Macrocyclic hepatitis C serine protease inhibitors

[0001] The present invention relates to new macrocycles having activity against hepatitis C virus (HCV) and useful in the treatment of HCV infections. More particularly, the invention relates to macrocyclic compounds for use in the treatment of HCV infection in a subject and compositions containing such compounds for use in the treatment of HCV infection in a subject.

BACKGROUND OF THE INVENTION [0002] HCV is a major cause of non-A, non-B hepatitis and is an increasingly serious health care problem in both the developed world and the developing world. It is estimated that the virus infects over 200 million people around the world, nearly five times the number of people infected with human immunodeficiency virus (HIV). HCV infected patients have an increased risk of developing cirrhosis due to hepatocellular carcinoma and terminal liver disease due to the high percentage of people affected by chronic infections. HCV is the most common cause of hepatocellular carcinoma and the reason why patients in the western world need a liver transplant.

[0003] There are significant barriers to the development of anti-HCV therapies that include, but are not limited to, virus persistence, genetic diversity of the virus during replication in the host, high incidence of the virus developing drug-resistant mutants, and the lack of repetitive infection culture systems and replication models and HCV pathogenesis on small animals. In most cases, due to the mild course of the infection and the complex liver biology, antiviral drugs that are likely to have significant side effects should be carefully considered.

[0004] Quinoxalinyl macrocyclic hepatitis C serine protease inhibitors and methods for their preparation are known from WO 2004/093798. Such compounds are useful for treating HCV infection.

[0005] WO 2007/001406 discloses macrocyclic compounds containing aryl and heteroaryl groups useful for the treatment of viral infection, in particular hepatitis C and SARS.

[0006] WO 2007/014926 discloses quinolinyl macrocyclic HCV inhibitors that are useful for the treatment of HCV infection.

[0007] Application EP 2222161 A1, which was filed December 3, 2008 and reserves the right to use the priority of application US60 / 992,584 filed December 5, 2007, discloses quinoxalinyl macrocyclic hepatitis C virus serine protease inhibitors and intermediates thereto.

SUMMARY OF THE INVENTION [0008] The present invention relates to new macrocyclic compounds and such compounds for use in the treatment of hepatitis C infection in a subject who needs such therapy with said macrocyclic compounds. Compounds of the present invention

EP 2 468 285 B1 disturb the hepatitis C virus life cycle and are useful as antiviral agents. The present invention further provides pharmaceutical compositions comprising the compounds of the present invention, or a pharmaceutically acceptable salt thereof, in combination with a pharmaceutically acceptable carrier or excipient, for use in treating HCV infection in a subject.

[0009] In one aspect, the invention relates to a compound of formula I or I ':

<img file="PL2468285T3_D0001.tif" />

or a pharmaceutically acceptable salt thereof, for use in treating HCV infection in a subject, wherein:

J is -C (O) - or -OC (O);

A represents group C₁-C₆alkyl, C₂-C₆alkenyl, C.₂-C₆alkynyl, aryl, heteroaryl, heterocyclic containing 5 to 10 ring atoms, or C₃-C₁₀carbocyclic, and A is optionally substituted with one or more R's₆;

Each of R₁ and R₂ is independently selected from the group consisting of (i) halogen, hydroxy, amino, -CN, -CF₃, -N₃, -NO₂, -OR₄, -SR₄, SOR₄, -SO₂R₄, -NR₃R₄, -C (O) OR₄, -C (O) R₄, -C (O) NR₃R₄, or -N (R₃) C (O) R₄;

(ii) a heterocyclic group containing 5 to 10 ring atoms and optionally substituted with one or more R's₇;

(iii) group C₃-C₁₀carbocyclic optionally substituted with one or more R's₇; or (iv) group C₁-C₆alkyl, C₂-C₆alkenyl, C.₂-C₆alkynyl, C₁-C₆haloalkyl, C.₂C₆haloalkenyl, or C₂-C₆-halogenoalkynyl;

G is -ER₅; where E is -NHS (O₂)-;

R₅ means group C₁-C₆alkyl, C₂-C₆alkenyl, C.₂-C₆alkynyl, C₃-C₁₀carbocyclic, heterocyclic having 5 to 10 ring atoms, or heteroaryl, and each with R₅ is optionally substituted with one or more R's₇;

each of R₃ and R₄ is independently selected at each occurrence from the group consisting of hydrogen, group C₁-C₆alkyl, C₂-C₆alkenyl or C₂-C₆alkynyl;

L is group C₃-C₆alkylene, C₃-C₆alkenylene or C₃-C₆alkynylene and is optionally substituted with one or more R's₇;

j = 0;

K = 0; m = 1;

n is 0, 1, 2, 3, or 4; and

373 is a single bond or a carbon-carbon double bond,

Y is -C (R ") -, R 'and R" taken together with the carbon atoms to which they are attached form an aryl or heteroaryl ring which is optionally substituted with one or more R₂;

each of R₆ and R₇ is independently selected at each occurrence from the group consisting of halogen, hydroxy, amino, -CF₃, -CN, -N₃, -NO₂, -C₁-C₆alkyl, C₂-C₆-alkenyl, C.₂-C₆alkynyl, C₁-C₆haloalkyl, C.₂-C₆haloalkenyl, or C₂-C₆-halogenoalkynyl.

In another aspect, the invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I or I ', or a pharmaceutically acceptable salt thereof, in combination with a pharmaceutically acceptable carrier or excipient for use in treating HCV infection in a subject.

DETAILED DESCRIPTION OF THE INVENTION [0010] The present disclosure relates to a compound of formula I or I ':

<img file="PL2468285T3_D0002.tif" />

G or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein:

J is absent or optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, -C (O) -, -OC (O) -, -N (R3) -C (O) -, -C ( S) -, -C (= NR4) -, -S (O) -, -S (O2) -, or N (R3) -;

A is an optionally substituted alkyl group, an optionally substituted alkenyl group, or an optionally substituted alkynyl group, each of which contains 0, 1, 2, or 3 heteroatoms selected from O, S, or N; optionally substituted aryl, optionally substituted arylalkyl, optionally substituted alkoxy, optionally substituted heteroaryl, optionally substituted heterocyclic, or optionally substituted carbocyclic;

Each of R₁ is independently selected from the group consisting of (i) halogen, hydroxy, amino, -CN, -CF₃, -N₃, -NO₂, -OR₄, -SR₄, SOR4, -SO2R4, -N (R3) S (O2) -R4, -N (R3) S (O2) NR3R4, -NR3R4, -C (O) OR4, -C (O) R4, 3

EP 2 468 285 B1

C (O) NR₃R₄, or -N (R₃) C (O) R₄;

(ii) an optionally substituted aryl;

(iii) optionally substituted heteroaryl;

(iv) an optionally substituted heterocyclic group;

(v) an optionally substituted carbocyclic group; or (vi) an optionally substituted alkyl group, an optionally substituted alkenyl group, or an optionally substituted alkynyl group, each of which contains 0, 1, 2, or 3 heteroatoms selected from O, S, or N;

G is -ER₅;

where E is absent; or is an optionally substituted alkylene group, optionally substituted alkenylene group, optionally substituted alkynylene group, each of which contains 0, 1, 2, or 3 heteroatoms selected from O, S, or N; or -O-,

-S-, -N (R3) -, -N (R3) S (Op) -, -N (R3) C (O) -, -N (R3) C (O) S (Op) -, -OS (Op) -, -C (O) S (Op) -, or

-C (O) N (R 3) S (Op) -; p is 0, 1 or 2;

R₅ is H; optionally substituted alkyl, optionally substituted alkenyl, or optionally substituted alkynyl, each containing 0, 1, 2, or 3 heteroatoms selected from O, S, or N; optionally substituted carbocyclic, optionally substituted heterocyclic, optionally substituted aryl, or optionally substituted heteroaryl;

R₃ and R₄ are each independently selected at each occurrence from the group consisting of the following: optionally substituted alkyl, optionally substituted alkenyl or optionally substituted alkynyl, each of which contains 0, 1, 2, or 3 heteroatoms selected from O, S, or N; optionally substituted aryl; optionally substituted heteroaryl; optionally substituted heterocyclic; optionally substituted carbocyclic; or a hydrogen atom;

L is absent or selected from the group consisting of an optionally substituted alkylene group, an optionally substituted alkenylene group or an optionally substituted alkynylene group, each of which contains 0, 1, 2, or 3 heteroatoms selected from O, S, or N;

Y is N or -C (R ") -;

where A, R₁, R 'and / or R "may be taken together to form a ring;

j = 0, 1, 2, 3, or 4;

k = 0, 1, 2, or 3;

m = 0, 1 or 2;

n is 0, 1, 2, 3, or 4; and is a carbon-carbon single or double bond (i.e., is

EP 2 468 285 B1

<img file="PL2468285T3_D0003.tif" />

where, if Y is N, then R 'is an optionally substituted heterocyclic group, optionally substituted heteroaryl group, optionally substituted aryl group or optionally substituted carbocyclic group, and includes two or more fused rings, and where R' is other than

<img file="PL2468285T3_D0004.tif" />

and

<img file="PL2468285T3_D0005.tif" />

where, if Y is -C (R ") -, then R 'and R" taken together with the carbon atoms to which they are attached form an aryl or heteroaryl ring, each of which is optionally substituted;

provided that said compound is other than (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -14- (cyclopropylsulfonylcarbamoyl) ) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate.

[0011] It is understood that the embodiments of the invention or disclosures discussed below with respect to preferred choices of variable parameters may be selected alone or in combination with one or more of other embodiments, or preferred choices of variables, according to the invention or disclosure, so as if each of the combinations was explicitly given herein.

[0012] In one aspect, the disclosure provides a compound of formula I or I ', or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein Y is CR ", and R' and R" taken together with the carbon atoms to which they are attached, form an optionally substituted aryl or an optionally substituted heteroaryl ring.

[0013] In another aspect, the disclosure provides a compound of formula I or I ', or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein Y is CR ", and R' and R" taken together with the carbon atoms to which they are attached form an optionally substituted aryl ring, preferably a phenyl group.

[0014] Alternatively or additionally, k = 3, j = 1 and L is absent.

[0015] Alternatively or additionally, R 'and R ", and the atoms to which each is attached form an aryl group which is substituted by (R₂)_xwhere each of R₂ is independently selected from the group consisting of halogen, hydroxy, amino, -CN, -CF₃, -N₃, -NO₂, -OR₄, -SR₄, -SOR₄, -SO₂R₄, -N (R₃) S (O₂) -R₄, -N (R₃) S (O₂) NR₃R₄, -NR₃R₄, -C (O) OR₄,

EP 2 468 285 B1

-C (O) R₄, -C (O) NR₃R₄, or -N (R₃) C (O) R₄; optionally substituted aryl; optionally substituted heteroaryl; optionally substituted heterocyclic; optionally substituted carbocyclic; or an optionally substituted alkyl group, an optionally substituted alkenyl group, or an optionally substituted alkynyl group, each of which contains 0, 1, 2, or 3 heteroatoms selected from O, S, or N; and x is 0, 1, 2, 3, or 4.

[0016] Alternatively or additionally, R₁ is absent (i.e., n = 0) or is halogen, hydroxy, amino, -CN, -CF₃, -N₃, -NO₂, -OR₄, -SR₄, -SOR₄, -SO₂R₄, -N (R₃) S (O₂) R4, -N (R3) S (O2) NR3R4, -NR3R4, -C (O) OR4, -C (O) R4, -C (O) NR3R4, or -N (R3) C (O) R4.

[0017] Alternatively or additionally, R 'and R ", and the atoms to which each is attached form an aryl group which is substituted by (R₂)_xwhere each of R₂ is independently absent (i.e., x = 0) or halogen.

[0018] Alternatively or additionally, R₁ is absent (i.e., n = 0) or is halogen.

[0019] Alternatively or additionally, E is -NH-, -NHS (O._p) -, or -NH (CO) S (O_p) -, ip is 2.

[0020] Alternatively or additionally, E is -NHS (O_p) -, ip is 2.

[0021] Alternatively or additionally, R₅ means a cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, pyridinyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, pyrrolidinyl, morpholinyl, piperidinyl, piperazinyl, or imidazolyl, group which is optionally substituted. In a further embodiment

R₅ is an optionally substituted cyclopropyl group or an optionally substituted thienyl group, preferably a cyclopropyl or thienyl group.

[0022] Alternatively or additionally, J is -C (O) -, -OC (O) -, -C (S) -, -C (= NR₄) -, -S (O) -, or -S (O₂) -. Preferably, J is -C (O) -.

[0023] Alternatively or additionally, m is 1.

[0024] Alternatively or additionally, each of R₃ is H.

[0025] Alternatively or additionally, A is an optionally substituted -C group₁-C₈ alkyl, containing 0, 1, 2, or 3 heteroatoms selected from O, S, or N; optionally substituted aryl, optionally substituted -C₁-C₈ alkoxy, optionally substituted heteroaryl, optionally substituted -C₃-C₁₂ cycloalkyl, or an optionally substituted -C group₃-C₁₂ heterocycloalkyl. In a further embodiment, A is selected from the group consisting of

<img file="PL2468285T3_D0006.tif" />

EP 2 468 285 B1

<img file="PL2468285T3_D0007.tif" />

Preferably, A is a 5-methyl-pyrazin-2-yl group.

[0027] In yet another aspect, the disclosure provides a compound of formula I or I ', or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein Y is CR ", and R' and R" taken together with the carbon atoms to which they are attached, form an optionally substituted heteroaryl ring. Other parameters have the meanings defined above, including alternative or preferred embodiments, as if they were repeated here.

[0028] The subject of the invention is a compound of formula I or I '(preferably of formula I), or a pharmaceutically acceptable salt thereof, for use in the treatment of an HCV infection in a subject, wherein Y is the group CR ", R' and R" taken together with the carbon atoms to which they are attached form an aryl or heteroaryl ring, preferably a phenyl ring, which is optionally substituted by one or more R₂;

k = 0, j = 0, m = 1, n = 0, 1, 2, 3 or 4, and L is a group C₃-C₆alkylene, C₃-C₆alkenylene or C₃-C₆alkynylene and is optionally substituted with one or more R's₇ (preferably a butylene group);

is a carbon-carbon single or double bond;

J is the group -C (O) - or -OC (O) - (preferably -C (O) -);

A represents group C₁-C₆alkyl, C₂-C₆alkenyl, C.₂-C₆alkynyl, C₃-C₁₀carbocyclic, aryl, heteroaryl or heterocyclic having 5 to 10 ring atoms, and A is optionally substituted with one or more R₆;

G is the group -ER₅, E represents the -NHS group (O₂) -; R₅ means group C₁-C₆alkyl, C₂C₆alkenyl, C.₂-C₆alkynyl, C₃-C₁₀carbocyclic (preferably cyclopropyl), heteroaryl (preferably thienyl) or heterocyclic having 5 to 10 ring atoms, and R₅ is optionally substituted with one or more R's₇;

each of R₁ and R₂ is independently selected from the group consisting of halogen, hydroxy, amino, -CN, -CF₃, -N₃, -NO₂, -OR₄, -SR₄, -S (O) R₄, -S (O₂) R₄, -NR₃R₄, -C (O) OR₄, C (O) R₄, -C (O) NR₃R₄, -N (R₃) C (O) R₄, C.₁-C₆alkyl, C₂-C₆alkenyl, C.₂-C₆alkynyl, C₁C₆haloalkyl, C.₂-C₆haloalkenyl, C.₂-C₆haloalkynyl, C₃-C₁₀carbocyclic optionally substituted with one or more R's₇, or heterocyclic having 5 to 10 ring atoms and optionally substituted with one or more R₇where each of R₆ and R₇ is independently selected at each occurrence from the group consisting of halogen, hydroxy, amino, -CF₃, -CN, -N₃, -NO₂, -C₁-C₆alkyl (preferably methyl), C₂-C₆alkenyl, C.₂-C₆alkynyl, C₁-C₆haloalkyl, C.₂-C₆haloalkenyl, or C₂-C₆haloalkynyl (and, preferably, R₁ and R₂ they are absent in each speech); and [0029] each of R₃ and R₄ is independently selected at each occurrence from the group consisting of hydrogen, group C₁-C₆alkyl, C₂-C₆alkenyl or C₂-C₆alkynyl and R₃ preferably represents a hydrogen atom.

The invention furthermore relates to a compound of formula I or I '(preferably of formula I) or a pharmaceutically acceptable salt thereof, for use in the treatment of HCV infection in a subject,

Wherein Y is the group CR ", R 'and R" taken together with the carbon atoms to which they are attached form a phenyl group optionally substituted with one or more R₂;

J is the group -C (O) - or -OC (O) -;

A represents group C₁-C₆alkyl, aryl, heteroaryl, C₅-C₆carbocyclic or heterocyclic having 5 to 6 ring atoms, and is optionally substituted with one or more R's₆; G is the group -ER₅, E represents the -NHS group (O₂) -; R₅ means group C₃-C₆carbocyclic or heteroaryl, and is optionally substituted with one or more R's₇; in one embodiment of R₅ is selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, pyridinyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, pyrrolidinyl, morpholinyl, piperidinyl, piperazinyl or any of which is substituted with or₇; preferably, R₅ is a cyclopropyl group;

each of R₁ and R₂ is independently selected from the group consisting of halogen, hydroxy, amino, -CN, -N₃, -CF₃, -NO₂, -OR₄, -SR₄, -S (O) R₄, -S (O₂) R₄, -NR₃R₄, C (O) OR₄, -C (O) R₄, -C (O) NR₃R₄, -N (R₃) C (O) R₄, C.₁-C₆alkyl, C₂-C₆alkenyl, C.₂-C₆alkynyl, C₁-C₆haloalkyl, C.₂-C₆haloalkenyl, or C₂-C₆-halogenoalkynyl;

R₃ is hydrogen; and each of R₄ is independently selected from the group consisting of hydrogen, group C₁-C₆alkyl, C₂-C₆alkenyl or C₂-C₆alkynyl;

R₆ and R₇ are independently selected at each occurrence from the group consisting of halogen, hydroxy, amino, -CF₃, -CN, -N₃, -NO₂, -C₁-C₆alkyl (preferably methyl), C₂C₆alkenyl, C.₂-C₆alkynyl, C₁-C₆haloalkyl, C.₂-C₆haloalkenyl, or C₂C₆-halogenoalkynyl;

A may be, for example, selected from the following groups, each group being optionally substituted by one or more R's₆:

<img file="PL2468285T3_D0008.tif" />

In yet another aspect, the disclosure provides a compound of formula I or I ', or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein R' is an optionally substituted heterocyclic group, optionally substituted heteroaryl group, optionally substituted aryl group or optionally substituted carbocyclic group , and includes two or more fused rings, and where R 'is other than

<img file="PL2468285T3_D0009.tif" />

EP 2 468 285 B1

<img file="PL2468285T3_D0010.tif" />

and and Y is N.

[0032] In yet another aspect, the invention provides a compound of formula I or I ', or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein R' is an optionally substituted heterocyclic group or an optionally substituted heteroaryl group, includes two or more fused rings, and where R 'is other than and and Y is N. Preferably, R 'is an optionally substituted fused bicyclic heterocyclic group or fused bicyclic heteroaryl group. Alternatively or additionally, R 'is optionally substituted with one or more R's₂and preferably via an alkyl or aryl group.

[0033] Other parameters have the meanings as defined above, including preferred and alternative embodiments.

[0034] In another aspect, the invention provides a compound of formula I or I ', or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein R' is an optionally substituted heterocyclic group, optionally substituted heteroaryl group, optionally substituted aryl group or optionally substituted carbocyclic group , and includes two or more fused rings, and where R 'is other than

<img file="PL2468285T3_D0011.tif" />

and and Y is N; where k = 3, j = 1 and L is absent.

[0035] Preferably, the compound has formula I.

[0036] Alternatively or additionally, m is 1.

[0037] Alternatively or additionally, each of R₃ is H.

[0038] Alternatively or additionally, R₁ and R₂ independently represent a hydrogen or halogen atom.

[0039] Alternatively or additionally, E is -NHS (O._p) -, ip is 2.

[0040] Alternatively or additionally, R₅ means a cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, pyridinyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, pyrrolidinyl, morpholinyl, piperidinyl, piperazinyl, or imidazolyl, group which is optionally substituted. In a further embodiment

R₅ is an optionally substituted cyclopropyl or an optionally substituted thienyl group, and preferably a cyclopropyl or thienyl group.

[0041] Alternatively or additionally, J is -C (O) -.

[0042] Alternatively or additionally, A is an optionally substituted -C group₁-C₈alkyl, containing 0, 1, 2, or 3 heteroatoms selected from O, S, or N; optionally substituted aryl, optionally substituted -C₁-C₈ alkoxy, optionally substituted heteroaryl, optionally substituted -C₃-C₁₂ cycloalkyl, or an optionally substituted -C group₃-C₁₂ heterocycloalkyl. In a further embodiment, A is selected from the group consisting of

<img file="PL2468285T3_D0012.tif" />

[0043] In another aspect, the invention provides a compound of formula I or I ', or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein N is N, and R' is

<img file="PL2468285T3_D0013.tif" />

\ Z \ -P or

<img file="PL2468285T3_D0014.tif" />

And is optionally substituted; provided that said compound is other than (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -14- (cyclopropylsulfonylcarbamoyl) ) -5.16-dioxo-1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2- a] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate.

[0044] A feature of the invention is also a compound of formula I or formula I '(preferably of formula I), or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein N is N, and R' is

<img file="PL2468285T3_D0015.tif" />

or and is optionally substituted with one or more R's₂;

k = 0, j = 0, m = 1, n = 0, 1, 2, 3, or 4, and L is a group C₃-C₆alkylene, C₃-C₆alkenylene or C₃-C₆alkynylene and is optionally substituted with one or more R's₇;

J is -C (O) - or -OC (O) -;

A represents group C₁-C₆alkyl, C₂-C₆alkenyl, C.₂-C₆alkynyl, C₃-C₁₀carbocyclic, aryl, heteroaryl, or heterocyclic having 5 to 10 ring atoms, and A is optionally substituted with one or more R₆;

G is -ER_5, E is -NHS (O₂) -; R₅ means group C₁-C₆alkyl, C₂-C₆alkenyl, C.₂C₆alkynyl, C₃-C₁₀carbocyclic, aryl, heteroaryl, or heterocyclic with 5 to 10 ring atoms, and R₅ is optionally substituted with one or more R's₇;

each of R₁ and R₂ is independently selected from the group consisting of halogen, hydroxy, amino, -CN, -CF₃, -N₃, -NO₂, -OR₄, -SR₄, -S (O) R₄, -S (O₂) R₄, -NR₃R₄, -C (O) OR₄, -C (O) R₄, -C (O) NR₃R₄, -N (R₃) C (O) R₄, C.₁-C₆alkyl, C₂-C₆alkenyl, C.₂-C₆alkynyl, C₁C₆haloalkyl, C.₂-C₆haloalkenyl, C.₂-C₆haloalkynyl, C₃-C₁₀carbocyclic optionally substituted with one or more R's₇or heterocyclic with 5 to 10 ring atoms and optionally substituted with one or more R₇where each of R₆ and R₇ is independently selected from the group consisting of halogen, hydroxy, amino, -CN, -CF₃, -N₃, -NO₂, -C₁-C₆alkyl, C₂-C₆alkenyl, C.₂-C₆alkynyl, C₁-C₆haloalkyl, C.₂-C₆haloalkenyl, or C₂-C₆-halogenoalkynyl; and each of R₃ or R₄ is independently at each occurrence selected from the group consisting of hydrogen, group C₁-C₆alkyl, C₂-C₆alkenyl or C₂-C₆alkynyl;

provided that said compound is other than (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -14- (cyclopropylsulfonylcarbamoyl) ) -5.16-diokso1,2,3,5,6, '7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a ] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate.

[0045] A further feature of the invention is a compound of formula I or formula I '(preferably of formula I), or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein Y is the N azome and R' is or

<img file="PL2468285T3_D0016.tif" />

and is optionally substituted with one or more R's₂;

k = 0, j = 0, m = 1, n = 0, 1, 2, 3, or 4, and L is a group C₃-C₆alkylene, C₃-C₆alkenylene or C₃-C₆alkynylene and is optionally substituted with one or more halogen;

J is -C (O) - or -OC (O) -; A represents group C₁-C₆alkyl, aryl, heteroaryl, C₃C₁₀carbocyclic or heterocyclic having 5 to 10 ring atoms, and is optionally substituted with one or more R's₆;

G is -ER₅, E is -NHS (O₂) -; R₅ means group C₃-C₁₀carbocyclic or heteroaryl, and is optionally substituted with one or more R's₇; each of R₁ and R₂ is independently selected from the group consisting of halogen, hydroxy, amino, -CN, -CF₃, -N₃, -NO₂, -OR₄, -SR₄, -S (O) R₄, -S (O₂) R₄, -NR₃R₄, -C (O) OR₄, C (O) R₄, -C (O) NR₃R₄, -N (R₃) C (O) R₄, C.₁-C₆alkyl, C₂-C₆alkenyl, C.₂-C₆alkynyl, C₁C₆haloalkyl, C.₂-C₆haloalkenyl, C.₂-C₆haloalkynyl, C₃-C₁₀carbocyclic optionally substituted with one or more R's₇or heterocyclic with 5 to 10 ring atoms and optionally substituted with one or more R₇where each of R₆ and R₇ is independently selected from the group consisting of halogen, hydroxy, amino, -CN, -CF₃, -N₃, -NO₂, -C₁-C₆alkyl, C₂-C₆alkenyl, C.₂-C₆alkynyl, C₁-C₆haloalkyl, C.₂-C₆haloalkenyl, or C₂-C₆-halogenoalkynyl; R₃ is hydrogen; and each of R₄ is independently selected from the group consisting of hydrogen, group C₁C₆alkyl, C₂-C₆alkenyl or C₂-C₆alkynyl;

provided that said compound is other than (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -14- (cyclopropylsulfonylcarbamoyl) ) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate.

[0046] In addition, a feature of the invention is a compound of formula I or formula I '(preferably of formula I), or a pharmaceutically acceptable salt, ester or prodrug thereof, wherein N is N, and R' is

EP 2 468 285 B1

<img file="PL2468285T3_D0017.tif" />

or

<img file="PL2468285T3_D0018.tif" />

and is optionally substituted with one or more R's₂;

k = 3, j = 1, m = 1, n = 0, 1, 2, 3, or 4, and L is absent; J is -C (O) - or -OC (O) -; A represents group C₅-C₆carbocyclic or heterocyclic having 5 to 6 ring atoms, and is optionally substituted with one or more R's₆;

G is -ER₅, E is -NHS (O₂) -; R₅ means group C₃-C₆carbocyclic or heteroaryl, and is optionally substituted with one or more R's₇;

each of R₁ and R₂ is independently selected from the group consisting of halogen, hydroxy, amino, -CN, -CF₃, -N₃, -NO₂, -OR₄, -SR₄, -S (O) R₄, -S (O₂) R₄, -NR₃R₄, -C (O) OR₄, C (O) R₄, -C (O) NR₃R₄, -N (R₃) C (O) R₄, C.₁-C₆alkyl, C₂-C₆alkenyl, C.₂-C₆alkynyl, C₁C₆haloalkyl, C.₂-C₆haloalkenyl, or C₂-C₆-halogenoalkynyl;

R₃ is hydrogen; and each of R₄ is independently selected from the group consisting of hydrogen, group C₁-C₆alkyl, C₂-C₆alkenyl or C₂-C₆alkynyl, and R₆ and R₇ have the meanings as defined above;

provided that said compound is other than (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -14- (cyclopropylsulfonylcarbamoyl) ) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14] 4a, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a ] [1,4] tert-butyl cyclopentadecin-6-ylcarbamate.

[0047] In one embodiment of R₅ is selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, pyridinyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, pyrrolidinyl, morpholinyl, piperidinyl, piperazinyl or any of which is substituted with or₇. Preferably, R₅ is a cyclopropyl group. A may be, for example, selected from the following groups and optionally substituted with one or more R's₆:

halls

<img file="PL2468285T3_D0019.tif" />

EP 2 468 285 B1

<img file="PL2468285T3_D0020.tif" />

[0048] Representative compounds of the invention include, but are not limited to, the following compounds:

(24) (2R, 6S, 13aS, 14aR, 16aS, Z) -14- (cyclopropylsulfonylcarbamoyl) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3,5,6,7 , 8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate;

(26) (2R, 6S, 13aS, 14aR, 16aS, Z) -14- (cyclopropylsulfonylcarbamoyl) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3,5,6,7 , 8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecin-cyclopentyl 6-ylcarbamate;

(33) (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -5,16-dioxo-2- (phenanthridin-6-yloxy) -6- (pyrimidine-4-carboxamido) -1.2 , 3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyno-14a-carboxamide;

(34) (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -6- (1-methyl-1H-pyrazole-3-carboxamido) -5,16-dioxo-2- (phenanthridine- 6-yloxy) -1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1, 4] diazacyklopentadecyno-14a-carboxamide;

(35) (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -6- (2-hydroxy-2-methylpropaneamido) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyno- 14a-carboxamide;

(37) (2R, 6S, 13aS, 14aR, 16aS, Z) -14- (cyclopropylsulfonylcarbamoyl) -2- (2-fluoro-phenanthridin-6-yloxy) -5,16-dioxo-1,2,3,5,6,7 , 8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecin-cyclopentyl 6-ylcarbamate;

(38) (2R, 6S, 13aS, 14aR, 16aS, Z) -14- (cyclopropylsulfonylcarbamoyl) -2- (2,9-difluorophenanthridin-6-yloxy) -5,16-dioxo-1,2,3,5 , 6,7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadechydrocyclopropa [e] pyrrolo [1,2-a] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate ;

(39) (2R, 6S, 13aR, 14aR, 16aS) -14- (cyclopropylsulfonylcarbamoyl) -5,16-dioxo-2- (phenanthridin-6-yloxy) octadecahydrocyclopropa [e] pyrrolo [1,2-a] [1 , 4] tert-butyl diazacyclopentadecin-6-ylcarbamate;

(40) (2R, 6S, 13aR, 14aR, 16aS) -14- (cyclopropylsulfonylcarbamoyl) -5,16-dioxo-2 (phenanthridin-6-yloxy) octadekahydrocyclopropa [e] pyrrolo [1,2-a] [1, 4] cyclopentyl diazacyclopentadecin-6-ylcarbamate;

(41) (2R, 6S, 13aR, 14aR, 16aS) -5,16-dioxo-2- (phenanthridin-6-yloxy) -14a- (thiophen-2-ylsulfonylcarbamoyl) octadechydrocyclopropa [e] pyrrolo [1,2- a] [tert-butyl diazacyclopentadecin-6-ylcarbamate;

[0049] In another aspect, the invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I or I 'according to the invention, or a pharmaceutically acceptable salt thereof, in combination with a pharmaceutically acceptable carrier or excipient for use in treating HCV infection in a subject.

[0050] According to another embodiment, the pharmaceutical compositions of the present invention may further comprise one or more other anti-HCV agents. Examples of anti-HCV agents include, but are not limited to, α-interferon; β-interferon;

Pegylated interferon-α; pegylated interferon-lambda; ribavirin; viramidine; R-5158; nitazoxanide; amantadine; Debio-025, NIM-811; HCV polymerase inhibitors such as R7128,

R1626, R4048, T-1106, PSI-7851, PF-00868554, ANA-598, IDX184, IDX102, IDX375, GS-9190, VCH-759, VCH-916, MK-3281, BCX-4678, MK-3281, VBY708, ANA598, GL59728 or GL60667; BMS-790052; BMS-791325; BMS-650032; inhibitors of HCV entry, helicase or internal ribosome entry site; or other HCV replication inhibitors such as GS-9132, ACH-1095, APH005, A-831, A-689, AZD2836. For further details, see S. Tan, A. Pause, Y. Shi, N. Sonenberg, Hepatitis C Therapeutics: Current Status and Emerging Strategies, Nature Rev. Drug Discov., 1, 867-881 (2002); WO 00/59929 (2000); WO 99/07733 (1999); WO 00/09543 (2000); WO 99/50230 (1999); US5861297 (1999); and US2002 / 0037998 (2002).

[0051] According to an additional embodiment, the pharmaceutical compositions of the present invention may further comprise another HCV protease inhibitor, such as telaprevir, boceprevir, ITMN-191, BI-201335, TMC-435, MK-7009, VBY-376, VX-500 , VX-813, PHX-B, ACH-1625, IDX136, or IDX316.

[0052] In other embodiments, the invention provides a pharmaceutical composition further comprising pegylated interferon, another anti-viral, anti-bacterial, anti-fungal or anti-cancer agent, or immune system modulator, and / or further comprising a cytochrome P450 monooxygenase inhibitor or a pharmaceutically acceptable salt thereof. In certain embodiments, the cytochrome P450 monooxygenase inhibitor is ritonavir.

[0053] In another aspect, the disclosure provides the use of a compound of the disclosure for the preparation of an agent for preventing or treating a viral infection. In another aspect, the disclosure provides a compound of the invention for the treatment of hepatitis C. The present disclosure also contemplates the use of a solvate (e.g., hydrate) of the compound of the disclosure in the manufacture of pharmaceutical compositions for the prevention or treatment of hepatitis C infection. The term "solvate" as used herein refers to the physical association of a compound of the disclosure with one or more solvent molecules, whether organic or inorganic. This physical association often includes hydrogen bonding. In some cases, the solvate is suitable for isolation, for example, when one or more solvate molecules are incorporated into the crystal lattice of the crystalline solid.

[0054] In another embodiment, the pharmaceutical compounds or compositions of the invention are administered with ritonavir, either simultaneously or sequentially. In certain embodiments, a compound or pharmaceutical composition of the invention is administered in the same composition as ritonavir. In another embodiment, the compound or pharmaceutical composition thereof of the invention is administered in a composition other than ritonavir.

[0055] According to yet another embodiment, the pharmaceutical compositions of the present invention may further comprise an inhibitor (s) of other HCV life cycle targets, including, but not limited to, helicase, polymerase, metalloprotease, CD81, NS5A, cyclophilin, and internal ribosome entry site (IRES).

[0056] In one aspect, the disclosure provides a compound of formula I or I 'described herein, or a pharmaceutically acceptable salt, ester or prodrug thereof, or a pharmaceutical composition containing it, for use in treating a viral infection in a subject by administering to the subject a therapeutically effective amount of said compound or composition.

[0057] The present disclosure includes a compound of formula I or I 'or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to the invention, for use in the treatment of hepatitis C infection in a subject in need of such treatment by administering to said subject an effective anti-HCV virus an amount or inhibitory amount of said compound or pharmaceutical composition of the present invention. [0058] According to another embodiment, the present invention includes a compound of formula I or I 'or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the invention, for use in the treatment of hepatitis C infection in a subject in need of such treatment by administering to said to the subject, said compound or said pharmaceutical composition of the present invention. The compounds for use may be further administered with an additional therapeutic agent, including another antiviral agent or anti-HCV agent, as described above. The additional agent may be co-administered (such as co-administered or sequentially administered) with a compound (its pharmaceutically acceptable salt) or pharmaceutical composition of the present invention. The additional agent (s) and the compound (or a pharmaceutically acceptable salt or ester thereof) of the present invention may be formulated in the same composition or in different compositions, but co-administered simultaneously or sequentially.

[0059] We also disclose the step of identifying that the subject needs treatment for hepatitis C infection. Identification can be carried out by subjective (e.g., determined by the healthcare provider) or objective (e.g., diagnostic test). [0060] In one aspect, the invention provides a compound or pharmaceutical composition of the invention for use in inhibiting hepatitis C virus replication by contacting the hepatitis C virus with an effective amount of a compound or pharmaceutical composition of the invention.

[0061] In another embodiment, the invention provides a compound or pharmaceutical composition of the invention for use as described above, further comprising administering an additional anti-hepatitis C virus agent. Examples of anti-hepatitis C virus agents include, but are not limited to, α-interferon; β-interferon; pegylated interferon-α; pegylated interferon-lambda; ribavirin; viramidine; R5158; nitazoxanide; amantadine; Debio-025, NIM-811; HCV polymerase inhibitors such as R7128,

R1626, R4048, T-1106, PSI-7851, PF-00868554, ANA-598, IDX184, IDX102, IDX375, GS-9190, VCH-759, VCH-916, MK-3281, BCX-4678, MK-3281, VBY708, ANA598, GL59728 or GL60667;

BMS-790052; BMS-791325; BMS-650032; inhibitors of HCV entry, helicase or internal ribosome entry site; or other HCV replication inhibitors such as GS-9132, ACH-1095, APH005, A-831, A-689, AZD2836. For further details, see S. Tan, A. Pause, Y. Shi, N. Sonenberg, Hepatitis C Therapeutics: Current Status and Emerging Strategies, Nature Rev. Drug Discov., 1, 867-881 (2002); WO 00/59929 (2000); WO 99/07733 (1999); WO 00/09543 (2000); WO 99/50230

EP 2 468 285 B1 (1999); US5861297 (1999); and US2002 / 0037998 (2002). Preferably, the compound or pharmaceutical composition of the present invention is co-administered with, or used in combination with, pegylated interferon (e.g., pegylated interferon alfa-2a or 2b) and ribavirin. Ritonavir or another cytochrome P450 monooxygenase inhibitor may also be used to enhance the pharmacokinetics of the compound of the present invention. Preferably, the patient being treated is infected with HCV genotype 1 (e.g., genotype 1a or 1b). Patients infected with HCV of a different genotype, such as genotypes 2, 3, 4, 5 or 6, may also be treated with the compound or pharmaceutical composition of the present invention.

[0062] In another embodiment, the invention provides a compound or pharmaceutical composition of the invention for use as described above, further comprising administering another HCV protease inhibitor, an HCV polymerase inhibitor, an HCV helicase inhibitor, or an internal ribosome entry (IRES) inhibitor, such as telaprevir, boceprevir, ITMN-191, BI-201335, TMC-435, MK-7009, VBY-376, VX-500, VX-813, PHX-B, ACH-1625, IDX136, IDX316, pegylated interferon, another antiviral, antibacterial, antifungal or anticancer agent, or immunomodulator, and / or further comprising a cytochrome P450 monooxygenase inhibitor or a pharmaceutically acceptable salt thereof. In certain embodiments, the cytochrome P450 monooxygenase inhibitor is ritonavir.

[0063] An additional embodiment of the present invention includes compounds of the present invention for use in the treatment of biological samples by contacting the biological samples with said compounds.

[0064] Still another aspect of the present disclosure is a method for preparing any of the compounds described herein using any of the synthetic agents described herein.

Definitions [0065] The following are definitions of various terms used to describe the present invention and disclosure. These definitions refer to terms that are used throughout this specification and claims, unless otherwise limited in specific cases, either alone or as part of a larger group. The number of carbon atoms in the hydrocarbon substituent may be indicated by the prefix "C_x-C_s"where x is the smallest and y is the largest number of carbon atoms in the substituent.

[0066] The prefix "halo" indicates that the substituent to which the prefix is ​​attached is substituted with one or more independently selected radicals derived from halogen atoms. For example, "haloalkyl" means an alkyl substituent in which at least one hydrogen radical is replaced by a radical derived from a halogen atom.

[0067] If the connecting element in the illustrated structure is "absent", then the left element in the illustrated structure is directly connected to the right element in the illustrated structure. For example, if the chemical structure is represented as XLY, where L is absent, then the chemical structure is XY.

[0068] The term "alkyl" as used herein, refers to a saturated, straight or branched chain hydrocarbon radical typically containing from 1 to 20 atoms

EP 2 468 285 B1. For example, group "C₁-C₆ alkyl "or" C₁-C₈ alkyl "contains from one to six, or from one to eight, carbon atoms, respectively. Examples of alkyl radicals include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tertbutyl, neopentyl, n-hexyl, heptyl, octyl, and the like.

[0069] The term "alkenyl" as used herein, means a straight or branched chain hydrocarbon radical having one or more double bonds and typically from 2 to 20 carbon atoms. For example, group "C₂-C₆ alkenyl "or" C₂-C₈ alkenyl "contains from two to six, or from two to eight carbon atoms, respectively. Alkenyl groups include, but are not limited to, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, heptenyl , octenyl and the like.

[0070] The term "alkynyl" as used herein, means a straight or branched chain hydrocarbon radical having one or more triple bonds and typically from 2 to carbon atoms. For example, group "C₂-C₆ alkynyl 'or' C.₂-C₈ alkynyl "contains from two to six, or from two to eight, carbon atoms, respectively. Representative alkynyl groups include, but are not limited to, for example, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl and the like.

[0071] The term "alkylene" refers to a divalent group derived from a straight or branched saturated hydrocarbon chain, typically containing from 1 to 20 carbon atoms, more typically from 1 to 8 carbon atoms, and even more typically from 1 to 6 carbon atoms . Representative examples of the alkylene group include, but are not limited to, the -CH group₂-, -CH₂CH₂-, -CH₂CH₂CH₂-, -CH₂CH₂CH₂CH₂-, their₂CH (CH₃) CH₂-.

[0072] The term "alkenylene" refers to a divalent unsaturated hydrocarbon group which may be linear or branched and which has at least one carbon-carbon double bond. An alkenylene group typically contains 2 to 20 carbon atoms, more typically 2 to 8 carbon atoms, and even more typically 2 to 6 carbon atoms. Non-limiting examples of alkenylene groups include -C (H) = C (H) -, -C ( H) = C (H) -CH₂-, -C (H) = C (H) -CH₂CH₂-, -CH₂-C (H) = C (H) -CH₂-, -C (H) = C (H) -CH (CH₃)-, their₂-C (H) = C (H) CH (CH₂CH₃)-.

[0073] The term "alkynylene" refers to a divalent unsaturated hydrocarbon group, which may be linear or branched, and which has at least one carbon-carbon triple bond. Representative alkynylene groups include, for example, -ChC-, CeC-CH₂-, -ChC-CH₂CH₂-, -CH₂-CHC-CH₂-, -ChC-CH (CH₃)-, their₂-CHC-CH (CH₂CH₃)-.

[0074] The term "cycloalkyl" means a monovalent group derived from a monocyclic or polycyclic saturated compound with a carbocyclic ring. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo [2.2.1] heptyl, and bicyclo [2.2.2] octyl and the like.

[0075] The terms "carbocyclic" or "carbocyclic" or "carbocyclyl" refer to saturated (e.g., "cycloalkyl"), partially saturated (e.g., "cycloalkenyl" or "cycloalkynyl") or completely unsaturated (e.g., "aryl")

EP 0 468 285 B1 containing zero ring heteroatoms and typically from 3 to 18 ring carbon atoms. The carbocyclyl group may be, without limitation, a single ring, or two or more rings fused, or bridged, or connected by a spiro moiety. The carbocyclyl group may contain, for example, from 3 to 14 ring members (i.e., group C₃C₁₄carbocyclyl, such as group C₃-C₁₄cycloalkyl), from 3 to 10 ring members (i.e., group C₃-C₁₀carbocyclyl, such as group C₃-C₁₀cycloalkyl), from 3 to 8 ring members (i.e., group C₃-C₈carbocyclyl, such as group C₃-C₈cycloalkyl), or from 3 to 6 ring members (i.e., group C₃-C₆carbocyclyl, such as group C₃-C₆cycloalkyl). The substituted carbocyclyl group may have either cis or trans geometry. Representative examples of carbocyclyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopentenyl, cyclopentadienyl, cyclohexadienyl, adamantyl, decahydronaphthylenyl, ochelenhelenyl, 1,2,3,4-tetrahydronaphthyl, indenyl, isoindenyl, bicyclodecanyl, anthracenyl, phenantrenyl, benzonaftenyl (also known as "phenalenyl"), decalinyl, and norpinanyl, and the like. The carbocyclyl group may be attached to the parent molecular moiety through any substitutable carbon atom of that group.

[0076] The term "aryl" refers to an aromatic carbocyclyl group containing from 6 to 14 ring carbon atoms. Non-limiting examples of aryl groups include phenyl, naphthalenyl, anthracenyl, and indenyl and the like. The aryl group may be attached to the parent molecular moiety through any substitutable carbon atom of that group.

[0077] The term "aralkyl" or "arylalkyl" refers to an alkyl residue attached to an aryl ring. Examples of the aralkyl group include, but are not limited to, benzyl, phenethyl, and the like.

[0078] The term "heteroaryl" means an aromatic heterocyclyl group typically containing from 5 to 18 ring atoms. The heteroaryl group may have a single ring, or two or more fused rings. Non-limiting examples of five-membered heteroaryl groups include an imidazolyl group; furanyl; thiophenyl (or thienyl or thiofuranyl); pyrazolyl; oxazolyl; isoxazolyl; thiazolyl; 1,2,3-, 1,2,4-, 1,2,5-, and 1,3,4-oxadiazolyl; and isothiazolyl. Non-limiting examples of six-membered heteroaryl groups include a pyridinyl group; pyrazinyl; pyrimidinyl; pyridazinyl; and 1,3,5-, 1,2,4-, and 1,2,3-triazinyl. Non-limiting examples of 6/5-membered fused ring heteroaryl groups include benzothiofuranyl, isobenzothiofuranyl, benzisoxazolyl, benzoxazolyl, purinyl, and anthranilyl. Non-limiting examples of 6/6-membered fused ring heteroaryl groups include a quinolinyl group; isoquinolinyl; and benzoxazinyl (including cinnolinyl and quinazolinyl).

[0079] The term "heteroaralkyl" or "heteroarylalkyl" refers to an alkyl residue attached to a heteroaryl ring. Examples include, but are not limited to, pyridinylmethyl, pyrimidinylethyl, and the like.

[0080] The term "heterocycloalkyl" refers to non-aromatic 3-, 4-, 5-, 6- or 7-membered rings or a bi- or tri-cyclic condensed group system, where (i) each the rings contain between one and three heteroatoms independently selected from oxygen, sulfur and nitrogen, (ii) each of the 5-membered rings has 0 to 1 double bonds and each of the 6-membered rings has 0 to 2 double bonds, (iii) nitrogen and sulfur heteroatoms may optionally be oxidized, (iv) the nitrogen heteroatom may optionally be quaternized, and (iv) any of the above rings may be fused to a benzene ring. Representative heterocycloalkyl groups include, but are not limited to, [1,3] dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, thiazylidinyl, thiazolidinyl

[0081] The terms "heterocyclic system" or "heterocyclic group" or "heterocyclyl group" refer to saturated ring systems (e.g., "heterocycloalkyl"), partially unsaturated (e.g., "heterocycloalkenyl" or "heterocycloalkynyl") or completely unsaturated (e.g., "heteroaryl") typically containing from 3 to 18 ring atoms, wherein at least one of the ring atoms is a heteroatom (i.e., nitrogen, oxygen or sulfur), wherein the remaining ring atoms are independently selected from the group consisting of carbon, nitrogen, oxygen and sulfur. The heterocyclyl group may be attached to the parent molecular moiety through any substitutable carbon atom or nitrogen atom in the group, provided that a stable molecule is formed. The heterocyclyl group may be, without limitation, a single ring which typically contains from 3 to 14 ring atoms, from 3 to 8 ring atoms, from 3 to 6 ring atoms, or from 5 to 6 ring atoms. Non-limiting examples of monocyclic heterocyclyl groups include furanyl, dihydrofuranyl, pyrrolyl, isopyrrolyl, pyrrolinyl, pyrrolidinyl, imidazolyl, izoimidazolilową, imidazolidinyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, triazolyl, tetrazolyl, ditiolilową, oksatiolilową, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, thiazolinyl , isothiazolinyl, thiazolidinyl, isothiazolidinyl, thiodiazolyl, oxatiazolyl, oxadiazoly, pyranyl, dihydropyranyl, pyridinyl, piperidinyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperazinyl, triazinyl, isoxazinyl, oxazolidinyl, isoxazolidinyl, oxynylepinylpynylidinyl. The heterocyclyl group may also include, without limitation, two or more rings fused together, such as, for example, a naphthyridinyl, thiazolopyrimidinyl, thienopyrimidinyl, pyrimidopyrimidinyl, or pyridopyrimidinyl group. A heterocyclyl group may contain one or more sulfur atoms as ring members; and in some cases, the sulfur atom (s) is oxidized to SO or SO₂. The nitrogen heteroatom (heteroatoms) in the heterocyclyl group may or may not be quaternized, and may or may not be oxidized to N-oxide. In addition, the terms "optionally substituted", "optionally substituted alkyl", "optionally substituted" "optionally substituted alkenyl", "optionally substituted alkynyl", "optionally substituted carbocyclic", "optionally substituted aryl", " optionally substituted heteroaryl "," optionally substituted heterocyclic ", and any other optionally substituted group refer to groups which are substituted or not 20

Substituted by independently substituting for one, two, or three or more hydrogen atoms for substituents including, but not limited to:

-F, -Cl, -Br, -I,

-OH, protected hydroxyl, alkoxy, oxo, thioxo group,

-WELL₂, -CN, CF₃, N₃,

NH₂, protected amino, -NH-alkyl, -NH-alkenyl, -NH-alkynyl, -NH-cycloalkyl, -NH-aryl, -NH-heteroaryl, -NH-heterocyclic, -dialkylamino, -darylamino, -diheteroarylamino,

-O-alkyl, -O-alkenyl, -O-alkynyl, -O-cycloalkyl, -O-aryl, -O-heteroaryl, -Oheterocyclic,

-C (O) -alkyl, -C (O) -alkenyl, -C (O) -alkynyl, -C (O) -cycloalkyl, -C (O) -aryl, -C (O) heteroaryl, -C ( O) -heterocykloalkilową,

-CONH₂, -CONH-alkyl, -CONH-alkenyl, -CONH-alkynyl, -CONH-cycloalkyl, -CONHaryl, -CONH-heteroaryl, -CONH-heterocycloalkyl,

-ABOUT WHAT₂-alkyl, -OCO₂-alkenyl, -OCO₂-alkynyl, -OCO₂-cycloalkyl, -OCO₂-aryl, OCO₂-heteroaryl, -OCO₂-heterocycloalkyl, -OCONH₂, -OCONH-alkyl, -OCONHalkenyl, -OCONH-alkynyl, -OCONH-cycloalkyl, -OCONH-aryl, -OCONH-heteroaryl, -OCONH-heterocycloalkyl,

-NHC (O) -alkyl, -NHC (O) -alkenyl, -NHC (O) -alkynyl, -NHC (O) -cycloalkyl, -NHC (O) aryl, -NHC (O) -heteroaryl, -NHC ( O) -heterocycloalkyl, -NHCO₂-alkyl, -NHCO₂alkenyl, -NHCO₂-alkynyl, -NHCO₂-cycloalkyl, -NHCO₂-aryl, -NHCO₂ heteroaryl, -NHCO₂-heterocycloalkyl, -NHC (O) NH₂, -NHC (O) NH-alkyl, -NHC (O) NHalkenyl, -NHC (O) NH-alkenyl, -NHC (O) NH-cycloalkyl, -NHC (O) NH-aryl, NHC (O) NH- heteroaryl, -NHC (O) NH-heterocycloalkyl, NHC (S) NH₂, -NHC (S) NH-alkyl, NHC (S) NH-alkenyl, -NHC (S) NH-alkynyl, -NHC (S) NH-cycloalkyl, -NHC (S) NH-aryl, NHC (S) NH -heteroaryl, -NHC (S) NH-heterocycloalkyl, -NHC (NH) NH₂, -NHC (NH) NHalkyl, -NHC (NH) NH-alkenyl, -NHC (NH) NH-alkenyl, -NHC (NH) NH-cycloalkyl, NHC (NH) NH-aryl, -NHC (NH) NH- heteroaryl, -NHC (NH) NH-heterocycloalkyl, -NHC (NH) alkyl, -NHC (NH) -alkenyl, -NHC (NH) -alkenyl, -NHC (NH) -cycloalkyl, -NHC (NH) aryl, - NHC (NH) -heteroaryl, -NHC (NH) -heterocycloalkyl,

-C (NH) NH-alkyl, -C (NH) NH-alkenyl, -C (NH) NH-alkynyl, -C (NH) NH-cycloalkyl, C (NH) NH-aryl, -C (NH) NH -heteroaryl, -C (NH) NH-heterocycloalkyl,

-S (O) -alkyl, -S (O) -alkenyl, -S (O) -alkynyl, -S (O) -cycloalkyl, -S (O) -aryl, -S (O) heteroaryl, -S ( O) -heterocycloalkyl, -SO₂NH₂, -SO₂NH-alkyl, -SO₂NH-alkenyl, SO₂NH-alkynyl, -SO₂NH-cycloalkyl, -SO₂NH-aryl, -SO₂NH-heteroaryl, -SO₂NHheterocykloalkilową,

NHSO₂-alkyl, -NHSO₂-alkenyl, -NHSO₂-alkynyl, -NHSO₂-cycloalkyl, -NHSO₂-aryl, -NHSO₂-heteroaryl, -NHSO₂-heterocykloalkilową,

CH₂NH₂, -CH₂SO₂CH₃, -alkyl, -alkenyl, -alkynyl, -aryl, -arylalkyl, -heteroaryl, -heteroarylalkyl, -heterocycloalkyl, -cycloalkyl, -carbocyclic, -heterocyclic, polyalkoxyalkyl, polyalkoxy, -methoxy-oxyethoxy

EP 2 468 285 B1

S-alkyl, -S-alkenyl, -S-alkynyl, -S-cycloalkyl, -S-aryl, -S-heteroaryl, -Sheterocycloalkyl, or methylthiomethyl.

[0083] It is understood that aryl, heteroaryl, carbocyclic, heterocyclic, alkyl and the like may be further substituted.

[0084] The terms "halo" and "halogen" as used herein refer to an atom selected from fluorine, chlorine, bromine and iodine.

[0085] The term "subject" as used herein refers to a mammal. Thus, the subject refers to, for example, dogs, cats, horses, cows, pigs, guinea pigs, and the like. Preferably, the subject being treated is human. When the treated person means human, the treated person can be either a patient or a healthy person.

[0086] As used herein, the term "hydroxyl activating group" refers to a labile chemical moiety that is known to activate the hydroxyl group so that it will go away during synthetic procedures such as substitution or elimination reactions. Examples of hydroxyl activating groups include, but are not limited to, mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate and the like.

[0087] The term "leaving group" or "LG" as used herein refers to any group that exits during a chemical reaction involving the group and includes, but is not limited to, for example, halogen, p-bromobenzenesulfonate, mesylate, tosylate, trifluoromethanesulfonate, p-nitrobenzoate, phosphonate.

[0088] The term "protected hydroxyl group," as used herein, refers to a hydroxyl group protected by a hydroxyl protecting group, as defined above, including, for example, benzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl.

[0089] The term "hydroxy protecting group," as used herein, refers to a labile chemical moiety that is known to protect a hydroxyl group against unwanted reactions during synthetic procedures. After said synthetic procedure (s), the hydroxyl protecting group as described herein can be selectively removed. Known hydroxyl protecting groups are described generally in TH Greene and PGM Wuts, Protective Groups in Organic Synthesis, ed. 3, John Wiley &amp; Sons, New York (1999). Examples of hydroxyl protecting groups include benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 4-bromobenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, methoxycarbonyl, tert-butoxycarbonyl, isopropoxycarbonyl, diphenylmethoxycycarbonyl, 2,2-tricycloxycarbonyl, 2,2-tricarbonylmethyl allyloxycarbonyl, acetyl, formyl, chloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, tert-butyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 1,1-dimethyl-2-propenyl, 3-methyl-3-butenyl, allyl, benzyl, para-methoxybenzyldiphenylmethyl, and triphenylmethyl) , tetrahydrofuryl, methoxymethyl, methylthiomethyl, benzyloxymethyl, 2,2,2-trichloroethoxymethyl, 2- (trimethylsilyl) ethoxymethyl, methanesulfonyl, para-toluenesulfonyl, trimethylsilyl, triethylsilyl, triisopropyl, and triisopropyl. Favorable groups

The hydroxyl protecting group for the present invention is an acetyl group (Ac or -C (O) CH₃), benzoyl (Bz or -C (O) C₆H₅), and trimethylsilyl (TMS or -Si (CH₃)₃).

[0090] The term "amino protecting group," as used herein, refers to a labile chemical moiety that is known to protect an amino group against unwanted reactions during synthetic procedures. After said synthetic procedure (s), the amino protecting group as described herein can be selectively removed. Known amino protecting groups are described generally in TH Greene and PGM Wuts, Protective Groups in Organic Synthesis, ed. 3, John Wiley &amp; Sons, New York (1999). Examples of amino protecting groups include, but are not limited to, tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyloxycarbonyl, and the like.

[0091] The term "protected amino" as used herein refers to an amino group protected by an amino protecting group as defined above.

[0092] The term "alkylamino" refers to a group having the structure -N (R_andR_b), where R_and and R_b are independently H or alkyl.

[0093] The term "acyl" includes residues derived from acids, including, but not limited to, carboxylic acids, carbamic acids, carbonic acids, sulfonic acids, and phosphoric acids. Examples include aliphatic carbonyl, aromatic carbonyl, aliphatic sulfonyl, aromatic sulfinyl, aliphatic sulfinyl, aromatic phosphate and aliphatic phosphate groups. Examples of aliphatic carbonyl groups include, but are not limited to, acetyl, propionyl, 2-fluoroacetyl, butyryl, 2-hydroxyacetyl, and the like.

[0094] As used herein, the term "pharmaceutically acceptable salt" refers to those salts of compounds of the method of the present invention that are, in the field of sound medical judgment, useful for use in contact with human and lower animal tissues without undue toxicity, irritation, response allergic and the like, and are commensurate with a reasonable benefit-risk ratio. Pharmaceutically acceptable salts are known in the art. For example, SM Berge et al. describe in detail the pharmaceutically acceptable salts in J. Pharmaceutical Sciences, 66: 1-19 (1977). Salts can be prepared in situ during the final isolation and purification of the compounds of the invention, or separately by reacting the free base with a suitable organic acid. Examples of pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts, or amino group salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or with organic acids such like acetic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid, or by using other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, campforate, camphorsulfonate, citrate, cyclopentanopropionate, dodecyl sulfate, dodecanoate,

Glucoheptonate, glycerophosphate, gluconate, hemisulphate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulphonate, lactobionate, lactate, laurate, lauryl sulphate, malate, mesilate, nitrate, malate, mesilate , palmitate, pamoate, pectate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, or magnesium salts, and the like. Further pharmaceutically acceptable salts contain, where appropriate, non-toxic ammonium, quaternary ammonium, and amine cations prepared using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl group having 1 to 6 carbon atoms, sulfonate and arylsulfonate .

[0095] The term "pharmaceutically acceptable ester," as used herein, refers to esters of compounds prepared according to the present disclosure that hydrolyze in vivo and include those that degrade readily in the human body to form the parent compound or a salt thereof. Useful ester groups include, for example, groups derived from pharmaceutically acceptable aliphatic carboxylic acids, especially alkane, alkene, cycloalkane and alkanedio acids, in which each alkyl or alkenyl moiety preferably has no more than 6 carbon atoms. Examples of individual esters include, but are not limited to, formates, acetates, propionates, butyrates, acrylates and ethyl succinates.

[0096] The term "pharmaceutically acceptable prodrugs," as used herein, refers to those prodrugs of the compounds of the method of this disclosure that are, in the field of sound medical judgment, useful for use in contact with human and lower animal tissues with inappropriate toxicity, irritation, response allergic, and the like, commensurate with a reasonable benefit-risk ratio, and effective for their intended use, as well as zwitterions, where possible, of the compounds described herein. The term "prodrug," as used herein, means a compound that can be transformed in vivo by metabolic agents (e.g., by hydrolysis) to give any compound represented by formulas I and I '. Various forms of prodrugs are known in the art, for example, as discussed in publications: Bundgaard, (ed.), Design of Prodrugs, Elsevier (1985); In idder, et al. (ed.), Methods in Enzymology, vol. 4, Academic Press (1985); Krogsgaard-Larsen, et al. (Ed.). Design and Application of Prodrugs, Textbook of Drug Design and Development, chapter 5, 113-191 (1991); Bundgaard, et al., Journal of Drug Deliver Reviews, 8: 138 (1992); Bundgaard, J. of Pharmaceutical Sciences, 77: 285 et seq. (1988); Higuchi and Stella (ed.) Prodrugs as Novel Drug Delivery Systems, American Chemical Society (1975); and Bernard Testa &amp; Joachim Mayer, "Hydrolysis In Drug And Prodrug Metabolism: Chemistry, Biochemistry And Enzymology", John Wiley and Sons, Ltd. (2002).

[0097] The present disclosure also includes pharmaceutical compositions comprising pharmaceutically acceptable prodrugs of the compounds of the disclosure and pharmaceutically acceptable prodrugs of the compounds of the disclosure for use in the treatment of viral infections by

Administration of said pharmaceutically acceptable prodrugs. For example, compounds of the disclosure having free amino, amide, hydroxyl or carboxy groups can be converted to prodrugs. Prodrugs include compounds in which an amino acid residue or polypeptide chain having two or more (e.g., two, three or four) amino acid residues is .. covalently bonded via an amide or ester bond to a free amino, hydroxyl or carboxylic acid group of the compounds of the invention. Amino acid residues include, but are not limited to, 20 amino acids commonly found in nature, indicated by three-letter symbols, and also include 4-hydroxyproline, hydroxylysine, demosine, isodemosine, 3-methylhistidine, norvaline, beta-alanine, gamma-aminobutyric acid, citrulline , homocysteine, homoserine, ornithine and methionine sulfone. Additional types of prodrugs are also disclosed. For example, free carboxyl groups can be derivatized as amides or alkyl esters. Free hydroxyl groups can be derivatized using groups including, but not limited to, hemisuccinates, phosphate esters, dimethylaminoacetates, and phosphoryloxymethyloxycarbonyl groups, as previously reported in Drug Delivery Reviews, 1996, 19, 115. Carbamate prodrugs of hydroxyl and amino groups are also included, as are carbonate prodrugs, sulfonate esters and sulfate esters of hydroxyl groups. Also included is derivatization of the hydroxyl groups as (acyloxy) methyl and (acyloxy) ethyl ethers, wherein the acyl group may be an alkyl ester optionally substituted with groups including, but not limited to, ether, amino and carboxylic acid functional groups, or where the acyl group is an amino acid ester as described above. Prodrugs of this type are described in J. Med. Chem. 1996, 39, 10. Free amines can also be derivatized as amides, sulfonamides or phosphonamides. All of these prodrug moieties may contain groups including, but not limited to, ether, amino and carboxylic acid functionalities.

[0098] Combinations of substituents and parameters envisioned by the present invention and disclosure are only those that result in the formation of stable compounds. The term "stable" as used herein refers to compounds that have sufficient stability to permit production, and which retain the entire compound for a period of time sufficient to be useful for the purposes specified herein (e.g., for therapeutic or prophylactic administration to a subject).

Pharmaceutical Compositions [0099] The pharmaceutical compositions of the present invention comprise a therapeutically effective amount of a compound of the present invention formulated together with one or more pharmaceutically acceptable carriers. As used herein, the term "pharmaceutically acceptable carrier" means a non-toxic, inert solid, semi-solid or liquid solid as a filler, diluent, encapsulating or auxiliary material for any type of formulation. The pharmaceutical compositions of the present invention can be administered to humans and other animals orally, rectally, parenterally, intra-arterially, vaginally, intraperitoneally, topically (as through powders, ointments, or drops), buccal, or as an oral or nasal spray.

[0100] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compounds, liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water, alcohol or other solvents, dissolving agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol , benzyl benzoate, propylene glycol, 1,3-butylene glycol, polysorbate, dimethylformamide, oils (in particular cottonseed, peanut, corn oils, (germ, olive, castor, and sesame), mono- or di-glycerides, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid and sorbitan esters, and mixtures thereof. In addition to inert diluents, oral compositions may also contain adjuvants such as wetting agents, emulsifying and suspending agents, antioxidants, sweetening, flavoring and flavoring agents. The liquid dosage form can also be enclosed in a gelatin capsule, where the compound of the present invention can be dissolved in a pharmaceutically acceptable carrier containing, for example, one or more dissolving agents (e.g., polysorbate 80 and mono- and diglycerides), and other useful excipients (e.g., antioxidants such as ascorbyl palmitate, or a sweetener or flavoring).

[0101] Injectable preparations, for example, sterile injectable aqueous or oily suspensions may be formulated to the best of the art's knowledge using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a non-toxic parenterally-acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Acceptable substrates and solvents that can be used include water, USP Ringer's solution, and isotonic sodium chloride. In addition, sterile, fixed oils are customarily used as the solvent or suspending medium. Any non-irritating fixed oil can be used for this purpose, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

[0102] To prolong the effect of the drug, it is often desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous material with poor water solubility. Then the rate of absorption of the drug depends on its dissolution rate, which, in turn, may depend on the size of the crystals and the crystal form. Alternatively, delayed absorption of a parenteral drug is achieved by dissolving or suspending the drug in an oil vehicle. The present invention also contemplates immediate release forms.

[0103] Compositions for rectal or vaginal administration are preferably suppositories that can be prepared by mixing the compounds of the present invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or suppository wax, which are solid at ambient temperature but liquid at body temperature, and therefore in the anus or vaginal cavity, they melt and release the active compound.

[0104] Solid compositions of a similar type may also be employed as fillers in soft and hard gelatin capsules using excipients such as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

[0105] The active compounds may also be in microencapsulated form with one or more excipients as mentioned above.

[0106] Solid dosage forms of tablets, dragees, capsules, pills, and granules can be made with coatings and coatings, such as enteric coatings, controlled release coatings and other coatings known in the field of pharmaceutical formulations. In such solid dosage forms, the active compound may be mixed with at least one inert diluent, such as sucrose, lactose or starch. Such dosage forms may also contain, as is normal practice, additives other than inert diluents, e.g., tabletting lubricants and other tableting auxiliaries, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0107] Dosage forms for topical or transdermal administration of a compound of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, aerosols, inhalants or patches. The active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulations, ear drops, eye ointments, powders and solutions are also contemplated as falling within the scope of the present invention.

[0108] In addition to the active compound of the present invention, ointments, pastes, creams and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid , talc and zinc oxide, or mixtures thereof.

[0109] In addition to the compounds of the present invention, powders and aerosols may contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Aerosols may additionally contain customary propellants such as chlorofluorocarbons.

[0110] Transdermal patches have the added benefit of providing controlled release of the compound into the body. Such dosage forms can be made by dissolving or dispensing the compound in the proper medium. Absorption enhancers may also be used to increase the flow of the compound through the skin. The rate can be controlled either by providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0111] According to the compounds for use in the treatment of the present invention, viral infections are treated or prevented in a subject, such as a human or other animal, by administering to the subject a therapeutically effective amount of a compound of the invention (or a pharmaceutically acceptable salt thereof) in such quantity and for such a period of time as is necessary to achieve the desired result. As used herein, the term "therapeutically effective amount" of a compound of the invention means an amount of compound sufficient to reduce the load

Viral in the subject and / or reduce HCV symptoms in the subject. As understood in the medical field, a therapeutically effective amount of a compound of the present invention will correspond to a reasonable benefit / risk ratio applicable to any medical treatment.

Antiviral Activity [0112] The inhibitory amount or dose of compounds for use according to the present invention may range from about 0.1 mg / kg to about 500 mg / kg, alternatively from about 1 to about 50 mg / kg. Inhibitory amounts or doses will also vary depending on the route of administration as well as the possibility of co-administration with other agents.

[0113] According to the present invention, viral infections are treated or prevented in a subject, such as a human or lower mammal, by administering to the subject an effective amount against hepatitis C virus or an inhibitory amount of a compound for use according to the present invention in such amounts and by such a period of time as necessary to achieve the desired result. The present invention also relates to compounds or compositions of the invention for use in treating biological samples with an inhibitory amount of a compound or composition of the present invention in such amounts and for such a period of time as is necessary to achieve the desired result.

[0114] As used herein, the "effective amount against hepatitis C virus" of a compound for use according to the invention means an amount sufficient to reduce viral load in a biological sample or in a subject. As understood in the medical field, an effective amount against the hepatitis C virus of the compound of the present invention will correspond to a reasonable benefit-risk ratio applicable to any medical treatment.

[0115] The term "inhibitory amount" of a compound for use according to the present invention means an amount sufficient to reduce hepatitis C virus load in a biological sample or in a subject. It is understood that when said inhibitory amount of a compound for use according to the present invention is administered to a subject, it will correspond to a reasonable benefit / risk ratio applicable to any medical treatment as determined by the physician. The term "biological sample (s)" as used herein, means a substance of biological origin intended for administration to a subject. Examples of biological samples include, but are not limited to, blood and blood components such as plasma, platelets, subpopulations of blood cells, and the like; organs such as the kidney, liver, heart, lung, and the like; semen and ova; bone marrow and its components, or stem cells.

[0116] Thus, another embodiment of the present invention is a compound or pharmaceutical composition of the present invention for use in the treatment of a biological sample by contacting said biological sample with an inhibitory amount of said compound or composition.

[0117] When improving the condition being treated, a maintenance dose of the compound, composition or combination for use of the present invention may be administered, if necessary. In turn,

Dosage or frequency of administration, or both, can be reduced as a function of symptoms to a level where the improved condition is maintained when the symptoms have been alleviated to the desired level, treatment should be discontinued. The subject may, however, require intermittent long-term treatment at any recurrence of disease symptoms.

[0118] However, it should be understood that the combined daily use of the compounds and compositions for use according to the present invention will be decided by the attending physician in the field of sound medical judgment. The proper inhibitory dose for any particular patient will depend on a variety of factors including the disorder being treated and the severity of the disorder; activity of the specific compound employed; the specific composition used; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the particular compound employed; duration of treatment; drugs used in combination or concurrently with the specific compound employed; and similar factors known in the medical field.

[0119] The total daily inhibitory dose of compounds for use according to the present invention administered to a subject in single or divided doses may be in amounts, for example, from 0.01 to 50 mg / kg or more, usually from 0.1 up to 25 mg / kg body weight. Single doses of the composition may contain such amounts or an aliquot thereof to obtain a daily dose. In one embodiment, the treatment regimens of the present invention comprise administering to a patient in need of such treatment from about 10 mg to about 1000 mg of the compound (s) for use according to the present invention per day in single or multiple doses. In another embodiment, the treatment regimen comprises administering to a patient in need of such treatment from about 25 mg to about 6000 mg of compound (s) for use per day in single or multiple doses, or with a cytochrome P450 monooxygenase inhibitor such as ritonavir, or without. A useful daily dose for a co-administered cytochrome P450 monooxygenase inhibitor (e.g., ritonavir) may range, without limitation, from 10 to 200 mg. Preferably, the compound (s) for use according to the present invention, or a combination of the compound (s) according to the invention and ritonavir, is administered once a day or twice a day to achieve the desired daily dose. For example, when used without ritonavir, the compound for use of the present invention can be administered to a patient twice a day at a total daily dose of 4,000, 4,200, 4,400, 4,600, 4,800 or 5,000 mg. Otherwise, when used in combination with ritonavir, the compound for use of the present invention may be administered to a patient once or twice a day at a total daily dose of 200, 400, 600 or 800 mg, where the amount of ritonavir may be 25, 50 or 100 mg for one application.

Synthetic Methods [0120] The compounds and methods described herein will be better understood in connection with the following synthetic schemes that illustrate the methods by which the compounds described herein can be prepared.

[0121] The definitions of the variables in the structures in the diagrams are commensurate with the definitions for the corresponding positions in the formulas described herein.

EP 2 468 285 B1

<img file="PL2468285T3_D0021.tif" />

[0122] Scheme 1 describes the synthesis of various compounds of the invention. The starting material was replaced on leaving groups by reaction with a nucleophile to form a nucleophile substituted macrocycle. The basic hydrolysis of the ester to the acid was followed by coupling of the sulfonamide derivative. Then the protected nitrogen was deprotected and substituted with another group.

In one aspect, this invention relates to a process for preparing a compound of formula I, comprising the step of subjecting a compound of formula II to:

<img file="PL2468285T3_D0022.tif" />

where,

J is absent, or is an optionally substituted alkylene group, optionally substituted alkenylene group, optionally substituted alkynylene group, -C (O) -, -OC (O) -, -N (R3) -C (O) -, -C ( S) -, -C (= NR4) -, -S (O) -, -S (O2) -, or -N (R3) -;

A is an optionally substituted alkyl group, an optionally substituted alkenyl group, or an optionally substituted alkynyl group, each of which contains 0, 1, 2, or 3 heteroatoms selected from O, S, or N; optionally substituted aryl, optionally substituted arylalkyl, optionally substituted alkoxy, optionally substituted heteroaryl, optionally substituted heterocyclic, or optionally substituted carbocyclic;

G is -ER₅;

where E is absent; or is an optionally substituted alkylene group, optionally

A substituted alkenylene group, optionally substituted alkynylene group, each of which contains 0, 1, 2, or 3 heteroatoms selected from O, S, or N; or -O-, -S-, -N (R₃)-,

-N (R3) S (Op) -, -N (R3) C (O) -, -N (R3) C (O) S (Op) -, -OS (Op) -, -C (O) S (Op) -, or -C (O) N (R3) S (Op) -;

p is 0, 1 or 2;

R₅ is H; optionally substituted alkyl, optionally substituted alkenyl, or optionally substituted alkynyl, each containing 0, 1, 2, or 3 heteroatoms selected from O, S, or N; optionally substituted carbocyclic, optionally substituted heterocyclic, optionally substituted aryl, or optionally substituted heteroaryl;

each of R₃ and R₄ is independently selected at each occurrence from the group consisting of: optionally substituted alkyl, optionally substituted alkenyl or optionally substituted alkynyl, each of which contains 0, 1, 2, or 3 heteroatoms selected from O, S, or N; optionally substituted aryl; optionally substituted heteroaryl; optionally substituted heterocyclic group; optionally substituted carbocyclic; or a hydrogen atom;

L is absent or selected from the group consisting of an optionally substituted alkylene group, an optionally substituted alkenylene group or an optionally substituted alkynylene group, each of which contains 0, 1, 2, or 3 heteroatoms selected from O, S, or N;

j = 0, 1, 2, 3, or 4;

k = 0, 1, 2, or 3;

m = 0, 1 or 2;

n is 0, 1, 2, 3, or 4; and τςςζ; is a carbon-carbon single or double bond; and LG is a leaving group;

reaction with a compound of formula III:

<img file="PL2468285T3_D0023.tif" />

where:

each of R₁ is independently selected from the group consisting of (i) halogen, hydroxy, amino, -CN, -CF₃, -N₃, -NO₂, -OR₄, -SR₄, SOR4, -SO2R4, -N (R3) S (O) 2-R4, -N (R3) (SO2) NR3R4, -NR3R4, -C (O) OR4, -C (O) R4, C (O) NO₃R₄, or -N (R₃) C (O) R₄;

(ii) an optionally substituted aryl;

(iii) optionally substituted heteroaryl;

(iv) an optionally substituted heterocyclic group;

(v) an optionally substituted carbocyclic group; or (vi) an optionally substituted alkyl group, an optionally substituted alkenyl group,

Or an optionally substituted alkynyl group, each of which contains 0, 1, 2, or 3 heteroatoms selected from O, S, or N;

R₃ and R₄ are each independently selected at each occurrence from the group consisting of the following: optionally substituted alkyl, optionally substituted alkenyl or optionally substituted alkynyl, each of which contains 0, 1, 2, or 3 heteroatoms selected from O, S, or N; optionally substituted aryl; optionally substituted heteroaryl; optionally substituted heterocyclic; optionally substituted carbocyclic; or a hydrogen atom;

Y is N or C (R ");

where if Y is N, then R 'is an optionally substituted heterocyclic group, optionally substituted heteroaryl group, optionally substituted aryl group or optionally substituted carbocyclic group, and includes two or more fused rings, and where R' is other than

<img file="PL2468285T3_D0024.tif" />

Ν vy · (and

<img file="PL2468285T3_D0025.tif" />

further provided that said compound is other than (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3 (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -14- (cyclopropylsulfonylcarbamoyl) ) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] Tert-butyl diazacyclopentadecin-6-ylcarbamate; where, if Y is -C (R ") -, then R 'and R" taken together with the carbon atoms to which they are attached form an aryl or heteroaryl ring, each of said rings being optionally substituted;

where A, R₁, R 'and / or R "may be taken together to form a ring; thereby forming a compound of formula I or I'.

[0124] The compound of formula I can also be prepared according to the method outlined in Scheme 2, where A, J, L, G, Y, R 'R₁, R₃, n, m, j, and k are as defined above, and means

<img file="PL2468285T3_D0026.tif" />

And wherein Q is a halogen atom or a leaving group, each of PG and PG_N is independently an amino protecting group, and PG_C is a carboxylic acid protecting group. Compound (b) can be produced by treatment

<img file="PL2468285T3_D0027.tif" />

reaction with a halogenating agent such as POCl₃. Non-limiting examples of an amino protecting group include a C group₁-C₆alkoxycarbonyl (e.g., tert-butoxycarbonyl or Boc), carboxybenzyl, p-methoxybenzylcarbonyl, 9-fluorenylmethyloxycarbonyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, p-methoxyphenyl, or other benzoyl. Non-limiting examples of the carboxylic acid protecting group include group C₁-C₆alkyl (e.g., tert-butyl, methyl or ethyl), benzyl, or silyl, all of which protect the carboxylic acid moieties in the form of esters.

Diagram 2

<img file="PL2468285T3_D0028.tif" />

[0125] In step 1, compound (a) reacts with compound (b) to form compound (c), where the reaction can be carried out, as a non-limiting example, in the presence of sodium tert-butoxide or potassium tert-butoxide. Preferably, the reaction is carried out in the absence of lanthanum chloride. Also preferably, the yield of this reaction is at least 50%. More preferably, the reaction yield is at least 60%, 70%, or 80%. Highly preferably, the reaction yield is at least 90% or 95%. A preferred PG is group C₁-C₆alkoxycarbonyl, such as tert-butoxycarbonyl or Boc.

[0126] Compound (c) can then be reacted with compound (d), or a salt thereof, such as the TsOH salt, to form compound (e) (step 2), and then deprotect the amino group to form compound (f) or a salt thereof salts (e.g., HCl salts) (step 3). Favorable PG_C includes, but is not limited to, group C₁-C₆an alkyl group such as an ethyl group. Compound (f) can then be reacted with compound (g) to form compound (h) (step 4), which in turn is subjected to amino protection to form compound (i) (step 5) followed by ring closing metathesis reaction to give compound (j) (step 6). Favorable PG_N includes, but is not limited to, group C₁-C₆alkoxycarbonyl, such as tert-butoxycarbonyl or Boc. General processes for ring-closing metathesis (RCM) are known in the art. Preferred processes include the use of transition metal catalysts such as those described in US Patent No. 6,921,753 and in US Patent Application Publication No. 20070043180. Non-limiting examples of useful catalysts include Zhana 1B (

<img file="PL2468285T3_D0029.tif" />

where Mes is 2,4,6-trimethylphenyl; also known as Zhan-B) and Zhan's 1C catalyst (

<img file="PL2468285T3_D0030.tif" />

where Cy is a cyclohexyl group), both of which are commercially available from Zannan Pharma, Ltd. (Shanghai, China). Deprotection of the amino moiety in compound (j) leads to compound (k) (or its free base) (step 7). In some cases, compound (h) may directly undergo ring-metathesis reaction to form compound (k) (or its free base), without the steps of protecting and deprotecting the amino group.

[0127] Then, the carboxylic acid moiety in compound (k) can be deprotected to give compound (1) (step 8), which reacts with compound (m) to give compound (n) (step 9). G in compound (m) is defined as -ER₅where E and R₅ have the meanings as defined above.

[0128] The compound of formula I 'as described herein can be similarly prepared according to Scheme 2.

[0129] The compounds described herein contain one or more asymmetric centers and thus give enantiomers, diastereomers, and other stereoisomeric forms that can be defined in the category of absolute stereochemistry as (R) - or (S) -, or as (D) - or (L) - for amino acids. The present invention and disclosure is intended to include all such possible isomers as well as their racemic and optically pure forms. Optical isomers can be prepared from their respective optically active precursors by the procedures described above, or by cleavage of the racemic mixture. The separation can be carried out in the presence of a resolving agent, by chromatography or by repeated crystallization or by some combination of techniques that are known to those skilled in the art. Further details regarding separation can be found in Jacques, et al., Enantiomers, Racemats, and Resolutions (John Wiley &amp; Sons, 1981). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless otherwise specified, the compounds are assumed to include both E and Z geometric isomers. Likewise, all tautomeric forms are also intended to be encompassed by the invention. The configuration of any carbon-carbon bond appearing herein is selected only for convenience, and is not intended to specify a particular configuration unless the text confirms it; and therefore the carbon-carbon double bond drawn here arbitrarily as trans can be cis, trans, or a mixture of the two in any proportion.

[0130] The synthesized compounds can be separated from the reaction mixture and further purified by a method such as column chromatography, high pressure liquid chromatography, or recrystallization. As a skilled specialist will appreciate, further methods for synthesizing compounds of the formulas set out herein will be apparent to those skilled in the art. Additionally, the various synthetic steps can be carried out in a different order or order to form the desired compounds. In addition, the solvents, temperatures, reaction times, etc. described herein are for illustration purposes only and one of ordinary skill in the art will recognize that changing reaction conditions may give the desired bridged macrocyclic products described herein. Synthetic chemistry transformations and protecting group methodologies (protection and deprotection) useful in synthesizing the compounds described herein are known in the art and include, for example, as described in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); TW Greene and PGM Wuts, Protective Groups in Organic Synthesis, ed. 2, John Wiley and Sons (1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995), and their subsequent editions.

[0131] The compounds described herein can be modified by attaching various functions to any of the synthetic agents described herein to enhance selective biological properties. Such modifications are known in the art and include those that increase biological penetration into a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral availability, increase solubility to allow injection, alter metabolism, and they change the rate of excretion.

[0132] The quotation of a list of chemical groups in any definition of a parameter used herein includes the definition of that parameter as any single group or combination of said groups. Citing an embodiment for a parameter used herein includes this embodiment as any single embodiment or in combination with any other embodiments or portions thereof.

Examples [0133] The compounds and methods described herein will become better understood in connection with the following examples, which are intended for illustration purposes only. The following examples can be prepared according to either Scheme 1 or Scheme 2 as described above. Various changes and modifications of the disclosed embodiments will be apparent to those skilled in the art and such changes and modifications include, without limitation, those that relate to chemical structures, substituents, derivatives, formulations and / or methods described herein.

[0134] In the following description, only Examples 24-30, 32-41, 49-51, 53 and 56 form part of the invention. Examples other than those mentioned above are not part of the invention and are given for comparative purposes.

Example 1. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -14a (cyclopropylsulfonylcarbamoyl) -5,16-dioxo -1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyn- Tert-butyl 6-ylcarbamate

Example 1a. (2S, 6S, 13aS, 14aR, 16AS, Z) -2- (4-bromofenylosulfonyloksy) -6- (tert-butoxycarbonylamino) -5.16-dioxo-1,2,3,5,6,7,8, 9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] ethyl diazacyclopentadecine-14a-carboxylate [0135] Solution (2S, 6S, 13aS, 14aR, 16AS, Z) -6- (tert-butoxycarbonylamino) -2-hydroxy-5,16diokso-1,2,3,5,6,7,8,9,10,11,13a, 14.14 , 15,16,16a-Hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxylate and DABCO in toluene were stirred at peace (rt). To this solution, a solution of 4-bromobenzene-1-sulfonyl chloride in toluene was added. After the addition, the reaction was quenched with 10% aqueous sodium carbonate solution and the mixture was stirred for 15 min. Tetrahydrofuran was added and the mixture was washed with 0.5 M HCl, water, followed by a saturated aqueous sodium chloride solution. The organic layer was dried over anhydrous magnesium sulfate, filtered, and evaporated under reduced pressure and dried to give the title compound.

Example 1b.

[0136] (2R, 6S, 13aR, 14aR, 16aS, Z) -6- (tert-butoxycarbonylamino) -2- (3-chloroquinoxalin-2-yloxy) -5,16-dioxo-1,2,3,5,6 , 7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] 1,4] diazacyclopentadecine-14a-carboxylate (1b) [0137 ] To a solution of compound 1a (15.0 g, 21.0 mmol) in NMP (55 ml) was added 3-chloroquinoxalin-2ol (4.56 g, 25.3 mmol) followed by Cs₂WHAT₃ (17.1 g, 52.6 mmol). The resulting mixture was heated to 70 ° C for 18 hours. The reaction mixture was cooled to room temperature and then partitioned between ethyl acetate (300 mL) and 1N HCl (100 mL). Organic layer

The solids were separated, washed with brine (100 ml), dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to obtain a crude product as a solid. The solid was purified by silica gel column chromatography (EtOAc-hexane gradient) to give the title compound as a solid (6.2 g, 45% yield); MS (ESI): m / z = 656.3 [M + H]

Example 1c.

[0138] (2R, 6S, 13aR, 14aR, 16aS, Z) -6- (tert-butoxycarbonylamino) -5,16-dioxo-2- (3-phenylquinoxalin-2-yloxy) -1,2,3,5 , 6,7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxylate (1c ) [0139] The product of Example 1b (700 mg, 1.07 mmol), 2 (tributylstannyl) benzo [d] thiazole (905 mg, 2.13 mmol), tetrakis (triphenylphosphine) palladium (113 mg, added to the microwave vessel 0.11 mmol) and dioxane (5 ml). The vessel was pumped out and nitrogen was introduced, which was repeated twice. The mixture was reacted in a microwave reactor at 110 ° C for 1 h. The reaction was diluted with MeCN and washed 3 times with hexane. The MeCN layer was evaporated and purified by silica gel column chromatography (CHCl gradient)₃EtOAc) to give the title compound (692 mg, 86% yield). MS (ESI): m / z = 755.2 [M + H].

Example 1d.

[0140] (2R, 6S, 13aR, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -6- (tert-butoxycarbonylamino) -5,16 dioxo-1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1, 4] diazacyclopentadecine-14a-carboxylic acid (1d) [0141] To a solution of the product of Example 1c (692 mg, 0.95 mmol) in a tetrahydrofuran (5 ml) / ethanol (2.5 ml) / water (2.5 ml) mixture ) lithium hydroxide monohydrate (154 mg, 3.7 mmol) was added. The resulting mixture was heated to 50 ° C for one hour and cooled to room temperature. Organic solvents were mainly removed under reduced pressure, EtOAc (100 mL) was added, followed by washing with 1N HCl (30 mL). The organic layer was separated, washed with brine (20 mL), dried over anhydrous MgSO₄, and concentrated under reduced pressure to obtain 666 mg of product 1d.

Example 1f.

[0142] To a solution of the product of Example 1d (666 mg, 0.92 mmol) in 1,2-dichloroethane (9 mL) was added 1,1'-carbonyldiimidazole (246 mg, 1.52 mmol). The reaction mixture was stirred at 40 ° C for 2 hours. Then cyclopropanesulfonamide (184 mg, 1.52 mmol) was added to the above solution, followed by DBU (0.23 mL, 1.52 mmol). The resulting mixture was stirred at 40 ° C for 1 hour. The reaction mixture was diluted with EtOAc (100 mL) and washed with 1N HCl (20 mL) followed by saturated sodium chloride (20 mL). The organic layer was separated, dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (gradient CHCl₃/ EtOAc) to give the title compound (322 mg, 38% yield). MS (ESI): m / z = 830.0 [M + H].

EP 2 468 285 B1

Example 2. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -6- (isonicotinamide) diokso1,2,3,5,6,7,8,9,10,11,13a--5.16, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [ 1,4] diazacyklopentadecyno-14a-carboxamide

Example 2a.

[0143] (2R, 6S, 13aS, 14aR, 16aS, Z) -6-amino-2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -5 16-dioxo-1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [ 1,4] diazacyclopentadecine-14a-carboxamide [0144] To a suspension of the product of Example 1 (320 mg, 0.39 mmol) in EtOAc (3 mL) was added a 4 M solution of HCl in dioxane (1.9 mL, 7.7 mmol ). The reaction mixture was stirred at room temperature for 20 h. The solvent was evaporated under reduced pressure and the obtained solid was dried under reduced pressure to obtain a hydrochloric acid salt of (2R, 6S, 13aR, 14aR, 16aS, Z) -6-amino-2- (3- (benzo [d] thiazol-2 yl) quinoxalin-2-yloxy) -N- (cyclopropylsulfonyl) -5.16-dioxo-1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15, 16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxamide (295 mg, quantitative yield).

Example 2b.

[0145] To a solution of the compound of Example 2a (28 mg, 0.037 mmol) in dichloromethane (0.5 mL) wasonicotinic acid (5.0 mg, 0.040 mmol), HATU (16.7 mg, 0.044 mmol) and diisopropylethylamine ( 0.021 mL, 0.12 mmol). The reaction mixture was stirred at 25 ° C for 2 h and evaporated. Purification of the crude material by reverse phase chromatography eluting with an acetonitrile / water / TFA mixture gave the title compound. MS (ESI): m / z = 835.0 [M + H].

Example 3. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -6- (2- fluorobenzamido) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a ] [1,4] diazacyclopentadecine-14a-carboxamide [0146] Example 3 was prepared according to the procedure used to prepare the compound of Example 2, replacing isonicotinic acid with 2-fluorobenzoic acid. Purification of the crude material by reverse phase chromatography eluting with an acetonitrile / water / TFA mixture gave the title compound. MS (ESI): m / z = 851.9 [M + H].

Example 4. N - ((2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -14a (cyclopropylsulfonylcarbamoyl) -5, 16-dioxo-1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4 ] diazacyclopentadecin-6-yl) -5-methylisoxazole-3-carboxamide [0147] Example 4 was prepared according to the procedure used to prepare the compound of Example 2, replacing isonicotinic acid with 5-methylisoxazole-3-carboxylic acid. Purification of the crude material by reverse phase chromatography eluting with an acetonitrile / water / TFA mixture gave the title compound. MS (ESI): m / z = 838.9 [M + H].

EP 2 468 285 B1

Example 5. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -6- (5- methylpyrazine-2-carboxamido) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1 , 2-a] [1,4] diazacyclopentadecine-14a-carboxamide [0148] Example 5 was prepared according to the procedure used to prepare the compound of Example 2, replacing isonicotinic acid with 5-methylpyrazine-2-carboxylic acid. Purification of the crude material by reverse phase chromatography eluting with an acetonitrile / water / TFA mixture gave the title compound. MS (ESI): m / z = 849.9 [M + H].

Example 6. N - ((2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -14a (cyclopropylsulfonylcarbamoyl) -5, 16-dioxo-1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4 ] diazacyclopentadecin-6-yl) isoxazole-5-carboxamide [0149] Example 6 was prepared following the procedure used to prepare the compound of Example 2, replacing isonicotinic acid with isoxazole-5-carboxylic acid. Purification of the crude material by reverse phase chromatography eluting with an acetonitrile / water / TFA mixture gave the title compound. MS (ESI): m / z = 824.9 [M + H].

Example 7. N - ((2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -14a (cyclopropylsulfonylcarbamoyl) -5, 16-dioxo-1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4 ] diazacyclopentadecin-6-yl) thiazole-4-carboxamide [0150] Example 7 was prepared according to the procedure used to prepare the compound of Example 2, replacing isonicotinic acid with thiazole-4-carboxylic acid. Purification of the crude material by reverse phase chromatography eluting with an acetonitrile / water / TFA mixture gave the title compound. MS (ESI): m / z = 840.9 [M + H].

Example 8. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -6- (1- methyl-1H-pyrazol-3-carboxamido) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e ] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxamide [0151] Example 8 was prepared according to the procedure used to prepare the compound of Example 2, replacing isonicotinic acid with 1-methyl-1H-pyrazole-3- carboxylic. Purification of the crude material by reverse phase chromatography eluting with an acetonitrile / water / TFA mixture gave the title compound. MS (ESI): m / z = 838.0 [M + H].

Example 9. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -5,16-dioxo -6- (pyrimidin-4-carboxamido) 1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1, 2-a] [1,4] diazacyclopentadecine-14a-carboxamide [0152] Example 9 was prepared according to the procedure used to prepare the compound of Example 2, replacing isonicotinic acid with pyrimidine-4-carboxylic acid.

[0153] Purification of the crude material by reverse phase chromatography eluting with an acetonitrile / water / TFA mixture gave the title compound. MS (ESI): m / z = 835.9 [M + H].

Example 10. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -6- (1, 3-dimethyl-1 H-pyrazole-4-carboxamido) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxamide [0154] Example 10 was prepared according to the procedure used to prepare the compound of Example 2, replacing isonicotinic acid with 1,3-dimethyl-1H- pyrazole-4-carboxylic acid. Purification of the crude material by reverse phase chromatography eluting with an acetonitrile / water / TFA mixture gave the title compound. MS (ESI): m / z = 852.0 [M + H].

Example 11. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -6- (3- fluorobenzamido) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a ] [1,4] diazacyclopentadecine-14a-carboxamide [0155] Example 11 was prepared according to the procedure used to prepare the compound of Example 2, replacing isonicotinic acid with 3-fluorobenzoyl chloride. Purification of the crude material by reverse phase chromatography eluting with an acetonitrile / water / TFA mixture gave the title compound. MS (ESI): m / z = 838.0 [M + H].

Example 12. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzofuran-2-yl) quinoxalin-2-yloxy) -14a (cyclopropylsulfonylcarbamoyl) -5,16-dioxo-1,2 , 3,5,6,7,8,9,10,11,13a, 14,14a, 15,16,16hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate [0156] Example 12a. (2R, 6S, 13aS, 14aR, 16AS, Z) -14a- (cyclopropylsulfonylcarbamoyl) -2-hydroxy-5,16-dioxo-1,2,3,5,6,7,8,9,10,11,13a. 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate.

[0157] To the solution (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (9H-fluoren-9-ylideneaminoxy) -14- (cyclopropylsulfonylcarbamoyl) -5,16-dioxo-1,2,3,5 , 6,7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadechydrocyclopropa [e] pyrrolo [1,2-a] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate (20.0 g,

26.8 mmol) in acetic acid (80 ml), zinc dust (10.52 g, 166 mmol) was added while stirring at 40 ° C. After the addition, the reaction mixture was stirred at 40 ° C for 1 hour. The mixture was then cooled to room temperature, diluted with toluene, and filtered through celite. The mother liquor was washed with water, 1 N HCl, and a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and filtered. Then, the filtrate was evaporated under reduced pressure to obtain the title compound 12a (14.8 g, 97% yield).

[0158] Example 12b. (2R, 6S, 13aS, 14aR, 16AS, Z) -2- (3-chloroquinoxaline-2-yloxy) -14a- (cyclopropylsulfonylcarbamoyl) -5.16-dioxo-1,2,3,5,6,7, 8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate.

[0159] A solution of compound 12a (10.0 g, 17.6 mmol), cesium carbonate (17.2, 52.8 mmol), and 2,3-dichloroquinoxaline (3.50 g, 17, 6 mmol) in dimethylformamide (175 ml) was heated to 70 ° C for 18 hours. An additional portion of 2,3-dichloroquinoxaline (0.70 g, 3.5 mmol) was added and the reaction mixture was stirred at 70 ° C for 18 hours. The reaction mixture was cooled to room temperature and then partitioned between ethyl acetate (300 mL) and 1N HCl (100 mL). The organic layer was separated, washed with brine (100 mL), dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to obtain a crude product as a solid. The solid was purified by silica gel column chromatography (EtOAc-hexane gradient) to give the title compound as a solid (4.7 g, 37% yield); MS (ESI): m / z = 731.1 [M + H].

Example 12c.

[0160] The product of Example 12b (0.40 g 0.547 mmol), benzofuran-2-yltributylstannane (0.245 g, 0.602 mmol), tris (dibenzylideneacetone) dipalladium (0) (50 mg, 0.055 mmol) was added to the microwave dish. , 3,5,7-tetramethyl-2,4,8-trioxa-6-phenyl-6-phosphate-adamantane (32 mg, 0.11 mmol), sodium bicarbonate (46 mg, 0.547 mmol) and dioxane (3 mL ). The vessel was pumped out and nitrogen was introduced. The mixture was reacted in a microwave reactor at 110 ° C for 1

h. The reaction mixture was diluted with ethyl acetate, washed with 1N HCl, followed by a saturated aqueous sodium chloride solution, and dried over anhydrous magnesium sulfate. The mixture was filtered through celite and evaporated under reduced pressure. The residue was dissolved in acetonitrile and washed with hexane (five times), followed by evaporation under reduced pressure. The residue was purified by silica gel column chromatography (gradient CHCl₃-EtOAc) to give the title compound (386 mg, 87% yield). MS (ESI): m / z = 813.0 [M + H].

Example 13. N - ((2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzofuran-2-yl) quinoxalin-2-yloxy) -14a (cyclopropylsulfonylcarbamoyl) -5,16-dioxo 1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyn-6 yl) -5-methyl-isoxazole-3-carboxamide.

Example 13a.

[0161] (2R, 6S, 13aS, 14aR, 16aS, Z) -6-amino-2- (3- (benzofuran-2-yl) quinoxalin-2-yloxy) -N- (cyclopropylsulfonyl) -5,16-dioxo hydrochloride -1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyno- 14α-carboxamide [0162] The product of Example 12 (0.386 g, 0.475 mmol) was dissolved in a mixture of ethyl acetate (2.4 mL) and 4 N HCl in dioxane (2.4 mL) and stirred at room temperature for one hour. Then the mixture was evaporated under reduced pressure to give the title compound (0.338 mg, 100% yield).

Example 13b.

[0163] A mixture of the product of Example 13a (30 mg, 0.040 mmol), 5-methylisoxazole-3-carboxylic acid (5.1 mg, 0.040 mmol), N-ethyl-N-isopropylpropan-2-amine (15.6 mg, 0 , 12 mmol), and HATU (18.3 mg, 0.048 mmol) in dichloromethane (0.5 mL) was stirred at room temperature for one hour and then evaporated. Raw material purification by road

Reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound (14 mg, 42% yield). MS (ESI): mlz = 822.0 [M + H].

Example 14. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzofuran-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -6- (5-methylpyrazine-2- carboxamido) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a ] [1,4] diazacyclopentadecine-14a-carboxamide [0164] Example 14 was prepared according to the procedure used to prepare the compound of Example 13, replacing 5-methylisoxazole-3-carboxylic acid with 5-methylpyrazine-2-carboxylic acid. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound. MS (ESI): m / z = 833.0 [M + H].

Example 15. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzofuran-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -6- (1-methyl-1H- pyrazole-3-carboxamido) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1 , 2-a] [1,4] diazacyclopentadecine-14a-carboxamide [0165] Example 15 was prepared according to the procedure used to prepare the compound of Example 13, replacing 5-methylisoxazole-3-carboxylic acid with 1-methyl-1H-pyrazole3- carboxylic. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound. MS (ESI): m / z = 821.1 [M + H].

Example 16. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzofuran-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -6- (1,5-dimethyl- 1 H-pyrazole-3-carboxamido) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxamide [0166] Example 16 was prepared according to the procedure used to prepare the compound of Example 13, replacing 5-methylisoxazole-3-carboxylic acid with 1,5-dimethyl- 1H-pyrazole-3-carboxylic acid. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound. MS (ESI): m / z = 835.0 [M + H].

Example 17. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzofuran-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -5,16-dioxo-6- ( pyrimidin-4-carboxamido) 1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxamide [0167] Example 17 was prepared according to the procedure used to prepare the compound of Example 13, replacing 5-methylisoxazole-3-carboxylic acid with pyrimidine-4-carboxylic acid. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound. MS (ESI): m / z = 818.9 [M + H].

Example 18. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [b] thiophen-2-yl) quinoxalin-2-yloxy) -14a (cyclopropylsulfonylcarbamoyl) -5,16-dioxo -1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a42

Hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate [0168] Example 18 was prepared according to the procedure used to prepare the compound of Example 12, replacing benzofuran-2- yllotributylstannane benzo [b] thiophen-2-yllotributylstannane. Purification of the crude material by chromatography on silica gel, eluting with hexane / ethyl acetate (1: 2), gave the title compound. MS (ESI): m / z = 829.1 [M + H].

Example 19. N - ((2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [b] thiophen-2-yl) quinoxalin-2-yloxy) 14- (cyclopropylsulfonylcarbamoyl) -5, 16 diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4 ] diazacyklopentadecyn-6-yl) -5-methyl-isoxazole-3-carboxamide

Example 19a.

[0169] (2R, 6S, 13aS, 14aR, 16aS, Z) -6-amino-2- (3- (benzo [b] thiophen-2-yl) quinoxalin-2-yloxy) -N- (cyclopropylsulfonyl) -5 hydrochloride 16-dioxo-1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1, 4] diazacyclopentadecine-14α-carboxamide [0170] The product of Example 18 (0.296 g, 0.357 mmol) was dissolved in a mixture of ethyl acetate (1.7 mL) and 4 N HCl in dioxane (1.7 mL) and stirred at room temperature for for one hour. Then the mixture was evaporated under reduced pressure to give the title compound (0.262 mg, 96% yield).

Example 19b.

[0171] A mixture of the product of Example 19a (30 mg, 0.039 mmol), 5-methylisoxazole-3-carboxylic acid (5.0 mg, 0.039 mmol), N-ethyl-N-isopropylpropan-2-amine (15.2 mg, 0.118 mmol), and HATU (17.9 mg, 0.047 mmol) in dichloromethane (0.5 mL) was stirred at room temperature for one hour and then evaporated. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound (18 mg, 53% yield). MS (ESI): m / z = 837.9 [M + H].

Example 20. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [b] thiophen-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -6- (5- methylpyrazine-2-carboxamido) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1 , 2-a] [1,4] diazacyclopentadecine-14a-carboxamide [0172] Example 20 was prepared according to the procedure used to prepare the compound of Example 19, replacing 5-methylisoxazole-3-carboxylic acid with 5-methylpyrazine-2-carboxylic acid. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound. MS (ESI): m / z = 848.7 [M + H].

Example 21. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [b] thiophen-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -6- (1- methyl-1H-pyrazol-3-carboxamido) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e ] pyrrolo [1,2-a] [1,4] diazacyklopentadecyno-14a-carboxamide

[0173] Example 21 was prepared according to the procedure used to prepare the compound of Example 19, replacing 5-methylisoxazole-3-carboxylic acid with 1-methyl-1H-pyrazole-3-carboxylic acid. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound. MS (ESI): m / z = 836.9 [M + H].

Example 22. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [b] thiophen-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -6- (1, 5-dimethyl-1 H-pyrazole-3-carboxamido) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxamide [0174] Example 22 was prepared according to the procedure used to prepare the compound of Example 19, replacing 5-methylisoxazole-3-carboxylic acid with acid 1, 5-dimethyl-1H-pyrazole-3-carboxylic acid. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound. MS (ESI): m / z = 851.0 [M + H].

Example 23. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [b] thiophen-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -5,16-dioxo -6- (pyrimidin-4-carboxamido) 1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1, 2-a] [1,4] diazacyclopentadecine-14α-carboxamide [0175] Example 23 was prepared according to the procedure used to prepare the compound of Example 19, replacing 5-methylisoxazole-3-carboxylic acid with pyrimidine-4-carboxylic acid. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound. MS (ESI): m / z = 835.1 [M + H].

Example 24. (2R, 6S, 13aS, 14aR, 16aS, Z) -14- (cyclopropylsulfonylcarbamoyl) -5,16-dioxo2- (phenanthridin-6-yloxy) -1,2,3,5,6,7,8 , 9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate

Example 24a.

[0176] (2S, 6S, 13aS, 14aR, 16aS, Z) -2- (4-bromophenylsulfonyloxy) -6- (tert-butoxycarbonylamino) -5,16-dioxo-1,2,3,5,6,7 , 8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxylate [0177] Solution (2S , 6S, 13aS, 14aR, 16AS, Z) -6- (tert-butoxycarbonylamino) -2-hydroxy-5,16diokso-1,2,3,5,6,7,8,9,10,11,13a. 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14α-carboxylate (22.1 g, 44.8 mmol) and DABCO (8.5 g. 76.7 mmol) in toluene (88 mL) was stirred at room temperature. To this solution, a solution of 4-bromobenzene-1-sulfonyl chloride 17.2 g, 67.2 mmol) in toluene (44 mL) was added. After the addition, the reaction was quenched with 10% aqueous sodium carbonate solution (110 mL) and the mixture was stirred for 15 min. Tetrahydrofuran (44 mL) was added and the mixture was washed with 0.5 M HCl, water, followed by a saturated aqueous sodium chloride solution. The organic layer was dried over anhydrous magnesium sulfate, filtered, and evaporated under reduced pressure and dried to give the title compound (27.7 g, 87% yield), which was used without further purification.

EP 2 468 285 B1

Example 24b (2R, 6S, 13aS, 14aR, 16aS, Z) -6- (tert-butoxycarbonylamino) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3,5,6 , 7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2a] [1,4] diazacyclopentadecine-14a-carboxylate [0179] To a solution of the compound from Example 24a (11.0 g, 15.4 mmol) in NMP (100 mL) added phenanthridin-6 (5H) -one (3.15 g, 16.2 mmol) followed by CS₂WHAT₃ (7.53 g, 23.1 mmol). The resulting mixture was heated to 55 ° C for four hours. The reaction mixture was cooled to room temperature and then partitioned between ethyl acetate (250 ml) and 5% aqueous sodium bicarbonate solution (200 ml). The organic layer was separated, washed with 5% aqueous sodium bicarbonate solution (200 mL) followed by brine (150 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product as a solid. Then the solid was dissolved in methyl tert-butyl ether (200 mL), the resulting suspension was stirred at room temperature for 1 hour and filtered. The filtrate containing the desired product was concentrated under reduced pressure to obtain 7.95 g of product 24b as a solid; MS-DCI / NH₃: 671 (M + H)⁺.

Example 24c (2R, 6S, 13aS, 14aR, 16aS, Z) -6- (tert-butoxycarbonylamino) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3, acid 5,6,7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxyl (24c ) [0181] To a solution of the product of Example 24b (7.8 g, 11.6 mmol) in tetrahydrofuran (40 mL) / ethanol (40 mL) was added an aqueous solution of lithium hydroxide (0.84 g of lithium hydroxide in 40 mL of H)₂ABOUT). The resulting mixture was heated to 50 ° C for two hours and cooled to room temperature. Organic solvents were mainly removed under reduced pressure, and the obtained residue was acidified with a 10% aqueous citric acid solution and extracted with ethyl acetate (200 mL). The organic layer was separated, washed with brine (200 ml), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a light yellow solid which was further dried in a vacuum oven at 45 ° C for 18 h to give 7.5 g of product 24c as a light yellow solid; MS-DCI / NH₃: 643 (M + H)⁺.

Example 24d.

[0182] (2R, 6S, 13aS, 14aR, 16aS, Z) -14- (cyclopropylsulfonylcarbamoyl) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3,5,6,7 , 8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate [0183] To A solution of the product of Example 24c (7.46 g, 11.6 mmol) in DMF (80 mL) was added with 1,1'-carbomyldiimidazole (5.64 g, 34.8 mmol). The reaction mixture was stirred at room temperature for 6 hours. Then cyclopropanesulfonamide (4.21 g, 34.8 mmol) was added to the above solution, followed by DBU (5.73 mL, 36.0 mmol). The resulting mixture was stirred at room temperature for 14 hours. EtOAc (200 mL), 10% aqueous citric acid (200 mL) and saturated aqueous sodium chloride (50 mL) were added to the reaction mixture. The organic layer was separated, washed with a saturated aqueous sodium chloride solution (100 ml), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure.

EP 2 468 285 B1

The residue was purified by silica gel column chromatography (ethyl acetate / heptane gradient) to give the title compound as a white solid (6.40 g, 74% yield). MS (ESI): m / z = 746.1 [M + H].

Example 25. (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -5,16-dioxo-2- (phenanthridin-6-yloxy) -6- (pyrazine-2-carboxamido) -1 , 2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyno-14A carboxamide

Example 25a [0184] (2R, 6S, 13aS, 14aR, 16aS, Z) -6-amino-N- (cyclopropylsulfonyl) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3 hydrochloride 5,6,7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-4a-carboxamide [0185 ] To a suspension of the product of Example 24 (0.35 g, 0.47 mmol) in acetonitrile (5 mL) was added a 4 M solution of HCl in dioxane (0.6 mL, 2.4 mmol). The reaction mixture was stirred at room temperature for 4 h. The solvent was evaporated under reduced pressure and the obtained solid was dried under reduced pressure to obtain the title compound (0.32 g, quantitative yield).

Example 25b.

[0186] (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -5,16-dioxo-2- (phenanthridin-6-yloxy) -6- (pyrazine-2-carboxamido) -1.2 , 3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyno-14A carboxamide [0187] To a solution of the compound of Example 25a (320 mg, 0.47 mmol) in dimethylformamide (5 mL) was added 2-pyrazinecarboxylic acid (0.065 g, 0.52 mmol), HATU (214 mg, 0.56 mmol) and diisopropylethylamine (0.2 mL, 1.18 mmol). The reaction mixture was stirred at 25 ° C for 2 ha then partitioned between 5% aqueous sodium bicarbonate solution and ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate, filtered, and evaporated. The residue was purified by crystallization from ethyl acetate / hexane to give the desired product (155 mg, 44% yield) as an off-white solid. MS (ESI): m / z = 752.0 [M + H].

Example 26. (2R, 6S, 13aS, 14aR, 16aS, Z) -14- (cyclopropylsulfonylcarbamoyl) -5,16-dioxo2- (phenanthridin-6-yloxy) -1,2,3,5,6,7,8 , 9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecin-cyclopentyl 6-ylcarbamate

Example 26a (2R, 6S, 13aS, 14aR, 16aS, Z) -6- (cyclopentyloxycarbonylamino) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3,5,6 , 7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2a] [1,4] diazacyclopentadecine-14a-carboxylate [0189] A compound of Example 26a was prepared according to the procedure used to prepare the compound of Example 24b, replacing the compound of Example 24a with (2S, 6S, 13aS, 14aR, 16aS, Z) -2- (4-bromophenylsulfonyloxy) -6- (cyclopentyloxycarbonylamino) -5,16-dioxo1,2,3,5,6,7, 8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxylate to obtain the title compound. MS (DCI / NH₃): m / z =

683.0 [M + H]

EP 2 468 285 B1

Example 26b (2R, 6S, 13a, 14aR, 16aS, Z) -6- (cyclopentyloxycarbonylamino) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3,5, acid 6,7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxylic [0191] Compound from Example 26b was prepared according to the procedure used to prepare the compound of Example 24c, replacing compound 24b with compound 26a to give the title compound.

MS (DCI / NH₃): m / z = 655.0 [M + H].

Example 26c.

[0192] (2R, 6S, 13aS, 14aR, 16aS, Z) -14- (cyclopropylsulfonylcarbamoyl) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3,4,6,7 , 8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] cyclopentyl diazacyclopentadecin-6-ylcarbamate [0193] A compound of the Example 26 was prepared according to the procedure used to prepare the compound of Example 24, replacing compound 24c with compound 26b to give the title compound.

MS (DCI / NH₃): mlz = 758.0 [M + H].

Example 27. (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -6- (5-methyl-1H-pyrazole-3-carboxamido) -5,16-dioxo-2- (phenanthridine-6- yloxy) 1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4 ] diazacyclopentadecine-14a-carboxamide [0194] The compound of Example 27 was prepared according to the procedure used to prepare the compound of Example 25, replacing 2-pyrazinecarboxylic acid with 5-methyl-1H-pyrazole-3-carboxylic acid. Purification of the crude material by reverse phase chromatography eluting with an acetonitrile / water / TFA mixture gave the title compound. MS (ESI): m / z = 754.2 [M + H].

Example 28. N - ((2R, 6S, 13aS, 14aR, 16aS, Z) -14- (cyclopropylsulfonylcarbamoyl) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3,5,6, 7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyn-6-yl) -5-metyloizoksazolo- 3-carboxamide [0195] The compound of Example 28 was prepared according to the procedure used to prepare the compound of Example 25, replacing 2-pyrazinecarboxylic acid with 5-methylisoxazole-3-carboxylic acid. Purification of the crude material by reverse phase chromatography eluting with an acetonitrile / water / TFA mixture gave the title compound. MS (ESI): m / z = 755.1 [M + H].

Example 29. (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -6- (5-methylpyrazine-2-carboxamido) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyno- 14a-carboxamide [0196] The compound of Example 29 was prepared according to the procedure used to prepare the compound of Example 25, replacing 2-pyrazinecarboxylic acid with 5-methylpyrazine-2-carboxylic acid. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound. MS (ESI): m / z = 766.1 [M + H].

EP 2 468 285 B1

Example 30. N - ((2R, 6S, 13aS, 14aR, 16aS, Z) -14- (cyclopropylsulfonylcarbamoyl) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3,5,6, 7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyn-6-yl) thiazole-5-carboxamide [0197] The compound of Example 30 was prepared according to the procedure used to prepare the compound of Example 25, replacing 2-pyrazinecarboxylic acid with thiazole-5-carboxylic acid. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound. MS (ESI): m / z = 757.1 [M + H].

Example 31. (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -6- (2-fluorobenzamido) 5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2, 3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyno-14a-carboxamide [0198] The compound of Example 31 was prepared according to the procedure used to prepare the compound of Example 25, replacing 2-pyrazinecarboxylic acid with 2-fluorobenzoic acid. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound. MS (ESI): m / z = 768.1 [M + H].

Example 32. (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -5,16-dioxo-2- (phenanthridin-6-yloxy) -6- (pyridazine-4-carboxamido) -1 , 2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyno-14A carboxamide [0199] The compound of Example 32 was prepared according to the procedure used to prepare the compound of Example 25, replacing 2-pyrazinecarboxylic acid with pyridazine-4-carboxylic acid. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound. MS (ESI): mlz = 752.1 [M + H].

Example 33. (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -5,16-dioxo-2- (phenanthridin-6-yloxy) -6- (pyrimidine-4-carboxamido) -1 , 2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyno-14A carboxamide [0200] The compound of Example 33 was prepared following the procedure used to prepare the compound of Example 25, replacing 2-pyrazinecarboxylic acid with pyrimidine-4-carboxylic acid. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound. MS (ESI): m / z = 752.1 [M + H].

Example 34. (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -6- (1-methyl-1H-pyrazole-3-carboxamido) -5,16-dioxo-2- (phenanthridine-6- yloxy) 1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4 ] diazacyclopentadecine-14a-carboxamide [0201] The compound of Example 34 was prepared according to the procedure used to prepare the compound of Example 25, replacing 2-pyrazinecarboxylic acid with 1-methyl-1H-pyrazole-3-carboxylic acid. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound. MS (ESI): m / z = 754.2 [M + H].

EP 2 468 285 B1

Example 35. (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -6- (2-hydroxy-2-methylpropaneamido) -5,16-dioxo-2- (phenanthridin-6-yloxy) 1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyno -14a-carboxamide [0202] The title compound of Example 35 was prepared according to the procedure used to prepare the compound of Example 25, replacing 2-pyrazinecarboxylic acid with 2-hydroxy-2-methylpropanoic acid. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound. MS (ESI):

mlz = 732.2 [M + H].

Example 36. (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -6- (1,5-dimethyl-1H-pyrazole-3-carboxamido) -5,16-dioxo-2- ( phenanthridin-6-yloxy) 1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxamide [0203] The title compound of Example 36 was prepared according to the procedure used to prepare the compound of Example 25, replacing 2-pyrazinecarboxylic acid with 1,5-dimethyl-1H-pyrazole-3-carboxylic acid. Purification of the crude material by reverse phase chromatography, eluting with an acetonitrile / water / TFA mixture, gave the title compound.

MS (ESI): m / z = 768.1 [M + H].

Example 37. (2R, 6S, 13aS, 14aR, 16aS, Z) -14- (cyclopropylsulfonylcarbamoyl) -2- (2-fluorophenanthridin-6-yloxy) -5,16-dioxo-1,2,3,5,6 , 7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecin-cyclopentyl 6-ylcarbamate. Example 37a.

[0204] 5'-fluoro-2'-nitrobiphenyl-2-carboxylate [0205] 2- (methoxycarbonyl) phenylboronic acid (63.4 mg, 0.352 mmol), 2-bromo-4-fluoro-1- nitrobenzene (77 mg, 0.35 mmol), diacetoxypalladium (0.93 mg, 4.1 μmol) and dicyclohexyl (2 ', 6'-dimethoxybiphenyl-2-yl) phosphine (3.47 mg, 8.45 μmol) . Ethanol (1760 μθ and sodium carbonate (176 pi, 0.352 mmol) were added and the mixture was reacted in a microwave reactor at 100 ° C for 30 min. The reaction mixture was diluted with dichloromethane, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative thin layer chromatography (eluent: 9: 1 hexane / ethyl acetate) to obtain methyl 5'-fluoro-2'-nitrobiphenyl-2-carboxylate (37a, 54.8 mg, 0.199 mmol, 56.6% yield) .

Example 37b.

[0206] To 2-fluoro-5-hydroxyphenanthridin-6 (5H) -one [0207] To the product solution of Example 37a (Methyl 5'-fluoro-2'-nitrobiphenyl-2-carboxylate, 56.79 mg, 0.206 mmol) in methanol (9 mL) 10% palladium on carbon (15.6 mg, 0.015 mmol) was added. A hydrogen balloon was attached to the flask and it was degassed with hydrogen three times. The reaction mixture was stirred, diluted with dimethylformamide, and filtered. The filtrate was concentrated to give 2-fluoro-5-hydroxyphenanthridin-6 (5H) -one (37b, 46.36 mg, 0.202 mmol, 98% yield).

EP 2 468 285 B1

Example 37c.

[0208] 2-Fluoro-phenanthridin-6 (5H) -one [0209] A mixture of the product of Example 37b (2-fluoro-5-hydroxyphenanthridin-6 (5H) -one, 46.4 mg, 0.202 mmol), acetic acid (3 mL), and zinc (99 mg, 1.517 mmol) was heated under reflux at 130 ° C for 1 h. The mixture was diluted with dimethylformamide and filtered, and the filtrate was concentrated to give a beige solid (100 mg). The solid was partitioned between dichloromethane / dimethylformamide (2/1, 50 mL) and sodium carbonate (10 mL). The organic layer was washed with water (2 x 10 mL) and concentrated to give 2-fluorophenanthridin-6 (5H) he (37c, 38.2 mg, 88% yield).

Example 37d.

[0210] (2R, 6S, 13aS, 14aR, 16aS, Z) -6- (cyclopentyloxycarbonylamino) -2- (2-fluorophenanthridin-6-yloxy) -5,16-dioxo-1,2,3,5,6,7 , 8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxylate [0211] The title compound Example 37d was prepared according to the procedure used to prepare the compound of Example 37b, substituting (2S, 6S, 13aS, 14aR, 16aS, Z) -2- (4-bromophenylsulfonyloxy) -6- (cyclopentyloxycarbonylamino) -5,16-dioxo1,2,3,5,6,7,8,9,10 , 11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxylate instead of 1a, and substituting 2-fluorophenanthridinid-6 (5H ) -on (80c) instead of phenanthridin-6 (5H) -one to give the title compound in 48% yield.

Example 37e.

[0212] (2R, 6S, 13aS, 14aR, 16aS, Z) -6- (cyclopentyloxycarbonylamino) -2- (2-fluorophenanthridin-6-yloxy) -5,16-dioxo-1,2,3,5 acid, 6,7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxylic [0213] Compound the title of Example 37e was prepared according to the procedure used to prepare the compound of Example 67c, substituting the product of Example 37d instead of the product of Example 24b.

Example 37f.

[0214] (2R, 6S, 13aS, 14aR, 16aS, Z) -14- (cyclopropylsulfonylcarbamoyl) -2- (2-fluorophenanthridinin-6-yloxy) -5,16-dioxo-1,2,3,5,6,7 , 8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] cyclopentyl diazacyclopentadecin-6-ylcarbamate [0215] Compound of Example 37 was prepared according to the procedure used to prepare the compound of Example 24, substituting the product from Example 37e for the product from Example 24c (14.6 mg, 78% yield).

MS (ESI): m / z = 776.1 [M + H].

Example 38. (2R, 6S, 13aS, 14aR, 16aS, Z) -14- (cyclopropylsulfonylcarbamoyl) -2- (2,9-difluorophenanthridin-6-yloxy) -5,16-dioxo-1,2,3,5 , 6,7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadechydrocyclopropa [e] pyrrolo [1,2-a] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate

EP 2 468 285 B1

Example 38a.

Methyl 5,5'-difluoro-2'-nitrobiphenyl-2-carboxylate [0217] Pd was added to 2-bromo-4-fluoro-1-nitrobenzene (185.16 mg, 0.842 mmol)₂cares₃ (23.12 mg, 0.025 mmol) and copper powder (271 mg, 4.26 mmol). Dimethyl sulfoxide (2.3 mL) and methyl 2-bromo-4-fluorobenzoate (0.122 mL, 0.842 mmol) were added and the mixture was stirred vigorously at 100 ° C for 2 h. The mixture was cooled to room temperature, diluted with ethyl acetate (20 mL), and filtered. The filtrate was washed with water and dried (anhydrous Na₂SO₄) and concentrated to give a yellow oil (279.8 mg). This oil was used without purification to prepare the compound of Example 38b.

Example 38b.

[0218] 2,9-Difluoro-5-hydroxyphenanthridin-6 (5H) -one [0219] To the product of Example 38a (279.8 mg) was added methanol (7.5 mL) and 10% palladium on carbon (76 mg , 0.071 mmol). A hydrogen balloon was attached to the flask and the mixture was degassed and back filled with hydrogen three times. The mixture was stirred under a hydrogen atmosphere for 16 h, diluted with dimethylformamide and filtered. The filtrate was concentrated to give a red solid. This material was triturated with dichloromethane / hexane (9/1) and filtered to give the title compound (Example 38b, 43.15 mg, 0.175 mmol, quantitative yield).

Example 38c.

[0220] 2,9-Difluorophenanthridin-6 (5H) -one [0221] The title compound of Example 38c was prepared according to the procedure used to prepare the compound of Example 37c, substituting the product of Example 38b instead of the product of Example 80b.

Example 38d.

[222, (2R, 6S, 13aS, 14aR, 16aS, Z) -6- (tert-butoxycarbonylamino) -2- (2,9-difluorophenanthridin-6-yloxy) -5,16-dioxo-1,2,3,5 , 6,7,8,9,10,11,13a, 14,14a, 5,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxylate [0223 ] The title compound of Example 38d was prepared following the procedure used to prepare the compound of Example 24c, substituting the product of Example 38c for phenanthridine 6 (5H) -one.

Example 38e.

[0224] (2R, 6S, 13aS, 14aR, 16aS, Z) -6- (tert-butoxycarbonylamino) -2- (2,9-difluorophenanthridin-6-yloxy) -5,16-dioxo-1,2 acid, 3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyno-14a-carboxylic acid [0225] The title compound of Example 38e was prepared according to the procedure used to prepare the compound of Example 24c, substituting the product of Example 38d instead of the product of Example 24b.

EP 2 468 285 B1

Example 38f.

[0226] (2R, 6S, 13aS, 14aR, 16aS, Z) -14- (cyclopropylsulfonylcarbamoyl) -2- (2,9-difluorophenanthridin-6-yloxy) -5,16-dioxo-1,2,3,5 , 6,7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadechydrocyclopropa [e] pyrrolo [1,2-a] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate [0227] The title compound of Example 38 was prepared according to the procedure used to prepare the compound of Example 24, substituting the product of Example 38e for the product of Example 24c. MS (ESI): m / z = 782.1 [M + H].

Example 39. (2R, 6S, 13aR, 14aR, 16aS) -14- (cyclopropylsulfonylcarbamoyl) -5,16-dioxo-2 (phenanthridin-6-yloxy) octadekahydrocyclopropa [e] pyrrolo [1,2-a] [1, 4] tert-butyl diazacyclopentadecin 6-ylcarbamate [0228] Ethanol degassed using argon (0.8 mL) was added to the product of Example 24 (79.1 mg, 0.106 mmol) and Crabtree catalyst (3.45 mg, 4.24 μmol ) (4 mol%) in a 4 mL pressure bottle. The vessel was purged three times with argon and then filled with hydrogen under pressure (50 psi, 345 kPa). The mixture was heated to 50 ° C under a hydrogen atmosphere and stirred for 4.5 h at 50 ° C.

[0229] The reaction mixture was concentrated and purified by reverse phase chromatography eluting with an acetonitrile (1% TFA) / water gradient to give the title compound as a white solid (70.41 mg, 0.094 mmol, 89% yield). MS (ESI): m / z = 748.2 [M + H].

Example 40. (2R, 6S, 13aR, 14aR, 16aS) -14- (cyclopropylsulfonylcarbamoyl) -5,16-dioxo-2 (phenanthridin-6-yloxy) octadekahydrocyclopropa [e] pyrrolo [1,2-a] [1, 4] cyclopentyl diazacyclopentadecin-6-ylcarbamate [0230] The title compound of Example 40 was prepared according to the procedure used to prepare the compound of Example 39, replacing the product of Example 24 with the product of Example 26. MS (ESI): m / z = 760.2 [M + H].

Example 41. (2R, 6S, 13aR, 14aR, 16aS) -5,16-dioxo-2- (phenanthridin-6-yloxy) -14- (thiophen-2-ylsulfonylcarbamoyl) octadechydrocyclopropa [e] pyrrolo [1,2- a] [1,4] tert-butyl diazacyclopentadecin-6-ylcarbamate [0231] The title compound of Example 41 was prepared according to the procedure used to prepare the compound of Example 24, replacing cyclopropanesulfonamide with thiophene-2-sulfonamide.

MS (ESI): m / z = 788.0 [M + H].

Example 42. (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -6- (2-methylpyrimidine-5-carboxamido) -5,16-dioxo-2- (3- (benzo [d] thiazole -2-yl) quinoxalin-2-yloxy) 1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1 , 2-a] [1,4] diazacyklopentadecyno-14a-carboxamide

Example 42a.

[0232] (S) -2- (2-methylpyrimidine-5-carboxamido) non-8-enoic acid.

[0233] Boc-2 (S) -amino-non-8-enoic acid dicyclohexylamine salt can be suspended in isopropyl acetate, washed several times with aqueous citric acid solution, and then once

EP 2 468 285 B1 with water. The washed, concentrated and then again diluted in isopropyl acetate can be reacted with HCl to give 2 (S) -amino-non-8-enoic acid HCl salt. 2-methylpyrimidine-5-carboxylic acid, N, N'-disuccinimidyl carbonate, and N, N-dimethylaminopyridine can be dissolved in N-methyl-2-pyrrolidone (NMP) and mixed. Subsequently, 2 (S) amino-non-8-enoic acid HCl salt is added, followed by triethylamine, and mixed to give the title compound of Example 42a, which can be crystallized by the addition of HCl followed by water.

Example 42b.

[0234] (1R, 2S) -1 - ((2S, 4R) -N- (tert-butoxycarbonyl) -1 - ((S) -2- (2-methylpyrimidine-5-carboxamido) non-8-enoyl) Ethyl -4- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) pyrrolidine-2-carboxamido) -2-vinylcyclopropanecarboxylate (2S, 4R) -N-Boc-4-Hydroxyproline can be subjected to reaction with 2-chloro-3- (thiophen-2-yl) quinoxaline in NMP, in the presence of sodium tert-butoxide, to give (2S, 4R) -1- (tert-butoxycarbonyl) -4- (3- (benzo [d ] thiazol-2-yl) quinoxalin-2-yloxy) -pyrrolidine-2-carboxylic acid.

Then tert-butyl methyl ether (MTBE) and water can be added. The aqueous layer is separated, washed, then HCl is added, followed by [0236] extraction with MTBE. The extracted product can be mixed with diisopropylethylamine (DIPEA) and HATU (CAS No. 148893-10-1) and then reacted with the tosylate salt (1R, 2S) ethyl-1-amino-2-vinylcyclopropanecarboxylate in dimethylformamide (DMF) and toluene . The reaction produces (2S, 4R) -2 - ((1R, 2S) -1- (ethoxycarbonyl) -2-vinylcyclopropylcarbamoyl) -4- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) pyrrolidine Tert-butyl carboxylate, which can be extracted with MTBE and washed with HCl, further extracted, washed, dried, and dissolved in 2-propanol.

[0237] HCl can be added to the solution in 2-propanol to give (1R, 2S) -1 - ((2S, 4R) -4- (3 (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) ethyl pyrrolidine-2-carboxamido) -2-vinylcyclopropane carboxylate, which can be crystallized by neutralization with NaOH.

[0238] (1R, 2S) -1 - ((2S, 4R) -4- (3- (Benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) pyrrolidine-2-carboxamido) -2-vinylcyclopropane carboxylate ethyl, the title compound of Example 42a, N-hydroxy-5-norbornene-2,3-dicarboximide, and N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride can be mixed and mixed in DMF for hours, then add N, N- dimethylethylenediamine. The reaction gives (1R, 2S) -1 - ((2S, 4R) -1 - ((S) -2- (2-methylpyrimidine-5-carboxamido) non-8-enoyl) -4 (3- (benzo [d ] thiazol-2-yl) quinoxalin-2-yloxy) pyrrolidine-2-carboxamido) -2-vinylcyclopropanecarboxylate, which can be dissolved in isopropyl acetate and extracted with an aqueous solution

H₃AFTER₄and then extract with aqueous K solution₂HPO₄. The product can be reacted with di-tert-butyl dicarbonate in the presence of dimethylaminopyridine and then extracted with a mixture of citric acid solution and sodium chloride solution to give the title compound of Example 42b.

Example 42c.

[0239] (2R, 6S, 13aS, 14aR, 16aS, Z) -6- (2-methylpyrimidine-5-carboxamido) -5,16 dioxo-2- (3- (benzo [d] thiazol-2-yl) hydrochloride quinoxalin-2-yloxy) -1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [ 1,4] ethyl diazacyclopentadecine-14α-carboxylate

[0240] The product of Example 42b can be subjected to a ring metathesis reaction in the presence of a Zhan B catalyst in toluene to give (2R, 6S, 13aS, 14aR, 16aS, Z) -6 (2-methylpyrimidine-2 carboxamido) -5.16-dioxo-2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) 2,3,5,6,7,8,9,10,11, 13a, 14.14a, 16.16a-tetradekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a, 15 (tert-butyl 14a-ethyl) dicarboxylate. The catalyst can be deactivated after reaction with imidazole.

[0241] The solvent for the ring closure product in toluene can be replaced with acetonitrile followed by the addition of hydrogen chloride in dioxane and heated to give the title compound of Example 42c.

Example 42d.

[224, (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -6- (2-methylpyrimidine-5-carboxamido) -5,16-dioxo-2- (3- (benzo [d ] thiazol-2-yl) quinoxalin-2-yloxy) 1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [[1,4] diazacyclopentadecine-14a-carboxamide. The separated product of Example 42c can be mixed with tetrahydrofuran, water and

LiOH ^ H2O, then heat and stir. The reaction mixture can be cooled later, an aqueous solution of H is added₃AFTER₄, aqueous NaCl solution and 2-methyltetrahydrofuran, and the organic layer is separated, washed and filtered. MeCN is added to the concentrated organic layer, heated and cooled, followed by addition of diethylamine. The suspension is heated and cooled to form (2R, 6S, 13aS, 14aR, 16aS, Z) -6- (2-methylpyrimidine-5-carboxamido) -5,16-dioxo-2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo Diethylamine [1,2-a] [1,4] diazacyclopentadecine-14α-carboxylate, which can be further washed and dried.

[0244] The diethylamine salt can be mixed with tetrahydrofuran, 2-methyltetrahydrofuran and an aqueous solution of H₃AFTER₄. The organic layer is separated, washed with aqueous NaCl solution and then concentrated and / or purified. The product can in turn be mixed with NMP, followed by the addition of carbonyldiimidazole (CDI) followed by 1,8-diazabicyclo [5.4.0] undec-7-ene (DBU). In turn, cyclopropylsulfonamide can be added. The reaction mixture is stirred for hours. Isopropyl acetate can then be added followed by an aqueous KH solution₂AFTER₄and then an aqueous solution of H₃AFTER₄. The organic layer can be separated, washed, and purified to give the title compound of Example 42d.

Example 43. N - ((2R, 6S, 13aS, 14aR, 16AS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) 14A (cyclopropylsulfonylcarbamoyl) -5.16-diokso1 , 2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyn- 6-yl) -3-methylisoxazole-5-carboxamide [0245] The compound of Example 43 can be prepared according to the procedure described for the preparation of the compound of Example 42, replacing 2-methylpyrimidine-5-carboxylic acid from Example 42a with 3-methylisoxazole-5-carboxylic acid in Example 43a to give (S) -2- (3-methylisoxazole-5-carboxamido) non-8-enoic acid. All subsequent steps should occur in a similar manner to Example 42 to prepare the compound of Example 43.

EP 2 468 285 B1

Example 44. N - ((2R, 6S, 13aS, 14aR, 16AS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) 14A (cyclopropylsulfonylcarbamoyl) -5.16-diokso1 , 2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyn- 6-yl) -5-methylisoxazole-3-carboxamide [0246] The compound of Example 44 can be prepared according to the procedure described for the preparation of the compound of Example 42, replacing 2-methylpyrimidine-5-carboxylic acid from Example 42a with 5-methylisoxazole-3-carboxylic acid in Example 44a to produce (S) -2- (5-methylisoxazole-3-carboxamido) non-8-enoic acid. All subsequent steps should occur in a similar manner to Example 42 to prepare the compound of Example 44.

Example 45. (2R, 6S, 13aS, 14aR, 16AS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -N- (cyclopropylsulfonyl) -6- (3-fluorobenzamido) diokso1,2,3,5,6,7,8,9,10,11,13a--5.16, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [ 1,4] diazacyclopentadecine-14a-carboxamide [0247] The compound of Example 45 can be prepared according to the procedure described for the preparation of the compound of Example 42, replacing the 2-methylpyrimidine-5-carboxylic acid of Example 42a with 3-fluorobenzoic acid in Example 45a to produce (S) -2- (3-fluorobenzamido) non-8-enoic acid. All subsequent steps should occur in a similar manner to Example 42 to prepare the compound of Example 45.

Example 46. (2R, 6S, 13aS, 14aR, 16AS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -N- (cyclopropylsulfonyl) -5.16-dioxo-6- (pyrimidin-4-carboxamido) 1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a ] [1,4] diazacyclopentadecine-14a-carboxamide. [0248] The compound of Example 46 can be prepared according to the procedure described for the preparation of the compound of Example 42, replacing 2-methylpyrimidine-5-carboxylic acid from Example 42a with pyrimidine-4-carboxylic acid in Example 46a to produce (S) -2 (pyrimidine-4-carboxamido) non-8-enoic acid. All next stages should take place in

Example 47. N - ((2R, 6S, 13aS, 14aR, 16AS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) 14A (cyclopropylsulfonylcarbamoyl) -5.16-diokso1 , 2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyn- 6-yl) isoxazole-5-carboxamide [0249] The compound of Example 47 can be prepared according to the procedure described for the preparation of the compound of Example 42, replacing 2-methylpyrimidine-5-carboxylic acid from Example 42a with isoxazole-5-carboxylic acid in Example 47a to produce (S) -2- (isoxazole-5-carboxamido) non-8-enoic acid. All subsequent steps should occur in a similar manner to Example 42 to prepare the compound of Example 47.

Example 48. (2R, 6S, 13aS, 14aR, 16aS, Z) -2- (3- (benzo [d] thiazol-2-yl) quinoxalin-2-yloxy) -N (cyclopropylsulfonyl) -6- (5- methylpyrazine-2-carboxamido) -5.16-diokso1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1 , 2-a] [1,4] diazacyklopentadecyno-14a-carboxamide

[0250] The compound of Example 48 can be prepared according to the procedure described for the preparation of the compound of Example 42, replacing 2-methylpyrimidine-5-carboxylic acid from Example 42a with 5-methylpyrazine-2-carboxylic acid in Example 48a to prepare (S) -2- (5-methylpyrazine-2-carboxamido) non-8-enoic acid. All subsequent steps should occur in a similar manner to Example 42 to prepare the compound of Example 48.

Example 49. (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -6- (5-methylpyrazine-2-carboxamido) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16aheksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyno- 14a-carboxamide

Example 49a.

[0251] (S) -2- (5-methylpyrazine-2-carboxamido) non-8-enoic acid.

[0252] The Boc-2 (S) -amino-non-8-enoic acid salt with dicyclohexylamine can be suspended in isopropyl acetate, washed several times with aqueous citric acid solution, and then once with water. The washed product, concentrated and then again diluted in isopropyl acetate, can be reacted with HCl to form 2 (S) -amino-non-8-enoic acid HCl salt. 5-methyl-2-pyrazinecarboxylic acid, N, N'-disuccinimidyl carbonate, and N, N-dimethylaminopyridine can be dissolved in N-methyl-2-pyrrolidone (NMP) and mixed. Subsequently, 2 (S) amino-non-8-enoic acid HCl salt is added, followed by triethylamine, and mixed to give the title compound of Example 49a, which can be crystallized by the addition of HCl followed by water.

Example 49b.

[0253] (2S, 4R) -N-Boc-4-hydroxyproline can be reacted with 6-chlorophenanthridine in NMP in the presence of sodium tert-butoxide to produce (2S, 4R) -1- (tert-butoxycarbonyl) - 4 (phenanthridin-6-yloxy) -pyrrolidine-2-carboxylic acid. Then methyl tert-butyl ether (MTBE) and water can be added. The aqueous layer is separated, washed, then HCl is added, followed by extraction with MTBE. The extracted product can be mixed with diisopropylethylamine (DIPEA) and HATU (CAS No. 148893-10-1) and then reacted with ethyl tosylate salt (1R, 2S) -1-amino-2-vinylcyclopropanecarboxylate in dimethylformamide (DMF) and toluene. The reaction gives tert-butyl (2S, 4R) -2 - ((1R, 2S) -1- (ethoxycarbonyl) -2-vinylcyclopropylcarbamoyl) -4- (phenanthridin-6-yloxy) pyrrolidine-1-carboxylate, which can be extracted with using MTBE and wash with HCl, further extract, wash, dry, and dissolve in 2-propanol.

[0254] To the solution in 2-propanol can be added HCl to produce (1R, 2S) -1 - ((2S, 4R) -4 (phenanthridin-6-yloxy) pyrrolidine-2-carboxamido) -2-vinylcyclopropanecarboxylate, which can be crystallized by neutralization with NaOH.

[0255] (1R, 2S) -1 - ((2S, 4R) -4- (Phenanthridin-6-yloxy) pyrrolidine-2-carboxamido) -2-vinylcyclopropanecarboxylate, title compound of Example 49a, N-hydroxy-5 -norbomene-2,3-dicarboximide, and N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride can be mixed and mixed in DMF, followed by the addition of N, N-dimethylethylenediamine. The reaction produces (1R, 2S) -1 ((2S, 4R) -1 - ((S) -2- (5-methylpyrazine-2-carboxamido) non-8-enoyl) -4- (phenanthridin-6-yloxy) ethyl pyrrolidine-2-carboxamido) -2-vinylcyclopropane carboxylate, which can be dissolved in isopropyl acetate and extracted with aqueous H₃AFTER₄and then extract with aqueous K solution₂HPO₄. The product can be reacted with di-tert-butyl dicarbonate in the presence of

Dimethylaminopyridine, followed by extraction with a mixture of citric acid solution and sodium chloride solution to obtain the title compound of Example 49b.

Example 49c.

[0256] (2R, 6S, 13aS, 14aR, 16aS, Z) hydrochloride -6- (5-methylpyrazine-2-carboxamido) -5,16 dioxo-2- (phenanthridin-6-yloxy) -1,2,3, 5,6,7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxylate [ 0257] The product of Example 49b can be subjected to a ring closing metathesis reaction in the presence of a Zhan B catalyst in toluene to produce (2R, 6S, 13aS, 14aR, 16AS, Z) -6- (5-methylpyrazine-2-carboxamido) -5.16-dioxo-2- (phenanthridin-6-yloxy) 2,3,5,6,7, 8,9,10,11,13a, 14,14a, 16,16a-tetradekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a, 15 (1H) -tert-butyl dicarboxylate 14a-acetate. The reaction catalyst can be deactivated with imidazole.

[0258] The solvent for the ring closure product in toluene can be replaced with acetonitrile followed by the addition of hydrogen chloride in dioxane and heated to give the title compound from

Example 49c.

Example 49d.

[0259] (2R, 6S, 13aS, 14aR, 16aS, Z) -N- (cyclopropylsulfonyl) -6- (5-methylpyrazine-2-carboxamido) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxamide [0260] The isolated product of Example 49c can be mixed with tetrahydrofuran, water and LiOH ^ H2O, then heated and stirred. The reaction mixture can be cooled later, aqueous H is added₃AFTER₄, aqueous NaCl and 2-methyltetrahydrofuran, and the organic layer is separated, washed and filtered. MeCN is added to the concentrated organic layer, heated and cooled, followed by addition of diethylamine. The suspension is heated and cooled to form (2R, 6S, 13aS, 14aR, 16aS, Z) -6- (5-methylpyrazine-2-carboxamido) -5,16-dioxo-2- (phenanthridin-6-yloxy) -1 salt, 2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyno 14a - diethylamine carboxylate, which can be further washed and dried.

[0261] The diethylamine salt can be mixed with tetrahydrofuran, 2-methyltetrahydrofuran and an aqueous solution of H₃AFTER₄. The organic layer is separated, washed with an aqueous NaCl solution and then concentrated and / or purified. The product can in turn be mixed with NMP, followed by the addition of carbonyldiimidazole (CDI) followed by 1,8-diazabicyclo [5.4.0] undec-7-ene (DBU). In turn, cyclopropylsulfonamide can be added. The reaction mixture is stirred for hours. Isopropyl acetate can then be added followed by an aqueous KH solution₂AFTER₄ and then an aqueous solution of H₃AFTER₄. The organic layer can be separated, washed, and purified to give the title compound of Example 49d. The isolated product can be further dissolved in isopropyl acetate and then the solution is diluted with ethanol. Water may be added in portions to the resulting solution in portions at appropriate holding times after each addition to ensure no supersaturation. The addition of water ends just when the ternary solvent system becomes binary due to the partial miscibility of the isopropyl acetate solvent system,

Ethanol, water. The suspension can be stirred for hours, and then the solid is isolated by filtration and drying to give the crystalline hydrate of the title compound.

Example 50. (2R, 6S, 13aS, 14aR, 16AS, Z) -N- (cyclopropylsulfonyl) -6- (1,5-dimethyl-1 H-pyrazole-3-carboxamido) -5.16-dioxo-2- (phenanthridin-6 yloxy) 1,2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1, 4] diazacyclopentadecine-14a-carboxamide. [0262] The compound of Example 50 can be prepared according to the procedure described for the preparation of the compound of Example 49, replacing 5-methylpyrazine-2-carboxylic acid from Example 49a with 1,5-dimethyl-1H-pyrazole-3-carboxylic acid in Example 50a to produce (S) 2- (1,5-dimethyl-1H-pyrazole-3- carboxamido) non-8-enoic acid. All subsequent steps should occur in a similar manner to Example 49 to prepare the compound of Example 50.

Example 51. (2R, 6S, 13aS, 14aR, 16AS, Z) -N- (cyclopropylsulfonyl) -6- (5-methyl-1H-pyrazole-3-carboxamide) -5.16-dioxo-2- (phenanthridin-6-yloxy) 1 , 2,3,5,6,7,8,9,10,11,13a, 14.14, 15,16,16a-heksadekahydrocyklopropa [e] pyrrolo [1,2-a] [1,4] diazacyklopentadecyno- 14a-carboxamide [0263] The compound of Example 51 can be prepared according to the procedure described for the preparation of the compound of Example 49, replacing 5-methylpyrazine-2-carboxylic acid from Example 49a with 5-methyl-1H-pyrazole-3-carboxylic acid in Example 51a to produce (S) -2 (5-methyl-1H-pyrazole-3-carboxamido) non- 8-enoic acid. All subsequent steps should occur in a similar manner to Example 49 to prepare the compound of Example 51.

Example 52. (2R, 6S, 13aS, 14aR, 16AS, Z) -N- (cyclopropylsulfonyl) -6- (2-fluorobenzamido) 5,16-dioxo-2- (phenanthridin-6-yloxy) -1,2,3,5 , 6,7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxamide [0264] The compound of Example 52 can be prepared according to the procedure described for the preparation of the compound of Example 49, replacing 5-methylpyrazine-2-carboxylic acid from Example 49a with 2-fluorobenzoic acid in Example 52a to produce (S) -2- (2-fluorobenzamido) non-8-enoic acid. All subsequent steps should occur in a similar manner to Example 49 to prepare the compound of Example 52.

Example 53. (2R, 6S, 13aS, 14aR, 16AS, Z) -N- (cyclopropylsulfonyl) -5.16-dioxo-2- (phenanthridin-6-yloxy) -6- (pyrazin-2-carboxamido) -1,2, 3,5,6,7,8,9,10,11,13a, 14,14a, 15,16,16ahexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxamide [0265 ] The compound of Example 53 can be prepared according to the procedure described for the preparation of the compound of Example 49, replacing 5-methylpyrazine-2-carboxylic acid from Example 49a with pyrazine carboxylic acid in Example 53a to produce (S) -2- (pyrazine-2-carboxamido) non-8-enoic acid. All subsequent steps should occur in a similar manner to Example 49 to prepare the compound of Example 53.

Example 54. (2R, 6S, 13aS, 14aR, 16aS, Z) -14- (cyclopropylsulfonylcarbamoyl) -5,16-dioxo2- (thiazolo [4,5-c] quinolin-4-yloxy) -1,2,3 , 5,6,7,8,9,10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecin-6-ylcarbamate tert butyl

[0266] A mixture of 5-bromothiazole-4-carboxylic acid methyl ester (0.521 g, 2.35 mmol), 2- (4,4,5,5-tetramethyl-1,3,2-dioxaborolane- 2-yl) aniline (0.514 g, 2.35 mmol), 1,1'bis (di-tert-butylphosphine) ferrocenopalladium chloride (0.060 g, 0.094 mmol), and sodium carbonate (1.17 mL of a 2M aqueous solution) in tetrahydrofuran (12 mL) was stirred under nitrogen at room temperature for 48 h. The reaction mixture was then heated at 50 ° C for an additional 16 h. The reaction mixture was then cooled to room temperature, diluted with dichloromethane (120 mL) and dimethylformamide (40 mL) and washed with water (20 mL). The resulting solid was isolated by filtration under reduced pressure to obtain the title compound (0.251 mg, 53% yield, thiazolo [4,5-c] quinolin-4 (5H) 10 on).

Example 55

Synthesis of the cyclic peptide precursor [0267]

<img file="PL2468285T3_D0031.tif" />

[0268] To a solution of Boc-L-2-amino-8-nonenoic acid 42a (1.36 g, 5 mol) and commercially available cis-L-hydroxyproline 42b methyl ester (1.09g, 6 mmol) in 15 ml DMF, DIEA (4 ml, 4eq.) And HATU (4g, 2eq) were added. Coupling was carried out at 0 ° C for 1 hour. The reaction mixture was diluted with 100 mL EtOAc and then washed with 2x 20 mL 5% citric acid, 2 x 20 mL water, 1M NaHCO, respectively₃ 4 x 20 ml and 2 x 10 ml brine. The organic phase was dried over anhydrous Na₂SO₄and then evaporated to give dipeptide 42c (1.91 g, 95.8%), which was identified by HPLC (retention time = 8.9 min, 30-70%, 90% B), and MS (found 421.37 , M + Na⁺).

[0269] Dipeptide 42c (1.91 g) was dissolved in 15 mL of dioxane and 15 mL of 1N aqueous solution

LiOH and the hydrolysis reaction were carried out at room temperature for 4 hours. The reaction mixture was acidified with a 5% citric acid solution and extracted with 100 mL EtOAc, followed by washing with 2 x 20 mL water and 2 x 20 mL brine, respectively. The organic phase was dried over anhydrous Na₂SO₄ and then removed under reduced pressure to give compound 42d as free carboxylic acid (1.79 g, 97%), which was used for the next synthetic step without the need for further purification.

[0270] To a solution of the above free acid (1.77, 4.64 mmol) in 5 ml DMF was added De-vinylcyclopropanoamic acid ethyl ester (0.95 g, 5 mmol), DIEA (4 ml, 4 eq.) And HATU (4 g, 2 eq). Coupling was carried out at 0 ° C for 5 hours. The reaction mixture was diluted with 80 mL EtOAc and then washed with 2x 20 mL 5% citric acid, 2 x 20 mL water, 1M NaHCO, respectively₃ 4 x 20 ml and 2 x 10 ml brine. The organic phase was dried over anhydrous Na₂SO₄and then evaporated. The residue was purified by flash chromatography on silica gel using different ratios of hexane: EtOAc as the eluent phase (5: 1 ^ 3: 1 ^ 1: 2 ^ 1: 2 ^ 1: 5). Linear tripeptide 42e was isolated as an oil after removal of elution solvents (1.59 g, 65.4%), identified by HPLC (retention time = 11.43 min) and MS (found 544.84, M + Na⁺).

[0271] A solution of linear tripeptide 42e (1.51 g, 2.89 mmol) in 200 mL dry DCM was deoxygenated by bubbling N₂. Then Hoveyda's first generation catalyst (5 mol% eq.) Was added as a solid. The reaction was heated to reflux under an atmosphere of N₂ for 12 hours. The solvent was evaporated and the residue was purified by flash chromatography on silica gel using different ratios of hexane: EtOAc as the eluent phase (9: 1 ^ 5: 1 ^ 3: 1 ^ 1: 1 ^ 1: 2 ^ 1: 5). The cyclic peptide precursor was isolated as a white powder after removal of the elution solvents (1.24 g, 87%), identified by HPLC (retention time = 7.84 min, 30-70%, 90% B), and MS (found 516, 28, M + Na⁺).

(2S, 6S, 13aS, 14aR, 16AS, Z) -2- (4-bromofenylosulfonyloksy) -6- (tert-butoxycarbonylamino) 5,16-dioxo-1,2,3,5,6,7,8,9 , 10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxylate [0272]

<img file="PL2468285T3_D0032.tif" />

[0273] Solution (2S, 6S, 13aS, 14aR, 16aS, Z) -6- (tert-butoxycarbonylamino) -2-hydroxy-5.16 dioxo-1,2,3,5,6,7,8,9, 10,11,13a, 14,14a, 15,16,16a-hexadekahydrocyclopropa [e] pyrrolo [1,2-a] [1,4] diazacyclopentadecine-14a-carboxylate (22.1 g, 44.8 mmol) and DABCO (8.5 g, 76.7 mmol) in toluene (88 mL) was stirred at room temperature. To this solution was added a chloride solution 460

Bromobenzene-1-sulfonyl 17.2 g, 67.2 mmol) in toluene (44 mL). After the addition, the reaction was quenched with 10% aqueous sodium carbonate solution (110 mL) and the mixture was stirred for 15 min. Tetrahydrofuran (44 mL) was added and the mixture was washed with 0.5M HCl, water, followed by saturated aqueous sodium chloride. The organic layer was dried over anhydrous magnesium sulfate, filtered, and evaporated under reduced pressure and dried to give the title compound (27.7 g, 87% yield), which was used without further purification.

Example 56

Measurement of inhibition potency of purified NS3 protease enzyme [0274] The activity of recombinant HCV NS3 proteases derived from isolates representing genotypes 1, 2, 3 or 4 is measured by testing the cleavage of the following peptide substrate:

<img file="PL2468285T3_D0033.tif" />

[0275] The substrate is labeled with fluorine and a fluorescent quencher. Cleavage releases the quencher and increases the fluorescence. NS3 protease is incubated with a series of dilutions of the inhibitor in 150 mM NaCl, 10% glycerol, 5 mM DTT, with or without 0.01% dodecyl maltoside for either 30 minutes or 300 minutes. The substrate is added at a concentration of 5 [mu] M to initiate the reaction, and fluorescence is measured at intervals of 2 minutes for 30 minutes. Enzyme concentrations range from 10 to 100 nM in the absence of detergent, or 10 times lower in the presence of detergent. Substrate peptides are labeled using either EDANS and DABCYL (355 nm excitation, 485 nm emission) or TAMRA and QSY (544 nm excitation, 590 nm emission). For routine IC50 determination, a series of 3-fold dilutions are used, starting from initial concentrations of 100 μΜ, 200 μΜ, or 2 mM. For compounds with K values_and close or lower than enzyme concentration, a tight binding calculation format is used, with 24 inhibitor dilutions covering a range of 0 to 100 nM inhibitor. K values_and is calculated using the tight binding test format according to the following equation:

V = A {[(K +1 - E)² + 4KE])^{I / 2} - (K + IE)}, where I = total inhibitor concentration, E = active enzyme concentration, K = apparent K value_and and A = [kcat) S / 2] [Km = (S)].

Replicon cell lines [0276] Two subgenomic replicon cell lines can be used to characterize compounds in cell culture: one derived from genotype 1a and one derived from genotype 1b. Both replicon constructs are bicistronic subgenomic replicons essentially similar to those described by Bartenschlager et al. (Lohmann et al., Science (1999) 285 (5424): 110-113). The replicon construct from genotype 1a contains the NS3-NS5B coding region

EP 2 468 285 B1 derived from strain H77 HCV (1a-H77) (Blight et al., J Virol (2003) 77 (5): 3181-3190). The first cistron of the construct consists of the first 36 nucleotides of the HCV 1a-H77 core gene condensed with the firefly luciferase reporter gene and the selective neomycin phosphotransferase (Neo) marker. Luciferase and Neo coding regions are separated by FMDV 2a protease. The second cistron contains the NS3-NS5B coding region derived from 1a-H77 with the addition of adaptive mutations E1202G in NS3, K1691R in NS4A, and K2040R and S22041 in NS5A. The 1b-Con-1 replicon construct is identical to the 1a-H77 replicon, except that the 5 'and 3' NTRs and the NS3-NS5B coding region may be derived from strain 1b-Con-1 (Blight et al., Science (2000 ) 290 (5498): 1972-1974), and adaptive mutations are E1202G and T1280I in NS3 and S2204I in NS5A.

Testing compounds on replicons [0277] Replicon cell lines can be maintained in Dulbecco's modified Eagle's medium (DMEM) containing 100 IU / ml penicillin, 100 mg / ml streptomycin (Invitrogen), 200 mg / ml G418 (Invitrogen) and % (vol) fetal calf serum (FBS). Cells containing replicon can be plated into 96-well plates at a density of 5000 cells per well in 100 μΐ DMEM containing 5% FBS. The next day, the compound may be initially diluted in dimethyl sulfoxide (DMSO) to give a 200x inhibitor stock solution in a series of 8 semi-logarithmic dilutions. The dilution series can then be diluted 100-fold in media containing 5% FBS. One hundred microliters of inhibitor medium can be added to each well of the previous day's culture plate already containing 100 μΐ DMEM with 5% FBS. In assays where the effect of protein binding on the inhibitor potency is assessed, the medium from the cell culture plates from the previous day can be replaced by 200 μΐ DMEM containing 40% human plasma (Innovative Research) plus 5% FBS as well as the compound. Cells can be grown for 4 days in tissue culture incubators. The inhibitory effect of compounds against replicons can be determined by measuring either the level of HCV luciferase or RNA. The luciferase assay can be performed using the Luciferase Assay System kit (Promega) according to the manufacturer's instructions. Briefly, cell culture media is removed and the wells are washed with 200 μΐ phosphate buffered saline. Passive Lysis buffer (Promega, WI) is added to each well and the plates are incubated for 30 min with rocking to lyse the cells. Luciferin solution (50 μΐ, Promega) is added, and luciferase activity is measured by a Victor II luminometer (Perkin-Elmer). To determine HCV RNA levels, RNA extractions can be performed using the CellsDirect kit (Invitrogen), and the number of HCV RNA copies can be measured using the SuperScript III Platinum One-Step qRT-PCR system (Invitrogen) and non-translated HCV 5 'specific primers . Cytotoxicity can be determined by colorimetric assay with 3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide (MTT) as follows. Replicon cells are placed in 96-well plates (4000 cells per well), the next day, compound dilutions are added as in the activity assay, and cells are cultured in the presence of inhibitors for 4 days. The MTT solution is diluted in DMEM containing 5% FBS and 60 μΐ of the solution is added to the cells. After 4 h, the cells are solubilized by the addition of 30 μΐ SDS (20% in 0.02 N HCl). Plates are incubated overnight and optical density at 570 nm can be measured. For EC determination₅₀ and TD₅₀ compounds, data for luciferase,

RNA and MTT inhibition can be analyzed using GraphPad Prism 4 software (equation: sigmoidal, variable dose-response slope).

Mutants in Transient Replicons [0278] Mutations detected in resistance selection studies can be introduced into wild type transient replicon constructs based on genotypes 1a-H77 and 1b-N. Both replicons are bicistronic subgenomic constructs containing firefly luciferase reporter gene similar to those described above, but which do not contain the Neo selective marker and are therefore only useful for transient replication assays. The 1a-H77 replicon for transient assays further differs from the replicon in a stable cell line in that it contains NS2 to NS5B in the second cistron. The 1b-N strain replicon contains NS3 to NS5B in a second cistron, with adaptation mutations E1202G in NS3 and S22041 in NS5A. Mutagenesis can be performed using the Stratagene QuikChange XL II site-directed mutagenesis kit. Mutant sequences can be confirmed, plasmids can be linearized using Xba I restriction enzyme and used as a template for in vitro transcription reactions to generate mutant replicon RNA for transient transfections. In vitro transcription can be performed using the T7 Megascript kit (Ambion).

[0279] Transfection with transient replicons can be performed essentially as described by Mo et al. (Antimicrob Agents Chemother (2005) 49 (10): 4305-4314) with minor modifications.

Fifteen micrograms of the RNA template can be used for 3 * 10 electroporation⁶ cells in a volume of 200 μl in a 0.2 cm cuvette. The cells used for transient transfections may be Huh7 cells obtained by treatment of replicon-containing cells with IFN (Mo et al., Supra). Electroporation can be performed using a Gene Pulser II instrument (Bio-Rad, CA) at 480 V and 25 μF, using two manual pulses. Transfected cells can be diluted to 7.5-10⁴ cells / ml and place in 96-well plates at a density of 7.5-10³ cells per well in DMEM with 5% FBS and 100 IU / ml penicillin, 100 mg / ml streptomycin (Invitrogen). Four hours after transfection, one plate is collected for luciferase measurement; this plate can give a measure of the amount of input RNA that can be translated and thus the transfection efficiency. Further dilutions of test compounds in DMSO (DMSO final concentration 0.5%) can be added to the remaining plates, and the plates are incubated for 4 days.

[0280] The exemplary compounds described herein were tested for their anti-HCV activity. Many of the compounds tested showed unexpected anti-HCV activities, including excellent activities in biochemical assays against HCV proteases representing various HCV genotypes, distinguishing activities in normal HCV replicon assays, including activity against HCV 1a-H77 and 1b-con1 strains in the absence or in the presence of 40% of human plasma, and / or excellent activities in replicon transient assays against drug-resistant mutants in a variety of different HCV genetic images.

[0281] Unless defined otherwise, all technical and scientific terms used herein correspond to those of a commonly known person in the art.

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