₆Alkyl radicals, such as methyl, ethyl, propyl, vinyl and-CHCHCH₃. Further preferred R⁴' groups include aryl or heteroaryl, such as optionally substituted phenyl, pyridyl, thiazolyl or benzimidazolyl or C₁-C₃Alkylaryl or C₁-C₃Alkylheteroaryl, wherein the alkyl moiety is methyl, ethyl, propyl, vinyl and-CHCHCH₃. Preferred aryl moietiesIncluding optionally substituted phenyl, benzothiazole and benzimidazole.

Preferred R⁴The radicals comprising-NH₂Fluorine or chlorine. Further preferred R⁴The group includes-OH and in particular ═ O.

Another embodiment of a is C (═ O) NHR³Wherein R is³Is optionally substituted C₀-C₃Alkylaryl group, C₀-C₃Alkylheteroaryl, OC₀-C₃Alkylaryl or OC₀-C₃An alkyl heteroaryl group. Suitable substituents appear in the definitions section below.

Another preferred structure for a is C (═ O) OR¹In particular wherein R¹Is C₁-C₆Alkyl groups such as methyl, ethyl or tert-butyl, and most preferably hydrogen.

A particularly preferred structure for a is C (═ O) NHSO₂R²In particular wherein R²Is optionally substituted C₁-C₆Alkyl (preferably methyl), or optionally substituted C₃-C₇Cycloalkyl (preferably cyclopropyl) or optionally substituted C₀-C₆Alkylaryl (preferably optionally substituted phenyl). Suitable substituents appear in the definitions section below.

substituent-W-R on cyclic P2 radical⁸Any proline substituent may be used, which is widely described in the following documents: WO/59929, WO/09543, WO/66103, WO/064455, WO/062228, WO/43339, WO/and WO 04165, and the like.

Preferred W functional groups include, W is-OC (═ O) NH-, -OC (═ O)O)-、-NH-、-NR⁸′-、-NHS(O)₂-or-NHC (═ O) -, in particular-OC (═ O) NH-or-NH-. Preferred R for the above W functional group⁸The radicals including optionally substituted C₀-C₃Alkyl carbocyclyl or C₀-C₃Alkyl-heterocyclyl including those described in WO0009543, WO0009558 and WO 00/174768. For example, an ester substituent on a cyclic P2 group, -W-R⁸Including those substituents disclosed in WO01/74768, such as C₁-C₆Alkanoyloxy group, C₀-C₃Alkylaryloxy, especially (optionally substituted) benzoyloxy or C₀-C₃Alkyl heterocyclic acyloxy, in particular the following groups:

this publication also describes the possibility of further-W-R⁸E.g. C₁-C₆Alkyl (e.g. ethyl, isopropyl), C₀-C₃An alkylcyclo group (such as cyclohexyl), 2-difluoroethyl, -C (═ O) NRc where Rc is C₁-C₆Alkyl radical, C₀-C₃Alkyl cyclopropyl, C₀ -C₃Alkylaryl or C₀-C₃An alkyl heterocyclic group.

Presently preferred W functional groups include-S-and in particular-O-. In said embodiment, R⁸Suitable meanings of (A) include C₀-C₃Alkylaryl or C₀-C₃Alkyl heteroaryl, each of which is optionally substituted by R⁹Mono-, di-, or tri-substituted, wherein:

R⁹is C₁-C₆Alkyl radical, C₁-C₆Alkoxy group, NO₂OH, halogen, trifluoromethyl, amino or amido (e.g. optionally substituted by C)₁-C₆Alkyl mono-or di-substituted amino or amido), C₀-C₃Alkylaryl group, C₀-C₃Alkylheteroaryl or carboxy, wherein aryl or heteroaryl is optionally substituted by R¹⁰Substitution, wherein:

R¹⁰is C₁-C₆Alkyl radical, C₃-C₇Cycloalkyl radical, C₁-C₆Alkoxy, amino (e.g. by C)₁-C₆Alkyl mono-or di-substituted amino), amido (such as C)₁-C₃Alkylamides), sulfonyl C₁-C₃Alkyl radical, NO₂OH, halogen, trifluoromethyl, carboxyl or heteroaryl.

In general, R⁸Is C₀-C₃Alkylaryl or C₀-C₃C in alkyl heteroaryl₀-C₃The alkyl moiety being methyl and being particularly absent, i.e. C₀. The aryl or heteroaryl moiety is as broadly described in the definitions section below.

Preferred R⁹Comprising C₁-C₆Alkyl radical, C₁-C₆Alkoxy, amino (e.g. di- (C)₁ -C₃Alkyl) amino), amido (such as-NHC (O) C₁-C₆Alkyl or C (═ O) NHC₁ -C₃Alkyl), aryl or heteroaryl, wherein the aryl or heteroaryl moiety is optionally substituted by R¹⁰Substitution; wherein: r¹⁰Is C₁-C₆Alkyl radical, C₃-C₇Cycloalkyl radical, C₁-C₆Alkoxy, amino (e.g. mono-or di-C)₁-C₆Alkylamino, amido (e.g., -NHC (O) C)₁ -C₃Alkyl or C (═ O) NHC₁-C₆Alkyl), halogen, trifluoromethyl or heteroaryl.

Preferred R¹⁰Comprising C₁-C₆Alkyl radical, C₁-C₆Alkoxy, amino, amido (e.g. -NHC (O) C₁-C₆Alkyl or C (═ O) NHC₁-C₆Alkyl), halogen or heteroarylAnd (4) a base.

Particularly preferred R¹⁰Including methyl, ethyl, isopropyl, t-butyl, methoxy, chloro, amino, amido (e.g., -NHC (O) C)₁-C₃Alkyl or C (═ O) NHC₁-C₆Alkyl) or C₁-C₃An alkyl thiazole.

R⁸Preferred embodiments include 1-naphthylmethyl, 2-naphthylmethyl, benzyl, 1-naphthyl, 2-naphthyl or quinolyl, each unsubstituted or substituted by R as defined above⁹Mono-or di-substituted, especially 1-naphthylmethyl, or unsubstituted, with R as defined above⁹Mono-or di-substituted quinolinyl.

Currently preferred R⁸Comprises the following steps:

wherein R is^9aIs C₁-C₆An alkyl group; c₁-C₆An alkoxy group; thio C₁-C₃An alkyl group; optionally is covered with C₁-C₆An alkyl-substituted amino group; c₀-C₃An alkylaryl group; or C₀-C₃Alkyl heteroaryl, C₀-C₃Alkylheterocyclyl, wherein said aryl, heteroaryl or heterocyclyl is optionally substituted with R¹⁰And wherein: r¹⁰Is C₁-C₆Alkyl radical, C₃-C₇Cycloalkyl radical, C₁-C₆Alkoxy, amino, amido, heteroaryl and heterocyclyl; and

R^9bis C₁-C₆Alkyl radical, C₁-C₆Alkoxy, amino, amido, NO₂OH, halogen, trifluoromethyl, carboxyl.

Suitable R^9aComprising aryl or heteroaryl, each optionally as defined aboveR of Yi¹⁰Substituted, especially wherein R^9aSelected from:

wherein R is¹⁰Is H, C₁-C₆Alkyl or C₀-C₃alkyl-C₃-C₆Cycloalkyl, amino (e.g. by C)₁-C₆Alkyl mono-or di-substituted amino), amido (e.g., -NHC (O) C₁-C₆Alkyl or C (═ O) NHC₁-C₆Alkyl), heteroaryl or heterocyclyl.

R^9aConveniently phenyl, and whereby R⁸Comprises the following steps:

wherein R is^10aIs H, C₁-C₆Alkyl radical, C₁-C₆Alkoxy or halogen; and R^9bIs C₁-C₆Alkyl radical, C₁-C₆Alkoxy, amino (e.g. C)₁-C₃Alkylamino), acylamino (e.g., -NHC (O) C₁-C₆Alkyl or C (═ O) NHC₁-C₃Alkyl), NO₂OH, halogen, trifluoromethyl or carboxyl.

Further preferred R⁸Comprises the following steps:

wherein R is^10aIs H, C₁-C₆Alkyl or C₀-C₃alkyl-C₃-C₆Cycloalkyl, amino (e.g. optionally substituted by C)₁-C₆Alkyl mono-or di-substituted amino), amido (e.g., -NHC (O) C₁-C₆Alkyl or C (═ O) NHC₁-C₃Alkyl or C (═ O) N (C)₁ -C₃Alkyl radical)₂) Heteroaryl or heterocyclyl; and R^9bIs C₁-C₆Alkyl radical, C₁-C₆Alkoxy, optionally substituted by C₁-C₆Alkyl mono-or di-substituted amino, amido (e.g. -NHC (O) C)₁-C₆Alkyl or C (═ O) NHC₁-C₃Alkyl or C (═ O) N (C)₁-C₃Alkyl radical)₂)、NO₂OH, halogen, trifluoromethyl or carboxyl.

In the embodiments described immediately above, R^9bConveniently is C₁-C₆-alkoxy, preferably methoxy.

Other R⁸A group, such as when W is an ether, is of the formula:

wherein W 'is N or CH, r is 0 or 1, Ra' is H, C₁-C₆Alkyl radical, C₀ -C₃Alkyl cycloalkyl radical, C₁-C₆Alkoxy, hydroxy or amine, and Rb' is H, halogen, C₁-C₆Alkyl radical, C₀-C₃Alkyl cycloalkyl radical, C₁-C₆Alkoxy radical, C₁-C₆Thioalkyl, cycloalkyl C₀-C₃Alkoxy radical, C₁-C₃Alkoxy radical C₁-C₃Alkyl radical, C₀-C₃Alkylaryl or C₀-C₃An alkyl heterocyclic group. A particularly preferred ether substituent is 7-methoxy-2-phenyl-quinolin-4-yloxy.

When W is a bond, then R⁸Preference is given to substituted or unsubstituted heterocyclic ring systems, as described in WO2004/072243 or WO 2004/113665.

When W is a bond, R⁸Representative examples include the following aromatic compounds which may optionally be substituted: 1H-pyrrole, 1H-imidazole, 1H-pyrazole, furan, thiophene, oxazole, thiazole, isoxazole, isothiazole, pyridine, pyridazine, pyrimidine, pyrazine, phthalazine, quinoxaline, quinazoline, quinoline, cinnoline, 1H-pyrrolo [2, 3-b ]]Pyridine, 1H-indole, 1H-benzimidazole, 1H-indazole, 7H-purine, benzothiazole, benzoxazole, 1H-imidazo [4, 5-c ]]Pyridine, 1H-imidazo [4, 5-b ]]Pyridine, 1, 3-dihydro-benzimidazol-2-one, 1, 3-dihydro-benzimidazol-2-thione, 2, 3-dihydro-1H-indole, 1, 3-dihydro-indolin-2-one, 1H-indole-2, 3-dione, 1, 3-dihydro-benzimidazol-2-one, 1H-pyrrolo [2, 3-c ] b]Pyridine, benzofuran, benzo [ b ]]Thiophene, benzo [ d ]]Isoxazoles, benzols [ d ]]Isothiazole, 1H-quinolin-2-one, 1H-quinolin-4-one, 1H-quinazolin-4-one, 9H-carbazole, 1H-quinazolin-2-one.

When W is a bond, R⁸Further representative examples include the following non-aromatic compounds which may optionally be substituted: aziridine, azetidine, pyrrolidine, 4, 5-dihydro-1H-pyrazole, pyrazolidine, imidazolidin-2-one, imidazolidin-2-thione, pyrrolidin-2-one, pyrrolidine-2, 5-dione, piperidine-2, 6-dione, piperidin-2-one, piperazine-2, 6-dione, piperazin-2-one, piperazine, morpholine, thiomorpholine-1, 1-dioxide, pyrazolidin-3-one, imidazolidine-2, 4-dione, piperidine, tetrahydrofuran, tetrahydropyran, [1, 4 ] pyrrolidine]Dioxane and 1, 2, 3, 6-tetrahydropyridine.

When W is a bond, R is preferably⁸The meaning of (a) includes tetrazole and derivatives thereof. The tetrazole moiety is attached to a cyclic P2 scaffold and is optionally substituted as shown below:

wherein Q^＊Selected from: absence of-CH₂-、-O-、-NH-、-N(R^1＊)-、-S-、-S(＝O)₂-and- (C ═ O) -; q^＊Selected from: absence of-CH₂-and-NH-; y is^＊Selected from: H. c₁-C₆Alkyl radical, C₀-C₃Aryl radical, C₀-C₃A heterocyclic group; and R^1＊Selected from: H. c₁-C₆Alkyl, carbocyclic radical, C₀-C₃Aryl radical, C₀-C₃A heterocyclic group.

Representative examples of substituted tetrazoles are described in Table 1 and subsequent structures in WO2004/072243 or in WO 2004/113665.

Further, when W is a bond, R is preferably⁸The meaning of (a) includes triazoles and derivatives thereof. The triazole moiety is attached to a cyclic P2 scaffold and is optionally substituted as shown below:

wherein X^＊And Y^＊Independently selected from: H. halogen, C₁-C₆Alkyl radical, C₀-C₃Carbocyclic radical, -CH₂-amino, -CH₂Arylamino, -CH₂-diarylamino, - (C ═ O) -amino, - (C ═ O) -arylamino, - (C ═ O) -diarylamino, C₀-C₃Aryl radical, C₀-C₃Heterocyclyl or, in addition, X^＊And Y^＊Together with the carbon atoms to which they are attached form a cyclic moiety selected from aryl and heteroaryl.

Representative examples of substituted triazoles are described in Table 2 and subsequent structures in WO2004/072243 or in WO 2004/113665.

Further, when W is a bond,preferably R⁸The meaning of (a) includes pyridazinones and derivatives thereof. The pyridazinone moiety is attached to a cyclic P2 scaffold and is optionally substituted as shown below:

wherein X^＊、Y^＊And Z^＊Independently selected from: H. n is a radical of₃Halogen, 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 and the carbon atoms to which they are attached are joined to form an aryl or heteroaryl ring moiety.

Representative examples of substituted pyridazinones are described in Table 3 and subsequent structures of WO2004/072243 or WO 2004/113665.

The P3 group is preferred, i.e. when m is 1, analogous to a natural or unnatural amino acid, especially an aliphatic amino acid, such as L-valyl, L-leucyl, L-isoleucyl or L-tert-leucyl. Further preferred P3 groups, as shown in WO02/01898, include C₀ -C₃Alkylcycloalkylalanines, in particular cyclohexylalanine, optionally coated with CO₂Rg is H, C₁-C₆Alkyl radical, C₀-C₃Alkylaryl group, C₀-C₃Alkyl heterocyclic group, C₀-C₃An alkylcycloalkyl or an amine; or N-acetylpiperidine or tetrahydrofuran. Thus, R is preferred¹¹The radicals including C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl (e.g. C)₀-C₃Alkyl radical C₃-C₇Cycloalkyl), C₀-C₃Alkylaryl or C₀-C₃Alkylheteroaryl, each of which is optionally substituted by hydroxy, halogen,Amino group, C₁-C₆Alkoxy radical, C₁-C₆Thioalkyl, C (═ O) OR¹⁴Carboxyl group, (C)₁-C₆Alkoxy) carbonyl, aryl, heteroaryl OR heterocyclyl, wherein the substituents are in particular hydroxy OR C (═ O) OR¹⁴。

Particularly preferred R¹¹Including t-butyl, isobutyl, cyclohexyl, phenethyl, 2-dimethyl-propyl, cyclohexylmethyl, benzyl, 2-pyridylmethyl, 4-hydroxy-phenylmethyl, or carboxypropyl. Most preferred R¹¹Meaning tert-butyl, isobutyl or cyclohexyl.

Embodiments of the invention include compounds wherein P4 is absent (i.e., n is 0) and wherein P3 is free of carbonyl groups (i.e., U is absent). Representative substructures include those of formula Ii:

wherein

Rx and Ry are as defined above, preferably H;

R¹¹' is C₁-C₆Alkyl, preferably C₃-C₅Branched alkyl groups such as the side chains of L-valyl, L-leucyl, L-isoleucyl, L-tert-leucyl; or C₀-C₂Alkyl radical C₃ -C₇Cycloalkyl groups such as cyclohexyl or cyclohexylmethyl;

R^16ais-Rba, -S (═ O)_pRba、-C(＝O)Rba；

Rba is C₁-C₆Alkyl radical, C₀-C₃Alkyl heterocyclic group, C₀-C₃An alkyl carbocyclyl group.

Alternatively, compounds with partial structure Ii may be in the appropriate R⁷The meaning and Rx, Ry or R¹¹One of which forms a macrocycle.

Representative examples of P3 groups that do not bear a carboxyl functionality (i.e., the variable U is absent) include the following formulas VIia-VIid:

wherein Ar is a carbocyclyl or heterocyclyl group, particularly an aryl or heteroaryl group, each of which is optionally substituted with R9. Although the partial structures of the formulae VIia-VIid have been elucidated in the context of the compounds of the invention in which k is 1 and q 'is 0, it is clear that the above-described VIi structure can also be applied to compounds at other values of q' and k. Similarly, although the partial structures of formulae VIic and VIid represent R corresponding to leucine¹¹Groups, but it is clear that these structures can also be used for other R' s¹¹Among the groups, in particular those analogous to the side chains of natural or unnatural L-amino acids, for example tert-butylalanine/tert-leucine.

In the compounds of the invention in which n is 1, R¹⁵Preferably optionally substituted C₁-C₆Alkyl or C₀-C₃Alkyl carbocyclyl (e.g. C)₀-C₃Alkyl radical C₃-C₇Cycloalkyl) which may each be optionally substituted. The P4 group is preferably a generic analogue of a natural or unnatural amino acid, in particular an aliphatic amino acid, such as L-valyl, L-leucyl, L-isoleucyl, L-tert-leucyl or L-cyclohexylalanine, and R is thus preferably R¹⁵The group includes cyclohexyl, cyclohexylmethyl, t-butyl, isopropyl or isobutyl.

Preferred G meanings include-NRy- (especially where Ry is methyl or preferably H) or hydrazine.

It is further preferred that G means O, thereby defining an ester with or in the carbonyl group of P4 (if present) or P3 (if present)Variants in which the group U is absent are defined as ethers. For R¹⁶Conventional pharmaceutically acceptable ether or ester end capping groups include C₁-C₆Alkyl (especially methyl or tert-butyl), C₀-C₃Alkylheterocyclyl (especially pyridyl, benzimidazolyl, piperidyl, morpholinyl, piperazinyl) or C₀-C₃An alkyl carbocyclyl (in particular phenyl, benzyl, 2, 3-indanyl), each of which is optionally substituted by hydroxy, halogen, amino or C₁-C₆Alkoxy groups.

Preferably, the compounds of the present invention may include hydrazine functional groups, for example where T is-NRd-and m is 1; while n is 0 or 1. In addition, particularly where m is 0, G may be-NRjNRj-such as-NHNH-. The compounds will generally not contain hydrazine on both G and T. Within compounds of formula VI wherein m and n are both 0, preferred hydrazines include compounds having the following partial structures VIja-VIjb:

in the formulae VIja and VIjb, R¹⁶' can be considered to be alkyl (or C)₁-C₃Alkyl heterocyclic radical or C₁-C₃Alkyl carbocyclyl) wherein the first alkyl carbon is substituted with an oxy group to produce a ketone functional group and R¹⁶' is the remaining alkyl, alkylheterocyclyl or alkylcarbocyclyl moiety. A variant is shown by the formula VIjb, in which R¹⁶Is a methylene group whose carbon is substituted by an oxo substituent and-ORb, wherein Rb is as defined above and is generally C₁-C₆Alkyl (e.g. tert-butyl), C₀-C₃Alkyl heterocyclic radical (e.g. pyridyl) or C₀-C₃An alkyl carbocyclyl (such as benzyl or phenyl), each of which is optionally substituted, as defined above. The compounds having the partial structures VIja and VIjb may be linear molecules as described above (both Rj are H), or preferably one of the Rj groups shown may be joined to the appropriate R via J⁷The group macrocyclates.

Further hydrazines of formula VI wherein m is 1 include those having the following partial structures VIjc and VIjd:

g, R therein¹⁵、R¹⁶RX, Rd, Rq, Rz and Ru are as defined above for formula VI. Compounds having the partial structures VIjc and VIjd may be linear molecules as described above (both Rx and Rd are H), or preferably one of the Rx or Rd groups shown may be reacted through J with the appropriate R⁷The group macrocyclates.

Although the formula VIja-VIjd is illustrated using a five-membered carbon ring as the P2 scaffold, it will be apparent that this aspect of the invention is equally applicable to other q' and k structures. Preferred embodiments within the formula VIja-VIjd include those wherein Rq and Rz are both H, or wherein Rz is an olefinic bond and Rq is C₁-C₃Those embodiments of alkyl.

When G is amino, and m and n are both 0, and R¹⁶When an N-linked unsaturated heterocycle as defined below (e.g. pyridyl or pyrimidinyl) or an N-linked saturated heterocycle as defined below (such as piperazinyl, piperidinyl and especially morpholinyl), another hydrazine-like structure results. Examples of the above embodiments include those with formulas VIjc and VIjd:

the compounds with partial structures VIjc and VIjd may be linear molecules as indicated above, or preferably Rx may be coupled via J with the appropriate R⁷The group undergoes macrocyclization. Although these partial structures are illustrated using five-membered rings as P2 scaffolding, it is clear thatHowever, this structure may extend to other values of q' and k. Similarly, these structures may be applied to others as R¹⁶The N-linked heterocycle of (1).

Returning now to formula VI, in general, R is preferred for the compounds of the present invention¹⁶The group includes 2-indanol, 2, 3-indanyl, 2-hydroxy-1-phenyl-ethyl, 2-thienylmethyl, cyclohexylmethyl, 2, 3-methylenedioxybenzyl, cyclohexyl, phenyl, benzyl, 2-pyridylmethyl, cyclobutyl, isobutyl, n-propyl, methyl or 4-methoxyphenylethyl.

Currently preferred R¹⁶The group includes 2-indanol, indane, 2-hydroxy-1-phenyl-ethyl, 2-thienylmethyl, 2, 3-methylenedioxybenzyl or cyclohexylmethyl.

Unnatural amino acids include L-amino acids, where the side chain is not one of the 20 naturally occurring amino acids. Examples of the unnatural amino acids include L- β -methanesulfonyl methylalanine, L-cyclohexylalanine, L-tert-leucine (L-tert-leucine), L-norleucine, L-norvaline, L-ornithine, L-sarcosine, L-citrulline, L-homophenylalanine, L-homoserine, L- β - (1-naphthyl) alanine, L- β - (2-naphthyl) alanine and the like. Unnatural amino acids also include D-amino acids corresponding to the 20 natural amino acids and D-amino acids with other side chains, such as those listed above.

"C" as used herein₁-C₆Alkyl "(also abbreviated as C)₁-C₆alk, or for compound expressions such as C₁-C₆Alkoxy, etc.) are meant to include straight or branched aliphatic carbon chains such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, isopentyl, hexyl, heptyl, and simple isomers thereof. The alkyl group may have an unsaturated bond. In addition, C₁-C₆Any C atom in the alkyl group may optionally be substituted with one, two or three halogen atoms as permitted by a valence bond and/or the alkylene chain is interrupted by a heteroatom S, O, NH. If the hetero atom is presentA sub-group is at the end of the chain, it may suitably be substituted by one or two hydrogen atoms. C1-C4 alkyl and C₁-C₅Alkyl having C adjusted as required by carbon number₁-C₆The corresponding meaning of alkyl.

"C" as used herein₁-C₃Alkyl "includes methyl, ethyl, propyl, isopropyl, cyclopropyl, each of which may be optionally substituted or interrupted by a heteroatom as described in the preceding paragraph, or at C₂Or C₃In the case of (B) with unsaturated bonds, e.g. CH₂＝CH。

"C" as used herein₁-C₃Alkylene "describes a divalent C₁-C₃Alkanediyl moieties include propylene, ethylene and in particular methylene. For J, the generally longer alkylene chain may include 1 to 3 unsaturations and/or be interrupted with heteroatoms as described above.

"amino" includes NH₂、NHC₁-C₆Alkyl or N (C)₁-C₆-alkyl groups)₂In particular C₁-C₃Alkyl variants.

"amido" includes C (═ O) NH₂And alkylamido radicals, such as C (═ O) NHC₁-C₆Alkyl, C (═ O) N (C)₁-C₆Alkyl radical)₂Especially C (═ O) NHC₁-C₃Alkyl, C (═ O) N (C)₁ -C₃Alkyl radical)₂or-NH (C ═ O) C₁-C₆Alkyl, for example-NHC (═ O) CHC (CH)₃)₃including-NH (C ═ O) C₁-C₃An alkyl group.

"halogen" as used herein is meant to include F, Cl, Br, I, especially chlorine and preferably fluorine.

"C" as used herein₀-C₃Alkylaryl "is meant to include aryl moieties such as phenyl, naphthyl or fused to C₃-C₇Phenyl of cycloalkyl (e.g. 2, 3-indanyl)) Wherein said aryl groups are directly bonded (i.e. C)₀) Or by C as above₁-C₃The middle methyl, ethyl or propyl or isopropyl linkage as defined for alkylene. Unless otherwise specified, the aryl group and/or its fused cycloalkyl moiety is optionally substituted with 1 to 3 substituents selected from the group consisting of: halogen, hydroxy, nitro, cyano, carboxy, C₁-C₆Alkyl radical, C₁-C₆Alkoxy radical, C₁-C₆Alkoxy radical C₁-C₆Alkyl radical, C₁-C₆Alkanoyl, amino, azido, oxo, mercapto, nitro, C₀ -C₃Alkyl carbocyclyl, C₀-C₃An alkyl heterocyclic group. "aryl" has the corresponding meaning, i.e. where C₀-C₃Alkyl linkages are absent.

"C" as used herein₀-C₃Alkyl radical C₃-C₇Cycloalkyl "is meant to include C₃-C₇Cycloalkyl radicals, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl, in which the cycloalkyl radicals are bonded directly (i.e. C)₀Alkyl) or by C as above₁-C₃The middle methyl, ethyl, propyl (proyl) linkage as defined by alkylene. The cycloalkyl group may include an unsaturated bond. Unless otherwise specified, the cycloalkyl moiety is optionally substituted with 1 to 3 substituents selected from the group consisting of: halogen, hydroxy, nitro, cyano, carboxy, C₁-C₆Alkyl radical, C₁-C₆Alkoxy radical, C₁-C₆Alkoxy radical C₁-C₆Alkyl radical, C₁-C₆Alkanoyl, amino, azido, oxo, mercapto, nitro, C₀-C₃Alkyl carbocyclyl, C₀-C₃An alkyl heterocyclic group.

"C" as used herein₀-C₃Alkylcarbocyclyl "is meant to include C₀-C₃Alkylaryl and C₀ -C₃Alkyl radical C₃-C₇A cycloalkyl group. Unless otherwise specified, the aryl or cycloalkyl group is optionally substituted with 1 to 3 substituents selected fromSubstituted by the following groups: halogen, hydroxy, nitro, cyano, carboxy, C₁-C₆Alkyl radical, C₁-C₆Alkoxy radical, C₁-C₆Alkoxy radical C₁-C₆Alkyl radical, C₁-C₆Alkanoyl, amino, azido, oxo, mercapto, nitro, C₀-C₃Alkyl carbocyclyl and/or C₀ -C₃An alkyl heterocyclic group. "carbocyclyl" has the meaning corresponding thereto, i.e. wherein C₀-C₃Alkyl linkages are absent.

"C" as used herein₀-C₃Alkylheterocyclyl "is intended to include monocyclic, saturated or unsaturated, heteroatom-containing rings, such as piperidinyl, morpholinyl, piperazinyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazinyl (thiazinoyl), isothiazolyl, thiazolyl, oxadiazolyl, 1, 2, 3-triazolyl, 1, 2, 4-triazolyl, tetrazolyl, furanyl, thienyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazolyl, or any of the foregoing groups fused to a phenyl ring, such as quinolinyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazinyl, benzisothiazinyl, benzothiazolyl, benzoxadiazolyl, benzo-1, 2, 3-triazolyl, benzo-1, 2, 4-triazolyl, benztetrazolyl, benzofuranyl, benzothienyl, benzopyridyl, benz-1, 2, 4-triazolyl, benz-thiazolyl, benz-azolyl, benz-thienyl, benz-pyridyl, Benzopyrimidinyl, benzopyrazinyl, benzopyrazolyl, etc., said rings being directly linked (i.e. C)₀) Or by C as above₁-C₃An intermediate methyl, ethyl, propyl or isopropyl linkage as defined by alkylene. Any of the above unsaturated rings having aromatic character may be referred to herein as heteroaryl. Unless otherwise specified, the heterocycle and/or its fused phenyl moiety is optionally substituted with 1 to 3 substituents selected from the group consisting of: halogen, hydroxy, nitro, cyano, carboxy, C₁-C₆Alkyl radical, C₁-C₆Alkoxy radical, C₁-C₆Alkoxy radical C₁-C₆Alkyl radical, C₁-C₆Alkanoyl, amino, azido, oxo, mercapto, nitro, C₀-C₃Alkyl carbocyclyl, C₀-C₃An alkyl heterocyclic group. "Heterocyclyl" and "heteroaryl" have the corresponding meanings, i.e. wherein C₀-C₃Alkyl linkages are absent.

Thus, general heterocyclyl and carbocyclyl moieties within the above definitions are monocyclic having 5 or, especially, 6 ring atoms, or bicyclic structures comprising a 6-membered ring fused to a 4-, 5-or 6-membered ring.

Typically the above groups include C₃-C₈Cycloalkyl, phenyl, benzyl, tetrahydronaphthyl, indenyl, 2, 3-indanyl, heterocyclyl, such as azepanyl, azocanyl, pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, indolinyl, pyranyl, tetrahydropyranyl, tetrahydrothiopyranyl, thiopyranyl, furanyl, tetrahydrofuranyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, tetrazolyl, pyrazolyl, indolyl, benzofuranyl, benzothienyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, quinolinyl, tetrahydroquinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, quinazolinyl, tetrahydroquinazolinyl, and quinoxalinyl, each of which may be optionally substituted, as defined herein.

Thus, the saturated heterocyclic moiety includes groups such as: pyrrolinyl, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, pyranyl, thiopyranyl, piperazinyl, indolinyl, azetidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydrofuranyl, hexahydropyrimidyl, hexahydropyridazinyl, 1, 4, 5, 6-tetrahydropyrimidinyl, dihydro-oxazolyl, 1, 2-thiazinyl-1, 1-dioxide, 1, 2, 6-thiadiazinyl-1, 1-dioxide, isothiazolidinyl-1, 1-dioxide, and imidazolidinyl-2, 4-dione, however, the unsaturated heterocycles include groups with aromatic character such as furanyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, and the like, Triazolyl, tetrazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, indolyl, isoindolyl. In each case, the heterocyclic ring may be fused to a phenyl ring, thereby forming a bicyclic ring system.

Synthesis of

The compounds of the invention can be synthesized in solution or in solid phase or a combination of both by different chemical strategies. The appropriately protected individual building blocks can be prepared first and subsequently coupled together, i.e., P2+ P1 → P2-P1. Alternatively, precursors of the building blocks may be coupled together and then altered in a later step in the sequential synthesis of the inhibitor. Additional building blocks, building block precursors or presynthesized larger fragments of the desired structure may then be coupled to the propagating chain, e.g., R¹⁶-G-P3+C(＝O)-P2-P1→R¹⁶-G-P3-C (═ O) -P2-P1 or R¹⁶-G-P4-P3+C(＝O)-P2-P1→R¹⁶-G-P4-P3-C(＝O)-P2-P1.

The coupling between two amino acids, between an amino acid and a peptide or between two peptide fragments can be carried out using standard coupling methods, such as the azide method, the mixed carbon-carboxylic anhydride (isobutyl chloroformate) method, the carbodiimide (dicyclohexylcarbodiimide, diisopropylcarbodiimide or water-soluble carbodiimide) method, the active ester (p-nitrophenyl ester, N-hydroxysuccinimide ester) method, the Woodward reagent K-method, the carbonyldiimidazole method, the phosphorous reagent or the redox method. Some of these methods, particularly the carbodiimide method, can be improved by adding 1-hydroxybenzotriazole or 4-DMAP. These coupling reactions can be carried out in solution (liquid phase) or in solid phase.

More specifically, the coupling step comprises dehydrative coupling of a free carboxyl group of one reactant with a free amino group of another reactant in the presence of a coupling agent to form an amide linkage. Descriptions of the above coupling agents are disclosed in textbooks of general Peptide Chemistry, such as m.bodanszky, "Peptide Chemistry", second revised edition, Springer-Verlag, Berlin, Germany, (1993), hereinafter abbreviated Bodanszky, the contents of which are incorporated herein by reference. 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 effective coupling agent is commercially available (benzotriazol-1-yloxy) tris- (dimethylamino) phosphonium hexafluorophosphate, used either alone or in the presence of 1-hydroxybenzotriazole or 4-DMAP. Another useful and effective coupling agent is the commercially available 2- (1H-benzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium tetrafluoroborate. Another useful and effective coupling agent is commercially available O- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate.

The coupling reaction is carried out in an inert solvent, such as dichloromethane, acetonitrile or dimethylformamide. An excess of a tertiary amine (e.g., diisopropylethylamine, N-methylmorpholine, N-methylpyrrolidine, or 4-DMAP) is added to maintain the pH of the reaction mixture at about 8. The reaction temperature is usually 0 ℃ to 50 ℃, and the reaction time is usually 15min to 24 h.

During the coupling reaction, the functional groups of the amino acid component must generally be protected to avoid the formation of undesirable bonds. Protecting groups that 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), hereinafter simply Greene, the disclosure of which is incorporated herein by reference.

The alpha-carboxy group of the C-terminal residue is typically protected as an ester, which can be cleaved to form the carboxylic acid. Protecting groups that may be used include 1) alkyl esters such as methyl, trimethylsilyl and tert-butyl esters, 2) aralkyl esters such as benzyl and substituted benzyl esters, or 3) esters such as trichloroethyl and acetophenone (ethanone) esters, which may be decomposed by weak bases or mild reduction methods.

The alpha-amino group of each amino acid to be coupled is generally protected. Any protecting group known in the art may be used. Examples of the above groups include: 1) acyl groups such as formyl, trifluoroacetyl, phthaloyl and p-toluenesulfonyl; 2) aromatic carbamate groups such as benzyloxycarbonyl (Cbz or Z) and substituted benzyloxycarbonyl and 9-fluorenylmethyloxycarbonyl (Fmoc); 3) aliphatic urethane groups such as a tert-butoxycarbonyl group (Boc), an ethoxycarbonyl group, a diisopropylmethoxycarbonyl group, and an allyloxycarbonyl group; 4) cyclic alkyl carbamate groups such as cyclopentyloxycarbonyl and adamantyloxycarbonyl; 5) alkyl groups such as trityl and benzyl; 6) trialkylsilyl groups such as trimethylsilyl; and 7) thiol-containing groups such as phenylthiocarbonyl and dithiosuccinyl. Preferably, the alpha-amino protecting group is Boc or Fmoc. Many amino acid derivatives are commercially available with appropriate protection for peptide synthesis.

The alpha-amino protecting group is cleaved prior to the next coupling step. When using a Boc group, the method of choice is trifluoroacetic acid alone or in dichloromethane, or HCl in dioxane or ethyl acetate. The resulting ammonium salt is then neutralized with an alkaline solution (such as aqueous buffer, or tertiary amine in dichloromethane or acetonitrile or dimethylformamide) either before coupling or in situ. When the Fmoc group is used, the reactant of choice is piperidine or substituted piperidine in dimethylformamide, but any secondary amine may be used. The deprotection is carried out between 0 ℃ and room temperature, and is usually 20-22 ℃.

During the preparation of the peptides using any of the above groups, any natural or unnatural amino acid having a side chain functionality is typically protected. It will be appreciated by those skilled in the art that the selection and use of appropriate protecting groups for these side chain functionalities will depend on the presence of amino acids and other protecting groups on the peptide. In selecting the above protecting groups, it is desirable that the groups are not removed during deprotection and coupling of the α -amino group.

For example, when Boc is used as the α -amino protecting group, the following side chain protecting groups are suitable: the p-toluenesulfonyl moiety can be used to protect amino acid side chains such as lysine and arginine; acetamidomethyl, benzyl (Bn) or tert-butylsulfonyl moieties can be used to protect the sulfide-containing side chain of cysteine; benzyl (Bn) ethers may be used to protect the hydroxyl-containing side chain of serine, threonine or hydroxyproline; and benzyl esters can be used to protect the carboxyl-containing side chains of aspartic acid and glutamic acid.

When Fmoc is chosen for alpha-amine protection, a protecting group based on typically a tert-butyl group is acceptable. For example, Boc may be used to protect lysine and arginine, t-butyl ether may be used to protect serine, threonine and hydroxyproline and t-butyl ester may be used to protect aspartic acid and glutamic acid. The Trityl (Trityl) moiety can be used to protect the sulfide-containing side chain of cysteine.

Once the sequential synthesis of the inhibitor is complete, any protecting groups are removed in any manner determined by the choice of protecting group. These methods are well known to those skilled in the art.

Introduction of P2 substituent

R⁸The groups may be coupled to the P2 scaffold in any suitable step of the synthesis of compounds according to the invention. One method is to firstly carry out R⁸The group is coupled to a P2 scaffold, followed by the addition of other desired building blocks, i.e. P1 and optionally P3 and P4. Another approach is to couple the P1, P2 and, if present, the P3 and P4 moieties using an unsubstituted P2 scaffold, followed by addition of R⁸A group.

Wherein W is O and R⁸Is alkyl, C₀-C₃Alkyl carbocyclyl, C₀-C₃Alkylheterocyclyl Compounds of the invention can be prepared according to the procedures described in E.M. Smith et al (J.Med.chem. (1988), 31, 875-885)The preparation was carried out as shown in scheme 1, which illustrates a process with a saturated P2 scaffold where q' is 0 and k is 1.

Scheme 1

Treatment of a compound (1a) containing the unsubstituted P2 structure (which compound may be prepared as described hereinafter) with a base such as sodium hydride or potassium tert-butoxide in a solvent such as dimethylformamide, followed by reaction of the resulting alkoxide with an alkylating agent R⁸-X, wherein X is a suitable leaving group such as halogen, mesylate, triflate or tosylate, to give the desired substituted derivative (1 b).

Alternatively, if X is OH or SH, then the P2 substituent may be introduced via a Mitsunobu reaction by reacting the hydroxyl group of compound 1a with the desired alcohol or thiol in the presence of triphenylphosphine and an activator such as diethyl azodicarboxylate (DEAD) or diisopropyl azodicarboxylate (DIAD), and the like. (Mitsunobu, 1981, Synthesis, month 1, 1-28; Rano et al, Tetrahedron Lett., 1995, 36, 22, 3779-.

Alternatively, the alcohol (1a) may be treated with phosgene to give the corresponding chloroformate, which is reacted with an amine R in the presence of a base such as sodium bicarbonate or triethylamine⁸NH₂Reacted to form the carbamate, i.e., W is-OC (═ O) NH-, and alcohol (1a) is reacted with acylating agent R⁸The reaction of-CO-X (such as an anhydride or acid halide (e.g. acid chloride)) may form an ester, i.e. W is-OC (═ O) -.

Plural alcohols R⁸-OH and alkylating agent R⁸X is described in WO00/09543 and WO 00/59929. Wherein R is⁸An example of the synthesis for substituted quinoline derivatives is shown in scheme 2.

Scheme 2

A commercially available or commercially available substituted aniline (2a) is subjected to Friedel-Craft acylation using an acylating agent such as acetyl chloride or the like in a solvent such as methylene chloride in the presence of boron trichloride and aluminum trichloride to give (2 b). Under basic conditions (such as in pyridine), in carboxylic ester group activators (e.g. POCl)₃) Coupling of (2b) to heterocyclic carboxylic acid (2c) in the presence of a base, followed by ring closure and dehydration in t-butanol under basic conditions such as potassium t-butoxide, yields quinoline derivative (2 e). In the Mitsunobu reaction, the quinoline derivative (2e) may be coupled to an alcohol as described above, or wherein the hydroxy group may be replaced by a suitable leaving group (such as a halogen, e.g. chlorine, bromine or iodine) by treating quinoline (2e) with a suitable halogenating agent (e.g. phosphoryl chloride or the like).

A variety of carboxylic acids having the general structure (2c) can be used in scheme 2. These acids are commercially available or available in the literature. An example of the preparation of 2- (substituted) -amino-carboxy-aminothiazole derivatives following the procedure of Berdikhina et al, chem.heterocyclic.Compd. (Engl. Transl.) (1991), 427-433, as shown in scheme 3 below.

Scheme 3

Thioureas (3c) having different alkyl substituents R' can be formed by: the appropriate amine (3a) is reacted 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. The thiourea derivative (3c) is then condensed with 3-bromopyruvic acid, thereby forming the acid (3 d).

Wherein R is⁸A P2 building block with substituents attached via an amine, amide, urea or sulfonamide can be prepared from an amino-substituted carbocyclic ring, for example, obtained by converting the hydroxy group of the corresponding hydroxy derivative to the azido group, for example, by converting the hydroxy group to a suitable leaving group such as mesylate or a halogen such as chlorine, and then substituting the leaving group with an azide or by using an azide transfer agent such as Diphenylphosphorylazide (DPPA). The azide is reduced by catalytic hydrogenation or any other suitable reduction method to form the amine⁸An alkylating agent of X, in which R is⁸And X is as described in scheme 1, thereby forming the P2 structural unit for preparing compounds of general formula VI, wherein W is-NH-. Reacting an amino-substituted carbocyclic ring with a compound of formula R⁸-COOH acid is reacted under standard amide coupling conditions to form a compound wherein R⁸Compounds in which the substituents are linked via an amide bond, so that the amino-substituted carbocyclic ring is coupled to the appropriate sulfonic acid derivative R⁸-S(O)₂-X, wherein X is a leaving group (e.g. chloro), in the presence of a base, thereby forming a sulfonamide. Wherein the ring-shaped scaffold and R⁸Compounds in which the link between the substituents consists of a urea group can be obtained, for example, by the following method: the amino-substituted carbocyclic ring is treated with phosgene to provide the corresponding chlorocarbamate, which is subsequently reacted with the desired amine. In addition, the amino-substituted carbocycle may be substituted with carbamoyl chloride or have the desired R⁸The isocyanates of the substituents react to form the urea linkage. Obviously, the corresponding reactions apply to P2 groups with other ring sizes and characteristics.

Wherein the heterocyclic ring R⁸Compounds of the invention in which the group is directly attached to a cyclic P2 scaffold, i.e. W is a bond in formula VI, can be prepared, for example, by a displacement reaction in which the group is on a P2 scaffoldR where a suitable leaving group (such as halogen or mesylate, etc.) is desired⁸Groups (such as heterocyclic groups). In addition, the R is⁸The groups may be introduced by way of a Mitsunobu reaction,

wherein the hydroxyl group of the P2 precursor is bound to the heterocycle R⁸The nitrogen atoms in the group react.

Compounds in which the tetrazole derivative is attached to one of its ring carbon atoms are desirably prepared by attaching the tetrazole moiety directly to the P2 precursor. This can be achieved, for example, by converting the hydroxyl group of the P2 precursor to a cyano group, which is then reacted with an azide reagent, such as sodium azide. Triazole derivatives can also be built directly on the P2 precursor, for example by converting the hydroxyl group of the P2 precursor to the azido group, followed by a 3+2 cycloaddition reaction of the provided azide with the appropriate alkyne derivative.

Structurally different tetrazoles for the above substitution reaction and Mitsunobu reaction can be prepared by reacting a commercially available nitrile compound with sodium azide. Triazole derivatives can be prepared by reacting an alkyne compound with trimethylsilylazide. The alkyne compounds used are either commercially available or they can be prepared, for example, according to the Sonogashira reaction, i.e.in PdCl₂(PPh)₃And CuI, as described in a.elangovan, y. -h.wang, t. -i.ho, org.lett., 2003, 5, 1841-. The heterocyclic substituent may also be altered when attached to the P2 building block, either before or after the P2 building block is coupled to other building blocks.

Preparation of a compound in which W is a bond and R⁸These methods for compounds that are optionally substituted heterocycles and their useIts process is broadly described in WO 2004/072243.

W-R with carbocyclic derivatives in scheme 1⁸Compounds according to the invention can also be prepared with additional ring sizes of substituents and/or with compounds thereof located elsewhere.

Synthesis and introduction of P1 building Block

The amino acids used for the preparation of the P1 fragment are either commercially available or are available in the literature, see for example WO00/09543 and WO00/59929 to Boehringer-Ingelheim or US2004/0048802 to BMS.

Scheme 4 shows an example of the preparation of sulfonamide derivatives that are used as P1 building blocks and subsequently coupled to P2 building blocks.

Scheme 4

By treating the amino acid with a coupling reagent (e.g., N' -Carbonyldiimidazole (CDI), etc.) in a solvent such as THF, followed by a strong base (e.g., 1, 8-diazabicyclo [5.4.0 ]]Undec-7-ene (DBU)) with the desired sulfonamide (4b) can introduce a sulfonamide group onto the appropriately protected amino acid (4 a). In addition, the amino acid is treated with the desired sulfonamide (4b) in the presence of a base such as diisopropylethylamine, followed by treatment with a base such as diisopropylethylamineThe coupling reagent of (a) is treated to effect the introduction of the sulfonamide group. The amino protecting group is removed by standard methods and then amide bond formation is performed using standard methods such as O- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexakis coupling reagent in a solvent such as dimethylformamide in the presence of a base such as diisopropylamineFluorophosphate (HATU), which was coupled to the P2 building block prepared as above, to give (4 e). Alternatively, the sulfonamide group may be introduced at a later step of the synthesis, for example as the last step. In this case, an amino acid having a reverse protected form, i.e., an amino acid having an unprotected amino function and a protected acid function, is coupled to the acid function of the P2 building block using, for example, standard peptide coupling conditions as described above. Compound 4e is obtained by removal of the acid protecting group using appropriate conditions for the existing protecting group, followed by coupling of the sulfonamide as described above.

The P1 building block for the preparation of compounds according to formula VI, wherein A is an ester or amide, can be prepared by reacting the amino acid (4a) with the appropriate amine or alcohol, respectively, under standard conditions for amide or ester formation. Wherein A is CR⁴R⁴' Compounds according to formula VI may be prepared by coupling an appropriate P1 building block to a P2 building block, as described in Oscarsson et al Bioorg Med Chem 200311(13)2955-2963 and in PCT/EP03/10595, filed on 09/23, 2003, the contents of which are incorporated herein by reference.

Compounds comprising the residue of the azapeptide P1, i.e. M being NRu in formula VI, can be prepared by using the appropriate P1 aza-aminoacyl moiety in the coupling to the P2 fragment. The preparation of aza-aminoacyl moieties is described by m.d. bailey et al in j.med.chem., 47, (2004), 3788-.

Scheme 5

The appropriate N-linked side chain Ru can be introduced onto commercially available tert-butylhydrazine, for example by reductive amination with the appropriate aldehyde or ketone as described in scheme 19 below, thereby yielding an N-alkylated carbazate (5 a). In the presence of, for example, triethylamine or diisoCondensation of 5a with the desired chloroformate in the presence of a base of propylethylamine in a solvent such as THF affords 5 b. Then, the utilization depends on the specific R¹Suitable conditions of¹' when it is benzyl, catalytic hydrogenation conditions are employed, optionally with R¹Partial removal thereby giving the corresponding acid. The resulting acid as described above is then reacted with the desired sulfonamide derivative as described in scheme 4 to give a sulfonamide-capped building block. In addition, the carbazate 5a is reacted with an isocyanate R³-N ═ C ═ O reaction, forming the structural units used to prepare the compounds according to general formula VI, where M is NRu and a is CONHR³。

Synthesis of end-capped P3 and P3-P4 building blocks

Structural unit R¹⁶-G-P3 and R¹⁶the-G-P4-P3 can be prepared by the general method shown in scheme 6.

Scheme 6

The appropriate N-protected amino acid (6a) may be reacted with an amino-capping group (R) in the presence of a base such as DIEA or DMAP, in a solvent such as dichloromethane, chloroform or dimethylformamide or mixtures thereof, by applying standard peptide coupling conditions (e.g.using coupling reagents such as HATU, DCC or HOBt) and ester formation conditions, etc¹⁶-NHRy) to form an amide, i.e. G is NHRy (6 b). Further, the amino acid (6a) is reacted with the general formula R in the presence of a base such as cesium carbonate or silver oxide (I)¹⁶-X compound reaction, wherein R¹⁶As defined above and X is a leaving group such as halogen, forming an ester, i.e. G is O (6 b). On the other hand, amino acid (6a) can be coupled to an appropriately O-protected secondary amino acid (6d) using standard peptide coupling conditions as described above, to form (6 e). Substitution of the ester group with a suitable end-capping group (6b) forms a group that can be used to prepare the hair according to the inventionFragment (6f) of the compound of formula (i), wherein m and n are both 1.

When G is N-Ry, the capped P3 or P2 building blocks can also be prepared on a solid support as illustrated in scheme 7.

Scheme 7

An appropriately N-protected (e.g. Boc-protected) amino acid (7a) may be immobilised on a solid support by reacting the amino acid with the desired solid support in a solvent such as dichloromethane and dimethylformamide in the presence of a coupling reagent such as N, N' -diisopropylcarbodiimide and a base such as DMAP, the latter being exemplified herein by the Agronaut resin PS-TFP. The above-mentioned immobilized amino acid (7b) can then be isolated from the support by using a suitable end-capping group (7c), thereby giving a fragment (7d) useful for the preparation of the compounds according to the invention, wherein m or n is 1. Optionally, the amino protecting group may be removed and subsequently coupled to the appropriate amino acid using standard methods, thereby forming a fragment useful in the preparation of compounds according to the invention, wherein m and n are 1.

Preparation and introduction of P2 building Block

A general route to synthesize compounds containing a 5-membered saturated P2 scaffold is shown in scheme 8.

Scheme 8

The cyclic scaffold (8b) can be prepared, for example, from 3, 4-bis (methoxycarbonyl) cyclopentanone (8a) as described by Rosenquist et al in Acta chem. Scand.46(1992)1127-1129Reduction of the ketone group with a reducing agent such as sodium borohydride in a solvent such as methanol, followed by hydrolysis of the ester and finally ring closure in acetic anhydride in the presence of pyridine. The bicyclic acid (8b) obtained above may then be coupled to the desired P3 fragment (8c), P3-P4 fragment or capping group R using HATU and diisopropylamine in a solvent such as dimethylformamide using conventional peptide coupling conditions¹⁶On the amine function of-NHRy, giving (8 d). The lactone (8d) is ring-opened using, for example, lithium hydroxide to form an acid, which is then coupled to the amino group of the P1 building block or precursor of the desired P1 fragment (8e) using conventional peptide coupling conditions. Carbocyclic ring R⁸The substituent may be introduced, for example, by a Mitsunobu reaction with an appropriate alcohol as described above or by any other suitable method as previously described. When R is⁷、R⁷Where "and A' comprise functional groups, these functional groups are optionally suitably protected by methods recognized by those skilled in the art, see, e.g., Bodanzky or Greene cited above.

Scheme 9 represents another method of synthesizing compounds of formula VI containing a saturated P2 scaffold, wherein the building blocks are introduced in the reverse order, i.e. the P1 fragment is introduced before the end capping group, the P3 or the P3-P4 building block is introduced.

Scheme 9

The acid group of (9a) is protected as tert-butyl ester by treatment with, for example, di-tert-butyl dicarbonate in a solvent such as dichloromethane in the presence of a base such as dimethylaminopyridine and triethylamine, to provide the ester (9 b). The lactone is ring-opened and coupled to the P1 building block (9c) as depicted in scheme 13 or directly through the amine group of the P1 fragment to form (9 d). Introduction of R as described above⁸Substituents, followed by subjecting the above esters to, for example, trifluoroethyl ether in a solvent such as dichloromethaneAcid and triethylsilane, removing the acid protecting group, and finally reacting it with P3 building block (9e), P3-P4 building block or blocking group R as described above¹⁶-NHRy coupling, thereby forming (9 f). When R is⁷、R⁷Where "and A' comprise functional groups, these functional groups are optionally suitably protected by methods recognized by those skilled in the art, see, e.g., Bodanzky or Greene cited above.

The unsaturated P2 scaffold to be used in the preparation of the compound of formula VI can be prepared as shown below, illustrated with cyclopentene.

The cyclopentene scaffold was generally prepared as described in scheme 10.

Scheme 10

3, 4-bis (methoxycarbonyl) cyclopentanone (10a) is subjected to bromination-elimination as described in J.org.chem.36(1971)1277-1285 by Dolby et al, followed by reduction of the ketone functional group with a reducing agent such as sodium borohydride to form the unsaturated hydroxy compound (10 b). In a mixed solvent such as dioxane and water, selective ester hydrolysis is carried out using, for example, lithium hydroxide to form the hydroxy-substituted monoester derivative (10 c).

Where Rq is not hydrogen, unsaturated P2 structural scaffolds such as methylated cyclopentene scaffolds can be prepared as shown in scheme 11.

Scheme 11

Commercial 3-methyl-3-butan-1-ol (11a) is oxidized by using an oxidizing agent such as pyridinium chlorochromate, followed by treatment with acetyl chloride, bromine and methanol to form the α -bromo ester (11 c). The resulting ester (11c) can then be reacted with an enolate (11e) by, for example, treating the corresponding tert-butyl ester with a base such as lithium diisopropylamide in a solvent such as tetrahydrofuran to give the alkylated compound (11 f). The tert-butyl ester (11e) can be prepared by treating the corresponding commercially available acid (11d) with di-tert-butyl dicarbonate in the presence of a base such as dimethylaminopyridine, wherein k' is 1 to 3. The cyclization of (11f) by the olefin metathesis reaction was carried out as described above to form the cyclopentene derivative (11 g). Stereoselective epoxidation of (11g) was carried out using the Jacobsen asymmetric epoxidation method to give epoxide (11 h). Finally, a base such as DBN (1, 5-diazabicyclo- [4.3.0] non-5-ene) is added to give alcohol (11 i). Optionally, the double bond of compound (11i) may be reduced, for example by catalytic hydrogenation using a catalyst such as palladium on carbon, to form the corresponding saturated compound.

The resulting circular scaffold can then be used to complete inhibitor synthesis as described above. This is illustrated in scheme 12.

Scheme 12

The amino group of the P1-building block or a suitable precursor thereof (12b) may be coupled to the cyclopentene-derivative acid (12a) using HATU using standard amide coupling conditions, such as in the presence of a base such as diisopropylaniline and the like, followed by introduction of R by Mitsunobu conditions, e.g., as described above⁸-a substituent, thereby forming (12 d). The remaining ester is hydrolyzed and the amide is then coupled to the desired P3 or P3-P4 building block (12e), optionally followed by treatment of the P1 moiety, to provide the cyclopentene-containing compound (12f) according to formula VI. When R is⁷、R⁷When "and A" comprise functional groups, these functional groups optionally pass through the present inventionMethods recognized by those skilled in the art are appropriately protected, see, e.g., Bodanzky or Greene, cited above.

Compounds with hydrazines containing a blocking group attached directly to the P2 moiety, i.e., P3 and P4 are absent and G is NRjNRj, can be prepared as shown in scheme 13.

Scheme 13

Tert-butyl carbazate (13a) optionally substituted with alkyl groups on one or both nitrogen atoms is reacted with an acid (13b) under peptide coupling conditions, for example using HATU and DIEA in a solvent such as DMF, to provide 9 Ac. The boc group is optionally removed by standard procedures, such as acid treatment with, for example, TFA in a suitable solvent such as dichloromethane, to provide the hydrazine-containing derivative (13 d). In addition, any suitable hydrazine derivative other than t-butyl carbazate derivatives, such as morpholin-1-ylamine or piperidin-1-ylamine, and the like, may be attached to the acid (13 b).

The resulting compound can then be further expanded by coupling the P3 or P4-P3 building blocks to the primary amine compound 13d, as shown in scheme 14.

Scheme 14

Alpha-amino compound (14a) (Yang et al, J. org. chem. (2001), 66, 7303-7312) was treated with sodium nitrite, potassium bromide and sulfuric acid to form the corresponding alpha-bromine compound (14b), which was reacted with the above-mentioned derivative (13d) to provide the hydrazine-containing derivative (14 c).

Compounds without a carboxyl group in the P3 unit can be prepared as shown in scheme 15, which is exemplified by the cyclopentane derivative as the P2 scaffold.

Scheme 15

Acid (15a) can be coupled to amino azide derivative (15b) prepared by literature known methods using standard peptide coupling conditions to give amide derivative (15 c). The azide functionality is reduced, for example by using polymer-bound triphenylphosphine in a solvent such as methanol or by any other suitable reduction method, to form intermediate (15d), which can then be reacted with an acid under peptide coupling conditions or an amine under reductive amination conditions to form an amide and a secondary amine, respectively.

Scheme 16 represents another route to compounds in which no carboxyl group is present in the P3 structural unit.

Scheme 16

Instead of using the azide derivative (15b) in scheme 15, the corresponding optionally protected hydroxy derivative (16b) is used for coupling with the acid (16a), thereby introducing the primary alcohol. The alcohol (16c) is then oxidized, after optional deprotection, with a suitable oxidizing agent, such as Dess-Martin periodinane (periodinane), to form the corresponding aldehyde. The aldehyde is reductively aminated with the desired amine in a solvent such as THF by using a reagent such as, for example, polystyrene to bind cyanoborohydride, to form the amine derivative (16 e).

In addition, under appropriate conditions, alcohol (16c) may react with a suitable acylating or alkylating agent to form ester and ether compounds, respectively, i.e., compounds of formula (I) wherein G is O.

Subsequently, ester and ether compounds, i.e. compounds of formula VI in which G is O, are formed, respectively, by reacting the formed alcohol with a suitable acylating or alkylating agent using appropriate conditions.

Although schemes 15 and 16 have been described with respect to cyclopentane derivatives, i.e. compounds of formula VI where q' is 0 and k is 1, it is clear that the corresponding methods can be used for the preparation of other compounds of formula VI.

When R is⁷、R⁷'and A' contain functional groups, which can be suitably protected by methods recognized by those skilled in the art, see, e.g., Bodanzky or Greene cited above.

Formation of macrocyclic Compounds

Wherein the alkylene chain is taken from R⁷/R⁷Extended cycloalkyl to Rx, Rd or R¹¹The compounds according to the invention, which thus form the macrocycle, can be prepared as described below. The appropriate P1, P2 and P3 building blocks or precursors thereof are coupled together using the strategy described above followed by a ring closure reaction (macrocyclization). The substituents W-R of said P2 structural unit before or after macrocycle formation⁸Which may be incorporated via the Mitsunobu reaction as described above, or the group may be treated with a desired substituted proline analogue or carbocyclic ring. For the slave R⁷/R⁷' cycloalkyl extends to R¹¹The macrocyclic structure of (a) P3 amino acids containing appropriate side chains may be prepared as described in WO 00/59929.

A general route for the synthesis of macrocyclic compounds is shown in scheme 17, which illustrates the application of the process to compounds having spiro-cyclopropyl P1, wherein the macrocycle contains a P3 side chain.

Scheme 17

Acid derivative (17a) is coupled with the appropriate acid-protected amino acid (17b) using standard peptide coupling conditions as described above, to form (17 c). Macrocycle formation is then carried out via olefin metathesis using a Ru-based catalyst such as that reported in Miller, s.j., Blackwell, h.e.; grubbs, r.h.j.am.chem.soc.118, (1996), 9606-; kingsbury, j.s., Harrity, j.p.a.bonitatebus, p.j., Hoveyda, a.h., j.am.chem.soc.121, (1999), 791-. It should also be recognized that catalysts containing other transition metals (such as Mo) may also be used for this reaction. The double bond is optionally reduced and/or the ethyl ester is hydrolyzed by standard hydrogenation methods and/or standard hydrolysis methods, respectively, well known in the art. Alternatively, the methyl ester may be selectively hydrolyzed and then coupled to R by standard peptide coupling conditions¹⁶-G-P4 building block coupling. The macrocyclization step described in scheme 17 can also be applied to the corresponding carbocyclic analogs as described above. When the linker contains a nitrogen atom, ring closure may be effected by reductive amination as described in WO 00/59929.

Macrocyclic compounds in which the cyclopropyl moiety is absent from the P1 moiety, i.e. the macrocycle is directly linked to R by a peptidic backbone⁷Compounds with adjacent carbon atoms extended can be prepared using the methods described herein. Examples of where 5-membered cycloalkyl derivatives are useful as P2 scaffolds are shown in scheme 18.

Scheme 18

The amide derivative (18c) was obtained by coupling the appropriate allylglycine derivative (18a) to the acid functionality of the P2 scaffold (18b) using standard peptide coupling conditions. The ester group is hydrolyzed and then subjected to a peptide coupling reaction with an olefin-substituted amino acid (18Ad), thereby providing an amide compound (18 e). The macrocyclic compound (18f) is then given by ring closure metathesis using, for example, Hoveyda-Grubbs catalyst.

Although scheme 18 represents a synthetic sequence using a P2 scaffold with unsubstituted hydroxyl groups, it is clear that R is⁸The substituent may be introduced at any convenient step of the synthesis, for example as described in schemes 9 and 10, or it may be introduced after the metathesis reaction, i.e. on compound 18f, using any of the methods described herein.

Building blocks for the preparation of compounds wherein the macrocycle extends from the amide nitrogen atom of the P3 fragment (i.e. Rx in formula VI is J) or for the preparation of compounds wherein the P3 and P4 fragments are absent (i.e. m and n are 0 and G is NRj in formula VI) may generally be prepared as outlined in scheme 18B.

Scheme 18B

Carbamate 18Ba, which is commercially available or can be readily prepared, for example, by reacting the desired alkylamine with di-tert-butyl dicarbonate, can be reacted with the appropriate omega-unsaturated alcohol under Mitsunobu conditions to form the alkylated carbamate (18 Bb). The 18Bb is subjected to acidic conditions, for example treatment with trifluoroacetic acid in a solvent such as dichloromethane, to give the free amine (18Bc) which can be attached to the P2 fragment using any of the previously described strategies.

Macrocyclic structures containing hydrazine groups, i.e. formula VI where T is NRd or m and N are 0 and G is NRjNRj, can be prepared by attaching an appropriate N-alkylated carbazate derivative to the P2 fragment. Alkylated carbazate derivatives may be prepared, for example, as described in scheme 19.

Scheme 19

The aldehyde (19b) is formed by oxidation of the appropriate alcohol (19a) by a suitable oxidation process, for example using N-methylmorpholine oxide and tetrapropylammonium ferulate in a solvent such as dichloromethane. Reductive alkylation of t-butyl carbazate with the resulting aldehyde provides the desired N-alkylated structural unit (19 c). In addition, any desired hydrazine derivative other than t-butyl carbazate may be used in the reaction with the aldehyde 19b, such as morpholin-1-ylamine or piperidin-1-ylamine, and the like.

Scheme 20 illustrates a synthetic sequence of building blocks suitable for preparing compounds in which the "external" nitrogen atom of the hydrazine group is alkylated with an omega-unsaturated alkyl chain suitable for subsequent macrocyclization, or any other suitable alkyl.

Scheme 20

An appropriately protected hydrazine derivative, such as tert-butyl (1, 3-dioxo-1, 3-dihydro-isoindol-2-yl) -carbamate (20a), which can be readily prepared by one skilled in the art, is reacted with the desired alcohol R-OH under Mitsunobu conditions to form the N-alkylated hydrazine compound (20 b). Removal of the phthalimido group may be achieved by treatment with hydrazine or a derivative thereof, such as hydrazine hydrate or hydrazine acetate, to form the carbazate ester (20 c). The resulting primary amine can then be attached to the desired P2 fragment using any of the methods previously described, to give 20d, or it can be further alkylated using, for example, the reductive amination method described in scheme 19, followed by coupling to the P2 fragment previously described, to give 20 e.

Scheme 21 illustrates the coupling of a hydrazine containing P3 building block to a cyclopentane scaffold, followed by macrocyclization.

Scheme 21

The carbazate derivative (21b) was coupled to the P2-P1 building block (21a) using standard peptide coupling conditions to form intermediate (21 c). (21c) is ring-closed by olefin metathesis as depicted in scheme 18, to give the macrocyclic compound (21 d).

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 the amino group from undesirable reactions during the synthetic process. Commonly used N-protecting Groups are disclosed in Greene, "Protective Groups in Organic Synthesis" (John Wiley & Sons, New York, 1981), which is incorporated herein by reference. The N-protecting group includes acyl groups such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthaloyl, o-nitrophenoxyacetyl, α -chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl and 4-nitrobenzoyl and the like; sulfonyl groups such as benzenesulfonyl and p-toluenesulfonyl, and the like; carbamate-form 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, 1- (p-biphenyl) -1-methylethoxycarbonyl, alpha-dimethyl-3, 5-dimethoxybenzyloxycarbonyl, diphenylmethoxycarbonyl, tert-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, trityl, benzyloxymethyl, and the like; and silyl groups such as trimethylsilyl and the like. Preferred N-protecting groups include Fmoc, formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, phenylsulfonyl, benzyl, t-Butoxycarbonyl (BOC), and benzyloxycarbonyl (Cbz).

As used herein, "hydroxyl-protecting group" refers to a substituent that protects a hydroxyl group from undesirable reactions during the synthetic process, such as those O-protecting Groups disclosed in Greene, "Protective Groups Inorganic Synthesis" (John Wiley & Sons, New York (1981)). Hydroxy protecting groups include substituted methyl ethers, for example, methoxymethyl, benzyloxymethyl, 2-methoxyethoxymethyl, 2- (trimethylsilyl) ethoxymethyl, tert-butyl and other lower alkyl ethers, lower alkyl such as isopropyl, ethyl and, in particular, methyl, benzyl and trityl; tetrahydropyranyl ether; substituted ethers, such as 2, 2, 2-trichloroethyl ether; silyl ethers such as trimethylsilyl ether, t-butyldimethylsilyl ether and t-butyldiphenylsilyl ether; and esters prepared by reacting a hydroxyl group with a carboxylic acid, such as acetates, propionates, and benzoates, among others.

In treating conditions caused by flaviviruses, such as HCV, the compounds of formula I are typically administered in amounts to achieve plasma concentrations of about 100-5000 nM, such as 300-2000 nM. This corresponds to a dosage rate of 0.01-10 mg/kg/day, preferably 0.1-2 mg/kg/day, of administration, depending on the bioavailability of the formulation. For normal adults, the dosage rate is typically about 0.05 to 5g, preferably 0.1 to 2g, such as 500 to 750mg, per day in one to four dosage units per day. As with all medicaments, the dose rate will vary with the weight and metabolic condition of the patient and the severity of the infection, and will need to be adjusted according to the accompanying medication.

Based on good prescription practices for antiviral therapy, the compounds of formula I are generally co-administered with other HCV therapeutic agents to avoid the generation of drug escape mutants. Examples of such additional HCV antiviral therapeutic agents include ribavirin, interferons (including pegylated interferons). In addition, a number of nucleoside analogs and protease inhibitors are in clinical or preclinical research settings and are suitable for co-administration with the compounds of the present invention.

Accordingly, another aspect of the present invention provides a composition comprising a compound of formula I and at least one other HCV antiviral agent in a universal dosage unit, such as any of the dosage forms described below, but in particular a tablet or capsule for oral administration or a liquid suspension or a solution for oral or injectable use. Another aspect of the present invention provides a method of treating or preventing flavivirus (such as HCV) infection, comprising: the compound of formula I and at least one other HCV antiviral agent are administered sequentially or simultaneously. A related aspect of the invention provides a patient pack comprising: a first pharmaceutical composition, preferably in unit dosage form, of a compound of formula I, and a second pharmaceutical composition, also typically in unit dosage form and typically in separate containers in a patient pack, of a second HCV antiviral agent. The patient pack is also suitably provided with instructions printed on the package or on 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 prone to superinfection with other infectious diseases. Accordingly, another aspect of the invention provides combination therapies comprising co-formulating the compound of the invention with at least one other anti-infective agent in the same dosage unit or co-packaged. The compounds of the present invention are administered simultaneously or sequentially with at least one other anti-infective agent, generally at dosages equivalent to the dosages of the agents involved in the monotherapy. However, certain anti-infective agents may induce a synergistic effect, which results in the administration of a lower dose of one or both active ingredients 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 be administered in lower doses.

Common co-infections or superinfections with HCV include hepatitis b virus or HIV. Accordingly, the compounds of the present invention are advantageously co-administered (either in the same dosage unit, or in co-packaged or separately prescribed dosage units) with at least one HIV antiviral agent and/or at least one HBV antiviral agent.

Representative HIV antivirals include NRTI such as alovudine (FLT), zidovudine (AZT, ZDV), stavudine (D4T, Zerit), zalcitabine (ddC), didanosine (ddI, Videx), abacavir (ABC, Zigen), lamivudine (3TC, Epivir), emtricitabine (FTC, Emtriva), racevir (racemic FTC), Adefovir (ADV), entacavir (BMS 200475), alovudine (FLT), tenofovir disoproxil (TNF, Viread), amdoxavir DAPD), D-D4FC (DPC-817),. dOTC (ShiSPD 754), elvucitabine (AchilliaceAchillion ACH-126443), BCH 36 (RV) set 756, Racivir, GS, INK 40, purine-7340, purine-3K-20, D-3 ' -D-3 ', 3 ' -D prodrug such as guanosine-3-D, pyr (FLD, D-210, adefovir-D-3 ', 3 ' -D-210, pharmasset DPC-817).

Representative NNRTIs include delavirdine (Rescriptor), efavirenz (DMP-266, Sutiva), nevirapine (BIRG-587, Viramune), (+) Cikara Nalimide A and B (advanced Life sciences), caspovirin (AG1549 fS-1153; Pfizer), GW-695634 (GW-8248; GSK), MIV-150(Medivir), MV026048 (R-1495; Medivir AB/Roche), NV-0522 (IdenixPharm), R-278474(Johnson & Johnson), RS-1588 (Idenixperm), TMC-120/125(Johnson & Johnson), TMC-125 (R-165335; Johnson & Johnson), Johnson-389, (Bioshim) and Yam 2152152152158.

Representative HIV protease inhibitors include PA-457(Panacos), KPC-2(Kucera Pharm.), 5HGTV-43(Enzo Biochem), amprenavir (VX-478, Agenerase), atazanavir (Reyataz), indinavir sulfate (MK-639, Crixivan), Lexiva (calcium fosamprenavir, 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 MiningHolding), PfAG-1859 (Pfizer), DPC-681/684 (GS, Germany 338), Ginelein I-224272), KNI-194272 (Min-P-100), Naphth-P-35 (Biophyr P-100P), Naringner P-6 (Naviivan), and so, P-1946(Procyon Biopharma), R-944(Hoffmann-LaRoche), RO-0334649(Hoffmann-LaRoche), TMC-114(Johnson & Johnson), VX-385(GW 640385; GSK/Vertex), VX-478 (Vertex/GSK).

Other HIV antivirals include entry inhibitors including fusion inhibitors, CD4 receptor inhibitors, CCR5 co-receptor inhibitors, and CXCR⁴A co-receptor inhibitor, or a pharmaceutically acceptable salt or prodrug thereof. Examples of entry inhibitors are AMD-070(AMD 11070; AnorMed), BlockAide/CR (ADVENTRXRX 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), PRO542(Progenics Pharm.), SCH-D (SCH-417690; Schering-Pluough), T-1249 (R724; Roche/Trimeris), TAK-220(Takeda Chem.Ind.), TNX-355(Tanox) and UK-427,857 (Uizer). An example of an integrase inhibitor is L-870810 (Merck)&Co.)、c-2507(Merck&Co.) and S (RSC) -1838 (shionogi/GSK).

Examples of HBV antiviral agents include adefovir dipivoxil (hepsera), and in particular lamivudine and 2 ', 3' -dideoxy-3 '-Fluoroguanosine (FLG) and prodrugs thereof, such as MIV-210, the 5' -O-valinyl-L-lactyl prodrug of FLG. These latter HBV antivirals are particularly suitable because they also have anti-HIV activity.

Although the active agent may be administered alone, it is preferably present as an integral part of a pharmaceutical formulation. Such formulations will comprise an active agent as defined above together with one or more acceptable carriers or excipients and optionally other therapeutic ingredients. The carrier must be acceptable in the sense that it is compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

The formulations include those suitable for rectal, nasal, topical (including buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration, but preferably the formulations are oral administration formulations. The formulations may conveniently be presented in unit dosage form, for example as tablets and sustained release capsules, and may be prepared by any of the methods well known in the art of pharmacy.

The above method comprises the step of bringing into association the active agent as defined above with a carrier. Generally, the formulation is prepared by the following method: the active agent is homogenized and intimately associated with liquid carriers or finely divided solid carriers or both, and the product is then shaped if necessary. The present invention provides a process for preparing a pharmaceutical composition comprising combining or associating a compound of formula VI or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable carrier or excipient. If the production of a pharmaceutical formulation involves homogeneous mixing of pharmaceutical excipients and the active ingredient in salt form, it is generally preferred to use excipients which are not basic in nature, i.e. acidic or neutral excipients. The formulations of the present invention for oral administration may be presented as discrete units, such as capsules, sachets or tablets each containing a predetermined amount of the active agent; as a powder or granules; as a solution or suspension of the active agent in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion, as well as a pill, and the like.

For compositions for oral administration (e.g., tablets and capsules), the term "suitable carrier" includes excipients such as common excipients, for example binding agents, e.g., syrup, acacia, gelatin, sorbitol, tragacanth, polyvinylpyrrolidone (Povidone), methylcellulose, ethylcellulose, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, sucrose and starch; fillers and carriers, such as 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 metal stearates, stearic acid, glyceryl stearate, silicone oil, talc, oils and silica colloids. Flavoring agents such as peppermint, methyl salicylate, or cherry flavoring, and the like, may also be used. Colorants may desirably be added to make the dosage form easily identifiable. Tablets may also be coated by methods well known in the art.

Tablets may be prepared by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active agent in a free-flowing form such as powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surface active or dispersing agent. Molded tablets may be made by molding in suitable equipment a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated so as to provide sustained or controlled release of the active agent.

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

The compounds of formula VI may form salts, which form a further aspect of the invention. Suitable pharmaceutically acceptable salts of the compounds of formula I include salts of organic acids, particularly carboxylates, including but not limited to acetates, trifluoroacetates, lactates, gluconates, citrates, tartrates, maleates, malates, pantothenate, isethionates, adipates, alginates, aspartates, benzoates, butyrates, digluconates, cyclopentonates, glucoheptonates, glycerophosphates, oxalates, heptanoates, caproates, fumarates, nicotinates, palmitates, pectinates, 3-phenylpropionates, picrates, pivalates, propionates, tartrates, lactobonates, pivalates, camphorates, undecanoates and succinates, organic sulfonates such as methanesulfonates, ethanesulfonates, 2-isethionates, camphorsulfonates, 2-naphthalenesulfonates, camphorsulfonates, and succinates, Benzene sulfonate, p-chlorobenzene sulfonate, and p-toluene sulfonate; and inorganic acid salts such as hydrochloride, hydrobromide, sulfate, bisulfate, hemisulfate, thiocyanate, persulfate, phosphoric acid, and sulfonate. In addition, the present invention provides salts of the compounds of formula I, which may or may not be pharmaceutically acceptable salts, but which may be useful as synthetic intermediates, with the salt moiety being replaced or substituted as desired.

The present invention includes prodrugs of the compounds of formula I. Prodrugs of compounds of formula VI are those which release the compound of formula VI in vivo upon administration to a patient, typically upon hydrolysis in the digestive tract, liver or plasma. Prodrugs are typically pharmaceutically acceptable ethers and especially esters (including phosphates), amides or carbamates, or esters, of hydroxy functional groups, or of carboxy functional groups. Preferred pharmaceutically acceptable esters include alkyl esters (including acetate, butyrate, t-butyrate, octadecyl ester, and pivalate), phosphate esters, and sulfonate esters (i.e., those derived from RSO2OH, where R is lower alkyl or aryl). Pharmaceutically acceptable esters include lower alkyl ethers as well as ethers disclosed in WO00/47561, in particular methoxyaminoacyl and ethoxyaminoacyl.

The compounds of the present invention have multiple stereocenters, and the present invention provides racemates and enantiomers at each of these stereocenters.

In general terms, the amount of the solvent to be used,corresponding to the P3 and P4 side chains (i.e., R)¹⁵And/or R¹¹) The stereochemistry of the groups of (a) will correspond to the L-amino acid configuration, although the invention also provides D-isomers at one or both of these centers. It should be noted that although the nature of the E moiety is that P3 and P4 are generally transferred with one atom and indeed flip the peptide residue relative to conventional polypeptides, the L configuration remains active, assuming that P3 and P4 subsequently tilt the amine acid side chain to the other side as compared to conventional peptide substrates.

The stereochemistry of the backbone components of the cyclic P2 group (i.e., the carbonyl group bridging the P1 amide bond and the carbonyl group extending from P3) will generally correspond to L-proline. The stereochemistry of the P2 ring atom bound to W is generally as follows:

in which R is⁷And R⁷' together as a spirocycloalkyl group in the compounds of the invention, the aforementioned spiro-cycloalkyl group will typically comprise R on the spiro-cyclopropyl ring, which is cis to A^7′aSubstituent(s):

or

Or in reverse to A:

or

Desirably, the spiro carbon atom of the aforementioned spiro-cyclopropyl ring has the R configuration:

desirably, in the absolute configuration below, R on the spiro-cyclopropyl ring adjacent to a^7＇aThe substituents are in the cis direction:

particular preference is given to the variable R^7＇aIncluding ethyl groups, whereby the asymmetric carbon atoms in the 1 and 2 positions have the R, R configuration. Further preferred is R^7＇aIncluding vinyl groups, whereby the asymmetric carbon atoms in the 1 and 2 positions have the R, S configuration.

In the case where the compound of the invention is a macrocycle comprising a J group, J is preferably a diastereomer represented by part of the structure (i) or (ii):

or

J and amide Forward (i) J and A Forward (ii)

In particular wherein J and A are in the forward direction.

Detailed description of the embodiments

Various embodiments of the present invention will now be described by way of illustration only with reference to the following non-limiting examples.

Example 1

7-methoxy-2-phenyl-quinolin-4-ol (1).

To a stirred round bottom flask charged with toluene (100mL) were added ethyl benzoylacetate (18.7g, 97mmol) and m-anisidine (12g, 97 mmol). 4M HCl in dioxane (0.5mL) was then added and the reaction mixture was refluxed for 6 hours (140 ℃). The resulting mixture was co-evaporated with toluene. To the resulting crude mixture was added diphenyl ether (50mL) and the resulting mixture was heated to 280 ℃ for 2 h. When the theoretical amount of ethanol (6mL) was collected in the Dean Stark trap, the heating was stopped and the mixture was cooled to room temperature. Dissolving the crude mixture in CH₂Cl₂(100mL) and stirred for 30 minutes. The precipitate formed was filtered off and dried, thus giving 1(4.12g, 16.4mmol, 17%): light yellow powder.

¹H(300MHz，DMSO-D₆)：δ3.8(s，3H)，6.24(s，1H)，6.88-6.96(dd，1H，J＝9.07Hz，J＝2.47Hz)，7.19(d，1H，J＝2.19Hz)，7.56(t，3H，J＝2.19Hz)，7.8(dd，2H，J＝7.14Hz，J＝2.19Hz)，8.0(d，1H，J＝9.06Hz)； ¹³C(75.5MHz，DMSO-D₆)：δ55.3，99.6，106.9，113.1，119.1，126.4，127.5，128.8，130.2，134.1，142.2，149.4，161.8，176.4。

Example 2

(rac) -4-Oxocyclopent-2-ene-1, 2-dicarboxylic acid dimethyl ester (2)

Dimethyl (1R, 2S) -4-oxo-cyclopentane-1, 2-dicarboxylate (4.8g, 23.8mmol) and CuBr₂(11.9g, 53.2mmol) was dissolved in anhydrous THF (70mL) and the mixture was refluxed at 90C for 2 hours. The CuBr formed is filtered off and the organic phase obtained is concentrated. Mixing CaCO₃(2.7g, 27.2mmol) and DMF (70mL) were added and the mixture was held at 100 ℃ for 1 hour. The dark brown mixture was poured onto ice (35g) and the precipitate formed was filtered off. The resulting aqueous layer was extracted with ethyl acetate (1X 300mL + 3X 150 mL). The resulting organic phase was dried, filtered and concentrated. Purification by flash chromatography (toluene/EtOAc 9: 1) gave compound 2 as yellow crystals (2.1g, 45%).

Example 3

((1S, 4R) & (1R, 4S)) -4-hydroxy-cyclopent-2-ene-1, 2-dicarboxylic acid dimethyl ester (3)

To a cold solution (-30 deg.C) of Compound 2(3.18g, 16.1mmol) in methanol (23mL) was added NaBH₄(0.66g, 17.5 mmol). After nine minutes, excess NaBH was added by addition of brine (80mL)₄And (4) destroying. The resulting mixture was concentrated and extracted with ethyl acetate (4 × 80 mL). The resulting organic phase was dried, filtered and concentrated to give compound 3 as a yellow oil (3.0g, 92%).

Example 4

((1S, 4R) & (1R, 4S)) -4-hydroxy-cyclopent-2-ene-1, 2-dicarboxylic acid 2-methyl ester (4)

To an ice-cold solution of compound 3(3.4g, 22mmol) dissolved in dioxane and water (1: 1, 110mL) was added LiOH (0.52g, 22 mmol). After two and a half hours, the mixture was co-evaporated with toluene and methanol. Purification by flash chromatography (toluene/ethyl acetate 3: 1+ 1% HOAc) gave the title compound as off-white crystals (1.0g, 27%).

¹H-NMR(300MHz，CD₃OD)：δ1.78-1.89(m，1H)，2.70-2.84(m，1H)，3.56-3.71(m，1H)，3.76(s，3H)，4.81-4.90(m，1H)，6.76-6.81(m，1H)；¹³C-NMR(75.5MHz，CDCl₃)：δ38.0，48.0，52.4，75.7，137.0，146.2，165.0 178.4。

Example 5

((3S, 5R) & (3R, 5S)) -5- ((S) -1-tert-butoxycarbonyl-butylcarbamoyl) -3-hydroxy-cyclopent-1-enecarboxylic acid methyl ester (5)

To an ice-cold solution of compound 4(0.20g, 1.1mmol) and tert-butyl 2-amino-pentanoate (0.24g, 1.4mmol) in DMF (7mL) was added DIPEA (0.18g, 1.4mmol) and HATU (0.53g, 1.4 mmol). After two hours the above solution was concentrated and purified by column chromatography (toluene/ethyl acetate 3: 1). This gave the title compound as a yellow oil (0.22g, 63%).

¹H-NMR(300MHz，CDCl₃)：δ0.84-0.96(m，3H)，1.14-1.39(m，2H)，[(1.44&1.49)s，9H]，1.50-1.60(m，1H)，1.61-1.85(m，1H)，1.97-2.10(m，1H)，2.11-2.28(m，1H)，3.57-3.68(m，1H)，[(3.73&3.76)s，3H]，4.30-4.50(m，1H)，4.63-4.73(m，1H)，6.80-6.95(m，1H)，6.95- 7.00(m，1H)。

Example 6

((3S, 5R) & (3R, 5S)) -5- ((S) -1-tert-butoxycarbonyl-propylcarbamoyl) -3-hydroxy-cyclopent-1-enecarboxylic acid methyl ester (6)

Compound 4(141mg, 76mmol) was reacted according to the procedure described for the preparation of compound 5, using tert-butyl L-2-amino-n-butyrate instead of tert-butyl 2-amino-pentanoate, thus giving the title compound as a pale yellow oil (171mg, 69%).

¹H-NMR(300MHz，CDCl₃)：δ0.89-0.98(m，3H)，[(1.42&1.44)s，9H]，1.60-1.78(m，1H)，1.79-1.95(m，1H)，1.99-2.11(m，1H)，2.18-2.30(m，1H)，3.58-3.65(m，1H)，[3.75&3.78)s，3H]，4.22-4.39(m，1H)，4.61-4.66(m，1H)，6.77-6.90(m，1H)，6.91-6.92(m，1H).

Example 7

((3S, 5R) & (3R, 5S)) -5- ((1R, 2S) -1-tert-Butoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -3-hydroxy-cyclopent-1-enecarboxylic acid methyl ester (7)

Compound 4(50mg, 37mmol) was reacted according to the procedure described for the preparation of compound 5, using tert-butyl (1R, 2S) -1-amino-2-vinylcyclopropanecarboxylate instead of tert-butyl 2-amino-pentanoate, to afford the title compound as a pale yellow oil (50mg, 38%).

¹H-NMR(300MHz，CDCl₃)：δ[(1.38&1.42)s，9H]，1.75-1.83(m，1H)，2.00-2.21(m，3H)，3.55-3.63(m，1H)，[(3.77&3.82)s，3H]，4.20-4.38(m，1H)，4.65-4.80(m，1H)，5.13-5.20(m，1H)，5.22-5.38 (m，1H)，5.60-5.82(m，1H)，6.95-6.96(m，2H)。

Example 8

((3R, 5R) & (3S, 5S)) -5- ((S) -1-tert-butoxycarbonyl-butylcarbamoyl) -3- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-1-enecarboxylic acid methyl ester (8)

To an ice-cold solution of compound 5(0.23g, 0.67mmol) in anhydrous THF was added 7-methoxy-2-phenyl-quinolin-4-ol (0.22g, 0.88mmol) and triphenylphosphine (0.23g, 0.88 mmol). DIAD (0.19g, 0.92mmol) was then dissolved in THF (2mL) and the solution added dropwise to the above solution. After one hour the mixture was concentrated and purified by flash chromatography (toluene/ethyl acetate 3: 1). This gave the title compound as a white powder (0.30g, 77%).

¹H-NMR(300MHz，CDCl₃)：δ0,88-1.00(m，3H)，1.18-1.43(m，2H)，[(1.45&1.50)s，9H]，1.53-1.65(m，1H)，1.66-1.85(m，1H)，2.29-2.43(m，1H)，3.10-3.25(m，1H)，[(3.79&3.83)s，3H]，3.97(s，3H)，4.05-4.20(m，1H)，4.38-4.50(m，1H)，6.03-6.13(m，1H)，6.65-6.90(m，1H)，7.04-7.18(m，3H)，7.40-7.56(m，4H)，8.00-8.12(m，3H)。

Example 9

((3R, 5R) & (3S, 5S)) -5- ((S) -1-tert-butoxycarbonyl-propylcarbamoyl) -3- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-1-enecarboxylic acid methyl ester (9)

Compound 6(132mg, 40mmol) was reacted according to the procedure described for the preparation of compound 8, thereby affording the title compound as a yellow oil (137mg, 61%).

¹H-NMR(300MHz，CDCl₃)：δ0.83-0.98(m，3H)，[(1.42&1.44)s，9H]，1.65-1.78(m，1H)，1.80-1.97(m，1H)，2.30-2.40(m，1H)，3.05-3.20(m，1H)，[(3.78&3.80)s，3H]，3.94(s，3H)，3.95-4.01(m，1H)，4.38-4.44(s，1H)，6.05-6.15(m，1H)，6.80-6.94(m，1H)，7.02-7.15(m，3H)，7.38-7.55(m，4H)，7.97-8.18(m，3H)。

Example 10

((3R, 5R) & (3S, 5S)) -5- ((1R, 2S) -1-tert-butoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -3- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-1-enecarboxylic acid methyl ester (10)

Compound 7(41mg, 116mmol) was reacted according to the procedure described for the preparation of compound 8 to form the title compound as a yellow oil.

¹H-NMR(300MHz，CDCl₃)：δ1.52-1.57(m，1H)，1.58(m，9H)，1.80-1.83(m，1H)，2.00-2.17(m，1H)，2.20-2.38(m，1H)，3.20-3.37 (m，1H)，3.80(s，3H)，3.81-3-3.98(m，1H)，3.99(s，3H)，5.12-5.20(m，1H)，5.22-5.40(m，1H)，5.63-5.80(m，1H)，6.05-6-20(m，1H)，7.00-7.21(m，4H)，7.40-7.58(m，4H)，8.02-8.18(m，3H)。

Example 11

((3R, 5R) & (3S, 5S)) -5- ((S) -1-tert-butoxycarbonyl-butylcarbamoyl) -3- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-1-enecarboxylic acid (11)

Methyl ester 8(0.35g, 0.61mmol) was dissolved in dioxane/water (1: 1, 7mL) and LiOH (0.031g, 1.3mmol) was added thereto. The reaction was stirred overnight and then co-concentrated. This gave compound 11 as a brown powder as lithium salt (0.32g, 90%).

Example 12

((3R, 5R) & (3S, 5S)) -5- ((S) -1-tert-butoxycarbonyl-propylcarbamoyl) -3- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-1-enecarboxylic acid (12)

Compound 9(225mg, 40mmol) was reacted according to the procedure described for the preparation of compound 11 to form the title compound as a yellow salt (157mg, 72%).

Example 13

((3R, 5R) & (3S, 5S)) -5- ((1R, 2S) -1-tert-butoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -3- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-1-enecarboxylic acid (13)

Compound 10(35mg, 59mmol) was reacted according to the procedure described for the preparation of compound 11 to form the title compound as a yellow salt (33mg, 97%).

Example 14

(S) -2- { [ ((1S, 4S) & (1R, 4R)) -2- { (S) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2-methyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -butyric acid tert-butyl ester (14)

Acid 12(38.4mg, 0.070mmol) and methyl (2-amino-3-methyl-butyrylamino) -cyclohexylacetate (26.6mg, 0.098mmol) were dissolved in DMF (1.5mL) and cooled in an ice bath. DIPEA (17.1. mu.L, 0.098mmol) and HATU (37.4mg, 0.098mmol) were added thereto. After ninety minutes the mixture was co-concentrated with toluene and methanol and then purified by flash column chromatography (toluene/ethyl acetate 6: 1). Further purification was performed on HPLC (90% methanol + 0.2% TEA). The diastereomeric mixture 14 was concentrated to give a pale yellow oil (20.6mg, 37%). After freeze-drying it, compound 14 was collected as a white powder.

¹H-NMR(300MHz，CDCl₃)：δ0.93-1.02(m，9H)，1.03-1.25(m，4H)，1.44(s，9H)，1.65-1.86(m，9H)，2.05-2.10(m，1H)，2.22-2.40(m，1H)，3.05-3.20(m，1H)，3.77(s，3H)，3.98(s，3H)，4.18-4.22(m，1H)，4.38-4.60(m，3H)，6.01-6.10(m，1H)，6.61-6.70(m，2H)，6.80-6.85(m，1H)，7.05-7.18(m，2H)，7.40-7.58(m，5H)，8.00-8.13(m，3H).¹³C-NMR(75.5MHz，CDCl₃)：δ9.7，18.4，19.2，[25.9&26.1]，[28.2&28.5]，29.6，32.0，37.3，41.0，46.2，50.7，52.4，54.4，55.8，57.2，58.5，82.0，82.8，98.4，110.2，118.4，120.1，123.2，127.9，128.2，128.9，129.5，131.2，135.1，135.2，142.7，144.2，161.6，164.3，164.7，170.9，171.4，172.4.MALDI-TOF m/z 821.56[(M+Na)⁺C₄₅H₅₈N₄NaO₉ ⁺Calculated value of 821.41]。

Example 15

(S) -2- { [ ((1R, 4R) & (1S, 4S)) -2- { (R) -1- [ ((R) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2-methyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -butyric acid tert-butyl ester (15)

Compound 12(20mg, 37mmol) was reacted according to the procedure described for the preparation of compound 14, using methyl (2-amino-3-methyl-butyrylamino) - (R) -cyclohexylacetate instead of methyl (2-amino-3-methyl-butyrylamino) - (S) -cyclohexylacetate, to give the title compound as a white powder (19mg, 66%).

¹H-NMR(300MHz，CDCl₃)：δ0.91-0.98(m，3H)，0.99-1.10(m，6H)，1.11-1.38(m，4H)，[(1.43&1.45)s，9H]，1-45-1.94(m，9H)，2.05-2.18(m，1H)，2.22-2.40(m，1H)，3.16-3.24(m，1H)，3.77(s，3H)，3.98(s，3H)，4.04-4.18(m，1H)，4.36-4.57(m，3H)，6.00-6.08(m，1H)，6.13-6.21(m，1H)，6.62-6.70(m，1H)，6.81-6.85(m，1H)，7.05-7.18(m，3H)，7.41-7.57(m，4H)，8.02-8.13(m，3H).¹³C-NMR(75.5MHz，CDCl₃)：δ9.3，18.2，19.0，[25.5&25.9]，[28.0&28.3]，29.4，31.4，32.1，35.7，40.7，50.4，52.2，54.2，55.5，57.0，58.2，81.8，82.4，98.2，107.5，115.0，118.1，122.9，127.6，128.7，128.8，128.9，129.2，135.1，140.4，142.2，151.4，161.3，163.9，170.4，170.9，171.2，172.0.MALDI-TOF m/z 821.60[(M+Na)⁺C₄₅H₅₈N₄NaO₉ ⁺Calculated value of 821.41]。

Example 16

(S) -2- { [ ((3R, 5R) & (3S, 5S)) -5- ((S) -1-tert-Butoxycarbonylpropylcarbamoyl) -3- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -3-methylbutanoic acid methyl ester (16)

Compound 12(24mg, 44mmol) was reacted according to the procedure described for the preparation of compound 14, using D-valine methyl ester instead of (2-amino-3-methyl-butyrylamino) cyclohexylacetic acid methyl ester, to give the title compound as a white powder (27mg, 97%).

¹H-NMR(300MHz，CDCl₃)：δ0.82-0.99(m，9H)，[(1.42&1.44)s，9H]1.65-1.95(m，2H)，2.18-2.25(m，1H)，2.26-2.40(m，1H)，3.20-3.25(m，1H)，3.75(s，3H)，3.97(s，3H)，4.15-4.19(m，1H)，4.36-4.43(m，1H)，4.64-4.75(m，1H)，6.03-6.15(m，1H)，6.80-6.85(m，2H)，7.10-7.20(m，3H)，7.42-7.58(m，4H)，8.0-8.10(m，3H).¹³C-NMR (75.5MHz，CDCl₃)：δ9.7，[18.2&19.1]，25.7，[28.1&28.2]，32.0，35.6，50.4，52.4，54.5，55.7，57.6，81.7，82.7，98.4，107.7，115.2，118.4，123.2，127.8，129.0，129.2，129.5，134.8，135.0，140.4，142.5，151.6，159.6，[161.1&161.5]，164.6，171.1，172.2.MALDI-TOF m/z 682.51[(M+Na)⁺ C₃₇H₄₅N₃NaO₈ ⁺Calculated value of 682.31]。

Example 17

(S) -2- { [ ((1R, 4R) & (1S, 4S)) -2- { (S) -1- [ (2, 5-dimethoxy-phenyl) -ethyl-carbamoyl ] -2-methyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -butyric acid tert-butyl ester (17)

Compound 17(28.6mg, 59%) was prepared from compound 12(33mg, 60mmol) according to the procedure described for the preparation of compound 14, using 2-amino-N- (2, 5-dimethoxy-phenyl) -N-ethyl-3-methylbutanamide without using methyl (2-amino-3-methyl-butyrylamino) -cyclohexylacetate. This gave the title compound as a white powder.

¹H-NMR(300MHz，CDCl₃)：δ0.75-0.95(m，9H)1.05-1.18(m，3H)，[(1.42&1.44)s，9H]，1.60-1.95(m，3H)，2.20-2.40(m，1H)，3.20-3.34(m，1H)，3.60-3.80(m，2H)，[3.62-3.65(m，3H)]，[3.79-3.82(m，3H)]，3.98(s，3H)，4.02-4-18(m，1H)，4.30-4.44(m，2H)，6.05-6.18(m，1H)，6.60-6.63(m，1H)，6.77-6.80(m，2H)，6.85-6.93(m，2H)，7.12-7.20(m，2H)，7.35-7.60(m，5H)，8.02-8.20(m，3H).¹³C-NMR(75.5MHz，CDCl₃)：δ[9.6&9.7]，[12.5&12.8]，[17.1&17.5]，[19.4&19.5]，25.6，[28.0&28.1]，32.4，35.8，43.0，44.3，[50.2&50.3]，54.3，[54.8&55.0&55.2&55.5]，[55.6&55.7&55.9&56.0]，81.7，82.8，98.4， 106.9，[112.4&112.5]，113.7，115.0，115.2，115.9，116.3，118.4，[123.0&123.1]，[127.7&127.8]，128.8，128.9，129.5，130.1，[134.1&134.2]，142.6，149.1，149.4，153.4，158.9，[161.4&161.6]，[163.2&163.5]，170.9，[171.3&171.5]，172.3.MALDI-TOF m/z 831.62[(M+Na)⁺C₄₆H₅₆N₄NaO₉ ⁺Calculated value of 831.39]。

Example 18

(S) -2- { [ ((1R, 4R) & (1S, 4S)) -2- { (S) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2, 2-dimethyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -butyric acid tert-butyl ester (18)

Compound 18(16.1mg, 26%) was prepared from compound 12(43.2mg, 0.077mmol) according to the procedure described for the preparation of compound 14, using (2-amino-3, 3-dimethyl-butyrylamino) -cyclohexyl-acetic acid methyl ester instead of (2-amino-3-methyl-butyrylamino) -cyclohexyl-acetic acid methyl ester. Flash column chromatography was performed on toluene/ethyl acetate 3: 1 instead of 6: 1, thereby giving the title compound as a white powder.

¹H-NMR(300MHz，CDCl₃)：δ0.77-0.83(m，3H)，[(0.92&0.93)s，9H]0.94-1.20(m，4H)，[(1.36&1.38)s，9H]，1.42-1.76(m，8H)，2.20-2.38(m，1H)，2.81-2.96(m，1H)，3.20-3.22(m，1H)，2.78(s，3H)，[(3.83&3.85)s，3H]，3.97-4.02(m，1H)，4.17-4.21(m，1H)，4.22-4.37(m，2H)，5.85-5.97(m，1H)，[6.76-6.78(m，0.5H)]，[6.80-6.82(m，0.5H)]，6.98-7.05(m，3H)，7.23-7.41(m，6H)，7.82-7.99(m，3H).¹³C-NMR(75.5MHz，CDCl₃)：δ[9.4&9.5]，[25.4&25.5]，25.8，[26.5&26.6]，[27.9&28.0]，[28.4&28.5]，29.3，[35.4&35.7]，[36.0&36.4]，[40.5&40.7]，[50.2&50.5]，[52.1&52.2]，[54.1&54.3]，55.5，[57.0& 57.3]，[60.4&60.7]，[81.8&82.0]，[82.4&82.5]98.1，107.5，115.0，118.1，123.0，127.5，128.7，128.8，129.2，134.9，135.8，141.9，142.5，151.3，159.4，[160.9&161.3]，[163.7&163.9]，[169.9&170.0][170.0&171.3]，[172.5&172.4].MALDI-TOF m/z 835.68[(M+Na)⁺C₄₆H₆₀N₄NaO₉ ⁺Calculated value of 835.43]。

Example 19

(S) -2- { [ (1R, 4R) -2- { (S) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2-methyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -pentanoic acid tert-butyl ester (19a) and

(S) -2- { [ (1S, 4S) -2- { (S) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2-methyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -pentanoic acid tert-butyl ester (19b)

Acid 11(0.051g, 0.087mmol) and methyl (2-amino-3-methyl-butyrylamino) -cyclohexylacetate (0.054g, 0.21mmol) were dissolved in DMF (1.5mL) and cooled in an ice bath. DIPEA (16mg, 0.12mmol) and HATU (47mg, 0.13mmol) were added thereto. After two and a half hours the mixture was co-concentrated with toluene and methanol and then purified by flash column chromatography (toluene/ethyl acetate 3: 1). Further purification was performed on HPLC (90% methanol + 0.2% TEA). Thus, after co-concentration, two diastereomers 19a (9.4mg, 13%) and 19b (5.3mg, 7%) were given as light yellow thick slurries. After freeze-drying 19a and 19b were collected as white powders.

¹H-NMR(300MHz，CDCl₃)：δ0.86-0.93(m，3H)，0.94-1.00(m， 6H)，1.00-1.41(m，7H)，1.46(s，9H)，1.50-1.88(m，8H)，2.05-2.20(m，1H)，2.20-2.37(m，1H)，3.12-3.25(m，1H)，3.73(s，3H)，3.97(s，3H)，4.05-4.20(m，1H)，4.40-4.55(m，3H)，6.02-6.18(m，1H)，6.30(d，J＝8.52Hz，1H)，6.63(s，1H)，6.76(d，J＝8.51Hz，1H)，7.06-7.16(m，2H)，7.42-7.56(m，5H)，8.00-8.12(m，3H)；¹³C-NMR(75.5MHz，CD₃OD)：δ14.0，18.4，19.3，26.1，28.3，28.5，29.7，31.9，34.9，36.0，41.0，50.7，52.4，53.3，55.7，57.2，58.6，82.0，82.7，98.4，105.7，107.7，115.2，118.4，123.2，125.3，127.9，129.0，129.1，135.1，138.0，142.4，151.6，159.4，161.6，164.3，170.7，171.2，172.3.19b：¹H-NMR(300MHz，CDCl₃)：δ0.90-1.04(m，9H)，1.04-1.43(m，7H)，1.47(s，9H)，1.50-1.87(m，8H)，2.10-2.27(m，1H)，2.33-2.45(m，1H)，3.10-3.20(m，1H)，3.73(s，3H)，3.96(s，3H)，4.02-4.10(m，1H)，4.36-4.53(m，3H)，6.00-6.16(m，1H)，6.30(d，J＝8.52Hz，1H)，6.73(s，1H)，6.86(d，J＝7.96Hz，1H)，7.08-7.16(m，2H)，7.36-7.56(m，5H)，8.03-8.11(m，3H).¹³C-NMR(75.5MHz，CD₃OD)：δ14.0，18.6，19.2，26.1，28.2，28.7，29.7，34.5，36.1，36.6，40.8，50.5，52.4，53.4，55.7，57.3，59.1，64.8，82.3，98.4，105.8，107.8，115.3，118.4，123.2，127.8，129.0，129.4，135.2，142.2，144.9，151.0，151.6，159.2，164.3，164.3，170.2，171.6，171.9。

Example 20

(S) -2- { [ (1R, 4R) -2- { (R) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2, 2-dimethyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -pentanoic acid tert-butyl ester (20a) and

(S) -2- { [ (1S, 4S) -2- { (R) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2, 2-dimethyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -pentanoic acid tert-butyl ester (20b)

The method A comprises the following steps: carboxylic acid 11(57mg, 0.10mmol) was dissolved in hot (50 ℃ C.) dry THF (2 mL). To this was added (2-amino-3, 3-dimethyl-butyrylamino) -cyclohexyl-acetic acid methyl ester (50mg, 0.12mmol), DIPEA (30mg, 0.23mmol), DCC (25mg, 0.12mmol) and HOBt (17mg, 13 mmol). After two hours the mixture was concentrated and added to a short column (toluene/ethyl acetate 1: 3+ 3% acetic acid). It was then further purified on HPLC using 90% methanol + 0.2% TEA. The diastereomeric products are not isolated. After HPLC, the resulting solution was co-concentrated with toluene and methanol to give compound 20(28mg, 34%).

The method B comprises the following steps: to an ice-cold solution of compound 11(60mg, 0.10mmol) and (2-amino-3, 3-dimethyl-butyrylamino) -cyclohexyl-acetic acid methyl ester (42mg, 0.15mmol) were added DIPEA (19mg, 0.15mmol) and HATU (62mg, 0.16 mmol). After two and a half hours, the mixture is concentrated and purified by column chromatography (toluene/ethyl acetate 3: 1). The resulting mixture of diastereomers was separated by HPLC (90% methanol + 0.2% TEA). This gave 20a (6mg, 6%) and 20b (9mg, 10%).

20a：¹H-NMR(300MHz，CDCl₃)：δ0.82-0.90(m，3H)，1.01(s，9H)，1.05-1.40(m，7H)，1.46(s，9H)，1.50-1.80(m，8H)，2.20-2.35(m，1H)，3.07-3.25(m，1H)，3.73(s，3H)，3.97(s，3H)，4.11(d，J＝7.96Hz，1H)，4.38-4.52(m，3H)，6.03-6.12(m，1H)，6.24(d，J＝8.79Hz，1H)，6.63(s，1H)，6.82(d，J＝9.06Hz，1H)，7.07-7.27(m，2H)，7.36(d，J＝7.96Hz，1H)，7.41-7.55(m，4H)，8.01-8.10(m，3H)；¹³C-NMR(75.5MHz，CD₃OD)：δ14.0，18.8，26.1，26.8，28.2，28.6，29.6，34.9，35.6，36.2，40.9，50.7，52.4，53.3，55.7，57.3，60.8，82.0，82.7，98.4，105.2，107.7，115.2，118.4，123.2，127.9，129.0，129.4，131.1，135.1，138.4，142.4，153.3，159.6，161.6，164.2，170.1，171.3，172.2.20b：¹H-NMR(300MHz，CDCl₃)：δ0.90-0.98(m，3H)，1.04(s，9H)，1.08-1.40(m，7H)，1.44(s，9H)，1.55-1.90(m，8H)，2.20-2.38(m，1H)，3.10-3.22(m，1H)，3.73(s，3H)，3.97(s， 3H)，4.02-4.15(m，1H)，4.35-4.48(m，3H)，6.00-6.08(m，1H)，6.72(s，1H)，6.90(d，J＝9.06Hz，1H)，7.09-7.20(m，3H)，7.44-7.55(m，5H)，8.03-8.11(m，3H)。

Example 21

(1R, 2S) -1- { [ ((1R, 4R) & (1S, 4S)) -2- { (S) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2, 2-dimethyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -2-vinyl-cyclopropanecarboxylic acid tert-butyl ester (21)

Acid 13(35mg, 0.060mmol) and methyl (2-amino-3, 3-dimethyl-butyrylamino) -cyclohexylacetate (22mg, 0.080mmol) were dissolved in anhydrous THF (1.5mL) and warmed to 50 ℃. HOBt (11mg, 0.080mmol) and DCC (31mg, 0.15mmol) were added thereto. After one hour the mixture was co-concentrated with toluene and methanol and then purified by flash column chromatography (toluene/ethyl acetate 1: 1). It was further purified on HPLC (80% methanol + 0.2% TEA). Diastereomeric mixture 21 was concentrated to give a pale yellow oil (26.4mg, 53%). After freeze-drying it, compound 21 was collected as a white powder.

¹H-NMR(300MHz，CDCl₃)：δ[(0.98&1.00)，s，9H]，1.01-1.38(m，5H)，[(1.39&1.40)s，9H]，1.52-1.63(m，4H)，1.65-1.80(m，4H)，1.90-2.05(m，1H)，2.20-2.40(m，1H)，3.02-3.20(m，1H)，[(3.66&3.67)s，3H)，3.98(s，3H)，3.99-4.02(m，1H)，4.30-4.45(m，2H)，5.05-5.11(m，1H)，5.20-5.30(m，1H)，5.60-5.81(m，1H)，6.03-6.17(m，1H)，6.77-6.82(m，1H)，6.95-7.22(m，5H)，7.40-7.50(m，4H)，8.01-8.10(m，3H).¹³C-NMR(75.5MHz，CDCl₃)：δ22.3，[25.7&25.8]，[26.4& 26.5]，[28.0&28.4]29.2，32.7，33.3，[35.3&35.4]，36.0，[40.2&40.3]，40.7，52.0，55.4，[57.2&57.4][60.4&60.5]，[87.6&87.7]，[82.3&82.5]，98.4，107.0，114.9，[117.4&117.5]，118.1，122.9，127.6，128.6，128.9，129.2，[133.6&133.8]，135.9，136.9，140.1，[141.4&141.6]，151.1，159.6，[160.9&161.3]，[164.2&164.6]，168.9，170.3，[172.1&172.6].MALDI-TOFm/z 859.77[(M+Na)⁺C₄₈H₆₀N₄NaO₉ ⁺Calculated value of 859.43]。

Example 22

(S) -2- { [ (1R, 4R) -2- { (R) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2, 2-dimethyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -pentanoic acid (22a) and (S) -2- { [ (1S, 4S) -2- { (R) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2, 2-dimethyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinoline- 4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -pentanoic acid (22b)

Tert-butyl ester 20(28mg, 0.034mmol), TES (8.7mg, 0.075mmol), DCM (1mL) and TFA (1mL) were combined in a round bottom flask. After two hours the mixture was concentrated and the resulting diastereomers were separated on HPLC using 65% methanol + 0.2% TEA as the mobile phase. This gave 22a (15mg, 55%) and 22b (12mg, 45%) as a pale yellow thick slurry. After freeze-drying it, the title compound was collected as a white powder.

22a：[α]²²D+155.8；¹H-NMR(300MHz，CD₃OD)：δ0.90-0.97(m，3H)，1.03(s，9H)，1.05-1.50(m，7H)，1.50-1.80(m，8H)，2.43-2.55(m，1H)，2.77-2.90(m，1H)，3.68(s，3H)，3.96(s，3H)，4.20-4.30(m，2H)，4.31-4.40(m，1H)，4.45-4.50(m，1H)，6.03-6.11(m，1H)，6.98(s， 1H)，7.12-7.19(m，1H)，7.36(s，1H)，7.41(d，J＝2.2Hz，1H)，7.50-7.60(m，3H)，8.03-8.10(m，3H)：¹³C-NMR(75.5MHz，CD₃OD)：δ13.1，19.1，26.1，28.7，28.9，29.5，34.3，34.8，35.9，40.1，50.8，51.2，54.8，55.0，57.9，60.7，83.5，99.1，106.0，115.2，118.2，123.3，127.8，128.0，128.7，128.8，129.7，135.2，139.8，143.7，150.6，160.1，162.2，165.2，171.7，172.2，173.4.22b：[α]²²D-72，3；¹H-NMR(300MHz，CD₃OD)：δ0.90-0.97(m，3H)，1.02(s，9H)，1.07-1.35(m，7H)，1.53-1.90(m，8H)，2.46-2.61(m，1H)，2.76-2.88(m，1H)，3.69(s，3H)，3.96(s，3H)，4.15-4.35(m，2H)，4.37-4.41(m，1H)，4.42-4.47(m，1H)，6.02-6.12(m，1H)，7.02(s，1H)，7.16(dd，J＝2.47，9.34Hz，1H)，7.32(s，1H)，7.40(d，J＝2.47Hz，1H)，7.48-7.58(m，3H)，8.03-8.12(m，3H)；¹³C-NMR(75.5MHz，CD₃OD)：δ13.0，18.8，25.9，26.0，28.8，29.4，34.2，34.8，36.3，39.9，48.8，50.5，51.1，54.8，57.9，60.5，82.8，99.0，106.0，115.1，118.2，123.1，127.8，127.9，128.7，129.0，129.5，136.7，139.8，142.8，150.6，160.1，162.0，162.2，164.7，172.1，173.5。

Example 23

(S) -2- { [ (1R, 4R) -2- { (R) -1- [ ((R) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2-methyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -butyric acid (23a) and (S) -2- { [ (1S, 4S) -2- { (R) -1- [ ((R) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2-methyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yl Oxy) -cyclopent-2-enecarbonyl ] -amino } -butyric acid (23b)

Compound 23a (6.6mg, 50%) and compound 23b (1.3mg, 10%) were prepared from compound 15(14mg, 0.018mmol) according to the procedure for preparing compounds 22a and 22 b. This gave the title compound as a white powder.

23a：¹H-NMR(300MHz，CD₃OD)：0.88-1.02(m，9H)，1.02-1.40(m，7H)，1.55-1.97(m，6H)，2.01-2.10(m，1H)，2.38-2.52(m，1H)，2.88-3.00(m，1H)，3.77(s，3H)，3.98(s，3H)，4.08-4.20(m，1H)，4.22-4.40(m，3H).6.03-6.18(m，1H)，6.86-6.99(m，1H)，7.08-7.20(m，1H)，7.23(s，1H)，7.40-7.43(m，1H)，7.45-7.70(m，3H)，8.02-8.20(m，3H). ¹³C-NMR(75.5MHz，CD₃OD)：δ9.0，17.6，18.2，24.5，25.3，28.1，28.8，30.9，35.4，39.4，49.6，51.1，54.7，57.2，58.0，82.4，98.5，105.5，114.5，117.7，122.7，127.2，127.3，128.2，129.0，135.6，136.4，141.7，149.9，159.5，161.2，161.4，164.0，171.0，171.7，172.4.23b：¹H-NMR(300MHz，CD₃OD)：δ0.9-1.20(m，9H)，1.21-1.53(m，7H)，1.55-1.93(m，6H)，2.05-2.20(m，1H)，2.41-2.50(m，1H)，2.96-3-05(m，1H)，3.77(s，3H)，4.00(s，3H)，4.05-4.40(m，4H)，6.05-6.18(m，1H)，6.90-6.95(m，1H)，7.05-7.22(m，2H)，7.50-7.65(m，4H)，8.01-8.16(m，3H)。

Example 24

(S) -2- { [ ((1R, 4R) & (1S, 4S)) -2- { (S) -1- [ ((S) -carboxy-cyclohexyl-methyl) -carbamoyl ] -2-methyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -butyric acid (24)

Tert-butyl 14(13.4mg, 0.017mmol), TES (4.83mg, 0.042mmol), DCM (2mL) and TFA (2mL) were combined in a round bottom flask. After one hour the mixture was concentrated and purified by HPLC using 65% methanol + 0.2% TEA as mobile phase. This gave 24 as a pale yellow thick syrup (4.3mg, 34%). After freeze-drying it, compound 24 was collected as a white powder.

¹H-NMR(300MHz，CD₃OD)：δ0.91-0.99(m，9H)，1.00-1.28(m，4H)，1.55-1.78(m，9H)，1.92-1.95(m，1H)，2.00-2.05(m，1H)，2.93-3.01(m，1H)，3.75(s，3H)，3.97(s，3H)，4.10-4.40(m，4H)，6.05-6.15(m，1H)，6.88-6.94(m，1H)，7.05-7.10(m，2H)，7.41-7.43(m，1H)，7.44-7.55(m，2H)，8.62-8.68(m，1H)，8.69-8.79(m，1H)，7.97-8.05(m，2H). ¹³C-NMR(75.5MHz，CD₃OD)：δ9.2，18.5，25.5，[29.0&29.2]，[30.0&30.5]，35.3，37.7，39.7，46.2，50.0，[51.4&51.5]，53.6，55.1，57.1，58.4，83.1，98.9，104.9，114.6，118.3，123.0，123.4，127.5，128.4，128.5，129.7，135.0，142.1，145.7，146.2，159.2，161.9，164.3，171.5，171.9，172.2.MALDI-TOF m/z 791.27[(M+K)⁺C₄₂H₄₈KN₄O₉ ⁺Calculated value of 791.31]。

Example 25

(S) -2- { [ ((3R, 5R) & (3S, 5S)) -5- ((S) -1-carboxy-propylcarbamoyl) -3- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -3-methylbutanoic acid methyl ester (25)

Compound 25(8.0mg, 60%) was prepared from compound 16(13.8mg, 0.022mmol) according to the method for preparing compound 24, thereby giving the title compound as a white powder.

¹H-NMR(300MHz，CD₃OD)：δ0.83-1.02(m，9H)，1.68-1.80(m，1H)，1.82-2.02(m，1H)，2.10-2.22(m，1H)，2.40-2.60(m，1H)，2.81-2.95(m，1H)，3.75(s，3H)，4.00(s，3H)，4.18-4.22(m，1H)，4.27-4.40(m，2H)，6.05-6.12(m，1H)，6.99-7.02(m，1H)，7.16-7.21(m，1H)，7.38(s，1H)，7.40-7.43(m，1H)，7.48-7.61(m，3H)，7.98-8.12(m，3H)。

Example 26

(S) -2- { [ ((1R, 4R) & (1S, 4S)) -2- { (S) -1- [ (2, 5-dimethoxy-phenyl) -ethyl-carbamoyl ] -2-methyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -butyric acid (26)

Compound 26(5.7mg, 36%) was prepared from compound 17(16.7mg, 0.021mmol) according to the method for preparing compound 24, thereby giving the title compound as a white powder.

¹H-NMR(300MHz，CD₃OD)：δ0.75-0.81(m，6H)，0.82-0.98(m，3H)，1.00-1.10(m，3H)，1.60-2.00(m，3H)，2.40-2.56(m，1H)，2.80-2.88(m，1H)，3.18-3.24(m，1H)，3.40-3.46(m，1H)，[3.67-3.80(m，6H)]，3.97(s，3H)，4.10-4.20(m，1H)，4.21-4.40(m，2H)，6.02-6.17(m，1H)，6.75-6.82(m，1H)，6.84-7.01(m，3H)，7.10-7.20(m，1H)，7.30-7.37(m，1H)，7.40-7.43(m，1H)，7.50-7.60(m，3H)，8.00-8.17(m，3H). ¹³C-NMR(75.5MHz，CD₃OD)：δ9.6，[11.8&12.0]，[17.2&17.4]，18.9，25.0，32.3，35.7，43.3，44.2，[50.3&50.5]，[54.5&54.8&54.9&55.0]，[55.1&55.2&55.3&56.0]，58.7，83.6，99.3，105.5，[112.5&112.7]，114.3，[15.1&115.2]，115.7，116.1，118.4，[123.3&123.4]，125.2，[128.0&128.1，128.8，129.1，129.8，[135.1&135.3]，139.2，[143.3&144.4]，149.2，[149.6&149.9]，153.8，159.9，162.4，[163.9&164.5]，172.1，172.8，[173.6&173.7].MALDI-TOF m/z 775.30[(M+Na)⁺C₄₂H₄₈N₄NaO₉ ⁺Calculated value of 775.33]。

Example 27

(S) -2- { [ ((1R, 4R) & (1S, 4S)) -2- { (S) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2, 2-dimethyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -butyric acid (27)

Compound 27(6.0mg, 72%) was prepared from compound 18(8.6mg, 0.011mmol) according to the procedure for preparing compound 24. Purification was performed by HPLC (60% methanol + 0.2% TEA) to give the title compound as a white powder.

¹H-NMR(300MHz，CD₃OD)：δ0.88-0.95(m，3H)，0.96(s，9H)，0.97-1.24(m，4H)，1.57-1.62(m，3H)，1.58-1.78(m，4H)，1.79-1.99(m，1H)，2.35-2.44(m，2H)，2.85-2.98(m，1H)，[(3.67&3.69)s，3H]，3.94(s，3H)，4.10-4.20(m，1H)，4.30-4.40(m，3H)，6.00-6.09(m，1H)，[6.80-6.82(m，0.5H)][6.85-6.87(m，0.5H)]，7.05-7.19(m，2H)，7.38-7.55(m，4H)，7.95-8.07(m，3H).¹³C-NMR(75.5MHz，CD₃OD)：δ[9.1&9.2]，[24.7&24.9]，[25.4&25.5]，[25.9&26.0]，[28.3&28.4]，28.9，[34.8&34.9]，[35.6&35.9]，[39.6&39.7]，[49.9&50.1]，[51.4&51.2]，[53.9&54.0]55.0，[57.2&57.4]，60.0，[82.1&82.5]，98.6，106.2，114.7，117.8，122.7，127.5，127.7，[128.4&128.5]，129.1，135.3，136.3，141.6，142.0，150.5，159.8，[161.0&161.3][164.0&164.1]，[171.6&171.9]，[172.2&172.3]，[173，0&173.2].MALDI-TOF m/z 779.43[(M+Na)⁺C₄₂H₅₂N₄NaO₉ ⁺Calculated value of 779.36]。

Example 28

(S) -2- { [ (1R, 4R) -2- { (S) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2-methyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -pentanoic acid tert-butyl ester (28)

Tert-butyl ester 19a (7.6mg, 0.0094mmol) and TES (2.4mg, 0.021mmol) were dissolved in DCM (1mL) and the mixture was cooled in an ice bath. TFA (1mL) was added. After two hours the mixture was concentrated and purified on HPLC using 60% methanol + 0.2% TEA as mobile phase. This gave 28(6.1mg, 86%) as a pale yellow thick syrup. After freeze-drying it, the title compound was collected as a white powder.

¹H-NMR(300MHz，CD₃OD+CDCl₃(1∶1))：δ0.90-1.00(m，9H)，1.00-1.30(m，7H)，1.50-1.90(m，8H)，2.00-2.10(m，1H)，2.40-2.50(m，1H)，2.85-2.98(m，1H)，3.65-3.72(s，3H)，3.99(s，3H)，4.15-4.22(m，1H)，4.24-4.35(m，2H)，4.38-4.44(m，1H)，6.10-6.20(m，1H)，6.95-6.96(m，1H)，7.16-7.23(m，1H)，7.31(s，1H)，7.42(d，J＝2.47Hz，1H)，7.53-7.72(m，3H)，7.97-8.16(m，3H)；¹³C-NMR(75.5MHz，CD₃OD+CDCl₃ 1∶1)：δ13.5，18.3，19.0，26.0，29.0，29.7，31.0，34.1，35.8，40.2，51.9，55.9，57.7，58.9，63.5，68.4，84.0，99.6，104.8，105.7，115.1，119.0，123.7，128.1，128.9，129.1，130.4，131.3，135.3，138.0，142.9，159.5，162.8，164.8，172.2，172.2，172.4

Example 29

(S) -2- { [ (1S, 4S) -2- { (S) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2-methyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -pentanoic acid tert-butyl ester (29)

Compound 29(1.3mg, 26%) was prepared from compound 19b (5.3mg, 0.065mmol) according to the procedure for preparation of compound 28. This gave the title compound as a white powder.

¹H-NMR(300MHz，CD₃OD)：δ0.85-1.00(m，9H)，1.00-1.23(m，7H)，1.50-1.78(m，8H)，2.05-2.23(m，1H)，2.50-2.66(m，1H)，2.70-2.85(m，1H)，3.69(s，3H)，3.92(s，3H)，4.02-4.16(m，1H)，4.20-4.25(m，1H)，4.35-4.40(m，2H)，6.09(m，1H)，7.00(s，1H)，7.12-7.18(dd，J＝2.47，2.19Hz，1H)，7.30(s，1H)，7.40(d，J＝2.42Hz，1H)，7.48-7.74(m，3H)，8.03-8.10(m，3H)；¹³C-NMR(75.5MHz，CDCl₃)：δ11.7，16.5，17.0，24.4，27.2，27.9，29.0，29.137.5，41.8，49.7，50.5，53.3，56.3，63.5，66.5，81.0，100.3，101.0，105.7，113.6，121.6，126.3，127.1，127.9，130.1，131.4，135.6，138.7，141.1，150.4，160.2，160.5，165.3，173.0，173.6，173.7

Example 30

(1R, 2S) -1- { [ (1R, 4R) -2- { (S) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2, 2-dimethyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -2-vinyl-cyclopropanecarboxylic acid (30a) and

(1R, 2S) -1- { [ (1S, 4S) -2- { (S) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2, 2-dimethyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopent-2-enecarbonyl ] -amino } -2-vinyl-cyclopropanecarboxylic acid (30b)

Compound 30a (6.3mg, 49%) and compound 30b (5.6mg, 43%) were synthesized from compound 21(13.8mg, 0.0016mmol) according to the procedure for preparing compounds 22a and 22 b. 30a and 30 b: white powder.

30a：¹H-NMR(300MHz，CD₃OD)：δ1.02(s，9H)，1.03-1.43(m，5H)，1.61-1.95(m，8H)，2.11-2.21(m，1H)，2.43-2.58(m，1H)，2.97-3.04(m，1H)，3，78(s，3H)，4.01(s，3H)，4.02-4.17(m，1H)，4.25-4.40(m，2H)，5.10-5-20(m，1H)，5.27-5.40(m，1H)，6.77-6.94(m，1H)，6.10-6.20(m，1H)，6.97(s，1H)，7.18(dd，J＝2.5，9.2Hz，1H)，7.22(s，1H)，7.46(d，J＝2.5Hz，1H)，7.52-7.65(m，3H)，8.00-8.18(m，3H).¹³C-NMR(75.5MHz，CD₃OD)：δ13.5，25.3，25.7，28.3，28.7，29.0，32.8，34.6，35.3，39.3，49.7，51.1，54.6，57.2，59.8，82.1，98.4，105.8，114.5，116.3，117.6，122.6，127.2，128.1，128.2，128.8，130.2，133.7，136.0，139.5，141.5，150.3，159.7，161.0，161.2，163，4，171.6，172.5.MALDI-TOF m/z 803.56[(M+Na)⁺C₄₄H₅₂N₄NaO₉ ⁺Calculated value of 803.36].30b：¹H-NMR(300MHz，CD₃OD)：δ1.03(s，9H)，1.04-1.42(m，5H)，2.60-2.90(m，8H)，2.17-2.22(m，1H)，2.40-2.55(m，1H)，2.96-3.10(m，1H)，3.77(s，3H)，4.01(s，3H)，4.05-4.16(m，1H)，4.30-4.40(m，2H)，5.15-5.20(m，1H)，5.25-5.40(m，1H)，5.78-5.95(m，1H)，6.10-6.20(m，1H)，6.98(s，1H)，7.17(dd，J＝2.5，9.1Hz，1H)，7.26(s，1H)，7.46(d，J＝2.5Hz，1H)，7.50-7.65(m，3H)，8.03-8.28(m，3H).¹³C-NMR(75.5MHz，CD₃OD)：δ13.7，26.0，26.3，28.8，29.4，29.6，34.0，35.2，35.8，40.1，50.6，51.7，55.3，57.8，60.6，83.0，99.1，106.3，115.2，117.0，118.3，123.2，127.9，128.0，128.8，129.6，130.6，134.4，136.1，140.0，142.5，150.8，160.3，161.8，162.0，165.7，172.3，173.0

Example 31

Trans- (3R, 4R) -bis (methoxycarbonyl) cyclopentanol (31)

Sodium borohydride (1.11g, 0.029mol) was added to a stirred solution of dimethyl (1R, 2S) -4-oxo-cyclopentane 1, 2-dicarboxylate (4.88g, 0.0244mol) in methanol (300mL) at 0 deg.C. After 1 hour the reaction was quenched with 90mL brine, concentrated and extracted with ethyl acetate. The organic phase was collected, dried, filtered and concentrated. The resulting crude product was purified by flash column chromatography (toluene/ethyl acetate 1: 1) to give compound 31(3.73g, 76%) as a yellow oil.

Example 32

3-oxo-2-oxa-bicyclo [2.2.1] heptane-5-carboxylic acid (32)

Sodium hydroxide (1M, 74mL, 0.074mol) was added to a stirred solution of compound 31(3.73g, 0.018mol) in methanol (105mL) at room temperature. After 4 hours, the reaction mixture was neutralized with 3M HCl, evaporated, and co-evaporated with toluene several times. Pyridine (75mL) and Ac₂O (53mL) was added and the reaction mixture was shaken overnight at room temperature. The resulting mixture was then co-evaporated with toluene and purified by flash column chromatography (ethyl acetate + 1% acetic acid) to give compound 32 as a yellow oil (2.51g, 88%).

Example 33

3-oxo-2-oxa-bicyclo [2.2.1] heptane-5-carboxylic acid tert-butyl ester (33)

DMAP (14mg, 0.115mmol) and Boc were added at 0 ℃ under an inert argon atmosphere₂O (252mg, 1.44mmol) was added to 2mL CH of stirred Compound 32(180mg, 1.15mmol)₂Cl₂In solution. The reaction was warmed to room temperature and stirred overnight. The reaction mixture was concentrated, and the resulting crude product was purified by flash column chromatography (toluene/ethyl acetate gradient 15: 1, 9: 1, 6: 1, 4: 1, 2: 1) to give compound 33(124mg, 51%) as white crystals.

¹H-NMR(300MHz，CD₃OD)δ1.45(s，9H)，1.90(d，J＝11.0Hz，1H)，2.10-2.19(m，3H)，2.76-2.83(m，1H)，3.10(s，1H)，4.99(s，1H)； ¹³C-NMR(75.5MHz，CD₃OD)δ27.1，33.0，37.7，40.8，46.1，81.1，81.6，172.0，177.7。

Example 34

(1R, 2R, 4S) -2- ((1R, 2S) -1-ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -4-hydroxy-cyclopentanecarboxylic acid tert-butyl ester (34)

Compound 33(56mg, 0.264mmol) was dissolved in dioxane/water 1: 1(5mL) and the mixture was cooled to 0 ℃.1M lithium hydroxide (0.52mL, 0.520mmol) was added and the mixture was stirred at 0 ℃ for 45 minutes, after which the mixture was neutralized with 1M hydrochloric acid, evaporated and co-evaporated with toluene. The resulting residue was dissolved in DMF (5mL) and (1R, 2S) -1-amino-2-vinyl cyclopropanecarboxylic acid ethyl ester hydrochloride (60mg, 0.313mmol) and Diisopropylethylamine (DIEA) (138L, 0.792mmol) were added thereto and the solution was cooled to 0 ℃. HATU (120mg, 0.316mmol) was added to it and the mixture was stirred at 0 ℃ for 0.5h and at room temperature for a further 2 h. The mixture was then evaporated and extracted with EtOAc, washed with brine, dried, filtered and concentrated. Purification was performed by flash column chromatography (toluene/EtOAc 1: 1) to provide compound 34 as a colorless oil (86mg, 89%).

Example 35

(1R, 2R, 4R) -2- ((1R, 2S) -1-ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarboxylic acid tert-butyl ester (35)

Compound 34(73mg, 0.199mmol) was dissolved in anhydrous THF (4mL) and 2-phenyl-7-methoxy-4-quinolinol (86mg, 0.342mmol) and triphenylphosphine (141mg, 0.538mmol) were added thereto. The mixture was cooled to 0 ℃ and DIAD (0.567mmol) dissolved in 1mL THF was added dropwise. The mixture was stirred at room temperature for 48 h. The solvent was evaporated and the crude product purified by flash column chromatography gradient elution (toluene/EtOAc 9: 1, 6: 1, 4: 1) to afford compound 35(81mg, 68%).

Example 36

Boc-L-tert-leucine-OH (36)

Triethylamine (890. mu.L, 6.40mmol) was added dropwise to a stirred 1: 1(8mL) solution of L-tert-leucine (300mg, 2.29mmol) and di-tert-butyl dicarbonate (599mg, 2.74mmol) in dioxane/water, and the solution was stirred overnight. The mixture was extracted with petroleum ether (2 ×), the aqueous phase was cooled to 0 ℃ and the solvent was removed by slow addition of 4M NaHSO₄·H₂O carefully acidify it to pH 3. The acidified aqueous phase was extracted with EtOAc (3 ×) and the combined organic phases were washed with brine (2 ×), then dried, filtered and concentrated to give compound 36 as a colourless powder (522mg, 99%). No further purification was required.

¹H-NMR(300MHz，CD₃OD)δ0.99(s，9H)，1.44(s，9H)，3.96(s， 1H)；¹³C-NMR(75.5MHz，CD₃OD)δ27.1，28.7，34.9，68.0，80.5，157.8，174.7。

Example 37

((S) -cyclohexyl-methylcarbamoyl-methyl) -carbamic acid tert-butyl ester (37)

Boc-Chg-OH (387mg, 1.50mmol) was coupled to methylamine hydrochloride (111mg, 1.65mmol) using the same HATU coupling conditions as in the synthesis of compound 34. The resulting crude product was extracted with EtOAc, washed with brine and concentrated. Purification was performed by flash column chromatography (EtOAc) to afford compound 37 as a colorless solid (307mg, 76%).

¹H-NMR(300MHz，CDCl₃)δ0.91-1.13(m，2H)，1.14-1.31(m，3H)，1.44(s，9H)，1.61-1.80(m，6H)，2.80(d，J＝4.7Hz，3H)，3.91(dd，J＝7.1，9.1Hz，1H)，5.23(b，1H)，6.52(bs，1H)；¹³C-NMR(75.5MHz，CDCl₃)δ25.9，26.0，26.1，28.3，28.5，29.6，40.5，59.5，79.7，155.9，172.4。

Example 38

{ (S) -1- [ ((S) -cyclohexyl-methylcarbamoyl-methyl) -carbamoyl ] -2, 2-dimethyl-propyl } -carbamic acid tert-butyl ester (38)

To a solution of compound 37(98mg, 0.362mmol) in dichloromethane (3mL) was added triethylsilane (115mL, 0.742mmol) and TFA (3 mL). The mixture was stirred at room temperature for 2h, then evaporated and co-evaporated with toluene. The deprotected amine was dissolved in DMF (5mL) and coupled to compound 36(84mg, 0.363mmol) using the same HATU coupling conditions as in the synthesis of compound 34. The resulting crude product was extracted with EtOAc, washed with brine, dried, filtered and concentrated. Purification was performed by flash column chromatography (toluene/EtOAc 1: 1) to provide compound 38 as a colorless solid (128mg, 92%).

¹H-NMR(300MHz，CDCl₃)δ0.99(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＝4.7Hz，3H)，4.11(d，J＝9.3Hz，1H)，4.33(app.t，J＝8.5Hz，1H)，5.65(b，1H)，7.25(b，1H)，7.39(b，1H)；¹³C-NMR(75.5MHz，CDCl₃)δ25.9，25.9，26.0，26.2，26.8，28.4，29.0，29.7，34.5，39.7，58.4，62.4，79.4，156.0，171.4，171.8。

Example 39

(1R, 2S) -1- { [ (1R, 2R, 4S) -2- { (S) -1- [ ((S) -cyclohexyl-methylcarbamoyl-methyl) -carbamoyl ] -2, 2-dimethyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarbonyl ] -amino } -2-vinyl-cyclopropanecarboxylic acid ethyl ester (39)

To a solution of compound 35(30mg, 0.050mmol) in dichloromethane (1.5mL) was added triethylsilane (21L, 0.132mmol) and TFA (1.5 mL). The mixture was stirred at room temperature for 2h, then evaporated and co-evaporated with toluene. Amine 38(1.3 equivalents) was deprotected in the same manner as compound and then coupled to deprotected compound 35 using the same HATU coupling conditions as in the synthesis of compound 34. The resulting crude product was extracted with EtOAc, washed with brine, dried, filtered and concentrated. Purification by HPLC (MeOH/water 9: 1+ 0.2% triethylamine) gave compound 39(30mg, 74%) as a colorless solid.

¹H-NMR(300MHz，CD₃OD) δ 0.81-1.14(m, 4H), 0.99(s, overlapping peak, 9H), 1.21(t, J ═ 7.1Hz, 3H), 1.35-1.51(m, 4H), 1.52-1.65(m, 3H), 1.66-1.72(m, 2H), 2.03-2.20(m, 2H), 2.24-2.39(m, 1H), 2.46-2.56(m, 1H), 2.66(s, 3H), 2.72-2.85(m, 1H), 3.39-3.48(m, 2H), 3.90(s, 3H), 4.03-4.15(m, 3H), 4.44(s, 1H), 5.09(dd, J ═ 1.9, 10.3Hz, 1H), 5.19-5.19 (m, 19, 5.15 (m, 3H), 5.7, 7, 7.7, 7H), 1, 7, j ═ 2.5Hz, 1H), 7.43-7.52(m, 3H), 7.86-7.98(m, 2H), 8.05(d, J ═ 9.3Hz, 1H);¹³C-NMR(75.5MHz，CD₃OD) δ 14.7, 23.4, 26.0, 26.9, 27.1, 27.3, 30.1, 30.7, 35.0, 35.4, 38.3, 38.8, 40.9, 41.0, 47.9, 55.9, 59.6, 62.0, 62.4, 79.8, 99.9, 107.3, 116.4, 118.0, 119.1, 124.4, 128.9, 129.8, 130.5, 135.3, 141.3, 152.1, 161.1, 162.4, 163.0, 171.6, 172.5, 173.7, 175.2, 176.8, Maldi-TOF-spectrum: (M + H)⁺Calculated values: 810.4, measurement: 810.5, respectively; (M + Na)⁺Calculated values: 832.4, measurement: 832.4, respectively; (M + K)⁺Calculated values: 848.5, measurement: 848.4.

example 40

(1R, 2S) -1- { [ (1R, 2R, 4S) -2- { (S) -1- [ ((S) -cyclohexyl-methylcarbamoyl-methyl) -carbamoyl ] -2, 2-dimethyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarbonyl ] -amino } -2-vinyl-cyclopropanecarboxylic acid (40)

To a 0 deg.C solution of compound 39(20mg, 0.025mmol) in THF/MeOH/water 2: 1(2mL) was added 1M LiOH (175. mu.L, 0.175mmol), the solution was allowed to warm to room temperature and stirred for 48 h. The above solution was acidified to pH3 with 1M HCl and then evaporated and co-evaporated with toluene. The resulting crude product was purified by HPLC (MeOH/water 6: 4+ 0.5% TFA, followed by MeOH/water 4: 1+ 0.2% TFA) to give compound 40 as a colorless solid (13mg, 67%).

¹H-NMR(300MHz，CD₃OD) δ 0.82-0.98(m, 1H), 1.01(s, 9H), 1.05-1.26(m, 3H), 1.34-1.43(m, 1H), 1.49-1.77(m, 8H), 2.10-2.21 (m, 1H), 2.28-2.42(m, 2H), 2.50-2.61(m, 1H), 2.64(s, 3H), 2.68-2.81(m, 1H), 3.36-3.45(m, 2H), 4.04-4.11(m, 1H), 4.06(s, overlap peak, 3H), 4.27(d, J ═ 8.8Hz, 1H), 5.10(dd, J ═ 1.8, 10.3Hz, 1H), 5.28(dd, 1.8, 17.8, 1H, 17, 17.8 Hz, 7, 7.7, 7H, 7, 7.7, 7H, 7H, 7, 7.69-7.78(m, 3H), 8.02-8.07(m, 2H), 8.39(d, J ═ 9.3Hz, 1H);¹³C-NMR(75.5MHz，CD₃OD) delta 23.5, 26.0, 26.9, 27.2, 27.3, 30.0, 30.7, 34.7, 35.3, 37.0, 38.7, 41.0, 41.3, 47.4, 56.9, 59.4, 62.7, 83.9, 100.4, 102.2, 116.2, 117.7, 121.7, 126.7, 129.8, 130.8, 133.4, 133.9, 135.6, 143.5, 158.0, 166.6, 168.6, 172.5, 173.4, 173.6, 175.4, 176.4. Maldi-TOF-spectrum: (M + H)⁺Calculated values: 782.4, measurement: 782.2, respectively; (M + Na)⁺Calculated values: 804.4, measurement: 804.2; (M + K)⁺Calculated values: 820.5, measurement: 820.2.

EXAMPLE 41

3-oxo-2-oxa-bicyclo [2.2.1] heptane-5-carboxylic acid methyl ester (41)

Compound 32(1.014g, 6.50mmol) was dissolved in acetone (35mL), to which methyl iodide (13.68g, 96.4mmol) and silver (I) oxide (1.61g, 6.95mmol) were then added. After stirring for 3 hours, the resulting mixture was filtered through celite and the filtrate was evaporated, followed by purification by flash column chromatography (toluene/ethyl acetate 4: 1) to give methyl ester 41 as white crystals (702mg, 64%).

¹H-NMR(300MHz，CDCl₃)：δ1.96(d，J＝10.7Hz，1H)，2.21-2.25(m，3H)，2.91-2.95(m，1H)，3.16(s，1H)，3.75(s，3H)，4.98(app.s，1H)。

Example 42

(1R, 2R, 4S) -2- ((S) -1-tert-Butoxycarbonyl-butylcarbamoyl) -4-hydroxy-cyclopentanecarboxylic acid methyl ester (42)

Compound 41(263mg, 1.55mmol) and H-Nva-OtBu (420mg, 2.42mmol) were dissolved in anhydrous THF (20 mL). DIEA (530. mu.L, 3.04mmol) and 2-hydroxypyridine (260mg, 2.73mmol) were added thereto, and the mixture was refluxed for five days. The solvent was evaporated and the resulting crude product was purified by flash column chromatography (toluene/EtOAc 1: 2) to give 42(510mg, 96%).

Example 43

(1R, 2R, 4R) -2- ((S) -1-tert-Butoxycarbonyl-butylcarbamoyl) -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarboxylic acid methyl ester (43)

Compound 42(249mg, 0.725mmol), 2-phenyl-7-methoxy-4-quinolinol (310mg, 1.23mmol) and PPh₃(580mg, 2.21mmol) was dissolved in anhydrous THF and its temperature was reduced to 0 deg.C. DIAD (435. mu.L, 2.21mmol) dissolved in 2mL of anhydrous THF was added to the above mixture over a period of 5 minutes. After two hours the temperature was raised to room temperature and the solution was stirred overnight. It was then evaporated and purified by flash column chromatography (toluene/EtOAc gradient 6: 1-4: 1) to give compound 43(324mg, 78%).

Example 44

(S) -2- { [ (1R, 2R, 4S) -2- { (S) -1- [ ((S) -cyclohexyl-methylcarbamoyl-methyl) -carbamoyl ] -2, 2-dimethyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarbonyl ] -amino } -pentanoic acid tert-butyl ester (44)

Compound 43(38mg, 0.066mmol) was dissolved in dioxane/water 1: 1(4mL) and the solution was cooled to 0 ℃ to which 1M LiOH (132. mu.L, 0.132mmol) was added. The temperature was raised to room temperature, the solution was stirred for 2 hours, after which it was neutralized by addition of 1M HCl, subsequently evaporated and co-evaporated with toluene. The resulting residue and deprotected amine 38(1.1 eq) were dissolved in DMF and coupled using standard HATU coupling conditions as used in the synthesis of compound 34. The resulting crude product was extracted with EtOAc, washed with brine, dried, filtered and concentrated. Purification by HPLC (MeOH/water 9: 1+ 0.2% TEA) gave compound 44 as a colorless solid (44mg, 81%).

¹H-NMR(CDCl₃300MHz) rotamers (5: 1) δ 0.79(t, J ═ 7.3Hz, 3H), 0.85-1.19(m, 3H), 0.93(s, overlapping peak, 9H), 1.20-1.35(m, 2H), 1.39(s, 1.5H), 1.43(s, 7.5H), 1.54-1.79(m, 6H), 2.06-2.28(m, 3H), 2.39-2.51(m, 2H), 2.66-2.78(m, 1H), 2.74(d, overlapping peak, J ═ 4.7Hz, 3H), 3.42-3.68(m, 2H), 3.84(s, 2.5H), 3.88(s, 0.5H), 4.19(t, J ═ 8.9, 1H), 4.59(m, 6H), 1.7H, 7H), 1.7H, 7H, 7.7H, 7H, 7H, 7, 1H) 7.85-7.97(m, 3H);¹³C-NMR(75.5MHz，CDCl₃) δ 13.7, 18.7, 25.6, 25.7, 26.0, 26.7, 28.0, 28.9, 29, 7, 34.5, 34.7, 37.7, 38.0, 39.2, 46.6, 47.7, 52.7, 55.3, 58.5, 60.3, 77.9, 81.7, 98.0, 107.4, 115.0, 117.9, 122.8, 127.4, 128.6, 129.0, 140.2, 151.2, 158.9, 160.6, 161.1, 170.9, 171.6, 171.8, 172.7, 173.3. Maldi-TOF-spectra: (M + H)⁺Calculated values: 828.5, measurement: 828.6, respectively; (M + Na)⁺Calculated values: 850.5, measurement: 850.6 of the carrier; (M + K)⁺Calculated values: 866.6, measurement: 866.6.

example 45

(S) -2- { [ (1R, 2R, 4S) -2- { (S) -1- [ ((S) -cyclohexyl-methylcarbamoyl-methyl) -carbamoyl ] -2, 2-dimethyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarbonyl ] -amino } -pentanoic acid (45)

Compound 44(21mg, 0.025mmol) was dissolved in CH₂Cl₂(1.5mL) and triethylsilane (10. mu.L, 0.063mmol) and TFA (1.5mL) were added thereto. The solution was stirred at room temperature for 2 hours, after which the solvent was evaporated and co-evaporated with toluene to give compound 45(20mg, 100%) as a colourless solid.

¹H-NMR(300MHz，CD₃OD) δ 0.93(t, overlapping peak, 3H), 0.98(s, 9H), 0.99-1.25(m, 4H), 1.30-1.49(m, 3H), 1.50-1.90(m, 8H), 2.25-2.39(m, 2H), 2.54-2.62(m, 1H), 2.64(s, 3H), 2.72-2.87(m, 1H), 3.34-3.57(m, 3H), 4.02-4.13(m, 1H), 4.06(s, overlapping peak, 3H), 4.27-4.36(m, 1H), 4.37-4.47(m, 1H), 5.57-5.66(m, 1H), 7.45(dd, J ═ 2.3, 9.2, 1H), 7.48(s, 1H), 7.54.7.7 (H, 7.7, 3H), 7.7.8 (m, 3H), 3H, 3Hz, 7.7.7.79 (m, 8H), 3H);¹³C-NMR(75.5MHz，CD₃OD)δ14.0，20.2，26.0，26.9，27.2，30.1，30.7，34.6，35.3，37.2，39.1，41.2，47.7，53.7，56.9，59.4，59.5，62.5，83.7，100.4，101.3，102.2，116.2，121.7，126.7，129.8，130.8，133.3，133.9，143.5，157.9，166.6，168.5，172.5，173.6，175.3，175.4，175.5.

Maldi-TOF-Spectroscopy: (M + H)⁺Calculated values: 772.4, measurement: 772.6, respectively; (M + Na)⁺Calculated values: 794.4, measurement: 794.6, respectively; (M + K)⁺Calculated values: 810.5, measurement: 810.6.

example 46

Hepta-6-enal (46)

To hept-6-en-1-ol (1mL, 7.44mmol) and N-methylmorpholine N-oxideTo a solution of material (1.308g, 11.17mmol) in DCM (17mL) was added the milled molecular sieve (3.5g,). The mixture was stirred at room temperature for 10 minutes under a nitrogen atmosphere, and then tetrapropylammonium ferulate (TPAP) (131mg, 0.37mmol) was added thereto. After stirring for a further 2.5 hours, the solution was filtered through celite. The solvent was then carefully evaporated and the remaining liquid was purified by fast column chromatography (DCM) to give the volatile aldehyde 46 as an oil (620mg, 74%).

Example 47

N' -hept-6-en- (E) -ylidene-hydrazinecarboxylic acid tert-butyl ester (47)

To a solution of compound 46(68mg, 0.610mmol) and tert-butyl carbazate (81mg, 0.613mmol) in methanol (5mL) was added ground molecular sieve (115mg,). The mixture was stirred for 3 hours, after which it was filtered through celite and evaporated. The resulting residue was dissolved in anhydrous THF (3mL) and AcOH (3 mL). Reacting NaBH₃CN (95mg, 1.51mmol) was added thereto and the solution was stirred overnight. With saturated NaHCO₃The reaction mixture was diluted with solution (6mL) and EtOAc (6 mL). The organic phase was washed with brine, saturated NaHCO₃Washed with brine and MgSO₄Dried and evaporated. The cyanoborane adduct was hydrolyzed by treatment with methanol (3mL) and 2M NaOH (1.9 mL). The mixture was stirred for 2 hours and the methanol was evaporated. H is to be₂O (5mL) and DCM (5mL) were added and the aqueous phase was extracted three times with DCM. The combined organic phases are dried and evaporated. Purification was performed by flash column chromatography (toluene/ethyl acetate 9: 1 with 1% triethylamine and toluene/ethyl acetate 6: 1 with 1% triethylamine) to give compound 47(85mg, 61%) as an oil.

Example 48

(1R, 2S) -1- { [ (1R, 2R, 4R) -2- (N' -tert-Butoxycarbonyl-N-hept-6-enyl-hydrazinocarbonyl) -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarbonyl ] -amino } -2-vinyl-cyclopropanecarboxylic acid ethyl ester (48)

Scaffold molecule 35(135mg, 0.225mmol) and triethylsilane (71 μ L, 0.447mmol) were dissolved in DCM (2mL), after which trifluoroacetic acid (TFA) (2mL) was added. The mixture was stirred for 2h and then co-evaporated with toluene to remove TFA. The resulting residue was dissolved in DMF (3mL) and compound 47(60mg, 0.263mmol) and DIEA (118. mu.L, 0.677mmol) were added to it. The temperature was lowered to 0 ℃ and the coupling reagent O- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium Hexafluorophosphate (HATU) (94mg, 0.247mmol) was added thereto. The cold solution was stirred for half an hour and then at room temperature for another 16 hours. The solvent was removed under reduced pressure by heating the reaction flask in a water bath. After this time, the resulting residue was dissolved in ethyl acetate, and the organic layer was washed three times with brine, dried, filtered and evaporated. By HPLC (methanol/H containing 0.2% triethylamine)₂O90: 10) gave compound 48 as an oil (140mg, 82%).

¹H-NMR(300MHz，CDCl₃，40℃)：δ1.22(t，J＝7.1Hz，3H)，1.28-1.42(m，6H)，1.46(s，9H)，1.52-1.62(m，2H)，1.82-1.91(m，1H)， 1.96-2.16(m，3H)，2.18-2.34(m，2H)，2.42-2.56(m，1H)，2.58-2.72(m，1H)，3.42(app.bs，3H)，3.66-3.84(m，1H)，3.92(s，3H)，4.15(q，J＝7.1Hz，2H)，4.88-5.02(m，2H)，5.07-5.18(m，2H)，5.20-5.32(m，1H)，5.63-5.84(m，2H)，6.62(bs，1H)，6.94(s，1H)，7.09(dd，J＝2.6，9.2Hz，1H)，7.36-7.51(m，4H)，7.99-8.10(m，3H)；¹³C-NMR(75.5MHz，CDCl₃)：δ14.3，23.0，26.4，26.6，28.3，28.6，33.2，33.5，35.6，37.6，40.6，44.7，47.1，48.6，55.5，61.5，81.9，98.4，107.9，114.5，115.6，118.1，123.2，127.6，128.3，128.7，129.1，133.5，138.7，140.7，151.5，154.5，159.2，160.9，161.5，170.5，174.2，176.3.

Example 49

(z) - (1R, 4R, 6S, 16R, 18R) -14-tert-Butoxycarbonylamino-18- (7-methoxy-2-phenyl-quinolin-4-yloxy) -2, 15-dioxo-3, 14-diaza-tricyclo [14.3.0.04, 6] nonadeca-7-ene-4-carboxylic acid ethyl ester (49)

A solution of compound 48(158mg, 0.209mmol) in dry DCM (25mL) was bubbled with argon for 5 min. Then, to this stirred solution was added a solution of Hoveyda-Grubbs catalyst second generation (11mg, 0.018mmol) in dry DCM (5mL) under an argon atmosphere. The mixture was stirred under argon at reflux for 16 hours. The solvent was evaporated and purified by HPLC (methanol/H containing 0.2% triethylamine)₂O90: 10) to yield compound 49(107mg, 70%) as a colorless solid.

¹H-NMR(300MHz，CD₃OD)：δ1.03-1.22(m，1H)，1.28(t，J＝7.1Hz，3H)，1.32-1.44(m，4H)，1.49(s，9H)，1.55-1.73(m，2H)，1.81-1.91(m，1H)，2.04-2.28(m，3H)，2.30-2.52(m，3H)，2.53-2.70(m，1H)，2.86-3.00(m，1H)，3.34-3.44(m，1H)，3.46-3.62(m，1H)，3.95(s，3H)， 4.19(q，J＝7.1Hz，2H)，4.32-4.48(m，1H)，5.20-5.33(m，1H)，5.34(bs，1H)，5.58-5.70(m，1H)，7.10(s，1H)，7.14(dd，J＝2.5，9.1Hz，1H)，7.39(d，J＝2.5Hz，1H)，7.45-7.55(m，3H)，8.00(d，J＝8.0Hz，2H)，8.17(d，J＝9.3Hz，1H)；¹³C-NMR(75.5MHz，CD₃OD): δ 14.6, 23.4, 27.5, 27.7, 28.0, 28.5, 30.7, 36.1, 38.1, 42.5, 45.6, 56.0, 62.7, 79.9, 82.8, 100.2, 107.4, 116.6, 119.1, 124.5, 126.5, 128.9, 129.8, 130.5, 135.8, 141.5, 152.2, 156.4, 161.3, 162.5, 163.1, 171.9, 175.8, 179.0 MALDI-TOF-spectroscopy: (M + H)⁺Calculated values: 727.4, measurement: 727.5.

example 50

(z) - (1R, 4R, 6S, 16R, 18R) -14-tert-Butoxycarbonylamino-18- (7-methoxy-2-phenyl-quinolin-4-yloxy) -2, 15-dioxo-3, 14-diaza-tricyclo [14.3.0.04, 6] nonadeca-7-ene-4-carboxylic acid (50)

To compound 49(27mg, 0.037mmol) in THF/MeOH/H₂To the O2: 1(5mL) solution was added 1M LiOH (300. mu.L, 0.300 mmol). The solution was stirred at room temperature for 24 hours and finally under reflux for 1 hour. After acidification to pH 3-4 with 1M HCl and evaporation, the residue was purified by HPLC (MeOH/H)₂O80: 20 and MeOH/H₂O90: 10) to yield compound 50(12mg, 46%) as a colorless solid.

¹H-NMR(300MHz，CD₃OD)：δ1.06-1.24(m，1H)，1.26-1.42(m，3H)，1.48(s，9H)，1.52-1.73(m，3H)，1.80-1.90(m，1H)，2.02-2.15(m，1H)，2.15-2.40(m，4H)，2.43-2.54(m，1H)，2.54-2.68(m，1H)，2.88-3.00(m，1H)，3.35-3.48(m，1H)，3.49-3.66(m，1H)，3.96(s，3H)，4.32-4.48(m，1H)，5.25-5.42(m，2H)，5.56-5.68(m，1H)，7.14(s，1H)， 7.17(dd，J＝2.5，9.1Hz，1H)，7.40(d，J＝2.2Hz，1H)，7.46-7.58(m，3H)，8.00(d，J＝8.0Hz，2H)，8.19(d，J＝9.1Hz，1H)；¹³C-NMR(75.5MHz，CD₃OD)：δ23.6，26.8，27.8，28.3，28.5，30.5，35.8，38.1，43.0，45.5，56.0，80.2，82.7，100.4，106.9，116.6，119.2，124.7，127.4，129.0，129.8，130.7，134.8，140.9，151.6，156.5，161.1，163.0，163.4，173.8，175.7，179.3.

Example 51

((S) -1-Cyclopentylcarbamoyl-2, 2-dimethyl-propyl) -carbamic acid tert-butyl ester (51)

To a cold solution of compound 36(133mg, 0.575mmol), cyclopentylamine (64 μ L, 0.648mmol) and DIEA (301 μ L, 1.73mmol) in DMF (3mL) was added the coupling reagent HATU (240mg, 0.631 mmol). The mixture was stirred for half an hour and then at room temperature for another two hours. Under reduced pressure, the solvent was removed by heating the reaction flask in a water bath, the residue was dissolved in ethyl acetate, after which the organic phase was washed three times with brine, dried, filtered and evaporated. Purification was performed by flash column chromatography (toluene/ethyl acetate 4: 1) to give 51 as colorless crystals (140mg, 82%).

¹H-NMR(300MHz，CDCl₃): δ 0.95(s, 9H), 1.28-1.48(m, overlap peak, 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)H)，5.91(b，1H)；¹³C-NMR(75.5MHz，CDCl₃)：δ23.6，23.7，26.5，28.3，32.6，33.1，34.5，51.0，62.2，79.4，155.9，170.3.

Example 52

(1R, 2S) -1- { [ (1R, 2R, 4S) -2- ((S) -1-cyclopentylcarbamoyl-2, 2-dimethyl-propylcarbamoyl) -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarbonyl ] -amino } -2-vinyl-cyclopropanecarboxylic acid ethyl ester (52)

Compounds 51(298mg, 0.048mmol) and 35(16mg, 0.054mmol) were deprotected and coupled according to the procedure for the preparation of compound 39. By HPLC (methanol/H containing 0.2% triethylamine)₂O90: 10) gave compound 52(22mg, 63%) as a colorless solid.

¹H-NMR(CDCl₃，300MHz)：δ0.97(s，9H)，1.21(t，J＝7.1Hz，3H)，1.26-1.37(m，1H)，1.38-1.46(m，2H)，1.48-1.58(m，4H)，1.78-1.85(m，1H)，1.86-2.02(m，3H)，2.03-2.19(m，1H)，2.28-2.40(m，2H)，2.41-2.54(m，1H)，2.64-2.78(m，1H)，3.10-3.24(m，1H)，3.30-3.44(m，1H)，3.95(s，3H)，4.04-4.21(m，3H)，5.12(dd，J＝1.7，10.3Hz，1H)，5.14-5.22(m，1H)，5.28(dd，J＝1.7，17.0Hz，1H)，5.59(b，1H)，5.75(ddd，J＝8.8，10.3，17.0Hz，1H)，6.66-6.82(m，2H)，6.99(s，1H)，7.09(dd，J＝2.5，9.1Hz，1H)，7.41-7.55(m，4H)，7.99-8.09(m，3H)；¹³C-NMR(75.5MHz，CDCl₃)：δ14.3，22.9，23.6，23.6，26.7，32.7，33.2，33.7，34.8，35.9，36.6，40.2，46.4，47.5，51.3，55.5，61.1，61.4，78.0，98.4，107.1，115.2，117.9，118.2，123.1，127.6，128.8，129.3，1335, 159.1, 161.4, 169.4, 169.9, 173.1, 174.0. MALDI-TOF-spectroscopy: (M + H)⁺Calculated values: 725.4, measurement: 725.6, respectively; (M + Na)⁺Calculated values: 747.4, measurement: 747.6, respectively; (M + K)⁺Calculated values: 763.3, measurement: 763.5.

example 53

(1R, 2S) -1- { [ (1R, 2R, 4S) -2- ((S) -1-cyclopentylcarbamoyl-2, 2-dimethyl-propylcarbamoyl) -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarbonyl ] -amino } -2-vinyl-cyclopropanecarboxylic acid (53)

To compound 52(14mg, 0.019mmol) in dioxane/H₂To the O1: 1(4mL) solution was added 1M LiOH (115. mu.L, 0.115 mmol). The solution was stirred at room temperature for 24 h. Thereafter, another portion of LiOH (75 μ L, 0.075mmol) was added thereto and the solution was stirred for a further 24 h. After acidification to pH about 3 with 1M HCl and co-evaporation with toluene, the resulting residue was purified by HPLC (MeOH/H)₂O70: 30, containing 0.2% TFA) to yield compound 53(8mg, 60%) as a colorless solid.

¹H-NMR(300MHz，CD₃OD): δ 0.98(s, 9H), 1.28-1.48(m, 3H), 1.49-1.76(m, 5H), 1.78-1.94(m, 2H), 2.10-2.24(m, 1H), 2.26-2.45(m, 2H), 2.50-2.62(m, 1H), 2.66-2.79(m, 1H), 3.35-3.48(m, 2H), 3.94-4.03(m, 1H), 4.06(s, 3H), 4.16-4.24(m, 1H), 5.10(dd, J ═ 1.8, 10.3Hz, 1H), 5.29(dd, J ═ 1.8, 17.2Hz, 1H), 5.62(b, 1H), 5.82(d, J ═ 9.1, 10.3H), 5.29(dd, 7.8, 7.7.7H), 7.7.7.7, 7.8 (dd, 7.8, 7H), 7.7.7.7, 7.7.7, 7H, 7.7H, 7H, j ═ 9.3Hz, 1H);¹³C-NMR(75.5MHz，CD₃OD): δ 24.7, 24.7, 27.3, 33.1, 33.6, 34.7, 35.4, 36.9, 38.7, 41.0, 47.4, 52.3, 56.9, 62.3, 83.9, 100.4, 102.3, 116.2, 117.7, 121.6, 126.7, 129.8, 130.8, 133.4, 133.8, 135.6, 143.5, 158.0, 166.5, 168.6, 171.9, 173.4, 175.2, 176.4 MALDI-TOF-spectroscopy: (M + H)⁺Calculated values: 697.4, measurement: 697.3, respectively; (M + Na)⁺Calculated values: 718.7, measurement: 719.3, respectively; (M + K)⁺Calculated values: 735.3, measurement: 735.3.

example 54

(S) -tert-Butoxycarbonylamino-cyclohexyl-acetic acid methyl ester (54)

To a solution of Boc-Chg-OH (53mg, 0.206mmol) in acetone (3mL) was added iodomethane (195. mu.L, 3.1mmol) and silver (I) oxide (53mg, 0.229 mmol). The mixture was stirred overnight in a reaction flask covered with aluminum foil. The solution was then filtered through celite and evaporated. Purification was performed by flash column chromatography (toluene/ethyl acetate 15: 1) to give methyl ester 54(56mg, 100%) as a colorless oil.

¹H-NMR(300MHz，CDCl₃)：δ1.00-1.34(m，5H)，1.44(s，9H)，1.54-1.82(m，6H)，3.73(s，3H)，4.20(dd，J＝2.8，5.0Hz，1H)，5.05(bs，1H)；¹³C-NMR(75.5MHz，CDCl₃)：δ26.0，28.2，28.3，29.5，41.1，52.0，58.3，79.7，155.6，172.9.

Example 55

(S) - ((S) -2-benzyloxycarbonylamino-3-methyl-butyrylamino) -cyclohexyl-acetic acid methyl ester (55)

Compound 54(93mg, 0.343mmol) was deprotected and coupled to Z-Val-OH (95mg, 0.378mmol) according to the procedure for preparation of compound 39. Flash column chromatography (toluene/ethyl acetate 4: 1) gave compound 55(131mg, 94%) as a colorless solid.

¹H-NMR(300MHz，CDCl₃)：δ0.92-1.30(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＝3.0，5.5Hz，1H)，5.12(s，2H)，5.49(bs，1H)，6.52(bs，1H)，7.34(s，5H)； ¹³C-NMR(75.5MHz，CDCl₃)：δ17.8，19.0，25.8，28.2，29.3，31.2，40.5， 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 56

(S) -2- { [ (1R, 2R, 4S) -2- { (S) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2-methyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarbonyl ] -amino } -pentanoic acid tert-butyl ester (56)

To a solution of compound 55(40mg, 0.099mmol) in ethanol (95%) (7.5mL) was added palladium on activated carbon (10%, 40mg) and the mixture was hydrogenated under pressure at room temperature for 2 h. The mixture was filtered through celite and evaporated. Compound 43(38mg, 0.083mmol) is dissolved in dioxane/H₂O 1∶1(3mL) and the mixture was cooled to 0 ℃ after which 1M LiOH (140 μ L, 0.140mmol) was added to the stirred solution above. After 1 hour the mixture was neutralized with 1M hydrochloric acid, the solvent was evaporated and co-evaporated with toluene. The resulting residue was coupled to deprotected 55 using the same HATU coupling conditions as in synthesis of compound 48. By HPLC (methanol/H containing 0.2% triethylamine)₂O90: 10) gave compound 56 as a colorless solid (56mg, 88%).

¹H-NMR(300MHz，CDCl₃): δ 0.82-0.96(m, 9H), 0.82-1.22(m, overlap peak, 6H), 1.23-1.40(m, 2H), 1.44(s, 9H), 1.50-1.69(m, 4H), 1.71-1.87(m, 2H), 1.95-2.06(m, 1H), 2.07-2.22(m, 1H), 2.28-2.54(m, 3H), 2.60-2.75(m, 1H), 3.08-3.28(m, 1H), 3.30-3.49(m, 1H), 3.70(s, 3H), 3.94(s, 3H), 4.28-4.38(m, 1H), 4.41-4.57(m, 2H), 5.17(b, 1H), 6.54-6.70(m, 6H), 6.74 (m, 6H), 7.9H), 7.7.7-7.9H, 7, 7.8 (ddh), 7.8-3H), 7.7.7H, 7H, 7.9H, 7H, 7H, 7;¹³C-NMR(75.5MHz，CDCl₃): δ 13.7, 18.1, 18.6, 19.2, 25.9, 28.0, 28.2, 29.6, 30.7, 34.6, 36.5, 37.6, 40.8, 47.4, 47.5, 52.1, 52.8, 55.5, 56.8, 58.9, 77.8, 82.0, 98.3, 107.5, 115.3, 118.1, 123.1, 127.5, 128.7, 129.1, 140.5, 151.4, 159.2, 160.7, 161.3, 171.0, 171.5, 172.3, 172.8, 173.0. MALDI-TOF-spectroscopy: (M + H)⁺Calculated values: 815.5, measurement: 815.7, respectively; (M + Na)⁺Calculated values: 837.4, measurement: 837.6, respectively; (M + K)⁺Calculated values: 853.4, measurement: 853.6.

example 57

(S) -2- { [ (1R, 2R, 4S) -2- { (S) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2-methyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarbonyl ] -amino } -pentanoic acid (57)

Tert-butyl ester 56(28mg, 0.034mmol) and triethylsilane (14 μ L, 0.088mmol) were dissolved in DCM (2mL), after which trifluoroacetic acid (2mL) was added and the mixture was stirred for 2 h. Coevaporated with toluene to give 57(26mg, 100%) as a colourless solid.

¹H-NMR(300MHz，CD₃OD): δ 0.86-1.00(m, 9H), 1.01-1.24(m, 4H), 1.36-1.46(m, 2H), 1.48-1.75(m, 8H), 1.70-1.89(m, overlap peak, 1H), 1.96-2.12(m, 1H), 2.22-2.40(m, overlap peak, 2H), 2.49-2.64(m, 1H), 2.72-2.91(m, 1H), 3.26-3.40(m, overlap peak, 1H), 3.50-3.68(m, overlap peak, 1H), 3.62(s, 3H), 4.05(s, 3H), 4.09-4.17(m, 1H), 4.17-4.25(m, 1H), 4.35-4.45(m, 1H), 5.62(b, 7.65, 7.7H), 7.7.7 (ddh, 7H), 7.7H, 7H, 1, 7.98-8.06(m, 2H), 8.41(dd, J ═ 2.8, 9.3Hz, 1H);¹³C-NMR(CD₃OD, 75.5 MHz): δ 13.9, 18.8, 19.7, 20.2, 27.0, 29.7, 30.5, 31.8, 34.6, 37.7, 38.9, 41.1, 47.8, 52.3, 53.6, 56.9, 58.8, 58.9, 60.3, 83.8, 100.4, 102.2, 116.2, 121.6, 126.7, 129.8, 130.8, 133.3, 133.8, 143.5, 157.9, 166.5, 168.5, 173.3, 173.9, 175.5, 175.5, 175.6. MALDI-TOF-spectrum: (M + H)⁺Calculated values: 759.4, measurement: 759.7, respectively; (M + Na)⁺Calculated values: 781.4, measurement: 781.7, respectively; (M + K)⁺Calculated values: 797.4, measurement: 797.7.

example 58

(S) -2- { [ (1R, 2R, 4S) -2- { (S) -1- [ ((S) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2-methyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarbonyl ] -amino } -butyric acid (58)

Prepared as described in example 42, but using tert-butyl L-2-amino-N-butyrate instead of H-Nva-OtBu. The resulting compound was then reacted as described in example 43 to give methyl (1R, 2R, 4R) -2- ((S) -1-tert-butoxycarbonyl-propylcarbamoyl) -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarboxylate. This compound was coupled with 55 as described in example 56 and subsequently esterolyzed as described in example 57 to give compound 58 as a colourless solid.

¹H-NMR(300MHz，CD₃OD): δ 0.82-0.99(m, 9H), 0.82-1.40(m, overlapping peak, 6H), 1.48-1.78(m, 6H), 1.80-1.95(m, 1H), 1.97-2.12(m, 1H), 2.22-2.40(m, overlapping peak, 2H), 2.51-2.64(m, 1H), 2.71-2.90(m, 1H), 3.16-3.39(m, overlapping peak, 1H), 3.49-3.59(m, 1H), 3.63(s, 3H), 3.95(s, 3H), 4.12-4.23(m, 2H), 4.28-4.38(m, 1H), 5.31(b, 1H), 7.43(dd, J ═ 2.2, 9.3, 1H), 7.43(dd, J ═ 2, 9.3, 1H), 7.47(s, 1H), 7.9.9, 9.9.9, 9H), 7.9.9, 7.9, 7.8 (m, 1H), 7.9, 9, 9.9.9 (H), 7.9, 9, 9.9, 9, 1H), 3H;¹³C-NMR(75.5MHz，CD₃OD): δ 10.7, 18.8, 19.7, 25.8, 27.0, 27.0, 29.7, 30.5, 31.8, 37.7, 38.9, 41.2, 47.9, 52.3, 55.3, 56.9, 58.8, 60.6, 83.6, 100.7, 102.2, 116.3, 121.5, 126.7, 129.8, 130.8, 133.7, 133.8, 143.9, 158.2, 166.4, 168.3, 173.3, 173.8, 175.2, 175.5, 175.6. MALDI-TOF-spectrum: (M + H)⁺Calculated values: 745.4, measurement: 744.9, respectively; (M + Na)⁺Calculated values: 767.4, measurement: 766.9, respectively; (M + K)⁺Calculated values: 783.5, measurement: 782.9.

example 59

(S) -2- { [ (1R, 2R, 4S) -2- { (R) -1- [ ((R) -cyclohexyl-methoxycarbonyl-methyl) -carbamoyl ] -2-methyl-propylcarbamoyl } -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarbonyl ] -amino } -butyric acid (59)

Preparation was carried out as described in example 54, but using Boc-D-cyclohexylglycine instead of Boc-L-cyclohexylglycine. The resulting compound was then reacted as described in example 55 followed by coupling with methyl (1R, 2R, 4R) -2- ((S) -1-tert-butoxycarbonyl-pentylcarbamoyl) -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarboxylate as described in example 56. The ester group was removed as described in example 57 to give compound 59 as a colorless solid.

¹H-NMR(CD₃OD, 300 MHz): δ 0.82-1.02(m, 9H), 1.04-1.42(m, 6H), 1.52-1.80(m, 6H), 1.80-1.96(m, overlapping peak, 1H), 2.00-2.14(m, 1H), 2.29-2.46(m, 2H), 2.51-2.65(m, 1H), 2.68-2.84(m, 1H), 3.24-3.39(m, overlapping peak, 1H), 3.47-3.60(m, 1H), 3.67(s, 3H), 4.07(s, 3H), 4.18-4.27(m, 2H), 4.28-4.38(m, 1H), 5.64(app.bs, 1H), 7.44(d, J ═ 2.3, 6.9, 1H), 7.42(s, 7.42H), 7.8 (m, 8H), 7.8 (d, 8H), 8H, 8(d, 8 Hz), 8.41 Hz);¹³C-NMR(CD₃OD, 75.5 MHz): δ 10.8, 18.5, 19.6, 25.7, 27.1, 27.1, 30.1, 30.6, 31.9, 37.3, 38.2, 41.1, 47.8, 52.3, 55.4, 56.9, 59.0, 59.1, 60.2, 83.8, 100.5, 102.2, 116.3, 121.6, 126.8, 129.8, 130.8, 133.6, 133.8, 143.7, 158.1, 166.5, 168.5, 173.4, 173.8, 175.4, 175.7. MALDI-TOF-spectroscopy: (M + H)⁺Calculated values: 745.4, measurement: 745.4, respectively; (M + Na)⁺Calculated values: 767.4, measurement: 767.4, respectively; (M + K)⁺Calculated values: 783.5, measurement: 783.3.

example 60

Resin bound 2-tert-Butoxycarbonylamino-3, 3-dimethylbutyric acid (60)

To Argonaut resin PS-TFP (1.38mmol/g, 10g) and 2-tert-butoxycarbonylamino-3, 3-dimethyl-butyric acid (4.5g, 20.7mmol) was added dichloromethane (40mL) and DMF (10 mL). DMAP (1g, 8.28mmol) was added to the mixture, followed by DIC (9.5mL, 60.7 mmol). After stirring at room temperature for 3 hours, the resin was filtered and washed sequentially with DMF, THF, DCM and diethyl ether, then dried in vacuo.

Example 61

[1- (2-hydroxy-indan-1-ylcarbamoyl) -2, 2-dimethyl-propyl ] -carbamic acid tert-butyl ester (61).

To portion 60(200mg) in DCM was added aminoindanol (0.14 mmol). The mixture was stirred for 2 hours. The liquid was filtered and the resin washed with 2 × DCM. The combined liquids were mixed and concentrated to dryness to give the title compound (20.5mg, 0.055 mmol). Purity > 95% by HPLC. M + H⁺363.15.

¹³C NMR δC(100MHz；CDCl₃；Me₄Si)27.0，28.5，34.2，39.8，50.8，57.9，68.2，73.7，124.8，125.6，127.4，128.5，140.4，171.6.¹H NMR δH(400 MHz；CDCl₃；Me₄Si)1.07(9H，s，CCH₃)，1.44(9H，s，OCCH₃)，2.93(1H，dd，J_gem16.4Hz，J_3，22.3Hz，CH₂)，3.15(1H，dd，J_gem16.4Hz，J_3，25.2Hz，CH₂)，

Example 62

2-amino-N- (2-hydroxy-indan-1-yl) -3, 3-dimethylbutanamide (62)

Compound 61 was held in DCM-TFA 2: 1(2mL) for 60 min at room temperature. The solution was co-evaporated to dryness with toluene.

Example 63

(2-tert-Butoxycarbonylamino-3, 3-dimethyl-butyrylamino) -cyclohexyl-acetic acid methyl ester (63)

To a solution of 2-tert-butoxycarbonylamino-3, 3-dimethylbutyric acid (500mg, 2.16mmol), amino-cyclohexyl-acetic acid methyl ester (444mg, 2.59mmol) and HATU (2g, 5.40mmol) in DMF (20mL) was added diisopropylethylamine (1.88mL, 10.8 mmol). The solution was stirred at room temperature for 1 hour. And diluted with dichloromethane (40 mL). NaHCO for the above solution₃The aqueous solution (saturated) and water (× 2) were washed, dried and concentrated. The purity of the obtained product is more than 95 percent. M + H⁺385.4。

Example 64

{1- [ (cyclohexyl-methylcarbamoyl-methyl) -carbamoyl ] -2, 2-dimethyl-propyl } -carbamic acid tert-butyl ester (64)

To compound 63 in EtOH-THF 1: 2 was added a large excess of methylamine (30% in water) and it was left at room temperature for 2 weeks. The above solution was concentrated to dryness and the resulting residue was passed through a short silica gel column eluting with 2% methanol in dichloromethane to give pure product (> 95%). M + H⁺384.5

Example 65

2-amino-N- (cyclohexyl-methylcarbamoyl-methyl) -3, 3-dimethyl-butyramide (65)

Compound 64 was kept in dichloromethane-trifluoroacetic acid 2: 1 for 1 hour at room temperature and concentrated to dryness. The resulting residue was dried in vacuo for 16 h. Reversed phase C18HPLC indicated > 95% purity. M + H⁺283.1。

Example 66

1- (2-amino-4-methoxyphenyl) ethanone (66)

M-methoxyaniline (10.0g, 82mmol) was dissolved in CH₂Cl₂(50mL) and the solution was cooled to-50 ℃. The BCl is added over a period of 20 minutes₃(in CH)₂Cl₂1M, 82mL, 82mmol) was added slowly thereto, after which the mixture was stirred at-50 ℃ for 30 minutes, and then AcCl (6.0mL, 84mmol) and AlCl were added sequentially thereto₃(11g, 82 mmol). The mixture was stirred at-50 ℃ for 1 hour, then allowed to warm to room temperature. After stirring at room temperature overnight, the above solution was heated at 40 ℃ for 4h, after which the resulting mixture was poured onto ice. The resulting aqueous mixture was basified with 10% NaOH (w/v) and extracted with EtOAc (4X 200 mL). The combined organic phases were washed with brine and dried (MgSO)₄) And evaporated to give a black solid which was purified by flash column chromatography (ether/CH)₂Cl₂20: 80) was purified. The resulting solid was recrystallized from ether/hexane to give compound 93(5.6g, 42%) as bright tan leaves.

Example 67

N- (tert-butyl) -N' -isopropylthiourea (67)

To tert-butyl isothiocyanate (5.0mL, 39mmol) in CH₂Cl₂(200mL) to the solution were added isopropylamine (4.0mL, 47mmol) and Diisopropylethylamine (DIEA) (6.8mL, 39mmol), and the resulting mixture was stirred at room temperature for 2 h. The resulting reaction mixture was diluted with EtOAc and diluted with 10% citric acid (2X), saturated NaHCO₃(2×)、H₂O (2X) and brine (1X). The resulting organic layer was dried (MgSO)₄) And evaporated to give the title compound as a white solid (3.3g, 52%) which was used without further purification.

Example 68

N-isopropylthiourea (68)

Compound 67(3.3g, 20mmol) was dissolved in concentrated HCl (45mL) and the solution was refluxed for 40 minutes. The mixture was cooled to room temperature, then cooled in an ice bath and quenched with solid NaHCO₃And saturated NaHCO₃Basified to pH 9.5, after which the product was extracted into EtOAc (3 ×). Combined organic phases with H₂O (2x) and brine (1x) were washed, dried (MgSO)₄) And evaporated to give the crude title compound (2.1g, 90%) which was used without further purification.

Example 69

2- (isopropylamino) -1, 3-thiazole-4-carboxylic acid hydrobromide salt (69)

A suspension of compound 68(2.1g, 18mmol) and 3-bromopyruvic acid (3.0g, 18mmol) in dioxane (180mL) was heated to 80 ℃. The mixture became clear when reaching 80 ℃ and the product began to precipitate as a white solid shortly thereafter. After heating for 2h, the reaction mixture was cooled to room temperature and the precipitate was filtered off and collected. The pure titled product was thus obtained (4.4g, 94%).

Example 70

N- (2-acetyl-5-methoxyphenyl) -2- (isopropylamino) -1, 3-thiazole-4-carboxamide (70)

A mixture of compound 69(4.4g, 16.5mmol) and aniline derivative 66(2.75g, 16.5mmol) in pyridine (140mL) was cooled to-30 deg.C (the clear solution partially became a suspension by cooling). During 5 minutes, POCl was added₃(3.3mL, 35mmol) was added slowly. The mixture was stirred at-30 ℃ for 1 hour and then allowed to reach room temperature. After stirring at room temperature for 1.5h, the resulting reaction mixture was poured onto ice and washed with solid NaHCO₃And saturated NaHCO₃Adjusting the pH value to about 9-10. Extracting the obtained crude product with CH₂Cl₂(3X) and the combined organic phases were dried (MgSO₄) And evaporation. The resulting crude dark beige solid was purified by flash column chromatography (hexanes/EtOAc 55: 45) to afford compound 70 as a pale yellow solid (5.6g, 76%).

Example 71

2- [2- (isopropylamino) -1, 3-thiazol-4-yl ] -7-methoxyquinolin-4-ol (71)

A solution of t.BuOK (2.42g, 21mmol) in dry t.BuOH (40mL) was heated to reflux. Compound 70(1.8g, 5.4mmol) was added thereto in portions over a period of 5 minutes and the resulting dark red solution was stirred under reflux for an additional 20 minutes. The mixture was cooled to room temperature and HCl (4M in dioxane, 8.0mL, 32mmol) was added, after which the resulting reaction mixture was concentrated in vacuo. To ensure that all HCl and dioxane were removed, the crude product was redissolved in CH₂Cl₂Twice and add itEvaporation was complete to give slightly impure compound 71 hydrochloride as a brown solid (1.62 g). Dissolving the above product in CH₂Cl₂Neutralized and saturated NaHCO₃Washing is carried out, after which the aqueous phase is washed with CH₂Cl₂And extracting for several times. The combined organic phases were dried (MgSO)₄) And evaporated to give compound 71 as a light brown solid (1.38g, 81%) (purity > 95% according to HPLC test).¹H-NMR(MeOH-d₄，400MHz)：δ1.30(d，J＝6.0Hz，6H)，3.93(s，3H)，3.95-4.07(m，1H)，6.73(s，1H)，6.99(dd，J＝2.4，9.2Hz，1H)，7.26(d，J＝2.4Hz，1H)，7.37(s，1H)，8.10(d，J＝9.2Hz，1H).

Example 72

(1R, 4R, 5R) -N- [ (1S) -1- [ [ [ (1S) -1-cyclohexyl-2- (methylamino) -2-oxoethyl ] amino ] carbonyl ] -2, 2-dimethylpropyl ] -3-oxo-2-oxabicyclo [2.2.1] heptane-5-carboxamide (72)

To a solution of compound 32(53mg, 0.34mmol) in DMF (9mL) was added compound 65(80mg, 0.28mmol) and DIEA (290L, 1.66 mmol). The solution was cooled to 0 ℃ and HATU (127mg, 0.33mmol) was added thereto. After stirring at 0 ℃ for 1h and at room temperature for 1h, the solvent was evaporated and the resulting crude product was purified by flash column chromatography (EtOAc/toluene 2: 1) to afford compound 72 as a white solid (110mg, 92%).

Example 73

(1R) -1- [ [ [ (1R, 2R, 4R) -2- [ [ [ (1S) -1- [ [ [ (1S) -1-cyclohexyl-2- (methylamino) -2-oxoethyl ] amino ] carbonyl ] -2, 2-dimethylpropyl ] amino ] carbonyl ] -4-hydroxycyclopentyl ] carbonyl ] amino ] -2-vinyl-cyclopropanecarboxylic acid ethyl ester (73)

Compound 72(60mg, 0.14mmol) was dissolved in dioxane (3.5mL) and H₂O (2.5mL) and the solution was cooled to 0 ℃. LiOH (1M, 280L, 0.28mmol) was added dropwise thereto over a period of 5 minutes, after which the reaction mixture was stirred at 0 ℃ for 40 minutes. The pH was adjusted to 7 with 1M HCl and the solvent was evaporated. The resulting residue was suspended in DMF (5mL) and 1-amino-2-vinyl-cyclopropanecarboxylic acid ethyl ester (32mg, 0.17mmol) and DIEA (146L, 0.84mmol) were added thereto. After cooling to 0 ℃, HATU (64mg, 0.17mmol) was added thereto, and the mixture was stirred at 0 ℃ for 1 hour and at room temperature for 1 hour. The solvent was evaporated and the resulting product was purified by flash column chromatography (EtOAc/methanol 9: 1) to afford compound 73 as a white solid (67mg, 82%).

Example 74

Tert-butyl (1R, 2R, 4R) -2- [ [ [ (1R) -1- (ethoxycarbonyl) -2-vinylcyclopropyl ] amino ] carbonyl ] -4- [ [2- [2- (isopropylamino) -1, 3-thiazol-4-yl ] -7-methoxyquinolin-4-yl ] oxy ] cyclopentanecarboxylate (74)

The title compound was prepared according to the procedure described in example 76 method a, but using compound 34 instead of compound 73. (Note: use of 4 equivalents of Ph₃P and DIAD. Chromatography eluent: toluene/EtOAc 1: 1)

Example 75

(1R, 2R, 4R) -2- [ [ [ (1R) -1- (ethoxycarbonyl) -2-vinylcyclopropyl ] amino ] carbonyl ] -4- [ [2- [2- (isopropylamino) -1, 3-thiazol-4-yl ] -7-methoxyquinolin-4-yl ] oxy ] cyclopentanecarboxylic acid (75)

To compound 74(20mg, 30. mu. mol) in CH₂Cl₂To the solution (2mL) was added TFA (2mL) and Et₃SiH (10. mu.L, 63. mu. mol). After 2 hours the volatiles were evaporated and the crude product was used without any purification steps. Compound 75: 18mg, quantitative, as a white solid.

Example 76

(1R) -1- [ [ [ (1R, 2R, 4S) -2- [ [ [ (1S) -1- [ [ [ (1S) -1-cyclohexyl-2- (methylamino) -2-oxoethyl ] amino ] carbonyl ] -2, 2-dimethylpropyl ] amino ] carbonyl ] -4- [ [ 7-methoxy-2- [2- [ (1-methylethyl) amino ] -4-thiazolyl ] -4-quinolinyl ] oxy ] cyclopentyl ] carbonyl ] amino ] -2-vinyl-cyclopropanecarboxylic acid ethyl ester (76)

The method A comprises the following steps:to a solution of compound 73(59mg, 0.10mmol) in anhydrous THF (4mL) was added quinoline 71(49mg, 0.16mmol) and Ph₃P (65mg, 0.25 mmol). After cooling to 0 deg.C, DIAD (50. mu.L, 0.25mmol) was added dropwise over a period of 5 minutes. The resulting solution was stirred at 0 ℃ for 1h and at room temperature for 48 h. Evaporating the solvent and utilizing the residue obtainedFlash column chromatography (CHCl)₃/2M NH₃In methanol, 95: 5) to give compound 76 as a white solid (9mg, 10%).

The method B comprises the following steps:compound 75 was coupled to compound 65 according to the procedure described in example 72 to give the title compound (82%).

Example 77

(1R) -1- [ [ [ (1R, 2R, 4S) -2- [ [ [ (1S) -1- [ [ [ (1S) -1-cyclohexyl-2- (methylamino) -2-oxoethyl ] amino ] carbonyl ] -2, 2-dimethylpropyl ] amino ] carbonyl ] -4- [ [ 7-methoxy-2- [2- [ (1-methylethyl) amino ] -4-thiazolyl ] -4-quinolinyl ] oxy ] cyclopentyl ] carbonyl ] amino ] -2-vinyl-cyclopropanecarboxylic acid (77)

Compound 76(8mg, 9. mu. mol) was dissolved in a mixture of methanol (150. mu.L) and THF (100. mu.L). LiOH (1mg, 42. mu. mol) in H₂O (25L) solution was added thereto, and the mixture was stirred at 50 ℃ overnight. The solution was neutralized with HOAc and evaporated. Suspending the residue in CH₂Cl₂And with H₂And O, washing the mixture. The organic phase was evaporated to give the title compound as a white solid (8mg, quantitative).

¹H-NMR(MeOH-d₄400MHz) (rotamer mixture): δ 0.60-1.33(m, 21H), 1.35-1.73(m, 12H), 1.90-2.42(m, 2H), 2.51-2.75(m, 6H), 3.20-3.38(m, 1H), 3.85(s, 3H), 3.95-4.28(m, 1H), 4.91-5.02(m, 1H), 5.12-5.23(m, 1H), 5.64-5.83(m, 1H), 7.01-7.11(m, 1H), 7.25-7.40(m, 1H), 7.42-7.57(m, 1H), 7.85-8.08(m, 1H).

Example 78

2-amino-3, 3-dimethyl-N-thiophen-2-yl-methyl-butyramide (78)

Prepared as described in example 61, but using thiophene-2-methylamine instead of aminoindanol, followed by removal of the Boc group as described in example 62, and the title compound was prepared.

Example 79

2-amino-N- (6-hydroxy-4, 5, 6, 7-tetrahydro-benzo [ b ] thiophen-5-yl) -3, 3-dimethylbutanamide (79)

The title compound was prepared as follows: as described in example 61, but using 2-amino-4, 5, 6, 7-tetrahydro-benzo [ b ] thiophen-5-ol instead of aminoindanol, the Boc group was subsequently removed as described in example 62.

Example 80

2-amino-N- (2-diethylamino-ethyl) -3, 3-dimethyl-butyramide (80)

The title compound was prepared as follows: the Boc group was subsequently removed as described in example 62, but using N, N-diethylethylenediamine instead of aminoindanol.

Example 81

2-amino-N- [2- (2-methoxy-phenoxy) -ethyl ] -3, 3-dimethyl-butyramide (81)

The title compound was prepared as follows: as described in example 61, but using 2-methoxyphenoxyethylamine instead of aminoindanol, the Boc group was subsequently removed as described in example 62.

Example 82

2-amino-1- (3-hydroxy-pyrrolidin-1-yl) -3, 3-dimethyl-butan-1-one (82)

The title compound was prepared as follows: the Boc group was subsequently removed as described in example 62, but using (R) -3-pyrrolidone instead of aminoindanol.

Example 83

2-amino-N- (1, 1-dioxo-tetrahydro-1-thiophen-3-yl) -3, 3-dimethyl-butyramide (83)

The title compound was prepared as follows: as described in example 61, but using 2-methoxyphenoxyethylamine instead of aminoindanol, the Boc group was subsequently removed as described in example 62.

Example 84

Carbamic acid, [ (1S) -1- [ [ (phenylsulfonyl) amino ] carbonyl ] butyl ] -, phenylmethyl ester (84)

To a stirred solution of Z-Nva-OH (150mg, 0.59mmol) in THF (6mL) was added CDI (400mg, 2.4 mmol). The slurry was stirred at room temperature for 30 minutes, to which was then added a solution of DBU (200. mu.L, 1.3mmol) and benzenesulfonamide (250mg, 1.59mmol) in THF (2 mL). The mixture was stirred at 60 ℃ for 48 hours and then concentrated to dryness. The resulting residue was dissolved in methanol and subjected to HPLC purification to give the title compound (118.5mg, 0.304 mmol). Purity > 95% by HPLC. M-H⁺389.0，+Na 412.96.

Example 85

(2S) -2-amino-N- (phenylsulfonyl) pentanamide (85)

Compound 84 was dissolved in methanol (5mL), followed by the addition of Pd/C and hydrogenation for 2 hours. The slurry was filtered through celite, washed with methanol and concentrated to dryness to give the title compound. The yield was 100%. M + H⁺257.3。

Example 86

4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentane-1, 2-dicarboxylic acid 1- { [1- (cyclohexylmethyl-carbamoyl) -2-methyl-propyl ] -amide }2- [ (1-benzenesulfonylaminocarbonyl-2-vinyl-cyclopropyl) -amide ] (86)

N- (tert-Butoxycarbonyl) -L-valine was attached to Argonaut resin PS-TFP as described in example 60, followed by reaction with cyclohexylmethylamine as described in example 61 and removal of the Boc group as described in example 62. The resulting amine was coupled with Compound 35 as described in example 39, followed by hydrolysis of the ethyl ester as described in example 40 to give 1- { [2- [1- (cyclohexylmethyl-carbamoyl) -2-methyl-propylcarbamoyl]-4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarbonyl]-amino } -2-vinyl-cyclopropanecarboxylic acid. The resulting acid was then treated as described in example 94, but using toluene sulfonamide instead of cyclopropyl sulfonamide, to give the title compound. The yield was 6%. Purity > 95% by HPLC. M + H⁺864.32。

Example 87

Acetic acid (1S, 2R) -1- ((2S) -2-amino-3, 3-dimethyl-butyrylamino) -indan-2-yl ester (87)

A solution of compound 61(4g) was held in pyridine-acetic anhydride 2: 1 for 30 minutes. DCM was added to the solution and the resulting solution was treated with citric acid (aq) and NaHCO₃(aqueous solution) washing was performed. The resulting organic layer was concentrated to dryness to give the acetylated product with a purity of > 90% by HPLC. The resulting compound was then placed in 30% TFA in DCM for 1.5 hours, and then concentrated to dryness. Coevaporation twice in toluene gave the title product with an HPLC purity of > 90%.

Example 88

(2S) -methanesulfonic acid 2-tert-butoxycarbonylamino-4-methyl-pentyl ester (88)

To a solution of ((1S) -1-hydroxymethyl-3-methyl-butyl) -carbamic acid tert-butyl ester (25g, 115mmol) in dichloromethane (500ml) cooled by an ice-water bath was added diisopropylethylamine (35.7g, 276mmol) and methanesulfonyl chloride (15.81g, 138mmol) in that order. The resulting solution was stirred overnight, during which time the mixture was allowed to warm gradually to ambient temperature. The resulting mixture was sequentially washed with water, 10% citric acid (aq), water and saturated NaHCO₃(aqueous solution) washing, then with Na₂SO₄Dried and concentrated to give a brown solid (32.6g, 96%) which was used in the next reaction without further purification.

Example 89

ii) ((1S) -1-Azidomethyl-3-methyl-butyl) carbamic acid tert-butyl ester (89)

The mesylate from example 88 (32.6g, 110mmol) was treated with sodium azide (21.45g, 330mmol) in DMF at 80 deg.C for 24 h. The solvent was evaporated, the resulting residue was taken up in DCM, filtered and washed with saturated NaHCO₃(aqueous solution) washing. The obtained solution was treated with Na₂SO₄Drying and concentration were carried out to give a brown oil, which was purified by flash chromatography using gradient ethyl acetate and hexane to give the title compound as a white solid (19.55g, 73%).

Example 90

(1S) -1-azidomethyl-3-methyl-butylamine (90)

((1S) -1-Azidomethyl-3-methyl-butyl) -carbamic acid tert-butyl ester (9.64g, 39.78mmol) was treated with TFA (30ml) in DCM (150ml) for 3h, the mixture was evaporated under reduced pressure, the resulting residue was dissolved in ethyl acetate and taken up with 1MK₂CO₃Washing with aqueous solution and adding Na₂SO₄Dried and concentrated to give a yellow liquid (4.55g, 80%).

Example 91

1- { [ 2-hex-5-enylcarbamoyl-4- (7-methoxy-2-phenyl-quinolin-4-yloxy) -cyclopentanecarbonyl ] -amino } -2-vinylcyclopropanecarboxylic acid ethyl ester (91)

The tert-butyl ester of compound 35 was removed by treatment with triethylsilane as described in example 39. The resulting acid (724mg, 1.33mmol), hex-5-enylamine hydrochloride (271mg, 2mmol) and diisopropylethylamine (1.85ml, 10.65mmol) were dissolved in DMF (20ml) and cooled to 0 ℃. After 30min, HATU (608mg, 1.6 mmol) was added thereto and the flask was removed from the ice bath. The reaction was followed by LC-MS. After 3 hours, the reaction mixture was extracted into EtOAc (100ml) and aqueous sodium bicarbonate (15 ml). The EtOAc phase was dried over magnesium sulfate, evaporated and purified by chromatography on silica gel (25% EtOAc in hexanes → 50% EtOAc in hexanes) to give the pure title product (726mg, 87%). MS (M + H)⁺)：525.8

Example 92

17- (7-methoxy-2-phenyl-quinolin-4-yloxy) -2, 14-dioxo-3, 13-diaza-tricyclo [13.3.0.0 [4, 6] octadec-7-ene-4-carboxylic acid ethyl ester (92)

Compound 91(363mg, 0.58mmol) was dissolved in degassed dichloromethane (100 ml). The Hoveyda-Grubbs catalyst second generation (26mg, 0.041mmol) was then added to it and the mixture was refluxed under an argon atmosphere overnight. The reaction mixture was evaporated on silica and purified by silica gel chromatography (50% EtOAc in hexanes → 70% EtOAc in hexanes) to give the pure title product (111mg, 32%). MS (M + H)⁺)：597.7

Example 93

17- (7-methoxy-2-phenyl-quinolin-4-yloxy) -2, 14-dioxo-3, 13-diaza-tricyclo [13.3.0.0^＊4，6^＊]Octadecan-7-ene-4-carboxylic acid (93)

Compound 92(95mg, 0.159mmol) was dissolved in tetrahydrofuran (10ml), methanol (5ml) and water (4 ml). Lithium hydroxide (40mg, 1.67mmol) was dissolved in water (1ml) and added to the above solution. The reaction mixture was heated to 65 ℃. After 3 hours the reaction mixture was cooled, acidified with aqueous HCl (pH 5), evaporated on silica and purified by silica gel chromatography (10% methanol in dichloromethane → 15% methanol in dichloromethane) to give the pure title product (65mg, 72%). MS (M + H)⁺)：569.8

Example 94

Cyclopropanesulfonic acid [17- (7-methoxy-2-phenyl-quinolin-4-yloxy) -2, 14-dioxo-3, 13-diaza-tricyclo [13.3.0.0^＊4，6^＊]Octadec-7-en-4-carbonyl]-amides (94)

Compound 93(65mg, 0.12mmol), DMAP (21mg, 0.17mmol) and EDAC (44mg, 0.23mmol) were dissolved in DMF (0.2 ml). The reaction mixture was stirred at room temperature for 5 h. Cyclopropyl sulfonamide (69mg, 0.57mmol) and DBU (80. mu.l, 0.57mmol) were then added thereto. In the roomAfter stirring overnight at room temperature, the reaction mixture was extracted into EtOAc (80ml) and aqueous citric acid (10%, 2 × 15 ml). The organic phase is MgSO₄Drying, evaporation on silica and purification by chromatography on silica gel (5% methanol in dichloromethane → 15% methanol in dichloromethane) twice gave a thick slurry. The thick slurry was dissolved in a small amount of acetonitrile and precipitated with ether to give the pure title product (19mg, 23%). MS (M + H)⁺)：673.2

Example 95

1- { [ 2-hex-5-enyl-methyl-carbamoyl) -4- (7-methoxy-2-phenyl-quinolin-4-yloxy) ] -cyclopentanecarbonyl ] -amino-2-vinyl-cyclopropanecarboxylic acid ethyl ester (95)

The tert-butyl ester of compound 35 was removed according to the procedure described in example 39. The resulting acid (850mg, 1.56mmol), N-methylhexan-5-enylamine hydrochloride (380mg, 2.5mmol) and diisopropylethylamine (2.3mL, 13.4mmol) were dissolved in DMF (60mL) and cooled to 0 ℃. After 30min, HATU (0.76mg, 2.0mmol) was added and the flask removed from the ice bath. The reaction was followed by TLC. After 2 hours, the reaction mixture was added to 5% citric acid and extracted three times with ethyl acetate. The organic phase obtained is dried over sodium sulfate and evaporated under reduced pressure. The resulting crude product was purified by silica gel chromatography to give the titled product (820mg, 82%).

Example 96

17- (7-methoxy-2-phenyl-quinolin-4-yloxy) -13-methyl-2, 14-dioxo-3, 13-diaza-tricyclo [13.3.0.0^＊4，6^＊]Octadecan-7-en-4-carboxylic acid ethyl ester (96)

Compound 95(648mg, 1.01mmol) was dissolved in degassed dichloroethane (500 ml). The Hoveyda-Grubbs catalyst second generation (35mg, 0.055mmol) was then added thereto and the mixture was refluxed under an argon atmosphere overnight. The reaction mixture was evaporated on silica and purified by chromatography on silica gel (30% EtOAc in toluene → 50% EtOAc in toluene) to give the pure title product (230mg, 37%). MS (M + H)⁺)：612.8

Example 97

17- (7-methoxy-2-phenyl-quinolin-4-yloxy) -13-methyl-2, 14-dioxo-3, 13-diaza-tricyclo [13.3.0.0^＊4，6^＊]Octadecan-7-ene-4-carboxylic acid ethyl ester (97)

Compound 96(260mg, 0.42mmol) was dissolved in 1, 4-dioxane (20mL), 1.0M lithium hydroxide (6.0mL) was added thereto, and the mixture was stirred at room temperature overnight and then at 60 ℃ for six hours. The reaction mixture was added to 5% citric acid, and extracted 3 times with ethyl acetate. The organic phase obtained is dried over sodium sulfate and evaporated under reduced pressure. The resulting crude product was purified by silica gel chromatography with DCM and 5% MeOH to give the title product (130mg, 53%). MS (M + H): 584,7

Example 98

Cyclopropanesulfonic acid [17- (7-methoxy-2-phenyl-quinolin-4-yloxy) -13-methyl-2, 14-dioxo-3, 13-diaza-tricyclo [13.3.0.0 ]^＊4，6^＊]Octadec-7-en-4-carbonyl]-amides (98)

Compound 97(58.3mg, 0.1mmol), DMAP (18.3mg, 0.15mmol) and EDAC (38.7mg, 0.2mmol) were dissolved in DMF (1.0 ml). The reaction mixture was stirred at room temperature overnight. Cyclopropyl sulfonamide (60.5mg, 0.5mmol) and DBU (76. mu.g, 0.5mmol) were then added thereto. After stirring at room temperature overnight, the reaction mixture was added to 5% citric acid and extracted three times with ethyl acetate. The organic phase obtained is dried over sodium sulfate and evaporated. The resulting residue was purified twice by silica gel chromatography to give the title product (20 mg). MS (M + H)687, 8.

Example 99

[ [ 4-cyclopropanesulfonylaminocarbonyl-17- (7-methoxy-phenyl-quinolin-4-yloxy) -2, 14-dioxo-3, 13-diaza-tricyclo [13.3.0.0 ]^＊4，6^＊]Octadec-7-en-13-yl]-carbamic acid tert-butyl ester (99)

N' -hex-5-en- (E) -ylidene-hydrazinecarboxylic acid tert-butyl ester was prepared according to the procedure described in examples 46 and 47, but starting from hex-5-en-ol instead of hept-6-en-ol. Compound 35 was treated as described in example 48, but using the above N' -hexane-5-ene- (E) -ylidene-hydrazinecarboxylic acid tert-butyl ester instead of the corresponding hept-6-ene derivative, followed by macrocyclization as described in example 49 and hydrolysis of the ethyl ester as described in example 50 to give the acid. The resulting acid (58mg, 0.0846mmol) was dissolved in anhydrous DMF (7mL) and DIEA was added dropwise over a period of 1 min. The resulting solution was stirred at room temperature for 1 hour, then a solution of cyclopropylsulfonamide (41mg, 0.338mmol), DMAP (41.3mg, 0.338mmol) and DBU (50. mu.L, 0.338mmol) in dry DMF (1.5mL) was added. The above solution was stirred at room temperature for 5 days. The resulting solution was diluted with EtOAc (50mL) and saturated NaHCO₃Washing is carried out. The resulting aqueous phase was extracted with DCM. The combined organic layers were dried, concentrated and purified by HPLC to give the title compound as a white solid (14.3mg, 0.018mmol) which was > 95% pure by HPLC. M + H⁺788.3。

Example 100

Cyclopropanesulfonic acid [ 13-amino-17- (7-methoxy-2-phenyl-quinolin-4-yloxy) -2, 14-dioxo-3, 13-diaza-tricyclo [13.3.0.0 ]^＊4，6^＊]Octadec-7-en-4-carbonyl]-amide trifluoroacetate salt (100)

Compound 99(2.4mg, 0.00304mmol) was held in TFA-DCM 1: 2(3mL) for 60 min at room temperature. Toluene (3mL) was added. The above sample was co-evaporated to dryness to form the title compound (2.1mg, 0.0026 mmol). Purity > 95% by HPLC. M + H⁺688.3.

Example 101

3-oxo-2-oxa-bicyclo [2.2.1] heptane-5-carboxylic acid hex-5-enyl-methylamide (101)

To HATU (2.17g, 5.7mmol) and N-methylhexan-5-enylamine hydrochloride (6.47mmol) in 5mL DMF in an ice bath under argon was added 1R, 4R, 5R-3-oxo-2-oxa-bicyclo [2.2.1] bicyclo [ 2.1] in 11mL DMF]Heptane-5-carboxylic acid (835.6mg, 5.35mmol) followed by DIEA (2.80mL, 16mmol) was added. After stirring for 40 minutes, the mixture was stirred at room temperature for 5 h. The solvent was evaporated, the residue was dissolved in EtOAc (70mL) and washed with saturated NaHCO₃(10mL) was washed. The aqueous phase was extracted with EtOAc (2X 25 mL). The organic phases were combined, washed with saturated NaCl (20mL), Na₂SO₄Dried and evaporated. Flash column chromatography (150g silica gel, 2/1 EtOAc-Petroleum Ether (PE), by KMnO₄TLC detection of the aqueous solution with Rf 0.55 in 4/1 EtOAc-PE) gave compound as a yellow oil (1.01g, 75%).

Example 102

4-Hydroxycyclopentane-1, 2-dicarboxylic acid 1- [ (1-cyclopropanesulfonylaminocarbonyl-2-vinylcyclopropyl) -amide ]2- (hex-5-enyl-methylamide (102)

LiOH solution (0.15M, 53mL, 8mmol) was added to lactonamide 101(996mg, 3.96mmol) in an ice bath and stirred for 1 h. The mixture was acidified with 1N HCl to pH 2-3 and evaporated, co-evaporated with toluene several times, and dried overnight under vacuum. Reacting (1R, 2S) -cyclopropanesulfonic acid (1-amino-2-vinylcyclopropane)Carbonyl) amide hydrochloride (4.21mmol) and HATU (1.78g, 4.68mmol) were added thereto. The mixture was cooled in an ice bath under argon, DMF (25mL) was added, and DIEA (2.0mL, 11.5mmol) was added. After stirring for 30 minutes, the mixture was stirred at room temperature for 3 hours. After evaporation of the solvent, the residue was dissolved in EtOAc (120mL), washed sequentially with 0.5N HCl (20mL) and saturated NaCl (2X 20mL), and Na₂SO₄It is dried. By flash column chromatography (200g of YMC silica gel in CH)₂Cl₂Medium 2-4% methanol) gave a white solid (1.25g, 66%).

Example 103

Cyclopropanesulfonic acid (17-hydroxy-13-methyl-2, 14-dioxo-3, 13-diazacyclo [ 13.3.0.0)^＊4，6^＊]Octadecan-7-en-4-carbonyl) -amide (103)

Cyclopentanol 102(52.0mg, 0.108mmol) was dissolved in 19mL of 1, 2-dichloroethane (argon bubbled before use). Hoveyda-Grubbs second generation catalyst (6.62mg, 10 mole%) was dissolved in DCE (2X 0.5mL) and added to the solution. The fresh solution was bubbled with argon for 1 minute. Aliquots (4mL each) were transferred to five 2-5 mL microwave tubes. To the last tube was added 0.8mL of a rinsing solvent (ringing with solvent). The tubes were heated by microwave (room temperature to 160 ℃ C. over 5 minutes). All aliquots were combined and the solvent was evaporated. By flash chromatography (silica gel, 3-7% methanol in CH)₂Cl₂In) gave 24.39mg of a solid (in 10% MeOH-CH)₂Cl₂Medium Rf 0.28 with two spots). The resulting solid was combined with 9.66mg of sample and passed through a second chromatography (2-8% methanol in EtOAc) to afford 80% of the desired compoundCream solid (23mg) (26% yield).

Example 104

Cyclopropanesulfonic acid {17- [2- (4-isopropyl thiazole-2-yl) -7-methoxy quinoline-4-base oxygen radical]-13-methyl-2, 14-dioxo-3, 13-diazacyclo [13.3.0.0^＊4，6^＊]Octadecyl-7-en-4-carbonyl) -amide (104)

DIAD (22. mu.L, 0.11mmol) was added to a mixture of metathesis product 103(23mg), 2- (4-isopropyl-1, 3-thiazol-2-yl) -7-methoxyquinolin-4-ol (24mg, 0.08mmol) and PPh in an ice bath₃(30mg, 0.11mmol) in 1mL anhydrous THF. The resulting mixture was stirred at room temperature overnight and then evaporated. The resulting residue (1.2 mL of 1.5mL MeCN solution) was purified by preparative HPLC (Hypercarb 7uL 100X 21.2mm, 40% to 99% aqueous MeCN solution over 10 minutes) to give 3.18mg MV062308 as a cream solid (13% yield).

¹H NMR (DMSO-d6) delta ppm: primary rotamers 0.99(m, 2H), 1.11(m, 2H), 1.20-1.30(m, 2H), 1.37 and 1.38(2d, J ═ 7.0Hz, 6H), 1.46-1.58(m, 2H), 1.70(m, 1H), 1.85(m, 1H), 1.90(dd, J ═ 8.5, 6.0Hz, 1H), 2.06(br, 1H), 2.26(m, 1H), 2.38(m, 1H), 2.52-2.62(m, 3H), 2.90-2.97(m, 2H), 3.06(s, 3H), 3.21(m, 1H), 3.40-3.56(m, 2H)3.97(s, 3H), 4.60(m, 1H), 5.04(m, 1H), 5.92 (m, 5.58H), 1H, 7.58 (m, 1H), 1H, 6(m, 1H), 1H, 6.8.58 (m, 1H), 1H

Example 105

N- {4- [4- (4-cyclopropanesulfonylaminocarbonyl-13-methyl-2, 14-dioxo-3, 13-diaza-tricyclo [13.3.0.0 ]^＊4，6^＊]Octadecyl-7-en-17-yloxy) -7-methoxy-quinolin-2-yl]-thiazol-2-yl } -3, 3-dimethylbutanamide (105)

Compound 103 was treated with 4-hydroxy-7-methoxy-2- [2- (2, 2-dimethylbutyryl) aminothiazol-4-yl ] quinoline as described in example 104 to afford the title compound.

LCMS: retention time 2.30min, gradient 30% -80% B in 3 min (flow rate: 0.8mL/min, UV 220nm, ACE C83X 50 mm; mobile phase A at 90% H₂10mM NH in O₄Ac, B10 mM NH in 90% ACN₄Ac)，(M+1)⁺＝807。

Example 106

1- { [2- (hex-5-enyl-methyl-carbamoyl) -4-hydroxy-cyclopentanecarbonyl ] -amino } -2-vinyl-cyclopropanecarboxylic acid ethyl ester (106)

Compound 101 was reacted as described in example 102, but using ethyl 1-amino-2-vinylcyclopropanecarboxylate instead of (1R, 2S) -cyclopropanesulfonic acid (1-amino-2-vinyl-cyclopropanecarbonyl) amide hydrochloride, thereby giving the title compound.

Example 107

1- { [4- (4-bromo-benzenesulfonyloxy-2- (hex-5-enyl-methyl-carbamoyl) -cyclopentanecarbonyl ] -amino } -2-vinyl-cyclopropanecarboxylic acid ethyl ester (107)

Compound 106(115mg, 0.286mmol) was dissolved in 5ml of toluene and 1ml of dichloromethane. DABCO (2.2.2-diazobicyclooctane) (96mg, 0.857mmol, 3 equiv.) was added to the above solution followed by BsCl (109mg, 0.428mmol, 1.5 equiv.). The reaction was stirred at room temperature overnight, diluted with toluene (+ 10% ethyl acetate), washed with saturated sodium bicarbonate, brine, dried over sodium sulfate and evaporated. The desired product was obtained by column chromatography (eluent EtOAc, Rf 0.25). The conversion was 80%. The yield was 106 mg.

Example 108

17- (4-bromo-benzenesulfonyloxy) -13-methyl-2, 14-dioxo-3, 13-diaza-tricyclo [13.3.0.0^＊4，6^＊]Octadecan-7-ene-4-carboxylic acid ethyl ester (108)

Compound 107(106mg, 0.169mmol) was dissolved in dichloromethane (40ml) and degassed by bubbling nitrogen into the solution for 20 minutes. The Hoveyda-Grubbs catalyst first generation (10mg, 0.017mmol, 10 mol%) was then added thereto and the mixture was refluxed under a nitrogen atmosphere overnight. The reaction mixture was then cooled to room temperature, and MP-TMT palladium scavenger (ca. 100mg) was added thereto and stirred for 2.5 hours. The scavenger was removed by filtration and washed with 50mL of dichloromethane. The resulting solution was concentrated by rotary evaporation. The resulting crude product was purified by column chromatography (EtOAc) to give 61mg of product. The yield was 60%.

Example 109

17- [2- (2-isopropylamino-thiazol-4-yl) -7-methoxy-quinolin-4-yloxy]-13-methyl-2, 14-dioxo-3, 13-diaza-tricyclo [13.3.0.0^＊4，6^＊]Octadecan-7-ene-4-carboxylic acid ethyl ester (109)

2- (isopropylamino-thiazol-4-yl) -7-methoxy-quinolin-4-ol (220mg, 0.7mmol), prepared as described in WO00/59929, was dissolved in 7ml of NMP (N-methylpyrrolidinone) and a spoon of Cs was added₂CO₃Added thereto, and stirred at 60 ℃ for 1.5 hours. Compound 108(150mg, 0.24mmol) was then added thereto. The reaction mixture was stirred at 80 ℃ overnight. The mixture was diluted with chloroform and washed with sodium bicarbonate, brine. The resulting aqueous phase was back extracted with chloroform. The combined organic layers were dried over sodium sulfate and evaporated. The crude product obtained is purified by preparative HPLC (Gilson) (MeOH-H)₂O, 65%) to give 21mg of product (yield 13%) and 12mg of isomer.

Example 110

17- [2- (2-isopropylamino-thiazol-4-yl) -7-methoxy-quinolin-4-yloxy]-13-methyl-2, 14-dioxo-3, 13-diaza-tricyclo [13.3.0.0^＊4，6^＊]Octadecan-7-ene-4-carboxylic acid (110)

To a solution of ester 109(21mg, 0.031mmol) in a mixture of THF (0.2ml) and methanol (0.3ml) was added a solution of LiOH (4mg, 0.17mmol) in 0.15ml water. The resulting mixture was stirred at 60 ℃ for 3.5 hours. After cooling to room temperature, acetic acid was added thereto (30 equivalents). The mixture was co-evaporated with toluene. The resulting residue was partitioned between chloroform and water, the aqueous phase was extracted three times with chloroform, the organic phases were combined, dried over sodium sulfate and evaporated, giving 20mg of pure product (yield 99%).

Example 111

Cyclopropanesulfonic acid {17- [2- (2-isopropylamino-thiazol-4-yl) -7-methoxy-quinolin-4-yloxy]-13-methyl-2, 14-dioxo-3, 13-diaza-tricyclo [13.3.0.0^＊4，6^＊]Octadecyl-7-en-4-carbonyl } amide (111)

Acid 110(20mg, 0.15mmol), DMAP (28mg, 0.225mmol) and EDAC (58mg, 0.3mmol) were dissolved in DMF (1.5 ml). The reaction mixture was stirred at room temperature overnight. Cyclopropyl sulfonamide (91mg, 1.125mmol) and DBU (114. mu.l, 0.75mmol) were then added thereto. After stirring at room temperature overnight, the reaction mixture was added to 5% citric acid and extracted three times with chloroform. The organic phase obtained is dried over sodium sulfate and evaporated. The resulting residue was purified by preparative HPLC to give the title product (5.6mg) (24% yield).

Measurement of

The activity of the compounds of the present invention against the flavivirus (e.g., HCV) NS3 protease is conveniently determined using conventional in vitro (enzyme) assays or cell culture assays.

One useful assay is the bartenshellager replicon assay disclosed in EP 1043399. Another replicon assay is described in WO 03064416.

Suitable enzyme assays involving inhibition of full-length hepatitis C NS3 are essentially described in Poliakov, 2002Prot Expression&Purification 25363371. Briefly, the depsipeptide matrix Ac-DED (Edans) EEAbu. psi [ COO]ASK(Dabcyl)-NH₂Hydrolysis of (Anaspec, SanJos é, USA) was determined by spectrofluorescence in the presence of the peptide cofactor KKGSVVVIVGRIVLSGK, as described by Landro, 1997Biochem 369340-9348. The enzyme (1nM) is incubated with 25. mu.M cofactor and the inhibitor in a buffer (such as 50mM HEPES, pH7.5, 10mM DTT, 40% glycerol, 0.1% n-octyl-beta-D-glucoside) for 10 minutes at about 30 ℃ where the reaction is initiated by the addition of a matrix, typically 0.5. mu.M matrix. Inhibitors are typically dissolved in DMSO, sonicated for 30s and spun. The above solutions are generally stored at-20 ℃ between measurements.

Another enzyme activity assay is described in WO 0399316, which utilizes the HCV NS3/4A protease complex FRET peptide assay. The purpose of this in vitro assay was to determine the inhibitory effect of the compounds of the invention on HCV NS3 protease complexes derived from BMS, H77C or J416S strains, as described below. This assay provides an indication of how effective the compounds of the present invention are in inhibiting HCV proteolytic activity.

Sera were taken from patients infected with HCV. Design of HCV genome (BMS strain) a full-length cDNA template was constructed from a DNA fragment obtained by reverse transcription-PCR (RT-PCR) of serum RNA, and using primers selected according to homology between other genotype la strains. Genotype Ia was determined for HCV isolates 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.Clin. Microbiol., 31(6), 1493-1503 (1993)). The amino acid sequence of the non-structural region NS2-5B was shown to be > 97% identical to HCV genotype Ia (H77C) and 87% identical to genotype Ib (J4L 6S). Infectious clones H77C (Ia genotype) and J4L6S (Ib genotype) can be obtained from r.purcell (NIH) and are disclosed in their sequence in Genbank (AAB67036, see Yanagi, m., Purcell, r.h., Emerson, s.u., and bukh.proc.natl.acad.sci.u.s.a.94(16) 8738-.

BMS, H77C, and J4L6S strains are conventional strains that produce recombinant NS3/4A protease complex. DNA encoding the recombinant HCV NS3/4A protease complex (amino acids 1027-1711) for these strains was manipulated as described in P.Gallinari et al (see Gallinari P, Paolini C, Brennan D, Nardi C, Steinkuhler C, De France sco R.biochemistry.38 (17): 562032, (1999)). Briefly, a lytic tail of tri-lysine was added to the 3' -end of the 30NS4A coding region. The cysteine at position P1 of the NS4A-NS4B cleavage site (amino acid 1711) was converted to glycine to avoid proteolytic cleavage of the lysine tag. In addition, a cysteine to serine mutation at amino acid 1454 can be introduced by PCR to prevent autolytic cleavage of the NS3 helicase domain. Variant DNA fragments can be cloned in pET21b bacterial expression vector (Novagen) and NS3/4A complex can be expressed in Escherichia coli strain BL21(DE3) (Invitrogen) as described in P.Galliari et al (see Galliari P, Brennan D, Nardi C, Brunettim, Tomei L, Steinkuhler C, De France sco R., J Virol.72 (8): 6758-69(1998)) and modifications thereof. Briefly, NS3/4A expression was induced by treatment with 0.5mM isopropyl-. beta. -D-thiogalactopyranoside (IPTG) for 22 hours at 20 ℃. Typically fermentation (101) produces about 80g of wet cell slurry. The cells were resuspended in 25mM N-2- (hydroxyethyl) piperazine-N' -2- (ethanesulfonic acid) (HEPES), pH7.5, 20% glycerol, 500mM sodium chloride (NaCl), 0.5% Triton-X100, 1. mu.g/mL lysozyme, 5mM magnesium chloride (MgCl)₂) mu.g/mL DnaseI, 5mM beta-mercaptoethanol (BME), protease inhibitor-ethylenediaminetetraacetic acid (EDTA) free (Roche) in lysis buffer (10mL/g) to homogenizeHomogenized and incubated in VC for 20 minutes. The homogenate was sonicated and clarified by ultracentrifugation at 235000g for 1 hour at 4 ℃.

Imidazole was added to the supernatant to a final concentration of 15mM and its pH was adjusted to 8. The crude protein extract was loaded onto a nickel nitrilotriacetate (Ni-NTA) column pre-equilibrated with buffer B (25n-tM 20HEPES, pH 820% glycerol, 500mM NaCl, 0.5% Triton-XIOO, 15mM imidazole, 5mM BMME). The sample was loaded at a flow rate of 1 mL/min. The column was washed with 15 column volumes of buffer C (same as buffer B except that it contained 0.2% Triton-X100). The protein was eluted with 5 column volumes of buffer D (same as buffer C except 200mM imidazole).

Fractions containing the NS3/4A protease complex were pooled and loaded onto a desalting column Superdex-S200 pre-equilibrated with buffer D (25MM HEPES, pH7.5, 20% glycerol, 300mM NaCl, 0.2% Triton-X100, 10mM BME). The sample was loaded at a flow rate of 1 mL/min. Fractions containing the NS3/4A protease complex 30 were collected and concentrated to approximately 0.5 mg/mL. The purity of the NS3/4A protease complex obtained from BMS, H77C and J4L6S strains was generally judged to be greater than 90% by SDS-PAGE and mass spectrometry analysis.

The enzyme is typically stored at-80 ℃, thawed on ice and diluted before use in assay buffer. The substrate used in the NS3/4A protease assay is suitably RET S1 (resonance energy transfer depsipeptide substrate; AnaPec, Inc. cat # 22991) (FRET peptide), described in anal. biochem.240 (2): 6067 (1996). The sequence of this peptide is presumably based on the natural cleavage site of NS4A/NS4B, except that an ester bond rather than an amide bond is present at the cleavage site. The peptide matrix was incubated with one of three recombinant NS3/4A complexes in the absence or presence of the compounds of the invention and the formation of fluorescent reaction products was followed in real time using Cytofluor Series 4000. The reagents used are listed below: HEPES and glycerol (ultrapure) are available from GIBCO-BRL. Dimethyl sulfoxide (DMSO) was obtained from Sigma. Beta-mercaptoethanol was obtained from Bio Rad.

Determination of buffer: 50mM HEPES, pH 7.5; 0.15M NaCl; 0.1% Triton; 15% of glycerol; 10mM BME. Matrix: mu.M final concentration (from 2mM stock solution 20 in DMSO, stored at-20 ℃ C.). HCV NS3/4 type A la (lb) at 2-3 nM final concentration (5. mu.M stock solution in 25mM HEPES, pH7.5, 20% glycerol, 300m.M NaCl, 0.2% Triton-X100, 10mM BME solution). For compounds whose potency is near the assay limit, the assay can be made more sensitive by adding 50 μ g/mL BSA to the assay buffer and/or reducing the terminal protease concentration to 300 pM.

The assay is desirably performed in a 96-well polystyrene blackboard (plate) from Falcon. Each well contained 25. mu.l of NS3/4A protease complex in assay buffer, 50. mu.l of the compound of the invention in 10% DMSO/assay buffer, and 25. mu.l of matrix in assay buffer. A control (no compound) was also prepared on the same assay plate. The enzyme complex is mixed with the compound or control solution, typically for 1 minute before the enzymatic reaction is initiated by the addition of the substrate. The assay plate is typically read instantaneously using a spectrophotometer such as the Cytofluor Series 4000 (persictive biosystems). The instrument is desirably set up to read a 340nm emission and 490nm excitation at 25 ℃. The reaction generally lasts about 15 minutes.

Percent inhibition can be calculated using the following equation.

100-[(dF_inh/dF_con)×100]

Where dF is the change in fluorescence over the linear range of the curve. Nonlinear curve fitting is applied to the inhibition-concentration data and 50% effective concentration (IC) is determined by using software such as Excel XI-fitting software using the following equation₅₀) And (3) calculating:

y＝A+((B-A)/(1+((C/x)＾D)))。

the enzyme assay desirably employs the principle of Fluorescence Resonance Energy Transfer (FRET) to produce a spectral response to the result of NS4A/4B cleavage catalyzed by the HCV NS3 serine protease. The activity is typically measured in a continuous fluorescence assay using an excitation wavelength of 355nm and an emission wavelength of 500 nm. The rate of initiation was determined from 10 minute consecutive readings of the enhanced fluorescence intensity resulting from the NS3 protease catalyzed cleavage event.

Another enzyme assay may be performed as follows:

material

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

The NS4A cofactor desirably has the amino acid sequence KKGSVVIVGRIVLSGK (commercially available), typically prepared as a 10mM stock in DMSO.

FRET-matrix (Ac-Asp-Glu-Asp (EDANS) -Glu-Glu-Abu--[COO)Ala-Ser-Lys(DABCYL)-NH₂MW1548.60, available from naspec RET S1, ca. usa) and is typically prepared as a 1.61mM stock in DMSO. Aliquots (50. mu.l/tube) should be covered with aluminum foil to prevent direct light and stored at-20 ℃.

Reference compound-1, N-1725 with the sequence AcAsp-D-Gla-Leu-Ile-Cha-Cys, MW 830.95, commercially available from BACHEM, Switzerland, is typically prepared as a 2mM stock DMSO solution and stored as an aliquot at-20 ℃.

1M HEPES buffer, commercially available from Invitrogen Corporation, was stored at 20 ℃.

Glycerol was purchased from Sigma, 99% pure.

CHAPS, 3- [ (3-cholamidopropyl) dimethylammonium ] -1-propanesulfonate: available from Research Organics, Cleveland, OH44125, USA. MW614.90

DTT, DL-dithiothreitol (Cleland Reagent: DL-DTT) 99% pure, MW.154.2. And (3) storage: +4 ℃.

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

TRIS, ultrapure (TRIS) is commercially available from ICN biomedicals inc.

Sodium chloride, available from KEBOlab AB.

N-dodecyl- β -D-maltoside, 98% minimum, available from Sigma, storage: at 20 ℃.

Device

Microtiter plate (Cliniplate, ThermoLab System scat No.9502890)

Eppendorf pipette

Biohit pipette, multiple dose

Ascent fluorometer, color filter pair ex 355nm, em 500nm

Method of producing a composite material

Test method

A10 mM stock solution of a compound of the present invention was prepared in DMSO. The stock solutions were stored at room temperature during the test, but were left at-20 ℃ during long-term storage.

Assay buffer a:

50mM HEPES buffer, pH7.5, 40% glycerol, 0.1% CHAPS

And (3) storage: at room temperature

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

Assay buffer B:

25mM TRIS pH7.5, 0.15M NaCl, 10% glycerol, 0.05% n-dodecyl-beta-D-maltoside

5mM DTT (stored in aliquots at-20 ℃ C. and fresh added for each experiment)

The test sequence is as follows:

preparation of reaction buffer (for one plate, 100 reactions) (buffer A)

1. Prepare 9500 μ l assay buffer (HEPES, pH7.5, 40% glycerol and 0.1% CHAPS) in deionized water. DTT was added to give a final concentration of 10mM (freshly prepared for each experiment).

2. Rapid melting of NS3 protease

3. 13.6. mu.l NS3 protease and 13.6. mu.l NS4A peptide were added and mixed appropriately. The mixture was left at room temperature for 15 minutes.

4. The enzyme stock solution was returned to liquid nitrogen or-80 ℃ as soon as possible.

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

5. Prepare 9500 μ l of assay buffer (TRIS, pH7.5, 0.15M NaCl, 0.5mM EDTA, 10% glycerol and 0.05% n-dodecyl β -D-maltoside) in deionized water. Addition of DTT gave a final concentration of 5mM (freshly prepared for each experiment).

6. Rapid melting of NS3 protease

7. Add 27.2. mu.l NS3 protease and 13.6. mu.l NS4A peptide and mix appropriately. The mixture was left at room temperature for 15 minutes.

8. The enzyme stock solution was returned to liquid nitrogen or-80 ℃ as soon as possible.

Preparation of inhibitor/reference Compound

Serial inhibitor dilutions were prepared and the inhibitor in DMSO diluted to 100 x final concentrations of 10, 1, 0.1, 0.01 and 0.001 μ M. The final DMSO concentration was 1% in the total reaction volume of 100. mu.l.

Serial dilutions of the reference compound were prepared and N-1725 was diluted to 100 x final concentrations of 120, 60, 30, 15, 7.5 and 3.75nM in DMSO.

Eight enzyme control wells were required for each experiment.

Blank wells contained 95 μ L buffer (no NS3PR present), 1 μ L DMSO, and 5 μ L matrix.

Preparation of FRET matrix

The stock matrix (1.61mM) was diluted with assay buffer to 40. mu.M working solution. Preventing exposure to light.

Order of measurement

A96-well format plate was used, and the total assay volume per well was 100. mu.l.

1. Add 95. mu.L of assay buffer to each well

2. Add 1. mu.l inhibitor/reference Compound

3. Preincubation for 30 minutes at room temperature

4. The reaction was started by adding 5. mu.L of 40. mu.M matrix solution (final concentration of 2. mu.M)

5. Readings were continued at ex-355 nm and em-500 nm for 20 minutes, and the increased fluorescence was monitored every minute.

6. Continuous curves (in the linear range, 8-10 time points) were plotted and the slope determined as the onset rate versus each individual inhibitor concentration.

7. % inhibition was calculated from enzyme control data.

Result processing

Results are expressed as% inhibition at a certain concentration (filter) or as Ki values in nM or μ M.

Calculation of% inhibition

The initial rate was determined from 10 minute consecutive readings of increased fluorescence intensity due to the NS3 protease catalyzed cleavage event. The change in the slope of the inhibitor compared to the enzyme control data gives the% inhibition at a certain concentration.

Calculation of Ki

All inhibitors were treated, assuming they all obeyed the competitive inhibition rules.

IC according to inhibition values at a series of inhibitor concentrations₅₀The value is calculated. The calculated values are used in the following equations:

Ki＝IC₅₀/(1+S/Km)

the plots were performed with the aid of two calculation procedures: grafit and Graphpad

The various compounds exemplified above in the present invention all showed IC's of 1nM to 6.9 micromolar in the above assays₅₀Value and sub-micromolar to micromolar ED₅₀The value is obtained.

Drug escape resistance forms and rates

Replicons cultured on microtiter plates can be used to determine the rate of resistance development and to select for drug escape mutants. The compounds tested were tested at about their ED₅₀Concentrations were added, 8 replicates per concentration. After an appropriate replicon incubation period, protease activity in the supernatant or lysed cells is assayed.

The following operations were performed in subsequent subcultures. Viruses produced at concentrations where the test compound showed > 50% protease activity on untreated infected cells (SIC), were passaged to fresh replicon cultures. Aliquots, say 15 μ l of supernatant, taken from each of the eight replicates were transferred to replicon cells without test compound (control) and to cells with the same concentration of test compound, and in addition, two concentrations were each five-fold higher. (see the following Table)

When the viral components of the replicon were allowed to multiply at the highest non-toxic concentration (5-40 μ M), e.g., as determined by HCV protease activity, 2-4 parallel wells were collected and expanded to yield material for sequence analysis and cross-resistance.

The key is as follows:

allowing viral growth

Inhibition of virus production

125xSIC

125xSIC 25xSIC→

25xSIC 5xSIC

25XSIC 5XSIC → Compound No

25XSIC 5XSIC → Compound No

5xSIC SIC

SIC → Compound No

SIC-free compounds

→

Pass1 Pass2 Pass3 Pass4 Pass5

Another method of assessing activity against drug escape mutants involves the preparation of mutant enzymes that produce specific mutations for use in standard Ki determinations as shown above. For example, WO04/039970 describes the structure of HCV proteases that can be directed to produce 155, 156 and/or 168 drug escape mutants that result from the selection pressure of BILN-2061 and VX-950. Thus, the above-described structures can be designed to replace the replicon vectors of natural-type proteases, thereby allowing for convenient evaluation of whether a given compound is active on a given drug escape mutant in a cellular assay.

P450 metabolism

The metabolic effects of the compounds of the invention via the major isoform of the human cytochrome system P450 are desirably determined in insect cells infected with baculovirus transfected with human cytochrome P450cDNA (supersomes) genttest corp.

Test compounds at concentrations of 0.5, 5 and 50 μ M were cultured in duplicate in the presence of supersomes overexpressing multiple cytochrome P450 isoforms including CYP1A2+ P450 reductase, CYP2A6+ P450 reductase, CYP2C9-Arg144+ P450 reductase, CYP2C19+ P450 reductase, CYP2D6-Val 374+ P450 reductase and CYP3A4+ P450 reductase. The culture includes a fixed concentration of cytochrome P450 (e.g., 50pmol) and is performed for 1 hour or more. The involvement of a given isoform in the metabolism of the test compound was determined by the disappearance of the parent compound as determined by UV HPLC chromatographic analysis.

Claims (82)

1. A compound of formula VI, or a pharmaceutically acceptable salt thereof:

wherein

A is C (═ O) OR¹Or C (═ O) NHSO₂R²(ii) a Wherein

R¹Is H, C₁-C₆Alkyl radical, C₀-C₃Alkyl carbonCyclic group, C₀-C₃An alkyl heterocyclic group;

R²is C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃An alkyl heterocyclic group;

wherein R is²Optionally substituted with 1 to 3 substituents independently selected from: halogen, oxo, nitrile, azido, nitro, C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃Alkyl heterocyclic group, NH₂C(＝O)-、Y-NRaRb、Y-O-Rb、Y-C(＝O)Rb、Y-(C＝O)NRaRb、Y-NRaC(＝O)Rb、Y-NHSO_pRb、Y-S(＝O)_pRb、Y-S(＝O)_pNRaRb, Y-C (═ O) ORb, and Y-NRaC (═ O) ORb;

y is independently a bond or C₁-C₃An alkylene group;

ra is independently H or C₁-C₃An alkyl group;

rb is independently H, C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl or C₀-C₃An alkyl heterocyclic group;

p is independently 1 or 2;

m is CR⁷R⁷′；

Ru is H or C₁-C₃An alkyl group;

R⁷is C₁-C₆Alkyl radical, C₀-C₃Alkyl radical C₃-C₇Cycloalkyl, or C₂-C₆Alkenyl, each of which is optionally substituted by 1 to 3 halogen atoms or by amino, -SH or C₀-C₃Alkyl cycloalkyl substituted; or R⁷Is J;

R⁷' is H or with R⁷Together form optionally substituted R^7′aSubstituted C₃-C₆A cycloalkyl ring, wherein;

R^7′ais C₁-C₆Alkyl radical, C₃-C₅Cycloalkyl radical, C₂-C₆Alkenyl, each of which may be optionally substituted with halogen; or R^7′aIs J;

q' is 0 or 1 and k is 0-3;

rz is H, or forms an olefinic bond with the asterisked carbon;

rq is H or C₁-C₆An alkyl group;

w is-CH₂-、-O-、-OC(＝O)NH-、-OC(＝O)-、-S-、-NH-、-NRa、-NHSO₂-, -NHC (═ O) NH — or-NHC (═ O) -, -NHC (═ S) NH — or a bond;

R⁸is a ring system comprising 1 or 2 saturated, partially saturated or unsaturated rings each having 4 to 7 ring atoms and each having 0 to 4 heteroatoms selected from S, O and N, optionally via C₁-C₃Alkyl and W are interrupted; any of the above R⁸All of which may be optionally substituted by R⁹Mono-, di-, or tri-substituted, wherein:

R⁹independently selected from: halogen, oxo, nitrile, azido, nitro, C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃Alkyl heterocyclic group, NH₂C(＝O)-、Y-NRa’Rb、Y-O-Rb、Y-C(＝O)Rb、Y-(C＝O)NRa’Rb、Y-NRaC(＝O)Rb、Y-NHSO_pRb、Y-S(＝O)_pRb、Y-S(＝O)_pNRaRb, Y-C (═ O) ORb, and Y-NRaC (═ O) ORb; wherein Ra' is Ra, with the proviso that when W is-S-or-O-, R⁸Is C₀-C₃Alkylaryl or C₀-C₃Alkyl heteroaryl, Y is a bond and Rb is H or C₁-C₆When alkyl is present, Ra' is Ra or C₁-C₆An alkyl group; and wherein said carbocyclyl or heterocyclyl moiety is optionally substituted with R¹⁰Substituted; wherein

R¹⁰Is C₁-C₆Alkyl radical, C₃-C₇Cycloalkyl radical, C₁-C₆Alkoxy, amino, sulfonyl, (C)₁-C₃Alkyl) sulfonyl, NO₂OH, SH, halogen, haloalkyl, carboxyl, amido;

rx is H or C₁-C₅An alkyl group; or Rx is J;

t is-CHR¹¹-or-NRd-, wherein Rd is H, C₁-C₃Alkyl or Rd is J;

R¹¹is H, or R¹¹Is C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃An alkylheterocyclic group, each of which may be substituted with 1 to 3 substituents independently selected from: halogen, oxo, nitrile, azido, nitro, C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃Alkyl heterocyclic group, NH₂CO-、Y-NRaRb、Y-O-Rb、Y-C(＝O)Rb、Y-(C＝O)NRaRb、Y-NRaC(＝O)Rb、Y-NHSO_pRb、Y-S(＝O)_pRb、Y-S(＝O)_pNRaRb, Y-C (═ O) ORb, Y-NRaC (═ O) ORb; or R¹¹Is J;

j, if present, is a single 3-to 10-membered saturated or partially unsaturated alkylene chain derived from R⁷/R⁷' cycloalkyl or from R⁷The carbon atom to which it is attached extends to Rd, Rj, Rx, Ry or R¹¹Is optionally substituted with one to three groups independently selected from-O-, -S-or-NR¹²-is interrupted by a heteroatom, and wherein 0 to 3 carbon atoms in the chain are optionally interrupted by R¹⁴Substitution; wherein:

R¹²is H, C₁-C₆Alkyl radical, C₃-C₆Cycloalkyl or COR¹³；

R¹³Is C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃An alkyl heterocyclic group;

R¹⁴independently selected from: H. c₁-C₆Alkyl radical, C₁-C₆Haloalkyl, C₁-C₆Alkoxy, hydroxy, halogen, amino, oxo, thio or C₁-C₆A thioalkyl group;

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

u is O or absent;

R¹⁵is H, C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃An alkylheterocyclic group, each of which may be substituted with 1 to 3 substituents independently selected from: halogen, oxo, nitrile, azido, nitro, C₁-C₆Alkyl radical, C₀-C₃Alkyl heterocyclic group, C₀-C₃Alkyl carbocyclyl, NH₂C(＝O)-、Y-NRaRb、Y-O-Rb、Y-C(＝O)Rb、Y-(C＝O)NRaRb、Y-NRaC(＝O)Rb、Y-NHSO_pRb、Y-S(＝O)_pRb、Y-S(＝O)_pNRaRb, Y-C (═ O) ORb, and Y-NRaC (═ O) ORb;

g is-O-, -NRy-, -NRjNRj-;

ry is H, C₁-C₃An alkyl group; or Ry is J;

one Rj is H and the other Rj is H or J;

R¹⁶is H, or R¹⁶Is C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃An alkylheterocyclyl group, each of which may be substituted with: halogen, oxo, nitrile, azido, nitro, C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃Alkyl heterocyclic group, NH₂CO-、Y-NRaRb、Y-O-Rb、Y-C(＝O)Rb、Y-(C＝O)NRaRb、Y-NRaC(＝O)Rb、Y-NHSO_pRb、Y-S(＝O)_pRb、Y-S(＝O)_pNRaRb、Y-C(＝O)ORb、Y-NRaC(＝O)ORb；

Wherein

At C₁-C₆Alkyl and C₁-C₃Any C atom in an alkyl group, unless otherwise specified, may be optionally substituted with one, two, or three halogens as valency permits;

at C₀-C₃Alkylaryl and C₀-C₃Alkyl radical C₃-C₇The aryl and cycloalkyl moieties in cycloalkyl, unless otherwise specified, are each optionally substituted with 1 to 3 substituents selected from: halogen, hydroxy, nitro, cyano, carboxy, C₁-C₆Alkyl radical, C₁-C₆Alkoxy radical, C₁-C₆Alkoxy radical C₁-C₆Alkyl radical, C₁-C₆Alkanoyl, amino, azido, oxo, mercapto and C₀-C₃An alkyl heterocyclic group;

at C₀-C₃Alkyl carbocyclyl and C₀-C₃The carbocyclyl and heterocyclyl portion of an alkylheterocyclyl, unless otherwise specified, are each optionally substituted with 1-3 substituents selected from: halogen, hydroxy, nitro, cyano, carboxy, C₁-C₆Alkyl radical, C₁-C₆Alkoxy radical, C₁-C₆Alkoxy radical C₁-C₆Alkyl radical, C₁-C₆Alkanoyl, amino, azido, oxo, mercapto, C₀-C₃Alkyl carbocyclyl and C₀-C₃An alkyl heterocyclic group;

each amino group is selected from NH₂、NHC₁-C₆Alkyl and N (C)₁-C₆Alkyl radical)₂(ii) a And

each amido group is selected from C (═ O) NH₂、C(＝O)NHC₁-C₆Alkyl, C (═ O) N (C)₁-C₆Alkyl radical)₂And NH (C ═ O) C₁-C₆An alkyl group.

2. A compound according to claim 1, wherein R⁷' is H, and R⁷Is ethyl, cyclopropylmethyl, cyclobutylmethyl or mercaptomethyl, or n-propyl or 2, 2-difluoroethyl.

3. A compound according to claim 2, wherein R⁷Is n-propyl or 2, 2-difluoroethyl.

4. A compound according to claim 1, wherein R⁷And R⁷' together form a spiro-cyclopropyl or spiro-cyclobutyl ring, optionally substituted by R^7′aMono-or di-substituted; wherein:

R^7′ais C₁-C₆Alkyl radical, C₃-C₅Cycloalkyl or C₂-C₆Alkenyl, each optionally substituted with halogen; or R^7aIs J.

5. A compound according to claim 4, wherein the ring is substituted with R^7′aA substituted spiro-cyclopropyl ring, wherein:

R^7′ais ethyl, vinyl, cyclopropyl, 1-or 2-bromoethyl, 1-or 2-fluoroethyl, 2-bromovinyl or 2-fluoroethyl.

6. A compound according to claim 1, wherein R⁷Is J and R⁷' is H.

7. A compound according to claim 1 having the partial structure:

8. a compound according to claim 1 having the partial structure:

9. a compound according to claim 8, wherein Rq is C₁-C₃An alkyl group.

10. A compound according to claim 9, wherein Rq is methyl.

11. A compound according to claim 1, wherein m is 0 and n is 0.

12. The compound according to claim 11, wherein G is-NRy-or-NRjNRj-.

13. A compound according to claim 12, wherein Ry or one Rj group is J, thereby defining a macrocyclic compound.

14. A compound according to claim 12, wherein R¹⁶Is H, C₁-C₆Alkyl or C₃-C₆A cycloalkyl group.

15. A compound according to claim 1, wherein m is 1.

16. The compound according to claim 15, wherein U is O.

17. The compound according to claim 15, wherein T is CHR¹¹。

18. A compound according to claim 17, wherein R¹¹Is C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C₀-C₃Alkylaryl or C₀-C₃Alkylheteroaryl, each of which is optionally substituted by hydroxy, halogen, C₁-C₆Alkoxy radical, C₁-C₆Thioalkyl, COOR¹⁴Carboxyl group, (C)₁-C₆Alkoxy) carbonyl, aryl, heteroaryl, or heterocyclyl.

19. A compound according to claim 18, wherein R¹¹Is C₁-C₆Alkyl radical, C₀-C₃Alkyl carbocyclyl, C⁰-C₃Alkylaryl or C₀-C₃Alkylheteroaryl, each of which is optionally substituted by hydroxy or COOR¹⁴And (4) substituting.

20. According toThe compound of claim 18, wherein R¹¹Is tert-butyl, isobutyl, cyclohexyl, phenethyl, 2-dimethyl-propyl, cyclohexylmethyl, benzyl, 2-pyridylmethyl, 4-hydroxy-benzyl or carboxypropyl.

21. A compound according to claim 20, wherein R¹¹Is tert-butyl, isobutyl or cyclohexyl.

22. The compound according to claim 15, wherein Rd, Rx or R¹¹One of which is J, thereby defining a macrocyclic compound.

23. The compound according to claim 15, wherein n is 1.

24. A compound according to claim 23, wherein R¹⁵Is C₁-C₆Alkyl or C₀-C₃An alkyl carbocyclyl, each of which is optionally substituted.

25. A compound according to claim 24, wherein R¹⁵Cyclohexyl, cyclohexylmethyl, tert-butyl, isopropyl or isobutyl.

26. The compound according to claim 15, wherein G is NRy or NRjNRj, wherein Ry or one Rj is H or methyl and the other Rj is H.

27. A compound according to claim 26, wherein R¹⁶Is H, C₁-C₆Alkyl is either a 5 or 6 membered heterocyclic ring.

28. A compound according to claim 27 wherein the 5 or 6 membered heterocyclic ring is selected from morpholine, piperidine or piperazine.

29. A compound according to claim 15, wherein R¹⁶Is C₁-C₆Alkyl radical, C₀-C₃Alkyl heterocyclic group, C₀-C₃Alkyl carbocyclyl, each of which is optionally substituted with hydroxy, halogen, or C₁-C₆Alkoxy groups.

30. A compound according to claim 29, wherein R¹⁶Is 2-indanol, 2, 3-indanyl, 2-hydroxy-1-phenyl-ethyl, 2-thienylmethyl, cyclohexylmethyl, 2, 3-methylenedioxybenzyl, cyclohexyl, benzyl, 2-pyridylmethyl, cyclobutyl, isobutyl, n-propyl or 4-methoxyphenylethyl.

31. A compound according to claim 1, wherein W is-OC (═ O) -, -NRa-, -nhs (O)₂-or-NHC (═ O) -; or-OC (═ O) NH-or-NH.

32. The compound according to claim 31, wherein W is-OC (═ O) NH-or-NH.

33. A compound according to claim 1, wherein W is-S-, a bond or-O-.

34. The compound according to claim 33, wherein W is-O-.

35. A compound according to claim 31 or 33, wherein R⁸Is optionally substituted C₀-C₃Alkyl carbocyclyl or optionally substituted C₀-C₃An alkyl heterocyclic group.

36. A compound according to claim 35, wherein C₀-C₃The alkyl moiety is methylene or a bond.

37. A compound according to claim 36, wherein C is₀-C₃The alkyl moiety is a bond.

38. A compound according to claim 36, wherein R⁸Is C₀-C₃Alkylaryl or C₀-C₃(ii) alkylheteroaryl, each of which is optionally mono-, di-, or tri-substituted with R9, wherein:

R⁹is C₁-C₆Alkyl radical, C₁-C₆Alkoxy group, NO₂OH, halogen, trifluoromethyl, optionally substituted by C₁-C₆Alkyl mono-or di-substituted amino or amido, C₀-C₃Alkylaryl group, C₀-C₃Alkyl heteroaryl, carboxy, optionally substituted by R¹⁰Substituted aryl or heteroaryl; wherein:

R¹⁰is C₁-C₆Alkyl radical, C₃-C₇Cycloalkyl radical, C₁-C₆Alkoxy, optionally substituted by C₁-C₆Alkyl mono-or di-substituted amino, C₁-C₃Alkylamides, sulphonyl C₁-C₃Alkyl radical, NO₂OH, halogen, trifluoromethyl, carboxyl or heteroaryl.

39. A compound according to claim 38, wherein R⁹Is C₁-C₆Alkyl radical, C₁-C₆Alkoxy, amino, di- (C)₁-C₃Alkyl) amino, C₁-C₃Alkylamides, aryls or heteroaryls optionally substituted by R¹⁰Substituted; wherein:

R¹⁰is C₁-C₆Alkyl radical, C₃-C₇Cycloalkyl radical, C₁-C₆Alkoxy, amino, mono-or di-C₁-C₃Alkylamino, acylamino, C₁-C₃Alkyl amides, halogens, trifluoromethyl or heteroaryl.

40. A compound according to claim 39, wherein R¹⁰Is C₁-C₆Alkyl radical, C₁-C₆Alkoxy, optionally substituted by C₁-C₃Alkyl mono-or di-substituted amino, acylamino, C₁-C₃-alkylamides, halogens or heteroaryls.

41. A compound according to claim 40, wherein R¹⁰Is methyl, ethyl, isopropyl, tert-butyl, methoxy, chlorine, optionally substituted by C₁-C₃Alkyl mono-or di-substituted amino, acylamino, C₁-C₃Alkyl amides or C₁-C₃An alkylthiazolyl group.

42. A compound according to claim 36, wherein R⁸Is 1-naphthylmethyl, 2-naphthylmethyl, benzyl, 1-naphthyl, 2-naphthyl or quinolyl, each unsubstituted or substituted by R as defined in claim 1⁹Mono-or di-substituted.

43. A compound according to claim 42, wherein R⁸Is 1-naphthylmethyl or quinolyl, each unsubstituted or substituted by R as defined in claim 1⁹Mono-or di-substituted.

44. A compound according to claim 43, wherein R⁸Comprises the following steps:

wherein R is^9aIs C₁-C₆An alkyl group; c₁-C₆An alkoxy group; thio C₁-C₃An alkyl group; optionally is covered with C₁-C₆An alkyl-substituted amino group; c₀-C₃An alkylaryl group; or C₀-C₃Alkyl heteroaryl, C₀-C₃Alkylheterocyclyl, said aryl, heteroaryl or heterocycle being optionally substituted by R¹⁰Substitution; wherein:

R¹⁰is C₁-C₆Alkyl radical, C₀-C₃Alkyl radical C₃-C₇Cycloalkyl radical, C₁-C₆Alkoxy, optionally substituted by C₁-C₆Alkyl mono-or di-substituted amino, acylamino, C₁-C₃An alkylamide; and

R^9bis C₁-C₆Alkyl radical, C₁-C₆Alkoxy, amino, di (C)₁-C₃Alkyl) amino, (C)₁-C₃Alkyl) amides, NO₂OH, halogen, trifluoromethyl, carboxyl.

45. A compound according to claim 44, wherein R^9aIs aryl or heteroaryl, each of which is optionally substituted by R as defined in claim 1¹⁰And (4) substituting.

46. A compound according to claim 45, wherein R^9aSelected from:

wherein R is¹⁰Is H, C₁-C₆Alkyl or C₀-C₃Alkylcycloalkyl, optionally substituted by C₁-C₆Alkyl mono-or di-substituted amino, acylamino, (C)₁-C₃Alkyl) amides.

47. A compound according to claim 45, wherein R^9aIs optionally substituted phenyl; c₁-C₆An alkoxy group; or a halogen.

48. According to the claimsThe compound of claim 47, wherein said optionally substituted phenyl is substituted with C₁-C₆Alkyl-substituted phenyl.

49. A compound according to claim 44, wherein R⁸Comprises the following steps:

wherein R is^10aIs H, C₁-C₆Alkyl or C₀-C₃Alkyl carbocyclyl, optionally substituted by C₁-C₆Alkyl mono-or di-substituted amino, acylamino, (C)₁-C₃Alkyl) amides, heteroaryls or heterocyclyls; and R^9bIs C₁-C₆Alkyl radical, C₁-C₆Alkoxy, amino, di (C)₁-C₃Alkyl) amino, (C)₁-C₃Alkyl) amides, NO₂OH, halogen, trifluoromethyl or carboxyl.

50. A compound according to claim 44, wherein R^9bIs C₁-C₆-alkoxy groups.

51. A compound according to claim 50, wherein R^9bIs methoxy.

52. The compound according to any one of claims 1-51, wherein A is C (═ O) NHSO₂R²。

53. A compound according to claim 52, wherein R²Is methyl, cyclopropyl or phenyl.

54. The compound according to any one of claims 1-51, wherein A is C (═ O) OR¹。

55. A compound according to claim 54, wherein R¹Is H or C₁-C₆An alkyl group.

56. A compound according to claim 55, wherein R¹Is hydrogen, methyl, ethyl or tert-butyl.

57. The compound according to claim 1, wherein J is optionally 1-2 independently selected from-O-, -S-or-NR¹²A 3 to 8 membered saturated or unsaturated alkylene chain of a heteroatom of (A) wherein R¹²Is H, C₁-C₆Alkyl, or-C (═ O) C₁-C₆An alkyl group.

58. A compound according to claim 57, wherein said C₁-C₆The alkyl group is a methyl group.

59. A compound according to claim 57, wherein-C (═ O) C₁-C₆The alkyl group is acetyl.

60. A compound according to claim 57 wherein J is a4 to 7-membered saturated or unsaturated all-carbon alkylene chain.

61. The compound according to claim 57, wherein J is saturated or monounsaturated.

62. The compound according to claim 57, wherein J is of a size to provide a macrocycle of 14 or 15 ring atoms.

63. The compound according to claim 1, having formula VIhe or VIhf:

64. the compound according to claim 1, having formula VIhe:

65. the compound according to claim 63, wherein J is a single 5 or 6 membered saturated or partially unsaturated alkylene chain.

66. The compound according to claim 64, wherein J is a single 5 or 6 membered saturated or partially unsaturated alkylene chain.

67. The compound according to claim 63, wherein J has one degree of unsaturation.

68. The compound according to claim 63, wherein W is O, R⁸Is aryl or heteroaryl, each of which is optionally substituted by R⁹Mono-, di-, or tri-substituted, wherein:

R⁹is C₁-C₆Alkyl radical, C₁-C₆Alkoxy group, NO₂OH, halogen, trifluoromethyl, amino or amido, C₀-C₃Alkylaryl group, C₀-C₃Alkylheteroaryl, carboxy, said aryl or heteroaryl being optionally substituted by R¹⁰Substitution; wherein:

R¹⁰is C₁-C₆Alkyl radical, C₃-C₇Cycloalkyl radical, C₁-C₆Alkoxy, amino, amido, sulphonyl C₁-C₃Alkyl radical, NO₂OH, halogen, trifluoromethyl, carboxyl or heteroaryl.

69. The compound according to claim 68, wherein R⁹Is selected from C₁-C₆Alkyl radical, C₁-C₆Alkoxy, amino, amido, aryl or heteroaryl, said aryl or heteroaryl moiety being optionally substituted by R¹⁰Substituted; wherein:

R¹⁰is C₁-C₆Alkyl radical, C₃-C₇Cycloalkyl radical, C₁-C₆Alkoxy, amino, amido, halogen, trifluoromethyl or heteroaryl.

70. A compound according to claim 63, wherein R⁸Comprises the following steps:

wherein R is^9aIs C₁-C₆An alkyl group; c₁-C₆An alkoxy group; thio C₁-C₃An alkyl group; optionally is covered with C₁-C₆An alkyl-substituted amino group; c₀-C₃An alkylaryl group; or C₀-C₃Alkyl heteroaryl, C₀-C₃Alkylheterocyclyl, said aryl, heteroaryl or heterocycle being optionally substituted by R¹⁰Substitution; wherein:

R¹⁰is C₁-C₆Alkyl radical, C₃-C₇Cycloalkyl radical, C₁-C₆Alkoxy, amino, amido, heteroaryl or heterocyclyl; and

R^9bis C₁-C₆Alkyl radical, C₁-C₆Alkoxy, amino, amido, NO₂OH, halogen, trifluoromethyl, carboxyl.

71. A compound according to claim 70, wherein R^9aThe method comprises the following steps:

wherein R is¹⁰Is H, C₁-C₆Alkyl radical, C₃-C₆Cycloalkyl, amino, amido, heteroaryl or heterocyclyl.

72. The compound according to claim 63, wherein A is C (═ O) NHSO₂R²。

73. A compound according to claim 72, wherein R²Is methyl, cyclopropyl or optionally substituted phenyl.

74. A compound according to claim 1 having a stereochemistry selected from the group consisting of the stereochemistry shown in the partial structures of the formulae:

75. a compound according to claim 1 having the formula:

76. a compound according to claim 1 having the formula:

77. a pharmaceutical composition comprising a compound as defined in any one of claims 1 to 76 and a pharmaceutically acceptable carrier therefor.

78. The pharmaceutical composition according to claim 77, further comprising an additional HCV antiviral selected from the group consisting of nucleoside analog polymerase inhibitors, protease inhibitors, ribavirin and interferons.

79. A compound as defined in any one of claims 1 to 76 for use in therapy.

80. Use of a compound as defined in any one of claims 1 to 76 in the manufacture of a medicament for the prevention or treatment of a flavivirus infection, said flavivirus comprising HCV.

81. A compound according to any one of claims 1 to 76 for use in the treatment or prevention of flavivirus infections, including HCV.

82. The compound of claim 81 for use in the treatment of HCV infection.