WO2009134624A1 - Hcv ns3 protease inhibitors - Google Patents

Abstract

The present invention relates to macrocyclic compounds of formula (I) that are useful as inhibitors of the hepatitis C virus (HCV) NS3 protease, their synthesis, and their use for treating or preventing HCV infections.

Description

TITLE OF THE APPLICATION HCV NS3 PROTEASE INHIBITORS

FIELD OF THE INVENTION The present invention relates to macrocyclic compounds that are useful as inhibitors of the hepatitis C virus (HCV) NS3 protease, the synthesis of such compounds, and the use of such compounds for treating HCV infection and/or reducing the likelihood or severity of symptoms of HCV infection.

BACKGROUND OF THE INVENTION

Hepatitis C virus (HCV) infection is a major health problem that leads to chronic liver disease, such as cirrhosis and hepatocellular carcinoma, in a substantial number of infected individuals, estimated to be 2-15% of the world's population. There are an estimated 3,9 million infected people in the United States alone, according to the U.S. Center for Disease Control, roughly five times the number of people infected with the human immunodeficiency virus (HIV). According to the World Health Organization, there are more than 170 million infected individuals worldwide, with at least 3 to 4 million people being infected each year. Once infected, about 20% of people clear the virus, but about 80% of those infected harbor HCV the rest of their lives. Ten to 20% of chronically infected individuals eventually develop liver- destroying cirrhosis or cancer. The viral disease is transmitted parenterally by contaminated blood and blood products, contaminated needles, or sexually and vertically from infected mothers or carrier mothers to their off-spring.

Current treatments for HCV infection are restricted to immunotherapy with recombinant interferon-α alone or in combination with the nucleoside analog ribavirin, and these treatments are of limited clinical benefit. Moreover, there is no established vaccine for HCV. Consequently, there is an urgent need for improved therapeutic agents that effectively combat chronic HCV infection. Treatment of HCV infection has been discussed in the following references: B. Dymock et al, "Novel approaches to the treatment of hepatitis C virus infection," 11 Antiviral Chem. & Chemotherapy 79-96 (2000); H. Rosen et al, "Hepatitis C virus: current understanding and prospects for future therapies," 5 Molec. Med. Today 393-399 (1999); D. Moradpour et al, "Current and evolving therapies for hepatitis C," 11 Euro. J. Gastroenterol. Hepatol. 1189-1202 (1999); R. Bartenschlager, "Candidate Targets for Hepatitis C Virus- Specific Antiviral Therapy," 40 Intervirology 378-393 (1997); GM. Lauer & B.D. Walker, "Hepatitis C Virus Infection," 345 N. Engl. J. Med. 41-52 (2001); B. W. Dymock, "Emerging therapies for hepatitis C virus infection," 6 Emerging Drugs 13-42 (2001); and C. Crabb, "Hard- Won Advances Spark Excitement about Hepatitis C," Science: 506-507 (2001). Several virally-encoded enzymes are putative targets for therapeutic intervention, including a metalloprotease (NS2-3), a serine protease (NS3), a helicase (NS3), and an RNA- dependent RNA polymerase (NS5B). The NS3 protease is located in the N-terminal domain of the NS 3 protein. Because it is responsible for an intramolecular cleavage at the NS 3/4 A site and for downstream intermolecular processing at the NS4A/4B, NS4B/5A and NS5 A/5B junctions, the NS3 protease is considered a prime drug target. Previous research has identified classes of peptides, such as hexapeptides as well as tripeptides discussed in U.S. Patent Application Publications US2005/0020503, US2004/0229818, and US2004/00229776, showing degrees of activity in inhibiting the NS3 protease. The aim of the present invention is to provide further compounds which exhibit activity against the HCV NS3 protease.

SUMMARY OF THE INVENTION

The present invention relates to macrocyclic compounds of formula (I) and pharmaceutically acceptable salts thereof. These compounds are useful in the inhibition of HCV (hepatitis C virus) NS3 (non-structural 3) protease, the prevention or treatment of one or more of the symptoms of HCV infection, either as compounds or their pharmaceutically acceptable salts, or as pharmaceutical composition ingredients. As pharmaceutical composition ingredients, these compounds and salts may be the primary active therapeutic agent, and, when appropriate, may be combined with other therapeutic agents including but not limited to other HCV antivirals, anti- infectives, immunomodulators, antibiotics or vaccines. More particularly, the present invention relates to a compound of formula (I) and a pharmaceutically acceptable salt thereof:

(Formula I) wherein:

is selected from the group consisting of:

wherein: each L is independently selected from the group consisting of N and CH, provided that the total number of L that are N is from 1 to 4;

R¹ is selected from the group consisting of -CO₂H, and -CONHSO₂(cyclopropyl); R² is selected from the group consisting of ethyl and ethenyl;

R³ is selected from the group consisting of cyclopentyl, cyclohexyl, and

-C(CH₃)₃; each R⁴ is independently selected from the group consisting of halogen atoms, C₁-C₄ alkoxy, C₁-C₇ alkyl, C₂-C₇ alkenyl, -CN, -CF₃, -OCF₃, SCH₃, -SO₂(CH₃), C₃-C₈ cycloalkyl, C₃-C₈ cycloalkoxy, C₁-C₆ haloalkyl, phenyl, naphthyl and heteroaryl groups, wherein each said R⁴ heteroaryl is selected from the group consisting of 5- and

6-membered aromatic rings having 1, 2 or 3 heteroatoms independently selected from N, O and S, and said R⁴ heteroaryl is attached through a ring atom selected from C or N, each said R⁴ phenyl, naphthyl and heteroaryl groups are substituted with 0 to 4 substituents independently selected from the group consisting of halogen atoms, -OR⁵, -SR⁵, -N(R⁵)₂, -N(C₁-C₆ alkyl)O(C₁-C₆ alkyl), C₁-C₆ alkyl, C₁-C₆ haloalkyl, (C₁-C₆ haloalkoxy), -NO₂, -CN, -CF₃, -SO₂(C₁-C₆ alkyl), -S(O)(C₁-C₆ alkyl), -NR⁵SO₂R⁶, SO₂N(R⁶)₂, -NHCOOR⁶, -NHCOR⁶, -NHCONHR⁶, -CO₂R⁵, -C(O)R⁵, and -CON(R⁵)₂, and 2 adjacent substituents of said R⁴ phenyl, naphthyl and heteroaryl groups may be taken together to form a 3- to ό-membered cyclic ring containing 0 to 3 heteroatoms independently selected from N, O and S; each R⁵ is independently H or C₁-C₆ alkyl; each R⁶ is independently H, C₁-C₆ alkyl, C₃-C₆ cycloalkyl, C₃-C₆ cycloalkyl(C₁- C₅)alkyl, aryl, aryl(C₁-C₄)alkyl, heteroaryl, heteroaryl (C₁-C₄ alkyl), heterocyclyl, or heterocyclyl(C₁-C₈ alkyl);

Y is selected from the group consisting of -C(O)-, -C(O)O- and -C(O)NH-; M is selected from the group consisting of C₄-C₇ alkylene and C₄-C₇ alkenylene, wherein said M is substituted with O to 3 substituents independently selected C₁-C₈ alkyl, provided that two adjacent substituents can together form a 3 to 6 membered ring.

The present invention also includes pharmaceutical compositions containing a compound of the present invention and methods of preparing such pharmaceutical compositions. The present invention further includes methods of treating or reducing the likelihood or severity of one or more symptoms of HCV infection.

Other embodiments, aspects and features of the present invention are either further described in or will be apparent from the ensuing description, examples and appended claims.

DETAILED DESCRIPTION OF THE INVENTION

The present invention includes compounds of formula (I), and pharmaceutically acceptable salts thereof. These compounds and their pharmaceutically acceptable salts are HCV protease inhibitors (e.g., HCV NS3 protease inhibitors). Preferred compounds are those with high activity (e.g., Ki of 5 nM or less, 1 nM or less, 0.5 nM or less, or 0.1 raM or less) against HCV NS3 genotype Ib Rl 55K. Rl 55K is a HCV Ib mutation that occurs in nature and which provides resistance against some NS3 protease inhibitors. Example 85 infra illustrates the ability of different compounds to provide high activity against such a mutation. Reference to formula I compounds throughout the present application includes reference to compounds within formula I including different embodiment and subgeneric formula (formula Ia and Ib).

Different embodiments for formula I compounds include the following:

In a first embodiment,

is selected from the group consisting of:

and the other substitutents are as provided for formula I above. In an aspect of the first embodiment, O or 1 R⁴ is present and, if present, is selected from the group consisting of -Br, -Cl, -CN, phenyl, -O-phenyl, -OCF₃, -OCH₃, -C(O)OH₃ -CH₃ and -C(O)CH₃.

In a second embodiment, R¹ is -CO₂H, and the other substituents are as provided in formula I above or in the above first embodiment.

In a third embodiment, R¹ is -C(0)NHSθ₂Cyclopropyl, and the other substituents are as provided in formula I above or in the first embodiment.

In a fourth embodiment, R² is -CH₂CH₃, and the other substituents are as provided in formula I above or in the first to third embodiments.

In a fifth embodiment, R² is -CH=CH₂, and the other substituents are as provided in formula I above or in the first to third embodiments.

In a sixth embodiment, R³ is cyclopentyl, and the other substituents are as provided in formula I above or the first through fifth embodiments.

In a seventh embodiment, R³ is cyclohexyl, and the other substituents are as provided in formula I above or in the first through fifth embodiments.

In a eighth embodiment R³ is -C(CH₃)₃, and the other substituents are as provided in formula I above or the first through fifth embodiments.

In a ninth embodiment, M is selected from the group consisting of

, where Z is

and the other substituents are as provided in formula I above or the first through eighth embodiments.

In another aspect of the invention, the formula I compound has the following structure:

formula Ia wherein "a" is an optionally present bond and R⁴ if present is selected from the group consisting of -Br, -Cl, -CN, phenyl, -O-phenyl, -OCF₃, -OCH₃, -C(O)OH, -CH₃ and -C(O)CH₃.

Different embodiments for formula Ia compounds include the following: In a first embodiment R⁴ is present and is selected from the group consisting of

-Br, -Cl, -CN, -OCF₃, -OCH₃, -C(O)OH, -CH₃ and -C(O)CH₃.

In a second embodiment R⁴ is not present.

In a third embodiment "a" is not present and R⁴ is as provided in the general formula Ia above, or in the first or second embodiments. In a fourth embodiment, "a" is present and R⁴ is as provided in the general formula Ia above, or in the first or second embodiments.

In another aspect of the invention, the formula I compound has the following structure:

Formula Ib wherein "a" is an optionally present bond and R⁴ if present is selected from the group consisting of -Br, -Cl, -CN, phenyl, -O-phenyl, -OCF₃, -OCH₃, -C(O)OH, -CH₃ and -C(O)CH₃. Different embodiments for formula Ib compounds include the following:

In a first embodiment R⁴ is present and is selected from the group consisting of -Br, -Cl, -CN, -OCF₃, -OCH₃, -C(O)OH, -CH₃ and -C(O)CH₃.

In a second embodiment R⁴ is not present.

In a third embodiment "a" is not present and R is as provided in the general formula Ia above, or in the first or second embodiments.

In a fourth embodiment, "a" is present and R⁴ is as provided in the general formula Ia above, or in the first or second embodiments,

In another embodiment of the invention, the compound of the invention is a compound provided in Examples 1 through 84 shown below or a pharmaceutically acceptable salt thereof.

Other embodiments of the present invention include the following:

(a) A pharmaceutical composition comprising an effective amount of a compound of formula (I) and a pharmaceutically acceptable carrier. (b) The pharmaceutical composition of (a), further comprising a second therapeutic agent selected from the group consisting of HCV antiviral agents, immunomodulators, and anti-infective agents.

(c) The pharmaceutical composition of (b), wherein the HCV antiviral agent is an antiviral selected from the group consisting of HCV protease inhibitors and HCV NS 5 B polymerase inhibitors.

(d) A pharmaceutical combination which is (i) a compound of formula (I) and (ii) a second therapeutic agent selected from the group consisting of HCV antiviral agents, immunomodulators, and anti-infective agents; wherein the compound of formula (I) and the second therapeutic agent are each employed in an amount that renders the combination effective for inhibiting HCV NS 3 protease, or for treating HCV infection and/or reducing the likelihood or severity of symptoms of HCV infection.

(e) The combination of (d), wherein the HCV antiviral agent is an antiviral selected from the group consisting of HCV protease inhibitors and HCV NS5B polymerase inhibitors.

(f) A method of inhibiting HCV NS 3 protease in a subject in need thereof which comprises administering to the subject an effective amount of a compound of formula (I).

(g) A method of treating HCV infection and/or reducing the likelihood or severity of symptoms of HCV infection in a subject in need thereof which comprises administering to the subject an effective amount of a compound of formula (I).

(h) The method of (g), wherein the compound of formula (I) is administered in combination with an effective amount of at least one second therapeutic agent selected from the group consisting of HCV antiviral agents, immunomodulators, and anti-infective agents.

(i) The method of (h), wherein the HCV antiviral agent is an antiviral selected from the group consisting of HCV protease inhibitors and HCV NS5B polymerase inhibitors. t (j) A method of inhibiting HCV NS3 protease in a subject in need thereof which comprises administering to the subject the pharmaceutical composition of (a), (b), or (c) or the combination of (d) or (e). (k) A method of treating HCV infection and/or reducing the likelihood or severity of symptoms of HCV infection in a subject in need thereof which comprises administering to the subject the pharmaceutical composition of (a), (b), or (c) or the combination of (d) or (e).

In the embodiments of the compound provided above, it is to be understood that each embodiment maybe combined with one or more other embodiments, to the extent that such a combination provides a stable compound and is consistent with the description of the embodiments. It is further to be understood that the embodiments of compositions and methods provided as (a) through (k) above are understood to include all embodiments of the compounds, including such embodiments as result from combinations of embodiments.

The present invention also includes a compound of the present invention for use (i) in, (ii) as a medicament for, or (iii) in the preparation of a medicament for: (a) inhibiting HCV NS 3 protease, or (b) treating HCV infection and/or reducing the likelihood or severity of symptoms of HCV infection. In these uses, the compounds of the present invention can optionally be employed in combination with one or more second therapeutic agents selected from HCV antiviral agents, anti -infective agents, and immunomodulators. Additional embodiments of the invention include the pharmaceutical compositions, combinations and methods set forth in (a)-(k) above and the uses set forth in the preceding paragraph, wherein the compound of the present invention employed therein is a compound of one of the embodiments, aspects, classes, sub-classes, or features of the compounds described above. In all of these embodiments, the compound may optionally be used in the form of a pharmaceutically acceptable salt.

As used herein, all ranges are inclusive, and all sub-ranges are included within such ranges, although not necessarily explicitly set forth. In addition, the term "or," as used herein, denotes alternatives that may, where appropriate, be combined; that is, the term "or" includes each listed alternative separately as well as their combination if the combination is not mutually exclusive,

As used herein, the term "alkyl" refers to any linear or branched chain alkyl group having a number of carbon atoms in the specified range. Thus, for example, "C^e alkyl" (or "C₁-Ce alkyl") refers to all of the hexyl alkyl and pentyl alkyl isomers as well as n-, iso-, sec- and t-butyl, n- and isopropyl, ethyl and methyl. As another example, "C₁^ alkyl" refers to n-, iso-, sec- and t-butyl, n- and isopropyl, ethyl and methyl. Alkyl groups may be substituted as indicated. The term "halogenated" refers to a group or molecule in which a hydrogen atom has been replaced by a halogen. Similarly, the term "haloalkyl" refers to a halogenated alkyl group. The term "halogen" (or "halo") refers to atoms of fluorine, chlorine, bromine and iodine (alternatively referred to as fluoro, chloro, bromo, and iodo), The term "alkoxy" refers to an "alkyl-O-" group. Alkoxy groups may be substituted as indicated.

The term "alkenylene" refers to any linear or branched chain alkenylene group containing a double and having a number of carbon atoms in the specified range. Thus, for example, "-C_2-6 alkenylene -" refers to any of the C₂ to C₆ linear or branched alkenylene. Alkenylene groups may be substituted as indicated.

The term "alkylene" refers to any linear or branched chain alkylene group (or alternatively "alkanediyl") having a number of carbon atoms in the specified range. Thus, for example, "-C_1-6 alkylene-" refers to any of the C₁ to C₆ linear or branched alkylenes. Examples of alkylenes include -(CH₂)_1-6-, -(CH₂)_1-4-, -(CH₂)_1-3-, -(CH₂)_1-2-, -CH₂- and -CH(CH₃)-. Alkylene groups may be substituted as indicated.

The term "cycloalkyl" refers to any cyclic ring of an alkane or alkene having a number of carbon atoms in the specified range. Thus, for example, "C_3-8 cycloalkyl" (or "C₃-C₈ cycloalkyl") refers to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycϊoheptyl, and cyclooctyl. The term "cycloalkoxy" refers to a "cycloalkyl-O-" group. Cycloalkyl groups may be substituted as indicated.

The term "carbocycle" (and variations thereof such as "carbocyclic" or "carbocyclyl") as used herein, unless otherwise indicated, refers to (i) a C₃ to C₈ monocyclic, saturated or unsaturated ring or (ii) a C₇ to C₁₂ bicyclic saturated or unsaturated ring system. Each ring in (ii) is either independent of, or fused to, the other ring, and each ring is saturated or unsaturated. Carbocycle groups maybe substituted as indicated, for example with C_1-6 alkyl, C_2-6 alkenyl, C_2-6 alkynyl, aryl, halogen, -NH2 or -OH. The carbocycle may be attached to the rest of the molecule at any carbon atom which results in a stable compound. The fused bicyclic carbocycles are a subset of the carbocycles; i.e., the term "fused bicyclic carbocycle" generally refers to a C₇ to C₁₀ bicyclic ring system in which each ring is saturated or unsaturated and two adjacent carbon atoms are shared by each of the rings in the ring system. A fused bicyclic carbocycle in which both rings are saturated is a saturated bicyclic ring system. Saturated carbocyclic rings are also referred to as cycloalkyl rings, e.g., cyclopropyl, cyclobutyl, etc. A fused bicyclic carbocycle in which one or both rings are unsaturated is an unsaturated bicyclic ring system. A subset of the fused bicyclic unsaturated carbocycles are those bicyclic carbocycles in which one ring is a benzene ring and the other ring is saturated or unsaturated, with attachment via any carbon atom that results in a stable compound. Representative examples

of this subset include

, and

Depicted ring systems include, where appropriate, an indication of the variable to

which a particular ring atom is attached. For example, in the structure

, the variable R⁴ is shown as a floating variable which can be attached to any ring atom, provided that such attachment results in formation of a stable ring.

The term "aryl" refers to aromatic mono- and poly-carbocyclic ring systems, also referred to as "arenes," wherein the individual carbocyclic rings in the polyring systems are fused or attached to each other via a single bond. Suitable aryl groups include phenyl, naphthyl, and biphenylenyϊ. Aryl groups may be substituted as indicated. The terms "heteroaryl" and "heteroaromatic ring" refer to a stable 5- or

6-membered monocyclic aromatic ring, a stable 7- to 12-membered bicyclic ring system, or a stable 11- to 15-mernbered tricyclic ring system, which consists of carbon atoms and one or more heteroatoms selected from N, O and S. In the case of substituted heteraromatic rings containing at least one nitrogen atom (e.g., pyridine), such substitutions can be those resulting in N-oxide formation. Representative examples of heteroaromatic rings include pyridyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, thienyl (or thiophenyl), thiazolyl, furanyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isooxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, and thiadiazolyl. Unless otherwise specifically noted a particular group such as alkyl, cycloalkyl, aryl and heteroaryl groups are unsubstituted. In different embodiments the alkyl, cycloalkyl, aryl and heteroaryl groups are substituted with one to three substitutents selected from the group consisting of: halo, C₁-C₂₀ alkyl, -CF₃, -NH₂, -N(C₁-C₆ alkyl)₂, -NO₂, oxo, -CN, -N₃, -OH, -0(C₁-C₆ alkyl), C₃-C]₀ cycloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, (C_ϋ-C₆ alkyl) S(O)₀^₂-, aiyl- S(O)_0-2-, (C₀-C₆ alkyl)S(0)o.2(C₀-C₆ alkyl)-, (C₀-C₆ alkyl)C(O)NH-, H₂N-C(NH)-, -0(C₁-C₆ alkyl)CF₃, (C₀-C₆ alkyl)C(O)-, (C₀-C₆ alkyl)OC(O)-, (C₀-C_SaIkYl)O(C₁-C₆ alkyl)-, (C₀-C₆ alkyl)C(0)_1-2(C₀-C₆ alkyl)-, (C₀-C₆ alkyl)OC(O)NH-, aryl, aralkyl, heteroaryl, heterocyclylalkyl, halo-aryl, halo-aralkyl, halo-heteroaryl, halo-heterocyclylalkyl, cyano-aryl, cyano-aralkyl, cyano-heterocycle and cyano-heterocyclylalkyl.

Unless expressly stated to the contrary, all ranges cited herein are inclusive. For example, a heteroaryl ring described as containing from "1 to 3 heteroatoms" means the ring can contain 1, 2, or 3 heteroatoms. It is also to be understood that any range cited herein includes within its scope all of the sub-ranges within that range. The oxidized forms of the heteroatoms N and S are also included within the scope of the present invention.

When any variable (e.g. L) occurs more than one time in any constituent or in formula (I) or in any other formula depicting and describing compounds of the invention, its definition on each occurrence is independent of its definition at every other occurrence. Also, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

Unless expressly stated to the contrary, substitution by a named substituent is permitted on any atom in a ring (e.g., aryl or a heteroaryl ring) provided such ring substitution is chemically allowed and results in a stable compound. A "stable" compound is a compound which can be prepared and isolated and whose structure and properties remain or can be caused to remain essentially unchanged for a period of time sufficient to allow use of the compound for the purposes described herein (e.g., therapeutic or prophylactic administration to a subject).

As a result of the selection of substituents and substituent patterns, certain of the compounds of the present invention can have asymmetric centers and can occur as mixtures of stereoisomers, or as individual diastereomers, or enantiomers. All isomeric forms of these compounds, whether isolated or in mixtures, are within the scope of the present invention.

Reference to a compound also includes stable complexes of the compound such as a stable hydrate.

As would be recognized by one of ordinary skill in the art, certain of the compounds of the present invention can exist as tautomers. For the purposes of the present invention a reference to a compound of formula (I) is a reference to the compound per se, or to any one of its tautomers per se, or to mixtures of two or more tautomers. The compounds of the present inventions are useful in the inhibition of HCV protease (e.g., HCV NS3 protease) and the treatment of HCV infection and/or reduction of the likelihood or severity of symptoms of HCV infection. For example, the compounds of this invention are useful in treating infection by HCV after suspected past exposure to HCV by such means as blood transfusion, exchange of body fluids, bites, accidental needle stick, or exposure to patient blood during surgery.

The compounds of this invention are useful in the preparation and execution of screening assays for antiviral compounds. For example, the compounds of this invention are useful for isolating enzyme mutants, which are excellent screening tools for more powerful antiviral compounds. Furthermore, the compounds of this invention are useful in establishing or determining the binding site of other antivirals to HCV protease, e.g., by competitive inhibition. Thus, the compounds of this invention may be commercial products to be sold for these purposes.

The compounds of the present invention may be administered in the form of pharmaceutically acceptable salts. The term "pharmaceutically acceptable salt" refers to a salt which possesses the effectiveness of the parent compound and which is not biologically or otherwise undesirable (e.g., is neither toxic nor otherwise deleterious to the recipient thereof). Suitable salts include acid addition salts which may, for example, be formed by mixing a solution of the compound of the present invention with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, sulfuric acid, acetic acid, trifluoroacetic acid, or benzoic acid. Many of the compounds of the invention carry an acidic moiety, in which case suitable pharmaceutically acceptable salts thereof can include alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., calcium or magnesium salts), and salts formed with suitable organic ligands such as quaternary ammonium salts. Also, in the case of an acid (-COOH) or alcohol group being present, pharmaceutically acceptable esters can be employed to modify the solubility or hydrolysis characteristics of the compound.

The term "administration" and variants thereof (e.g., "administering" a compound) in reference to a compound of the invention mean providing the compound or a prodrug of the compound to the individual in need of treatment. When a compound of the invention or a prodrug thereof is provided in combination with one or more other active agents (e.g., antiviral agents useful for treating HCV infection), "administration" and its variants are each understood to include concurrent and sequential provision of the compound or salt and other agents.

As used herein, the term "prodrug" is intended to encompass an inactive drug form or compound that is converted into an active drag form or compound by the action of enzymes, chemicals or metabolic processes in the body of an individual to whom it is administered.

As used herein, the term "composition" is intended to encompass a product comprising the specified ingredients, as well as any product which results, directly or indirectly, from combining the specified ingredients, By "pharmaceutically acceptable" is meant that the ingredients of the pharmaceutical composition must be compatible with each other and not deleterious to the recipient thereof.

The term "subject" (alternatively referred to herein as "patient") as used herein refers to an animal, preferably a mammal, most preferably a human, who has been the object of treatment, observation or experiment.

The term "effective amount" as used herein means that amount of active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue, system, animal or human that is being sought by a researcher, veterinarian, medical doctor or other clinician. In one embodiment, the effective amount is a "therapeutically effective amount" for the alleviation of one or more symptoms of the disease or condition being treated. In another embodiment, the effective amount is a "prophylactically effective amount" for reduction of the severity or likelihood of one or more symptoms of the disease or condition. The term also includes herein the amount of active compound sufficient to inhibit HCV NS 3 protease and thereby elicit the response being sought (i.e., an "inhibition effective amount"). When the active compound (i.e., active ingredient) is administered as the salt, references to the amount of active ingredient are to the free acid or free base form of the compound.

For the purpose of inhibiting HCV NS 3 protease and treating HCV infection and/or reducing the likelihood or severity of symptoms of HCV infection, the compounds of the present invention, optionally in the form of a salt, can be administered by means that produces contact of the active agent with the agent's site of action. They can be administered by conventional means available for use in conjunction with pharmaceuticals, either as individual therapeutic agents or in a combination of therapeutic agents. They can be administered alone, but typically are administered with a pharmaceutical carrier selected on the basis of the chosen route of administration and standard pharmaceutical practice. The compounds of the invention can, for example, be administered by one or more of the following routes: orally, parenterally (including subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques), by inhalation (such as in a spray form), or rectally, in the form of a unit dosage of a pharmaceutical composition containing an effective amount of the compound and conventional non-toxic pharmaceutically-acceptable carriers, adjuvants and vehicles. Liquid preparations suitable for oral administration (e.g., suspensions, syrups, elixirs and the like) can be prepared according to techniques known in the art and can employ any of the usual media such as water, glycols, oils, alcohols and the like. Solid preparations suitable for oral administration (e.g., powders, pills, capsules and tablets) can be prepared according to techniques known in the art and can employ such solid excipients as starches, sugars, kaolin, lubricants, binders, disintegrating agents and the like. Parenteral compositions can be prepared according to techniques known in the art and typically employ sterile water as a carrier and optionally other ingredients, such as solubility aids. Injectable solutions can be prepared according to methods known in the art wherein the carrier comprises a saline solution, a glucose solution or a solution containing a mixture of saline and glucose. Further guidance for methods suitable for use in preparing pharmaceutical compositions of the present invention and of ingredients suitable for use in said compositions is provided in Remington's Pharmaceutical Sciences, 20^th edition (ed. A. R. Gennaro, Mack Publishing Co., 2000).

The compounds of this invention can be administered orally in a dosage range of 0.001 to 1000 mg/kg of mammal (e.g., human) body weight per day in a single dose or in divided doses. One dosage range is 0.01 to 500 mg/kg body weight per day orally in a single dose or in divided doses. Another dosage range is 0.1 to 100 mg/kg body weight per day orally in single or divided doses. For oral administration, the compositions can be provided in the form of tablets or capsules containing 1.0 to 500 mg of the active ingredient, particularly 1, 5, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, and 500 mg of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. The specific dose level and frequency of dosage for any particular patient may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy.

As noted above, the present invention also relates to a method of inhibiting HCV NS3 protease, inhibiting HCV replication, or preventing or treating HCV infection with a compound of the present invention in combination with one or more therapeutic agents and a pharmaceutical composition comprising a compound of the present invention and one or more therapeutic agents selected from the group consisting of a HCV antiviral agent, an immunomodulator, and an anti-infective agent. Such therapeutic agents active against HCV include ribavirin, levovirin, viramidine, thymosin alpha-1, interferon-β, interferon-α, pegylated interferon-α (peginterferon-α), a combination of interferon-α and ribavirin, a combination of peginterferon-α and ribavirin, a combination of interferon-α and levovirin, and a combination of peginterferon-α and levovirin. Interferon-α includes recombinant interferon-α2a (such as ROFERON interferon available from Hoffmann-LaRoche, Nutley, NJ), pegylated interferon-α2a (PEGASYS), interferon-α2b (such as INTRON-A interferon available from Schering Corp., Kenilworth, NJ), pegylated interferon-α2b (PEGINTRON), a recombinant consensus interferon (such as interferon alphacon-1), and a purified interferon-α product. Amgen's recombinant consensus interferon has the brand name INFERGEN. Levovirin is the L-enantiomer of ribavirin which has shown immunomodulatory activity similar to ribavirin, Viramidine represents an analog of ribavirin disclosed in WO 01/60379. The individual components of the combination can be administered separately at different times during the course of therapy or concurrently in divided or single combination forms.

For the treatment of HCV infection, the compounds of the present invention may also be administered in combination with an agent that is an inhibitor of HCV NS3 serine protease. HCV NS3 serine protease is an essential viral enzyme and has been described to be an excellent target for inhibition of HCV replication. Both substrate and non-substrate based inhibitors of HCV NS3 protease inhibitors are disclosed in International Patent Application Publications WO 98/22496, WO 98/46630, WO 99/07733, WO 99/07734, WO 99/38888, WO 99/50230, WO 99/64442, WO 00/09543, WO 00/59929, WO 02/48116 and WO 02/48172, British Patent No. GB 2 337 262, and U.S. Patent No. 6,323,180. Ribavirin, levovirin, and viramidine may exert their anti-HCV effects by modulating intracellular pools of guanine nucleotides via inhibition of the intracellular enzyme inosine monophosphate dehydrogenase (IMPDH). DVIPDH is the rate-limiting enzyme on the biosynthetic route in de novo guanine nucleotide biosynthesis. Ribavirin is readily phosphorylated intracellularly and the monophosphate derivative is an inhibitor of IMPDH. Thus, inhibition of EVIPDH represents another useful target for the discovery of inhibitors of HCV replication. Therefore, the compounds of the present invention may also be administered in combination with an inhibitor of IMPDH, such as VX-497, which is disclosed in International Patent Application Publications WO 97/41211 and WO 01/00622; another IMPDH inhibitor, such as that disclosed in WO 00/25780; or mycophenolate mofetil. See A.C. Allison and E.M. Eugui, 44 (Suppl.) Agents Action 165 (1993).

For the treatment of HCV infection, the compounds of the present invention may also be administered in combination with the antiviral agent amantadine (1 -aminoadamantane). For a comprehensive description of this agent, see J. Kirschbaum, 12 Anal. Profiles Drug Subs. 1-36 (1983).

For the treatment of HCV infection, the compounds of the present invention may also be administered in combination with the antiviral agent polymerase inhibitor R7128 (Roche).

The compounds of the present invention may also be combined for the treatment of HCV infection with antiviral 2'-C-branched ribonucleosides disclosed in R. E. Harry-O'Kuru et al., 62 J. Ore. Chem. 1754-59 (1997); M. S. Wolfe et al, 36 Tet. Lett. 7611-14 (1995); U.S. Patent No. 3,480,613; and International Patent Application Publications WO 01/90121, WO 01/92282, WO 02/32920, WO 04/002999, WO 04/003000 and WO 04/002422; the contents of each of which are incorporated by reference in their entirety. Such 2'-C-branched ribonucleosides include, but are not limited to, 2'-C-methyl-cytidine, 2'-C-methyl-uridine, 2'-C- methyl- adenosine, 2'-C-methyl-guanosine, and 9-(2-C-methyl-β-D-ribofuranosyl)-2,6- diaminopurine, and the corresponding amino acid ester of the ribose C-2', C-3', and C-5' hydroxyls and the corresponding optionally substituted cyclic 1,3 -propanediol esters of the 5'- phosphate derivatives.

The compounds of the present invention may also be combined for the treatment of HCV infection with other nucleosides having anti-HCV properties, such as those disclosed in International Patent Application Publications WO 02/51425, assigned to Mitsubishi Pharma Corp.; WO 01/79246, WO 02/32920, WO 02/48165 and WO2005/003147 (including Rl 656, (2'/?)-2'-deoxy-2'-iluoro-2'-C-methylcytidine, shown as compounds 3^6 on page 77); WO 01/68663; WO 99/43691; WO 02/18404 and WO2006/021341, and U.S. Patent Application Publication US 2005/0038240, including 4'-azido nucleosides such as Rl 626, 4'-azidocytidine; U.S. Patent Application Publications US 2002/0019363, US 2003/0236216, US 2004/0006007 and US 2004/0063658; and International Patent Application Publications WO 02/100415, WO 03/026589, WO 03/026675, WO 03/093290, WO 04/011478, WO 04/013300 and WO 04/028481 ; the content of each is incorporated herein by reference in its entirety.

For the treatment of HCV infection, the compounds of the present invention may also be administered in combination with an agent that is an inhibitor of HCV NS5B polymerase. Such HCV NS5B polymerase inhibitors that may be used as combination therapy include, but are not limited to, those disclosed in International Patent Application Publications WO 02/057287, WO 02/057425, WO 03/068244, WO 2004/000858, WO 04/003138 and WO 2004/007512; U.S. Patent No. 6,777,392 and U.S. Patent Application Publication US2004/0067901 ; the content of each is incorporated herein by reference in its entirety. Other such HCV polymerase inhibitors include, but are not limited to, valopicitabine (NM-283; Idenix) and 2'-F-2'-beta-methylcytidine (see also WO 2005/003147). In one embodiment, nucleoside HCV NS5B polymerase inhibitors that are used in combination with the present HCV NS3 protease inhibitors are selected from the following compounds: 4-amino-7-(2-C-methyl-β-D-arabinofuranosyl)-7H-pyrrolo[2,3-d]pyriniidine; 4- amino-7-(2-C-methyl-β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine; 4-methylamino-7-(2-C- methyl-β-D-riboruranosyl)-7H-pyrrolo[2,3-d]pyrimidine; 4-dimethylamino-7-(2-C-methyl-β-D- ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine; 4-cyclopropylamino-7-(2-C-methyl-β-D- ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine; 4-amino-7-(2-C-vinyl-β-D-ribofuranosyl)-7H- pyrrolo[2,3-d]pyrimidine; 4-amino-7-(2-C-hydroxymethyl-β-D-ribofuranosyl)-7H- pyrrolo[2,3-d]pyrimidine; 4-amino-7-(2-C-fluoromethyl-β-D-ribofuranosyl)-7H- pyrrolo[2,3-d]pyrimidine; 4-amino-5-methyl-7-(2-C-methyl-β-D-ribofιιranosyl)-7H- pyrrolo[2 , 3 -d\ pyrimi dine ; 4-amϊno-7 -(2-C-methy 1 - β -D-ribofuranosyl)-7H- pyrrolo[2,3-d]pyrimidine-5-carboxylic acid; 4-amino-5-bromo-7-(2-C-methyl-β-D- ribofuranosyl)-7H-pyrrolof2,3-d]pyrimidine; 4-amino-5-chloro-7-(2-C-methyl-β-D- ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine; 4-amino-5-fluoro-7-(2-C-methyl-β-D- ribofuranosyl)-7H-pyπ:olo[2,3-d]pyrimidine; 2_;4-diamino-7-(2-C-methyl»β-D--ribofuranosyl)- 7H-pyrrolo[2,3-d]pyrimidine; 2-amino-7-(2-C-methyl-β"-D-ribofuranosyl)-7H- pyrrolo[2 , 3-d]pyrimidine ; 2-amino-4-cyclopropyl amino-7-(2-C-methyI - β-D-ribofuranosyl)-7H- pyrrolo[2 , 3-d]pyrimidine ; 2- amino-7 -(2-C-methyl- β-D-ribofuranosy 1)-7H- pyrrolo[2,3-d]pyrimidin-4(3H)-one; 4-amino--7-(2-C-ethyl-β-D-ribofuranosyl)-7H- pyrrolo[23-d]pyriinidme; 4-amino-7-(2-C,2-0-dimethyl-β--D-riboftιranosyl)-7H- pyrrolo[2,3-d]pyrimidine; 7-(2-C-methyl-β-D-ribofiH-anosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)- one; 2-amino-5-methyl-7-(2-C, 2-0-dimethyl-β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin- 4(3H)-one; 4-amino-7-(3-deoxy-2-C-methyl-β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine; 4-amino-7-(3-deoxy-2-C-methyl-β-D-arabinofuranosyl)-7H-pyrrolo[2,3-d]pyrimidme; 4-amino- 2-fluoro-7-(2-C-methyl-β-D-ribofuranosyl)-7H-pyrrolo[23-d]pyriniidine; 4-amino-7-(3-C- methyl-β-D-ribofuranosyl)-7H-pyrτolo[2,3-d]pyrimidine; 4-amino-7-(3-C-methyl-β-D- xylofeanosyl)-7/i-pyiτolo[2,3-d]pyrimidine; 4-amino-7-(2,4-di-C-methyl-β--D-ribofuranosyl)- 7H-pyrrolo[2,3-d]pyrimidine; 4-ammo-7-(3-deoxy-3-fluoro-2-C-methyl-β-D-ribofuranosyl)-7H- pyrrolo[2,3-d]pyrimidine; and the corresponding 5 '-triphosphates; or a pharmaceutically acceptable salt thereof.

The compounds of the present invention may also be combined for the treatment of ΗCV infection with non-nucleoside inhibitors of ΗCV polymerase such as those disclosed in International Patent Application Publications WO 01/77091 ; WO 01/47883 ; WO 02/04425 ; WO 02/06246; WO 02/20497; WO 2005/016927 (in particular JTK003); the content of each is incorporated herein by reference in its entirety; and ΗCV-796 (Viropharma Inc.).

In one embodiment, non-nucleoside HCV NS5B polymerase inhibitors that are used in combination with the present HCV NS3 protease inhibitors are selected from the following compounds: 14-cyclohexyl-6-[2-(dimethylamino)ethyl]-7-oxo-5,6,7,8- tetrahydroindolo[2,1-α][2,5]benzodiazocine-l 1-carboxylic acid; 14-cyclohexyl-6-(2-morpholin- 4-ylethyl)-5,6,7,8-tetrahydroindolo[2,1-α][2,5]benzodiazocine-l 1-carboxylic acid; 14- cyclohexyl-6-[2-(dimethylammo)ethyl]-3-methoxy"5,6,7,8-tetrahydroindolo[2,1-α] [2,5]benzodiazocine-l 1-carboxylic acid; 14-cyclohexyl-3-methoxy-6-methyl-5,6,7,8- tetrahydroindolo[2,l-β][2,5]benzodiazocine-l 1-carboxylic acid; methyl ({[(14-cyclohexyl-3- methoxy-6-methyl-5,6,7₎8-tetrahydroindolo[2,1-α][2,5]benzodiazocin-l l- yl)carbonyl]amino}sulfonyl)acetate; ({[(14-cyclohexyl-3-methoxy-6-methyl-5,6,7,8- tetrahydroindolo[2,1-α][2,5]benzodiazocin-l l-yl)carbonyl]amino}sulfonyl)acetic acid; 14- cyclohexyl-7V^r-[(dimethylamino)sulfonyl]-3-methoxy-6-methyl-5,6,7,8-tetrahydroindolo[2,1-α] [2 ,5]benzodiazocine- 11 -carboxamide; 3-chloro-l 4-cyclohexyl-6-[2-(dimethylamino)ethy!]-7- oxo-5,6_j7,8-telrahydroindolo[2,1-α][2,5]benzodiazocine 11 -carboxyϊic acid; N-(l l-carboxy-14- cyclohexyl-7,8-dihydro-6H-indoIo[l ,2-e][ 1,5]benzoxazocin-7-yl)-N,N-dimethylethane-l ,2- diaminium bis(trifluoroacetate); 14-cyclohexyl-7,8-dihydro-6H-indolo[l ,2-e][l ,5] benzoxazocine-11-carboxylic acid; 14-cyclohexyl-6-methyl-7-oxo-5,6,7,8-tetrahydroindolo [2,1-α)[2,5]benzodiazocine-l 1-carboxylic acid; 14-cyclohexyl-3-methoxy-6-methyl-7-oxo- 5 ,ό,7,8-tetrahydroindolo[2, 1 -a] [2,5]benzodiazocine-l 1 -carboxylic acid; 14-cyclohexyl-6-[2- (dimetliylamino)ethyl]-3-niethoxy-7-oxo-5,6,7,8-''^:etrahydroindolo[2, 1 -α][2,5]benzodiazocine- 11 -carboxylic acid; 14-cyclohexyl-6-[3-(dimethylamino)propyl]-7-oxo-5,6,7,8-tetrahydroindolo [2,1-α][2,5]benzodiazocine-l 1-carboxylic acid; 14-cyclohexyl-7-oxo-6-(2-piρeridin-l-ylethyl)- 5,6,7,8-tetrahydroindolo[2,1-α][2,5]benzodiazocine-l 1-carboxylic acid; 14-cyclohexyl-6-(2- morpholin-4-ylethyl)-7-oxo-5,6,7,8"tetrahydroindolo[2, 1 -α][2,5]benzodiazocine- 11 -carboxylic acid; 14-cyclohexyl-6-[2-(diethylammo)ethyl]-7-oxo-5,6,7,8-tetrahydroindolo[2,l-c]

[2, 5]benzodiazocine-l 1-carboxylic acid; 14-cyclohexyl-6-(l-methylpiperidin-4-yl)-7-oxo- 5,6,7,8-tetrahydroindolo[2,1-α][2,5]benzodiazocine-l 1 -carboxylic acid; 14-cyclohexyl-N- [ (dimethyl amino)sulfonyl] -7-oxo-6-(2-piperidin- 1 -ylethyl)-5 ,6 ,7,8-tetrahydroindolo[2 , 1 -a] [2,5]benzodiazocine-l 1 -carboxamide; 14-cyclohexyl-6-[2-(dimethylamino)ethyl]-N- [(dimethylamino)sulfonyl]-7-oxo-5,6,7,8-tetrahydroindolo[2,1-α][2,5]benzodiazocine-l 1 - carboxamide; 14-cyclopentyl-6-[2-(dimethylammo)ethyl]-7-oxo-5,6,7,8"tetxahydroindolo[2,1-α] [2, 5]benzodiazocine-l 1-carboxylic acid; 14-cyclohexyl-5,6,7,8-tetrahydroindolo[2,1-α] [2,5]benzodiazocine-l 1-carboxylic acid; 6-allyl-14-cyclohexyl-3-methoxy-5,6,7,8- tetrab.ydroindolo[2,1-α][2,5]benzodiazocine-l 1-carboxylic acid; 14-cyclopentyl-6-[2- (dimethylamino)ethyl]-5,6,7,8-tetrahydroindolo[2 , 1-α] [2,5]benzodiazocine-l 1 -carboxylic acid; 14-cyclohexyl-6-[2-(dimethylamino)ethyl]-5,6,7,8-tetrahydroindolo[2,1-α][2,5]benzodiazocine- 1 1-carboxylic acid; 13-cyclohexyl-5-methyl-4,5,6,7-tetrahydrofuro[3',2':6,7][l ,4]diazocino[l ,8- α]indole- 10-carboxylic acid; 15-cyclohexyl-6-[2-(dimethylamino)ethyl]-7-oxo-6,7 ,8,9- tetrahydro-5/J-indolo[2,l-β][2,6]benzodiazonine-12-carboxylic acid; 15-cyclohexyl-8-oxo- 6,7,8,9-tetrahydro-5H-indolo[2,1-α][2,5]benzodiazomne-12-carboxylic acid; 13-cyclohexyl-6- oxo-6,7-dihydro-5H-indolo[1,2-d][1,4]benzodiazepine-l O-carboxylic acid; and pharmaceutically acceptable salts thereof.

The ΗCV NS3 protease inhibitory activity of the present compounds may be tested using assays known in the art. One such assay is ΗCV NS 3 protease time-resolved fluorescence (TRF) assay as described below and in International Patent Application Publication WO 2006/102087. Other examples of such assays are described in e.g., International Patent Application Publication WO 2005/046712. A NS3 protease assay can be performed, for example, in a final volume of 100 μl assay buffer containing 50 niM HEPES, pH 7.5, 150 mM NaCl, 15 % glycerol, 0.15 % TRITON X- 100, 10 mM DTT, and 0.1 % PEG 8000. NS3 and NS4A protease is pre-incubated with various concentrations of inhibitors in DMSO for 30 minutes. The reaction is initiated by adding the TRF peptide substrate (final concentration 100 nM). NS3 mediated hydrolysis of the substrate is quenched after 1 hour at room temperature with 100 μl of 500 mM MES, pH 5.5. Product fluorescence is detected using either a VICTOR V2 or FUSION fluorophotometer (Perkin Elmer Life and Analytical Sciences) with excitation at 340 nm and emission at 615 nm with a 400 μs delay. Testing concentrations of different enzyme forms are selected to result in a signal to background ratio (S/B) of 10-30. IC₅₀ values are derived using a standard four- parameter fit to the data. K_i values are derived from IC₅₀ values using the following formula,

IC₅₀ - Ki (I + [S] / KM), E_qn (I), where [S] is the concentration of substrate peptide in the reaction and KM is the Michaelis constant. See P. Gallinari et al., 38 BlOCHEM. 5620-32(1999); P. Gallinari et al., 72 J. VIROL. 6758-69 (1998); M. Taliani et al, 240 ANAL. BlOCHEM. 60-67 (1996).

The present invention also includes processes for making compounds of formula (I). The compounds of the present invention can be readily prepared according to the following reaction schemes and examples, or modifications thereof, using readily available starting materials, reagents and conventional synthesis procedures. In these reactions, it is also possible to make use of variants which are themselves known to those of ordinary skill in this art, but are not mentioned in greater detail Furthermore, other methods for preparing compounds of the invention will be readily apparent to the person of ordinary skill in the art in light of the following reaction schemes and examples. Unless otherwise indicated, all variables are as defined above. The following reaction schemes and examples serve only to illustrate the invention and its practice.

Olefin metathesis catalysts include the following Ruthenium based species: F. Miller et al., 118 J. AM. CHEM. Soc. 9606 (1996); G. Kingsbury et al., 121 J. Am. Chem. Soc. 791 (1999); H. Scholl et al., 1 ORG. LETT. 953 (1999); U.S. Patent Application Publication US2002/0107138; K. Furstner et al., 64 J. ORG. CHEM. 8275 (1999). The utility of these catalysts in ring closing metathesis is well known in the literature (e.g. Trnka and Grubbs, 34 ACC. CHEM. RES. 18 (2001).

The following examples serve only to illustrate the invention and its practice. The examples are not to be construed as limitations on the scope or spirit of the invention.

List of Abbreviations

BOC (also Boc) t-Butyloxycarbonyl

B(OMe)₃ Trimethyl borate

BOP Benzotriazole-1 -yl-oxy-tris-(dimethylamino)-phosphonium hexafluorophosphate Brosyl chloride 4-Bromophenyl sulfonylchloride tBuOH t-Butanol

BuLi Butyl lithium

CAN C eric ammonium nitrate

CDCl₃ Deuterio-trichloromethane

CDI N,N'-Carbonyl diimidazole

CH₃CN Acetonitrile mCPBA m-Chloroperbenzoic acid

Cs₂CO₃ Cesium carbonate

CuI Copper iodide

Cu(I)Br-SMe₂ Copper (I) bromide dimethyl sulfide complex

DABCO 1 ,4-diazabicyclo[2.2.2)octane

DBA (also dba) Dϊbenzylidene acetone

DBU 1,8-Diazabicyclo[5.4.0]undec-7-ene

DCC Dicyclohexylcarbodiimide

DCE Dichloroethane

DCM Dichloromethane

DEAD Diethyl azodicarboxylate

DIAD Diisopropyl azodicarboxylate

DIEA Diethylamine

DFA Diethylpropylamine

DIPEA Diisopropyl ethyl amine

DMAP 4-DimethyIammo pyridine

DMF Dimethylformamide

DMSO Dimethyl Sulfoxide

DPPF (also dppf) 1 , 1 ^r-bid(Diphenylphosphino)ferrocene

EDC iV-(3-Dimethylaminoρroρyl)-iV^J-ethylcarbodiimide

ESI Electrospray ionization

Et₂O Diethyl ether

EtOAc Ethyl Acetate

EtOH Ethanol

H₂ Hydrogen or hydrogen atomosphere

HATU 0-(7-Azabenzotriazol- 1 -yl)-N,N,N',N'-tetrainethyluronium hexafluorophosphate

HBr Hydrobromic acid

HCl Hydrochloric acid

HMPA Hex amethylphosphorami de

HOAc Acetic acid

HOAt 1 -Hydroxy-7-azabenzotriazole HOBT 1 -Hydroxy benzotriazole

H₂O Water

H₂O₂ Hydrogen peroxide

HPLC High performance liquid chromatography

I₂ Iodine

KHSO₄ Potassium bisulfate

K₂SO₄ Potassium sulfate

K₂CO₃ Potassium carbonate

KOH Potassium hydroxide

LAH Lithium aluminium hydride

LCMS High performance liquid chromatography - mass spectrometry

LiOH Lithium hydroxide

LiOH-H₂O Lithium hydroxide monohydrate

LRMS Low resolution mass spectrometry

Me₃Al Trimethylalumimum

MeLi Methyllithium

MeOH Methanol

MgSO₄ Magnesium Sulfate

MsCl Mesyl chloride

N₂ Nitrogen or nitrogen atomosphere

NH₄Cl Ammonium chloride

NH₄OH Ammonium hydroxide

NIe Norleucine

NMP N-Methyl pyrrolidinone

NaH Sodium hydride

NaHCO₃ Sodium hydrogen carbonate (sodium bicarbonate)

NaHSO₃ Sodium bisulfite

NaOH Sodium hydroxide

NaOMe Sodium methoxide

Na₂SO₃ Sodium sulfite

Na₂S₂O₃ Sodium thiosulfate

Na₂SO₄ Sodium sulfate (anhydrous) PCy₃ Tricyclohexyl phosphine

POBr Phosphoryl bromide

POBr₃ Phosphoryl tribromide

P₂O₅ phosphorus pentoxide (P₄O₁₀)

Pd/C Palladium on carbon

PhMe Toluene

PPh₃ Triphenylphosphine

RT Room temperature, approximately 25 C

Ru/C Ruthenium on carbon

SiO₂ Silica or silica gel

TBAF Tetrabutylammonium fluoride

TBTU O-Benzotriazol-l-yl-iV,7V,N^'',iV'-tetramethyluronium tetrafluoroborate

TEA Triethyl amine

TFA Trifluoroacetic acid

THF Tetrahydofuran

TJPSOTf Triisopropylsilyl triflate

TMSC3 Chlorotrimethyl silane

TsCl p-Toluenesulfonyl chloride

Zn(CN)₂ Zinc cyanide

Synthesis of Intermediates Intermediates A

Intermediate A3 : (1 R,2R)- 1 - Amino-N-(cyclopropylsulfbnyl)-2 -ethylcyclopropanecarboxamide hydrochloride

Step 1 ; t-Butyl (Y 1 R2R)- 1 - ([ (cyclopropylsulfonyl)aminolcarbonyl} -2- ethyl c yclopropyl)carb amate

A hydrogenation vessel was charged with a MeOH (1000 mL) slurry of ϊ-butyl (( IR,2S)- 1 - { [(cyclopropylsulfonyljamino] carbonyl } -2»vinylcyclopropyl)carbamate ( 164 g, 0.50 mol) (US 6,995,174) and 5% Ru/C (dry, 7.5 wt%, 12.4 g) and stirred. The vessel was placed under N₂ (20 psi) and vented to atmospheric pressure (3x) to remove residual oxygen. The vessel was then placed under H₂ (50 psi). After 20 hours, the vessel was vented to atmospheric pressure. The reaction slurry was then transferred out of the reaction vessel and filtered through SOLKA FLOK (34 g, wetted with 100 mL MeOH) to yield a clear, light brown solution. The SOLKA FLOK was rinsed with MeOH (200 mL x 2). The combined MeOH solutions were concentrated under reduced pressure to yield crude product as a white solid (153 g). The crude product was slurried in EtOAc (800 mL), wanned to 40°C and aged 30 minutes. The solution was then seeded, aged 30 minutes, and heptane (500 mL) was added via addition funnel over 30 minutes. The partially crystallized solid was cooled to RT and aged overnight, after which additional heptane (500 mL) was added. After 1 hour, additional heptane (250 mL) was added via addition funnel, and the white slurry aged for 1 hour. The solution was filtered, and the solid was rinsed with heptane/EtOAc (500 mL, 4:1) and dried under reduced pressure to give t-butyl ((1R,2R)-1 -{[(cyclopropylsulfonyl)amino] carbonyl}-2- ethylcyclopropyl)carbamate (125.9 g). Step 2: (IR2R)-1-Aniino-N-(cvclopropylsυlfonyl)-2-ethylcyclopropanecarboxamide hydrochloride (Intermediate A3)

A solution of the product from Step 1 (92 g, 0.28 mol) in DCM (1200 mL) was cooled to 0°C, and HCl was bubbled through the solution for 10 minutes. The cooling bath was then removed, and the reaction mixture stirred for 2 hours. N₂ was bubbled through the reaction mixture for 5 minutes, and the volatiles evaporated. The residue was azeotroped with DCM (3x) to give an off-white powder (75 g). LRMS (M+H)⁺ Calcd. = 233; found 233.

Intermediate A4 : Trans-4-pent-4-en- 1 - yltetrahvdrofuran-3 -ol

To a mixture of CuI (1.66 g, 8.71 mrnol) in THF (100 mL) at -5°C, a 0.5M solution of bromo(pent4-en-1-yl)magnesium (116 mL, 5.81 mmol) was added. The solution was stirred for 1 hour and cooled to -20°C. 3,6-Dioxabicyclo[3.1.0]hexane (5.0 g, 58.1 mmol) was added dropwise, and the reaction mixture was slowly warmed to RT and stirred for 15 hours. The reaction mixture was quenched with NH₄Cl(_aq) and extracted with Et₂O (3x). The combined organics were washed with H₂O and brine, dried over Na₂SO₄, filtered and concentrated. The crude product was purified on SO2 (gradient elution, 10-100% EtOAc/hexanes). ¹H NMR (500 MHz, CDCl₃) δ 5.83 - 5.75 (m, 1 H); 5.04 - 4.95 (m, 2 H); 4.14 - 4.07 (m, 3 H); 3.85 (m, 1 H); 3.70 (m, 1 H); 3.44 (m, 1 H); 2.07 (m, 3 H); 1.45 (m, 3 H) ppm.

Intermediate A5: Trans-2-pent-4--en-lylcyclopentanol

A solution of 5-bromopent-1-ene (11.81 mL, 100 mmol) in Et₂O (100 mL) was added to magnesium (2.43 g, 100 mmol) over 20 minutes. The resulting suspension was heated under reflux for 40 minutes, then cooled to 20°C, taken up in a syringe, and added dropwise at -5°C to a stirred suspension of CuI (3.17 g, 16.6 mmol) in THF (160 mL). The resulting solution was stirred for 30 minutes at -5°C, then cooled to -20°C. Cycϊopentene oxide (7.21 mL, 83 mmol) was added dropwise, and the resulting mixture was warmed to 20°C over 2 hours, then stirred for 48 hours. The reaction was quenched by addition of NHUClfaq_.); then the layers were separated, and the aqueous layer was extracted with Et₂O. The combined organic phases were washed with H₂O and brine, then dried over Na₂SO₄. Filtration and removal of the volatiles gave a residue that was purified by column chromatography on SiO₂ (gradient elution, 1-100% EtOAc/petroleum ether) to afford the title compound (7.92 g, 62%) as a liquid, ¹H NMR (400 MHz, CDCl₃) δ 5.88-5.76 (m, 1H), 5.01 (d, J = 17.2 Hz, 1H), 4.95 (d, J - 10.6 Hz, 1H), 3.83 (br S₃ 1H), 2.12-2.00 (m, 2H), 1.99-1.84 (m, 2H), 1.76-1.30 (m, 7H)₃ 1.24-1.11 (m, 2H).

Intermediate A6: Trans-2-but-3-en-lylcyclopentanol

The title compound was prepared in a similar manner to Intermediate A5, using but-3-enylmagnesium bromide.

Intermediate A7: Trans-2-allyl-lylcyclopentanol

The title compound was prepared in a similar manner to Intermediate A5, usin| allylmagnesium bromide.

Intermediate A8: (1R,2R)-2-pent-4-en-1-ylcyclopentyl acetate

AMANO LIPASE PS (7.0 g, 64.7 mmol) was added to a solution of ^rαrø-2-pent-

4-en-1-ylcyclopentanol (10.0 g, 64.7 mmol) and vinyl acetate (19.5 g, 129.4 mmol) in Et₂O (275 mL). The mixture was stirred for 16 hours, then filtered through CELITE. The filtrate was concentrated to afford a residue that was purified by column chromatography on SiO₂ (gradient elution, 0-100% Et₂θ/petroleum ether) to afford in the first fractions the title compound (5.43 g, 43%). ¹H NMR (300 MHz, CDCl₃) δ 5.89-5.72 (m, 1H), 5.00 (d, J= 18.1 Hz, 1H), 4.95 (d, J = 11.0 Hz, 1H), 4.82-4.73 (m, 1H)₃ 2.11-1.98 (m, 2H)₃ 2.03 (s, 3H), 1.98-1.85 (m, 3H), 1.71-1.60 (m, 3H), 1.504.35 (in, 3H), 1.29-1.14 (m, 2H); [α]_D - -36.1 (c=0.73 in CHCl₃). The later fractions contained enantio-enriched (15',2S)-2-pent-4-en-1-ylcyclopentanol.

Intermediate A9: (li?,2jg)-2-pent-4-en- 1 -ylcyclopentanol

A stirred solution of (l/ϊ,2J?)-2-pent-4-en-1-ylcyclopentyl acetate (3,79 g, 19.3 iranol) in MeOH (320 ml) was treated with methanolic NaOMe (25%, 8.1 ml, 35.4 mmol) and stirred for 15 hours at 20°C. DOWEX 50WX8-100 ion-exchange resin (washed with MeOH) was added portionwise until the pH was neutral, then the mixture was filtered through CELITE. The filtrate was concentrated in vacuo, and the residue was partitioned between

EtOAc and H₂O. The organic layer was separated, washed with brine, and dried over Na₂SO₄. Filtration and removal of the volatiles afforded the title compound (2.61 g, 88%) as a liquid that was used directly in the subsequent reactions. [α]D -37.3 (c - 0.65, CHCl₃).

Intermediate AlO flJ?.2fiV2-but-3-en-1-ylcvclopentanol

The title compound was prepared from Intermediate A6 using the method described for the preparation of Intermediate A9.

Intermediate Al l : πJg.2jgV2-allvl-1-ylcvclopentanol

The title compound was prepared from Intermediate A7 using the method described for the preparation of Intermediate A9. Intermedi ate A 12 : 2-but-3 -en- 1 -ylcyclppropanol

A solution of 1.0 M hexenylmagnesium bromide (113 ml, 113 mmol) was added slowly to a mixture of methyl formate (3.53 ml, 56.5 mmol) and chlorotitanium trisisopropoxide (56.5 ml, 56.5 mmol) in THF (210 mL) at -78°C. The reaction was allowed to stir at -78°C for 10 min and then allowed to warm to -40°C and age 2 hrs. The reaction was allowed to warm to room temperature and then warmed to 40°C for 1 hr. The reaction was quenched into ice cold 10% sulfuric acid (500 mL). The product was extracted with diethyl ether (3X 100 mL) washed with saturated NaHCO_{3(aq )}, brine, dried over magnesium sulfate, filtered and concentrated to give ~15 gm oil. The oil was chromatographed on silica using 5-15% ethyl acetate/hexane to afford the title compound (1.0 g, 8 %) as an oil ¹H NMR (400 MHz, CDCl₃) δ 5.80 (m, 1H), 5.02 (d, J = 17.2 Hz, 1H), 4.95 (d, J = 10.6 Hz, 1H), 3.21 (m, 1H), 2.20-2.00 (m, 2H), 1.88 (s, 1H), 1.45-1.15 (m, 4H), 0.92 (m, 1H), 0.70 (m, 1H), 0.36 (m, 1H).

Intermediates B

Intermediate Bl : (2,SVCvclopentyl|Y {[( li?,2i?)-2-pent-4-en-1-ylcvcloρentvlloxv)carbonyl) amino] acetic acid

Step. l : Methyl (25^r)-cyclopentyl(isocyanato)acetate

A suspension of methyl (25)-amino(cyclopentyl)acetate hydrochloride (3.21 g, 16.57 mmol) in DCM (69 mL) and saturated NaHCO_3(aq > (132 mL) was cooled to 0°C and treated with triphosgene (2.21 g, 7.46 mmol). The mixture was stirred at 0°C for 3 hours, then warmed to 20°C and diluted with DCM. The layers were separated, and the aqueous phase was re-extracted with DCM. The combined organic phases were washed with brine and dried over Na₂SO₄. Filtration and removal of the volatiles afforded the title compound (2.95 g, 97%) as an oily solid that was used directly in subsequent steps. ¹H NMR (400 MHz₃ DMSO-J₆) δ 4.36 (d, J= 4.8 Hz, 1H), 3.73 (s, 1H), 2.35-2.24 (m, 1H), 1.76-1.24 (m, SH).

Step 2: Methyl (2_i5^f)-cvclopentyl[(i{^'πi?.2/gV2-ρent-4-en-1-vlcvclot)entyl]oxylcarbonvl) amino] acetate

A solution of methyl (2_*S)-cyclopentyl(isocyanato)acetate (1.57 g, 8.56 mmol) and (lJ?,2Λ)-2-pent-4-en-1-ylcycloρentanol (Intermediate A9)(1.20 g, 7.78 mmol) in PhMe (56 mL) was treated portionwise with DMAP (0.95 g, 7.78 mmol). The resulting mixture was stirred for 5 hours at 85°C, then cooled to 20°C, and diluted with EtOAc and HCl_(aq,_} (IN). The organic layer was separated, washed with brine, and dried over Na₂SO₄. Filtration and removal of the volatiles gave a residue that was purified by column chromatography (gradient elution, 4-40% Et₂O/petroleum ether) to afford the title compound (1.97 g, 76%) as an oil LCMS (ES+) m/z 338 (M+H)⁺.

Step 3 : (2S)- Cyclop entyl [ ( { [ ( 1 R2R)-2 -p ent-4-en- 1 -yl cyclopent ylj oxy } carbonyl)amino] ac.eti c acid

A mixture of methyl (25)-cyclopentyl[({[(li?,2Λ)»2-pent-4-en-1- ylcyclopentyl]oxy}carbonyl)amino]acetate (1.97 g, 5.84 mmol) and LiOH¹H₂O (0.74 g_f 17.51 mmol) in a 1 :1 mixture of THF:H₂θ (60 mL) was heated to 40°C. The solution was stirred for 4 hours, then cooled to 20°C. The THF was removed under reduced pressure, and the residual aqueous solution was extracted with EtOAc. The organic phase was washed with H₂O and brine, then dried over Na₂SO₄. Filtration and removal of the volatiles afforded the title compound (1.85g, 98 %) as an oil that was used directly in subsequent steps. ¹H NMR

(400 MHz, DMS(W₆) δ 12.25 (br s, 1H), 7.28 (d, J= 8.1 Hz, 1H), 5.84-5.71 (m, 1H), 4.98 (d, J = 17.4 Hz₃ 1H), 4.92 (d, J=ICl Hz, 1H), 4.60-4.52 (m, 1H), 3.78 (t, J= 8.0 Hz, 1H), 2.18-2.05 (m, 1H), 2.04-1.94 (m, 3H), 1.91-1.74 (m, 3H), 1.70-1.06 (m, 15 H). Intermediate B2: ^^-[({[(lJU^.Sj-g-allylcyclopentvljoxvlcarbonynaminolfcyclohexyl)acetic acid

Step 1 : Methyl (2S)-[({[^"f ljf?,2jy)-2-allylcyclopentyl]oxy}carbonyl)amino](cyclohexyl)acetate

NjN'-disuccininiidyl carbonate (8.28 g, 32.3 mmol) and triethylamine (4.51 ml, 32.3 mmol) were added to a solution of Intermediate Al 1 (3.4 g, 26.9 mmol) in acetonitrile (70 ml) and left stir under a N₂ atmosphere for 6 hours. Methyl (2S)-amino(cyclohexyl)acetate hydrochloride (8.39 g, 40.4 mmol) and triethylamine (7.51 ml, 53.9 mmol) were added to the reaction mixture and left stir under a N₂ atmosphere for 12 hours. Reaction was concentrated in vacuo, diluted with EtOAc, and washed organics with IN HCl₃ brine, sat'd NaHCθ₃, and brine. The organics were dried over Na₂SO₄, filtered, and removal of the volatiles under reduced pressure gave a residue that was purified by column chromatography (gradient elution, 10-30% EtOAc/Hexane) to afford the title compound (5.2 g, 60%) as an oil. LCMS (ES+) m/z 324.08 (M+H)⁺.

Step 2: (2S)-\({\( lJ?,2₁S^f)-2-allylcvclopentyl]oxy}carbonyl)amino](cyclohexyl)acetic acid

LiOH*H₂O (2.02 g, 48.2 mmol) was added to a solution of the product from Intermediate B2, Step 1 (5.2 g, 16.1 mmol) in a 2:1:1 mixture of THF:MeOH:H₂O (100 mJL) and stirred for 12 hours. The organics were removed under reduced pressure; the residual aqueous solution was acidified with dilute HCl and extracted with EtOAc. The organic phase was washed with brine and dried over Na₂SO₄. Filtration and removal of the volatiles afforded the title compound (4.9g, 100 %) as an oil that was used directly in subsequent steps. ¹H NMR (400 MHz, CDCl₃): δ 5.77 (ddt, J - 17.0, 10.2, 6.8 Hz, 1 H); 5.08 (d, J = 8.8 Hz, 1 H); 5.07-4.91 (m, 2 H); 4.75 (s, 1 H); 4.30 (dd, J = 9.0, 4.9 Hz, 1 H); 2.27-2.19 (m, 1 H); 2.05-1.71 (m, 10 H); 1.81-1.48 (m, 6 H); 1.36-1.04 (m, 8 H). Intermediate B3: Methyl (2S)-Ff {rfl/J.2.SV2-allvlcyclopentylloxy>carbonyl>amino1 ( c yclopentyl) acetate

The title compound was prepared from methyl (2S)-amino(cyclopentyl)acetate hydrochloride and Intermediate Al 1 using the method described for the preparation of Intermediate B2. ¹H NMR (400 MHz, CDCl₃): δ 5.85-5.71 (m, 1 H); 5.08 (d, J - 8.7 Hz, 1 H); 5.07-4.93 (m, 2 H); 4.75 (s, 1 H); 4.35-4.27 (m, 1 H); 2.31-2.19 (m, 2 H); 2.10-1.75 (m, 4 H); 1.93-1.59 (m, 2 H); 1.92-1.33 (m, 8 H); 1.49-1.22 (m, 3 H).

Intermediate B4: Methyl N-(IFf li?.,2S)-2-allvlcvclopentvlloxy)carbonyl)-3-metfavl-L-valinate

The title compound was prepared from methyl 3-methyI-L-valinate hydrochloride and Intermediate Al 1 using the method described for the preparation of Intermediate B2. ¹H NMR (400 MHz, CDCl₃): δ 5.83-5.71 (m, 1 H); 5.17 (d, J = 9.7 Hz, 1 H); 5.04 (s, 1 H); 5.03- 4.91 (m, 1 H); 4.74 (s, 1 H); 4.19 (d, J = 9.5 Hz, 1 H); 2.32-2.14 (m, 1 H); 2.02-1.82 (m, 4 H); 1.68-1.61 (m, 3 H); 1.35-1.13 (m, 2 H); 1.03 (s, 9 H).

Intermediate B5 : N- { [(2-but-3 -en- 1 -ylcyclopentyl)oxy] carbonyl } -3-methyl-L- valine

The title compound was prepared from methyl 3-methyl-L-valinate hydrochloride and Intermediate A6 using the method described for the preparation of Intermediate B2. ¹H NMR (400 MHz, CDCl₃) δ 5.80 (m, 1H), 5.17 (d, J- 8.9 Hz, 1H), 5.00 (d, J = 17.1 Hz, 1H), 4.94 (d, J = 10.6 Hz₃ 1H), 4.72 (brs, 1H), 4.19 (d, J= 8.9 Hz, 1H), 2.08 (m, 2H), 1.92 (m, 3H), 1.64 (m, 4H), 1.38-1.12 (m, 2H), 1.02 (s, 9H). Intermediate B6: (2S)-CIf (2-but-3 -en- 1 -ylcyclppropy^oxy] carbonyl } amino) (cyclopentyl)acetic acid

The title compound was prepared from methyl (2 S)-amino(cyclopentyl) acetate hydrochloride and Intermediate Al 2 using the method described for the preparation of

Intermediate Bl. ¹H NMR (400 MHz, CDCl₃) δ 5.82 (rn, 1H), 5.20 (m, 1H), 5.02 (d, J = 17.1 Hz, 1H), 4.95 (d, J - 10.6 Hz, 1H), 4.40-3.60 (m, 4H), 2.404.50 (m, 8H), 1.504.20 (m, 4H), 1.02 (in, 1H), 0.82 (m, 1H), 0.51 (m, 1H).

Intermediate B7 : (2SHf ( Ff 1 R.2S)-2-but-3 -en- 1 -ylcyclopentyl] oxy } carbonypamino]

(cyclopentyl)acetic acid

The title compound was prepared from methyl (2S)-amino(cyclopentyl)acetate hydrochloride and Intermediate AlO using the method described for the preparation of Intermediate Bl. LCMS (ES+) m/z 310.0 (M+H)+.

Intermediates C

Intermediate C 1 : 2-But-3-en- 1 -ylisoquinoline- 1 ,3(2H,4H)-dione

Homophthalϊc anhydride (1.5 g, 9.25 mmol), 3-buten-l -amine (1.129 ml, 11.10 mmol), and 4A molecular sieves (300mg) were combined in toluene (15 ml) and heated to 160°C in a microwave for 40 minutes. The reaction mixture was diluted with CH₂Cl₂ and the slurry was filtered over CELITE. The filtrate was concentrated in vacuo and the resulting residue was purified by column chromatography (gradient elution, 10-20% EtOAc/Hexane) to afford the title compound (0.75 g, 36%) as an oil. LCMS (ES+) m/z 216.03 (M+H)+.

Intermediate C2: 2-But-3-en- 1 -yl-6-methoxyisoquinoline-l ,3f2H,4HVdione

Step 3 : 2-(Carboxymethyl)-4-metfaoxybenzoic acid

A solution of diisopropylamine (3.43 ml, 24.07 mmol) in THF (3mL) was cooled to -78°C and n-butyllithium (9,63 ml, 24.07 mmol) was added dropwise and allowed the solution to warm to 0°C for 5 min and then re-cooled to -78°C. 4-methoxy-2-methylbenzoic acid (1 g, 6.02 mmol) and dimethyl carbonate (1.013 ml, 12.04 mmol) in THF (3mL) was added dropwise. After the addition is complete, the reaction is allowed to slowly warm to RT and stir for 4h. The reaction was quenched with 10 mL of H₂O and allowed to stir for 16h. The layers were separated, washed the organics with H₂O (2x), combined all the aqueous extractions and adjusted the pH to 2-3 with 6N HCl while in an ice bath. The resulting in a white precipitate was filtered and dried under vacuum to afford the title compound (1.09g, 86%) as a white solid. LCMS (ES+) m/z 211.05 (M+H)⁺.

Step 2: 2-But-3-en-l -yl-6-methoxyisoquinoline-l ,3(2H,4H)-dione The title compound was prepared according to the procedure for Intermediate CI, using 2-(carboxymethyl)-4-methoxybenzoic acid in place of homophthalic anhydride. LCMS (ES+) m/z 246.04 (M+H)⁺. Intermediate C3 : 2-but-3-en- 1 -yl-ό-chloroisoquinoline- 1 ,3(2H,4H)-dio:ne

The title compound was prepared according to the procedure for Intermediate C2, using 2-(carboxymethyl)-4-chlorobenzoic acid. LCMS (ES+) m/z 250.01 (M+H)⁺.

Intermediate C4: 2-but-3-en-l -yI-7-fluoroisoquinoline-L3f2H,4H)-dione

The title compound was prepared according to the procedure for Intermediate C2, using 2-(carboxymethyl)-5-fluorobenzoic acid. LCMS (ES+) m/z 234.08 (M+H)⁺.

Intermediate C5: 2-allylisoquinoline4,3f2H,4H)-dione

The title compound was prepared according to the procedure for Intermediate Cl , using proρ-2-en- 1 -amine in place of 3-buten- 1 -amine. LCMS (ES+) rn/z 202.04 (M+H)^÷.

Intermediates D

Intermediate Dl : 3-aHvlquinazoline-2,4(1H,3H)-dione

The title compound was prepared according to Chern, Ji-Wang, J. Heterocyclic Chem., 27, 1467 (1990). Intermediate D2: 3-Allyl-7-bromoquinazoline-2,4πH,3H)-dione

Step 1: 4-Bromo-2-[(emoxycarbonyl)ammo]benzoic acid

Elhyl chloroformale (5.00 ml, 52.1 mmol) was added to a solution of 2-amino-4- bromobenzoic acid (3,75 g, 17.36 mmol) in THF (70 ml) at room temperature. The mixture was heated to 70°C and let stir for 24 hrs. The reaction was concentrated Io give an oily solid. The resulting solid was diluted with toluene (50 mL), concentrated and then triturated with 5% ether/hexanes (10OmL) to give upon filtration and subsequent washing of the solid with hexane the title compound (4.6 gm, 92% ) as a solid. LCMS (ES⁺) m/z 287.88 (M+H)⁺.

Step 2: 3-Myl-7-bromoouinazoIine-2,4f1H,3Hydione

BOP (8.47 g, 19.16 mmol) was added to a solution of 4-bromo-2-

[(ethoxycarbonyl)amino]benzoic acid (4.6 g, 15.97 mmol), N-methylmorpholine (3.51 ml, 31.9 mmol) and allylamine (1.434 ml, 19.16 mmol) in DMF (50 ml) at room temperature. The mixture was allowed to stir at room temperature for 4 hrs. DBU (24.07 ml, 160 mmol) was added to the reaction and the temperature was raised to 60°C. The mixture let was allowed to stir at 60°C for Ihr. The mixture was quenched into 0.5 N HCL (20OmL), and the pH was adjusted to 1.5 with 3N HCL. Thick white solids were observed. The reaction was diluted to -400 mL with water let stir 10 min and solids filtered. The solids were washed with 0.5 N HCL (50 mL) and then slurry washed with water and sucked dry overnight to give the title compound (4.5 gm, 100% ) as a solid. . LCMS (ES⁺) m/z 280.87 (M+H)⁺. Intermediate D3: 3-Allyl-7-chlorooquinazolin&-2,4(1H.3H)-dione

The title compound was prepared utilizing a similar procedure for Intermediate D2, using 2-amino-4-chIorobenzoic acid. LCMS (ES⁺) m/z 236.98 (M+H)⁺.

Intermediate D4: 3-Allyl-7-(^'trifluoromethyl)qtιinazoline-2,4(1H,3//)-dione

The title compound was prepared utilizing a similar procedure for Intermediate D2, using 2-amino-4-trifluoromethylbenzoic acid. LCMS (ES⁺) m/z 271.25 (M+H)⁺.

Intermediate D5 : 3-Allyl-7-(methylsulfonyl)quinazoline-2,4(1H,3H)-dione

The title compound was prepared utilizing a similar procedure for Intermediate D2, using 2-amino-4-methylsulfonylbenzoic acid. LCMS (ES⁺) m/z 281.3 (M+H)⁺.

Intermediate D6: 3-Allyl-7-(meth.oxγ)qυinazoline-2,4f \H3H)-άϊone

The title compound was prepared utilizing a similar procedure for intermediate D2, using 2-amino-4-methoxybenzoic acid. LCMS (ES⁺) m/z 233.1 (M+H)⁺. Intermediate D7: 3-Allyl-8-methoxyquinazoline-2,4(TH,3H)-dione

The title compound was prepared utilizing a similar procedure for Intermediate D2, using 2-amino-3-methoxybenzoic acid. LCMS (ES⁺) m/z 233.3 (M+H)⁺.

Intermediate D8: 3-Allyl-5-methoxyquinazoline-2,4(l/1,3H)-dione

The title compound was prepared utilizing a similar procedure for Intermediate D2, using 2-amino-ό-methoxybenzoic acid. LCMS (ES⁺) m/z 233.02 (M+H)⁺.

Intermediate D9; 3-Aliyl-2-hydroχy-4H-ρyrido[1,2-a]ρyrimidin-4-one

2-aminoρyridine (996 mg, 10.58 mmol) and diethyl allylmalonate (2098 μl, 10.58 mmol) were combined and heated to 160°C neat for 9 hours. The brown solution solidified upon cooling to room temperature. The solid was filtered and washed with Et₂O to yield the title compound as a tan solid (580 mg, 27%) LCMS (ES+) m/z 203.02 (M+H)+.

Intermediate DlO: 3-allyl-2-hvdroxy-8-methoxy-4H-pyrido[ 1 ,2-a1pyrimidin-4-one

Bis(pentachlorophenyl) allylmalonate (905 mg, 1.412 mmol) and triethylamine

(0.359 ml, 2.58 mmol) were added to a slurry of 2-amino-4-methoxypyridine (160 mg, 1.289 mmol) in acetone (6.5 ml). The slurry was stirred for 16 hours, filtered, and washed with acetone to yield the title compound as a white solid (194 mg, 65%). LCMS (ES⁺) m/z 233.0

(M+H)⁺.

Example 1: (3a^J^lQ^12-R,23g,25a^-7-cvclopentyl-N-(fli?,2^-1- {[(cycloρrgpylsulfonyl)amino3carbonyl}-2-vinylcyclopropyl)-5,8J9-trioxo- L2,3,3a,5_>6J,8.11J2,21.22,25,25a-tetradecahvdro40H.19H-9J2- methanocyclopentaf 18, 19][ ϊ , 10,3,6, ϊ 2]dipxatriazacyclononadecinoj^' 12 ,11 -bjisoquinoline- 10- carboxamide

Step 1 : 1 -tert-Butyl 2-methvl f2ff.4R)-4-|Y2-but-3-en-l -yl-1 -oxo- 1 ,2-dihydroisoqumolin-3- yl)oxy]pyrrolidine- 1 ,2-dicarboxylate

Cs₂CO₃ (1.1 g, 3.39 mmol) was added to a solution of 2-but-3-en-1- ylisoquinoline-1,3(2H,4H)-dione (0.49 g, 2.26 mmol) and 1-t-butyl 2-methyl (25,46)-4-{[(4- bromophenyljsulfonyljoxylpyrrolidine-l^-dicarboxylate (1.05 g, 226 mmol) in NMP (10 mL), and the reaction mixture was stirred for 16 hours at 40°C. An additional portion of ϊ-t-butyl 2- methyl (25¹ ₃4_J5)-4-{[(4-bromophenyl)sulfonyl]oxy}pyrrolidine-1,2-dicarboxylate (1.0 g, 2.16 mmol) was added, and the reaction mixture was stirred at 40°C for 16 hours. The reaction mixture was cooled and poured onto a mixture of EtOAc and H₂O, and the layers were separated. The organic layer was washed with H₂O (2x), IN HCl (1 x) and brine, dried over Na₂SO₄, filtered and concentrated in vacuo. The resulting residue was purified by column chromatography (gradient elution, 10-40% EtOAc / Hexane) to afford the title compound (0.83 g, 83%) as an oil. LCMS (ES+) m/z 443.0 (M+H)⁺. Step 2: Methyl (^'4J?)-4-r(2-but-3-en-1-yl-1-oxo-1,2-dihydroisoqmnplin-3-yl>oxyl-L-prolinate

A portion of the product from Step 1 (150 mg, 0.34 mmol) was dissolved in CH₂Cl₂ (4 ml), cooled in an ice bath, and then added TFA (4 ml). The reaction was allowed to warm to room temperature and stir for 1.5 hours. The reaction was concentrated and the resulting oil was partitioned between CH₂Cl₂ and sat'd NaHCO3, extracted with CH₂Cl₂ (2x), combined organics, washed with brine, dried organics over Na₂SO₄, filtered, concentrated in vacuo to yield the title compound as a oil (113 mg, 97%). LCMS (ES+) mlz 342.9 (M+H)⁺.

Step 3: Methyl C4i?)-1-{(26^f)-2-Ffl^lJ?,2^-2-allvIcvclopentvl]oxy)carborjyl)amino1-2- cyclopentylacetyl)-4-[(2-but-3-en-1-yl-1-oxo-L2-dihydrpisoqιiinolin-3-yl)oxyl-L-prolinate

Intermediate B3 (356 mg, 1.2 mmol), HATU (424 mg, 1.115 mmol), and DIEA (0.487 ml, 2.79 mmol) were added to a solution of the product from Step 2 (31 S mg, 0.929 mmol), in DMF (8 ml). After stirring 16 hours, the reaction mixture was diluted with EtOAc and washed with IN HCl (2x), brine, sat'd NaHCO₃ (2x), brine. The organics were dried over Na₂SO₄, filtered, concentrated in vacuo, and the resulting residue was purified by column chromatography (gradient elution, 15-50% EtOAc/Hexane) to afford the title compound (0.276 g, 48%) as an oil. LCMS (ES+) m/z 620.2 (M+H)⁺.

Step 4: Methyl (SaRJS, 1 OSΛ 2&23&25a5V7-cvclopentyl-5,8.19-trioxo- 1 ,2,3 ,3a,5,6.7,8, 1 U 2,21 ,22,25,25a-tetradecahvdro- 1 OH, 19H-9 J 2- me thanpcvclopentaf 18 J 911- 1 ,10 ,3 ,6 J 2]dioxatriazacyclononadecino [ 1241 -bϋsoquinoline- 10- carboxylate

To a degassed solution of the product from Step 3 (390 mg, 0.63 mmol) in DCE (100 mL) was added p-benzoquinone (20.4 mg, 0.19 mmol) and Zhan Ib catalyst (73 mg, 0.094 mmol). Afterlό hours, the reaction was concentrated in vacuo and the crude product was purified by column chromatography (gradient elution, 15-40% EtOAc/hexanes) to yield the title compound as a gray foam (215 mg, 53%). LCMS (ES+) m/z 592.1 (M+H)⁺.

Step 5: OaRJSΛ0SΛ2R23E25aS)-7-cγc\θOQntγl-5$Λ9-tήoxo- 1.2.3.3a.5.6.7,8,l l,12.21^2,25.25a-tetradecahvdiO-10H,19H-9.12- methanocyclopentaf 18 , 19] [ 1 , 10,3 ,6, 12] dioxatriazacyclononadecino[ 12,11 -b]isoquinoline- 10- carboxylic acid

Lithium hydroxide monohydrate (5.77 μl, 0.208 mmol) was added to a solution of a portion of the product from Step 4 (40 mg, 0.068 mmol) in THF:MeOH:H₂O 2: 1 : 1 (2 ml) and left stir 2h. The reaction mixture was treated with 6N HCl (35 μL). The organics were removed under reduced pressure and the residual aqueous solution was extracted with EtOAc. The organics were washed with brine and dried over Na₂SO₄. Filtration and removal of the solvent afforded the title compound (39 mg, 100 %) as a white solid. LCMS (ES+) m/z 578.1 (M+H)⁺. Step 6: (3aRJSΛ0SΛ2R23E25BS)-7-cvdcφeaty\-ϋ-((ΪR2S)-U

{ [ (cycloprop ylsulfonyl) amino] carbonyl I -2- vinylcyclopropyl)-5 ,8 J 9-trioxo-

1.2.3.3a,5,6J,8.U,12,21.22,25,25a-tetradecahvΦo-10H,19H-9.12- methanocyclopentaf 18 , 191 [ 1,10,3 ,6 , 12]dioxatriazacγclononadecmo[ 12,11 -b] isoquinoline- 10- carboxamide

Intermediate Al (21.61 mg, 0.081 mmol), HATU (30.8 mg, 0.081 mmol), DffiA (0.047 ml, 0.270 mmol) were added to a solution of the product from Step 5 (39 mg, 0.068 mmol) in DMF (1 ml), and left stir 16 hours. The reaction was directly purified by reverse phase chromatography, and the resulting product was concentrated in vacuo to give the title compound as a white solid (30 mg, 56%). LCMS (ES+) m/z 790.2 (M+H)⁺.

By following the procedures outlined in Example 1 and using the appropriate Intermediates A, B, and C, the following compounds were prepared.

Example 13: (3aRJSΛQSΛ2R25&RY7-cγdθOmiyl-^~N-((lR2S)A- { [(cydopropylsulfonyl)amino] carbonyl I -2- vinylcyclopropyl)-5 ,8 , 19-trioxo- 1,2,3,3a,5,6,7,8,l U2,2U2,23,24,25,25a-hexadecahvdro-10H.19H-9.12- methanocyclopentaf 18 , 19] [ IJ 0,3 ,6 J 2] dioxatriazacyclononadecino [ 12, 11 -blisoquinoline- 1 Q- carboxaroide

Step 1: Methyl (3aR.75.105.12Je,25aR)-7-cγcloρeαtvi-5,8.,l 9-trioxo-

1,2,3,3a,5,6,7,8J U2,21,22,23,24,25,25a-hexadecahvdro-10H.19H-9J2- methanocyclopenta[ 18 J 9] [ 1,10,3 ,6 J 2] dioxatriazacyclononadecinof 12, 11 -b] isoqυmoh^'ne- 10- carboxylate

Pd/C (21.58 mg, 0.020 mmol) was added to a solution of the product from Example 1, Step 4 (80 mg, 0.135 mmol) in THF (4 ml) and stirred under a hydrogen atmosphere for 16 hours. The reaction was filtered through a syringe filter and the organics were removed under reduced pressure. The resulting residue was purified by column chromatography (gradient elution, 20-45% EtOAc/hexanes) to yield the title compound as a gray foam (66 mg, 82%). LCMS (ES⁺) m/z 594.1 (M+H)⁺. Rh/C, EtOAc-MeOH (50:50).

Step 2: (3aRJSΛ0SΛ2R25aR)-l-cydovQτAyl-5.8Λ9-tήoxo- l ,2,3,3a,5,6,7,8J lJ2,21,22,23,24,25,25a-hexadecahvdro-10HJ9H-9J2- methanocycloρenta[ 18, 19] [ 1 ,10,3 ,6, 12]dioxatriazacyclononadecino[ 12,11 -b]isoquinoline- 10- carboxylic acid

Using the product from Step 1 , the title compound was prepared according to the procedure in Example 1 Step 5. LCMS (ES⁺) m/z 580.1 (M+H)⁺.

Step 3: (3aRJSΛ0SΛ2R25aR)-7-oychy&ntvl-^'N'({lR2^-l-

{[(cyolopropylsulfonyl)amino]carbonvl}-2-vinvlcyclopropyl)-5_^8,19-trioxo- L2,3,3a,S,6J,8,11.12.21,22,23,24.25.25a-hexadecahvdro-lQH.19H-9,12- methanocyclopenta[ 18 J 9][ 1 , 10,3 ,6, 121dioxatri.azacyclononadecino[^" 12,1 1 -b]isoquinoh^'ne- 10- carboxamide Using the product from Step 2, the title compound was prepared according to the procedure in Example 1 Step 6. LCMS (ES⁺) m/z 792.2 (M+H)⁺.

By following the procedures outlined in Example 1, Steps 1-4 through

Example 13 and using the appropriate A, B, and C Intermediates, the following compounds were prepared.

Example 37: (3aR.75.1Q_ly.l2/g.25a/gV7-cyclopentyl-N-ff lJe^.?)-1-

{r(cyclopropylsulfonyl)ammo1carbonyl)-2-vinylcvclopropylV5,8.19-trioxo- l ,2,3.3a,5,6J.8.11.12.2 L21a,22,22a.23.24.25,2Sa-octadecahvdro-10HJ9H-9.12- methanocvcloρe3ita[19,201cγclopropa [ 14,151F1, 10,3 ,6 J21dioxatnazacycloicosino[l 2,11- blisoquinoline-l O-carboxamide

Step 1 : Methyl (3&RJSΛQSΛ2R,22E26aR)-7-cvdoyentv\~5$Λ9~tnoxo- 2.3.3a.5.6J,8.11.12^1.24.25.26,26a-tetradecahvdro-1H.10H,19H-9,12- methanocyclopenta[ 19,20] [ 1 J0,3,6,12]dioxatnazacycloicosmo[l 2,1 l-b]isoquinolme-10- carboxylate

The title compound was prepared utilizing a similar procedure for Example 1 utilizing Intermediates C5 and Bl LCMS (ES⁺) m/z 606.2 (M+H)⁺

Step 2- Methyl (3aRJSΛ0SΛ2R25aR)-7-cγdopentv\-5£Λ9-tnoxo- UΛ3a,5A7,8α U22Ula,22,22a,2324.25.25a-octadecahydro40HJ9H-9J2- methanocyclopentaf 19,20] cyclopropa[ 14, 151 f l , 10,3 ,6 , 121 dioxatπazacycloicosino [12,11- bi isoq umohne- 10-carbox ylate

PdOAc₂ (6.67 mg, 0 030 mmol) was added to a mixture of methyl (3aR,lS,\0S,l2R22E,26aR)-7-cydopmty\-5β,l 9-tnoxo-2,3,3a,5.6,7,8,l 1,12,21 ,24,25,26,26a- tetradecahydro-1H,l OH, 19H-9.12-methanocyclopenta[ 19,20][ 1 , 10,3 ,6,12] dioxatnazacycloicosino[12,l l-b]isoqumohne-10-carboxylate (90 mg, 0 149 mmol) m diethyl ether/THF (2 ml), and the mixture allowed to stir for 5 mm To this mixture was added dropwise a solution of diazomethane (0.97 mmol, 10 eq) in diethyl ether (2 mL) The reaction was stirred for 10 mm, and an additional chaige of diazomethane (2 mL) and catalyst added. An additional 2 charges of diazomethane solution (2 mL) and catalyst were added over 1 hr resulting in -95% conversion. The reaction mixture was allowed to evaporate with a nitrogen bleed and the residue purified by preparatory HPLC to give the title compound. (60 mg, 65 %). LCMS

(ES⁺) m/z 620.1 (M+H)⁺.

Step 3: (3aRJSΛ0SΛ2R25aR)-7-cγc\opQntγh53Λ9-tήoxo- 123 ,3aΛ6 ,7,8 , 1 U 2,21 ,21 a.22,22a.23.24.25.25a^ctadecahvdro- 1 QH, 19H-9, 12- methanoc yclopenta[ 19,20] cyclopropaf 14, 15] [ IJ 0,3 ,6, 121 dioxatriazacycloicosino[ 12,11- blisoqumoline-10-carboχylic acid

The title compound was prepared in a similar manner as Example 1 Step 5 utilizing methyl (3aΛ,75, 105,12#,25ajR)-7-cycIopentyl-5,8, 19-trioxo- 1 ,23,3^5,6,7,8,1 l,12,21 ₎21a,22₃22a,23,24,25_;25a-octadecahydro-10H,19H-9,12- methanocyclopenta[19,20]cyclopropa[14, IS][I,10,3,6, 12]dioxatriazacycloicosino[12_: 11- bjisoquinolme-10-carboxylate in 300 % yield. LCMS (ES⁺) m/z 606.1 (M+H)⁺.

Step 4: (3aR.lSΛ0SΛ2R25aRY7-cγc\oγentyl-^'N-(aR2S)~\' {ffcyclopropylsulfonyl)aminolcarbonvU-Z-vinylcyclopropylVS^_^^-trioxo- 1,23.3a3,6.7.8.11.12,2L21a,22,22a,23.24,25.25a-octadecahvdro-lQH,19H-9J2- methanocvclopenta[19,20]cyclopropa[14J5][l J0,3,6,121dioxatriazacycloicosino[12J l- biisoquinoline-1 O-carboxamide

The title compound was prepared in a similar manner as Example 1 Step 6, utilizing (3^,75,105,12Λ,25ai?)-7-cyclopentyl-5,8,l 9-trioxo-

1 ,2,3,3^5,6,7,8,1 l,12,21,21a,22,22a,23,24,25,25a-octadecahydrθ"10H,19H-9₃12- methanocyclopenta[19,20]cyclopropa[14,15][l ,10,3,6,12]dioxatriazacycloicosino[12,l 1- b]isoquinoline-10-carboxylic acid in 66 % yield. LCMS (ES⁺) m/z 818.4 (M+H)⁺.

Example 38: r3ai;,75.1Q>S,12i?,24aS^r)-16-chloro-7-cvcloυentyl-N-fflj;,2y>4- {[fcyclopropylsulfonyl)amino]carbonyl}-2-vinylcγclopropyl)-5,8,19-trioxo- 2,3.3a.5,6J,8.11.12.21.22,22a_t23,23a^4.24a-hexadecahvdro-1H,10H,19H-9,12- methanocyclopenta[l 8,191cyclopropa[l 5,16][1 ,10,3i6,121dioxatriazacyclononadecinιθ[12,l 1 - blisoquinoline-10-carboxamide

The title compound was prepared in a similar manner as Example 37, utilizing Intermediates C3 and B3. LCMS (ES+) m/z 838.2 (M+H)+. Example 39: (3aRJSA0SA2R22E25aSh7-cvclOO&Ayl-N-<aR2S)-l- { [ (cyclopropyisulfonyl)amino] carbonyl ) -2- vinylcyclopropyl)-5 ,8,19-trioxo- 1.2.3.3a.5,6J.8.11.12.21 , 24,25,25a-tetradecahydro-I0HJ9H-9,12- methanocyclopentaf 18 , 19] F IJ 0,3 ,6, 12] dioxatriazacyclononadecinof 11 ,12-blqτj.inazoline- 10- carboxamide

Step 1: 1-fert-butvl 2-methyl (2»y.4J?^')-4-rf3-allyl-4-oxo-3.4-dihydroqυinazolin-2- yl)oxy1pyrrolidine- 1 ,2-dicarboxylate

The title compound was prepared in a similar manner as Example 1 Step 1 utilizing 3-allylqumazolme-2,4(1H,3H)-dione in 83% yield. LCMS (ES⁺) m/z 430.0 (M+H)⁺.

Step 2: Methyl (4J?)-4-f(3-allyl-4-oxo-3,4-dihydroquinazolin-2-yl^')oxy1-L-prolinate

The title compound was prepared utilizing a similar procedure for Example 1 Step 2, utilizing 1-tert-butyl 2-methyl (2S,4R)-4-[(3-allyl-4-oxo-3,4-dihydroquinazolin-2- yl)oxy]pyrrolidine-l ,2-dicarboxylate in 99% yield. LCMS (ES⁺) m/z 330.0 (M+H)⁺.

Step 3: Methyl (4JϊM4f3-allyl-4-oxo-3,4-dihvdroqυi^pazolin-2-vnoxyl-lWf25V2-r(f [(1R.2SV2- but-3-en-1-ylcyclopentyl1oxy)carbonyl)amino1-2-cycloτ>entylacetvU-L-prolinate

The title compound was prepared in a similar manner as Example 1 Step 3, utilizing methyl (4R)-4-[(3-aIlyl-4-oxo-3,4-dihydroquinazolin-2-yl)oxy]-L-prolinate and Intermediate B7 in 64 % yield. LCMS (ES⁺) m/z 621.1 (M+H)⁺.

Step 4: Methyl f3aRJ-?,105,12f1,22g,25a^-7-cyclopentyl-5_<8,19-trioxo- 123,3a,5,6.7,8J U2,21,24.25,25a4etradecahydro-10HJ9H-9J2- methanocvclopentaf 18, 19][ 1 ,10,3,6 J2]dioxatriazacyclpnonadecino[ 11,12-b]quinazoline-l O- carboxylate

The title compound was prepared in a similar manner as Example 1 Step 4, utilizing methyl (4Λ)-4-[(3-allyl-4-oxo-3 ,4-dihydroquinazolin-2-yl)oxy]-l - {(2S)-2-[({[(lR,2S)- 2-but-3-en-1-ylcyclopentyl]oxy}carbonyl)amino]-2-cyclopentylacetyI}-L-prolinate in 93 % yield. LCMS (ES+) m/z 593.1 (M+H)+.

Step 5: (3&RJSA0SA2R22E25aS)-7'Cvc\opQntγ\-S$A9-tήoxo- 1.233a,5,6j,8,l lJ2.2L24.25,25a4etradecahvdro-10H,19H-9,12- methanocyciopenta[l 8 ,19-[[I A 0,3,6,12Jdioxatriazacvclononadeeino[ 1 l J2-b]quinazoline-l 0- carboxylic acid

The title compound was prepared in a similar manner as Example 1 Step 5 utilizing methyl (3aR,7S,\0S,l2R,22E,25aS)-7-cydofentyl-5,SA9-tήoxo- 1,2,3,3^5,6,7,8,1 l^^l^^S^Sa-tetradecahydro-lOH^H^π- methanocyclopentaf 18,19][1 , 10,3 ,6,12]dioxatriazacyclononadecino[ 11 ,12-b]quinazoline- 10- carboxylate in 100 % yield. LCMS (ES+) m/z 579.1 (M+H)+.

Step 6: OaRJS. I OS, 12i?,22£,25a£)-7-cyclopentvl-N-( (IR2SY- 1 - ( [(cyclopropylsulfonyl)aroinoi carbon yl i -2- vinylcyclopropyl)- 5 , 8.19-trioxo- 1.2.3,3a.S.6.7.8.11.1221,24.25.25a-tetradecahvdro-10HJ9H-9.12- methanocyclopentaf 18 , 19] [ 1 , 10,3 ,6 J 2]dioxatri azac yclononadecino [ 11 , 12-b] quinazoϊine- 10- carboxamide

The title compound was prepared in a similar manner as Example 1 Step 6, utilizing OaRJS, 1 OS, 12i?,22£,25a₁S)-7-cγclopenty}-5_;8, 19-trioxo- 1,2,3,3a,5,6₎7,8,l l,12^1,24,25,25a-te(xadecahydro-10H,19H-9,12- methanocyclopenta[ 18.19] [ 1 , 10,3 ,6, 12]dioxatriazacyclononadecino[ 11,12-b]quinazoline- 10- carboxylic acid in 64 % yield. LCMS (ES+) m/z 791.2 (M+H)+.

By following the procedures outlined in Example 39 and using the appropriate A, B, and D intermediates, the following compounds were prepared.

Example 59: (3aR.7.S',10^12/?,22g,25a5)-7-cvclopentyl-N-ffli;,25)-1- {[fcyclopropylsιdfonyl)amino]caτbonyl}-2-vinylcvclopropyπ-5,8,19-trioxo-16-phenyl- 1,233a,5.6.7,8J L122K24,25,25a-tetradecahvdro-10HJ9H-9.12- methanocyclopenta[l 8, 19] [ 1 ,10,3,6, 12]dioxatriazacyclononadecino[ 11 J2-b-]quinazoline-l 0- carboxamide

Step 1: Methyl Gai?J5U05a2i?,22£25a5yi6-bromo-7-cyclopentyl-5,8a94rioxo-

1.2.3.3a,5,6J,8.1 L12,2L24,25,25a-tetradecahvdro-10HJ9H-9J2- methanocyclopenta[ 18, 19] [ 1 , 10,3 ,6, 12] ctøoxatriazacyclononadecino[ 11 ,12-b] quinazoline- 10- carboxylate

The title compound was prepared in a similar manner as Example 39 starting from intermediate D2. LCMS (ES+) m/z 671.0 (M+H)+.

Step 2: Methyl (3siRJSJ0SΛ2R22E25aS)-7-cyclopentγl-5SΛ9-tήoxo-16-phenγl~

U,3,3a,5.6.7.8,π.l2.21.24,25,25a-tetradeciihvdro-10H,19H-9a2- methanocycloρenta[ 18, 19] ϊ ΪΛ 03 ,6 Λ 21dioxatriazacyclononadecmo[ 11,12-b]qBinazoline- 10- carboxylate

Aqueous sodium carbonate (2 M, 0.223 ml, 0.447 mmol) and Pd(Ph₃P)₄ (17.21 rag, 0.015 mmol) were added to a solution of methyl (3ai?,7^^r,l0_iS',12Λ,22_Jδ',25a5}-16-bromo-7- cyclopentyl-S^.^-trioxo-lAS^a.S.aj^^ l.^^l^^S^Sa-tetradecahydro-lOH.^H-^^- methanocyclopenta[ 18, 19][ 1 , 10,3 ,6, 12]dioxatriazacyclononadecino[ 11 , 12-b]quinazoline-l 0- carboxylate (100 mg. 0.149 mmol) in toluene (1.5 mL) in a sealed tube. Phenylboronic acid

(27.2 mg, 0.223 mmol) was added and the mixture heated to 90°c. The reaction was let stir for 2 hours, allowed to cool and diluted with aqueous. KHSO4. The product was extracted into ethyl acetate, washed with aqueous sodium bicarbonate, dried over sodium sulfate, filtered and concentrated to a foam. The foam was chromatographed on silica using 20-50% ethyl acetate/hexane to give 81 mg foam. LCMS (ES+) m/z 669.0 (M+H)+.

Step 3: ( 3 aRJSΛ0SΛ2 R22E25aS)-7- cyclopentyl-N-fd R2S)-\-

{ [( cyclopropylsulfonypamino^'lcarbonyl ) -2-vinylcyclopropyl)-5 ,8 , 19-trioxo- 16-phenyl-

1.2.3.3a.5.6J.8.11,12.21.24,2S.25a-tetradecahvdro-10H.19H-9.12- methanocyclopenta[l 8,19]fl J0,3,6,12]dioxatri azacyclononadecinofl l_^^-biqmnazolme-lO- carboxamide

The title compound was prepared in a similar manner as Example 39, utilizing methyl (3 ΆRJS, 1 OS, 12R ,22E,25a5)-7-cyclopentyl-5 , 8 , 19-trioxo- 16-phenyl- 1,2,3,3^5,6,7,8,1 l,12,21,24,25,25a4etradecahydro-10H,19H-9,12- methanocyclopenta[ 18,19] [ 1 ,10,3 ,6, 12]dioxatriazacyclononadecino[ 11,12-b]quinazoline-l 0- carboxylate. LCMS (ES+) m/z 867.37 (M+H)+.

Example 60: (3aR.7S,10SJ2R.22E,25aSV7-cvclopeatyl-N-f(lR,2RVl-

{ [(cyclopropylsulfonyl)amino] carbonyl } -2-ethylcyclopropyl)-5 ,8 , 19-trioxo- 16-phenyl-

1,2,3,3a,5,6,7.8.11.12.21.24.25.25a-tdτadecahv(faO-10H,19H-9a2- methanocyclopenta[l 8 J9iriJ0,3,6,12]dioxatriazacyclononadecino[l l J2-b]qumazoline-10- carboxamide

The title compound was prepared in a similar manner as Example 39 utilizing methyl (3aΛ,7.S,l O-S.l 2J?,22£,25aS)-7-cyclopentyl-5 ,8, 19-trioxo- 16-phenyl- 1,2,3,3a,5,6,7,8₉l l,12,21,24_}25,25a-tetradecahydro-10H₎19H-9,12- methanocyclopenta[l 8,19][ 1 ,10,3 ,6,12]dioxatriazacyclononadecino[l 1 ,12-b]quinazoline-l 0- carboxylate. LCMS (ES+) m/z 869.39 (M+H)+.

Example 61: (3dιRJSΛ0SΛ2R22E25aS)-7-cγchOQntγ\-Η-((ΪR2S)-l- {|Tovclopropyls^'ulfonyl)amino]carbonyl} -2-vinylcyclopropyl)- 16-methyl-5,8, 19-trioxo- 1,2.3 ,3a,S.6,7.8,l 1,12,2 L24,25.2Sa-tetradecahvdro-10R19H-9,12- methanocyclopentaf 18 J 9][1 J 0,3,6,121dioxatriazacvclononadecinof 11 ,12-biquinazoline-l 0- carb ox amide

Step 1 : MethvIOa^J^lOS.U^^lE^Sayy-y-cvclopentyl-lό-methyl-S^jg-trioxo- 1,2,3,3a.5,6J,8,11.12.21.24,25,25a-tetradecahvdro-10H.19H-9,12- methanocvclopenta[18J91f l J0,3,6J2]dioxatriazacyclononadecino[m2-blqιtma2oline-10- carboxylate

Tetramethyltin (0.041 ml, 0.298 mmol), lithium chloride (50.5 mg, 1.191 πraiol), triphenylphosphine (15.62 mg, 0.060 mmol) and bis(triphenylphosphine)palladium(II) chloride (15.68 mg, 0.022 mmol) were added to a nitrogen purged solution of methyl (3ai?/7S,l 05,12/ζ22£,25aS> 16-bromo-7-cyclopentyl-5.8, 19-trioxo- 1,2,3,38,5,6,7,8,11,12,21,24,25,258-16^3(1608117^0-1011,1913-9,12- methanocyclopentaf 18 , 19] [ 1 , 10 , 3 ,6 , 12] di ox atri azacycl ononadecino [ 11 , 12 -b ] quinazoline- 10- carboxylate, Example 59 Step 1 (100 mg, 0.149 mmol) in DMF (2 ml). The mixture was heated to 90°C for 3 hr. The reaction was diluted with aqueous. KHSO₄ and product extracted into ethyl acetate. The organic extract was washed with aqueous sodium bicarbonate, dried over sodium sulfate, filtered, concentrated and chromatographed on silica using 20-50% ethyl acetate/hexane to give the title compound as a foam, (74 mg, 82 %). LCMS (ES+) m/z 607,3 (M+H)+.

Step 2: (3aJ?.7^105,12i;,22£,25a5^f)-7-cvclopentyl-N-αi^.2^)-1-

{[(cvclop^opylsulfonyl)amino]carbonyl}-2-vmylcyclopropyl)-16-methyl-5,8J9-trioxo-

1.2.3.3a.5.6.7.8.11.12,2L24,2S,2Sa-tetradecahydro-10HJ9H-9,12- methanocyclopenta[ 18 J 91 [1 , 10,3.6, 12] dioxatriazacyclononadecinof 11 , 12-b] quinazoline- 10- carboxamide

The title compound was prepared in a similar manner as Example 39, utilizing methyl(3ai?,7S, 105,12i?,22£,25aS)-7-cyclopentyl- 16-methyl-5,8, 19-trioxo- 1,2,3,38,5,6,7,8,1 l,12,21,24.25,25a-tetradecahydro-10H,19H-9,12- methanocyclopenta[ 18,19][l,10,3,6,l 2]dioxatriazacyclononadecino [ 1 1 , 12-b] quinazoline- 10- carboxylate. LCMS (ES+) m/z 805.3 (M+H)+.

Example 62: (ΪRΛ4EΛ 8^.22 J?,26^29^)-26-cvclopentyl-iV-(( li?,2i?)-l - {j-fcvclopropylsulfonyl)amino1carbonγl)-2-ethylcvclopropyl)-7-methyl-l K24,27-trioxo-2,23- dioxa-4J225,28-tetraazaρentacvclo[^'26.2, 1.0³'¹²,0^5;1().0^l8'²²]hentriaconta-3,5,7,9.14-pentaene-29- carboxamide

The title compound was prepared in a similar manner as Example 39 utilizing methyl(3 aR,7S, 1 OS, 12Λ,22J^25a5)-7-cyclopentyl- 1 ό-methyl-5 ,8, 19-trioxo- 1 ,2,3,3^5,6,7,8,1 lJ2,21,24,25,25a4etradecahydro-10H,19H-9_;12- methanocyclopentat 18, 19] [1,10, 3, 6,12]dioxatriazacyclononadecino[ 11 ,12-b]quinazoline- 10- carboxylate. LCMS (ES+) m/z 807.3 (M+H)+.

Example 63: (3&RJSΛ0SΛ2R22E25aS)-l6-cvaxιo-7-cγclopentyl-N-(αR2S)~l- { [(cyclopropylsulfonyl)aminol carbonyl I -2-vinylcyclopropyl)-5 ,8,19-trioxo- 1.2.3,3a,5,6J.8.11.12.21 ,24.25.25a-tetradecahvdro-10H.19H-9.12- methanocyclopenta[ 18 , 19] [ IJ 0,3 ,6, 12]dioxatriazacyclononadecino[^' 11,12-b]quina2oline- 10- carboxamide

Step 1: Methyl (3ai?JSJ0£J2i^2£.2Sa5)-16-cyano-7-cvclopentyl-5,8J94rioxo-

L2Λ3a,5,6J,8J U22U4,25.25a-tetradecahydro-10HJ9H-9J2- methanocyclopenta[ 18,19] [ IJ 0,3 ,6 J 2]dioxatriazacyelononadecino[ 1 IJ 2-b] quinazoline- 10- carboxylate

Zinc cyanide (5.20 μl, 0.082 mniol) was added to a nitrogen-purged solution of methyl (3aR,7S,l OS, 12R,22E,25s£)- 16-bromo-7-cycloρentyl-5,8, 19-trioxo- 1,2,3_;3a_;5,6,7,8J l,12,21,24,25,25a-tetradecahydro-10HJ9H-9J2- methanocyclopenta[l 8 J 9] [ 1 ,10,3,6 J2]dioxatriazacyclononadecino[ 11 J 2-b]quinazoline-l 0- carboxylate, Example 59 Step 1 (50 mg, 0.074 mmol) in DMF (1 ml), followed by Pd(Ph₃P)₄ (8.60 mg, 7.45 μmol), and the mixture was heated at 100°C for 3 hrs. The reaction was filtered and purified by reverse phase chromatography to give the title compound. (36 mg, 78%) LCMS (ES+) m/z 618.3 (M+H)+.

Step 2: (3aRJSΛ0SΛ2R22E25aS)-l6-cγano-7-CΥc}opentγl-N-((W2S)-l-

{ [(cyclopropylsulfonyl)aminoJcarbonyl) -2-vinylcvclopropyl)-5,8,l 9-trioxo-

1,2,3,3aΛ6J,8,l U2,2U4,25,25a4etradecahydro-10HJ9H-9,12- methanocyclopentafl 8,19] [U 0,3,6,12] dioxatriazacvclononadecmof 11 ,12-b]quinazoline-l 0- carboxamide

The title compound was prepared in a similar manner as Example 39, utilizing methyl (3aR,7S,WS,l2R,22E,25aS)- 16-cyano-7-cyclopentyl-5,8, 19-trioxo- 1, 2,3,38,5,6/7,8,11,12,21, 24,25,25a-tetradecahydro-10H,19H-9_;l 2- methanocyclopenta[ 18, 19] [ 1 , 10,3,6, 12]dioxatriazacyclononadecino[ 11 ,12-b]quinazoline- 10- carboxylate. (ES+) m/z 816.3 (M+H)+.

Example 64:(3aRJSΛ0SΛ2R22E25aS)-l 6-cvano-7-cvclopentyl-N-((lJ;,2J;)-1- (rCcvclopropylsulfonyl)amino]carbonyl}-2-ethylcvclopropyl)-5,8,19-trioxo- 1,2,3,3^5,6,7,8,1 U2,21,24,2S.25a-tetradecahvdro-10H.19H-9,12- methanocvclopentafl8J9][U0,3,6_ι,12]dioxatria2acvclononadecino[l l,_ι12-b]quinazolirse-10- carboxamjde

The title compound was prepared in a similar manner as Example 39, utilizing methyl (3&R,1S, 1 OS, 12R,22E, 2SaS)-16-cy∞o-7-cycIαpentyl-5,8,l 9-trioxo- 1,2,3,3a,5,6,7,8,l l,12^1,24,25,25a-tetradecahydro-10H,19H-9,12- methanocyclopenta[ 18, 19] [1,10, 3, 6,12]dioxatriazacyclononadecino[ 11 , 12-b]quinazoline- 10- carboxylate. (ES+) m/z 818.3 (M+H)+.

Example 65: (38^,75.10^12J?,25aR)-7-cvclopeαtvi-N-f(lJ?,25>-1-

{[(cvclopropylsulfonyl)aminQlcarbonyl}-2-vinylcγclopropyl)-5,8J9-trioxo--

1.2.3,3a.5,6,7,8,l l,12,21 ,22JZ3,24.25,2Sa-hexadecahydro-10H.19H-9.12- methanocyclopentaf 18 , 191 [ 1 , 10,3 ,6, 121dioxatriazacyclononadecino[ 1 IJ 2-bi quinazoline- 10- carboxamide

Step 1 : Methyl (3eJtJSΛOSΛ2R22E25aS)-7<vclop&ityl-5,8Λ9-tάoxo-

1 ,2.3.3a,S.6,7.,8 J 1 J 2.2U4.25.25a-tetradecahvdro-l 0H.19H-9 J 2- methanoc yclopentaf 18 J 91 IT J 0.3 ,6 J 2] dioxatriazac yclononadecino [ 11 J 2-b] quinazoline- 10- carboxylate

The title compound was prepared in a similar manner as Example 13, utilizing methyl (4R)-4-[(3-allyl-4-oxo-3,4-dihydroquinazolin"2-yl)oxy]-1-{(2S)-2-[({[(lR,2S)-2-but-3- en- l-ylcyclopentyl]oxy}carbonyl) amino] -2-cyclopentyl acetyl }-L-prolinate and 5% Rhodium on carbon as catalyst in 1 :1 EtOAcMeOH. LCMS (ES+) m/z 595.1 (M+H)+.

Step 2: (3aRJSJ0-?,12jR.22Jg,25aS)-7-cvclopentvϊ-5_<8J9-trioxo- 1.2.3.3a.5,6,7,8.11 J2.21.24,25.25a-tetradecahvdro-10H,19H-9J2- methanocyclopentafl8J91|^'l J0,3.6J21dioxatriazacyclononadecino[l l J2-b]quinazoline-10- carboxylic acid

The title compound was prepared in a similar manner as Example 1 Step 5, utilizing methyl (3aR,lS,ϊ OS, 12Λ,22£,25aS)-7-cyclopentyl-5,8,l 9-trioxo- 1,2,3 ,3a,5,6,7,8,ll,12,2U4,25,25a-tetradecahydro-10H,19H-9,12- methanocyclopenta[l 8_jl 9][1 ,10,3,6,12]dioxatriazacyclononadecino[l 1 ,12-b]quinazoline-l 0- carboxylate in 90 % yield. LCMS (ES+) m/z 581.1 (M+H)+.

Step 3; (3aRJSΛ0SΛ2R25aR)-7-cγc\opmtv\~N-((\R2S)-\-

{ [(cyclopropylsulfonyl)aminol carbonyl } -2-vinylcycloproρyl V5 ,8 , 19-trioxo-

1,2,33a,5,6J.8,πj2,21,22,23,2425.25a-hexadecahydro-10H,19H-9J2- methanocycloρenta[ 18,19] [1,10,3.6,121dioxatriazacyclononadecino[ 11,12-b] quinazoline- 10- carboxamide

The title compound was prepared in a similar manner as Example 1 Step 6, utilizing (3ai?₎7₁S^r _f105,12J?,22£',25aS)-7-cycloρentyl-5₃8,19-trioxo- 1,2,3,3a,5,6_?7,8,l l,12,21,24,25,25a-tetradecahydro-10H,19H-9,12- methanocyclopenta[18,19][l ,10,3,6,12]dioxatriazacyclononadecino[l 1 ,12-b]quinazoline-10- carboxylic acid in 60 % yield. LCMS (ES+) m/z 793.1 (M+H)+.

By following the procedures outlined in Examples 59- 65 and using the appropriate A, B, and D intermediates, the following compounds were prepared.

Example 81 f 1 R Λ OE, and 10ZΛ4S, I %R22S25S)-22-Cyclopmtyl-N-( (I R25D-1 - {[(cyclopropvl- sulfonyl)aminolcarbonvU -2-vinylcyclopropyl)-7,2(K23-trioxo-2, 19-dioxa-8,21 ,24- triazatetracvclo[22.2.1 ,0³'⁸.0¹⁴'ⁱ⁸lheptacosa-3,5J0-triene-25-carboxamide

Step 1 : 2-(BenzyloxyV6-fluoropyridine

An oven-dried 3 -neck 2L round bottom flask under nitrogen was charged with benzyl alcohol (9.04 ml, 87 mmol) and tetrahydrofuran (75 ml). Attached an addition funnel containing a solution of 2,6-diffuoropyridine (10.0Og, 87 mmol) in tetrahydrofuran (30 ml). Sodium hydride (3.82 g, 96 mmol) was added in small portions with stirring. Heated in a 50°C oil bath for 45 minutes; cooled to room temperature. Added contents of addition runnel dropwise. After 18 hours poured into water. Extracted three times with ethyl acetate, washed combined organic portions with brine, dried with anhydrous magnesium sulfate, filtered and rotary evaporated filtrate. The crude product was subjected to flash column chromatography eluting with hexane. Evaporation of fractions gave the title compound as a colorless oil. (17.7Og, 87 mmol, 100 % yield). LCMS (ES+) m/z 204.3 (M+H)⁺.

Step 2: (4J?V4-{j^"6-fBenzyloxy)ρyridin-2-ylloxyl-1-ffert-butoxycarbonyl)-L-ρroline

A 50OmL 3-neck round bottom flask under nitrogen was charged with Boc-L-4-hydroxyproline (Chcm Impex) (2.0Og, 8.65 mmol) and DMSO (80 ml). Attached an addition funnel containing a solution of the product of Step 1 (1.758 g, 8.65 mmol) in DMSO (10 ml). Cooled reaction in an ice bath to +18°C. Added potassium-t-butoxide (2.91 g, 25.9 mmol) slowly keeping the internal temperature <20°C, Stirred 45 minutes at room temperature. Cooled to 15°C. Added contents of addition funnel dropwise. Warmed to room temperature. Stirred 1.5 hours. Poured into 2.5% potassium bisulfate. Extracted three times with ethyl acetate, washed combined organic portions with water then brine, dried with anhydrous magnesium sulfate, filtered and rotary evaporated filtrate to give the title compound as a white foam. (3.58 g, 8.65 mmol, 100 % yield). LCMS (ES+) m/z 415.3 (M+H)⁺. Step 3: 1-fert-ButyI 2-methyl (25',4J?)-4-{r6-fbenzyloxy^')pyridin-2-yl1oxy>pyrrolidiiie-1.2- dicarboxylate

A 50OmL round bottom flask under nitrogen was charged with the product of Step 2 (7.17g, 17.30 mmol), methanol (50 ml), and dichloromethane (50 ml). Attached an addition funnel containing trimethylsilyldiazomethane in diethyl ether (2,0M) (25.9 ml, 51.9 mmol). Cooled reaction flask in an ice bath. Added contents of addition funnel dropwise. Bubbles. HPLC/MS looks good after 0,5 hours. Quenched by dropwise addition of 2.5% potassium bisulfate. Evaporated to remove methanol. Added water. Extracted three times with ethyl acetate, dried with anhydrous magnesium sulfate, filtered and rotary evaporated filtrate. Subjected to flash column chromatography eluting with 80 hexaiie / 20 ethyl acetate. Evaporation of fractions containing product gave the title compound as a colorless oil. (6.5Og, 15.17 mmol, 88 % yield). LCMS (ES+) m/z 429.3 (M+H)⁺.

Step 4: 1 -/erf-Butyl 2-methyl f2-?_<4J?)-4-[f6-hvdroχypyridin-2-yl)oxy1ρvrrolidine-l 2- dicarboxvlate

A Parr jar was charged with the product of Step 3 (6.50 g, 15.17 mmol), EtOH (100 ml), and 10% Pd/C (0.25O g, 2.349 mmol). Hydrogenated at 48psi. Reaction complete after 18 hours. Filtered through celite. Evaporation of the filtrate gave the title compound as a white foam. (4.95g, 14,63 mmol, 96 % yield). LCMS (ES+) m/z 339.3 (M+H)⁺. Step 5: 1-tert -Butyl 2-methyl (25.4J?M-{[6-(allγloxy>ρyridin-2-ylloxy}ρyrrolidine--1.2- dicarboxylate

A 50OmL round bottom flask under nitrogen was charged with the product of Step 4 (4.95g, 14.63 mmol), tetrahydrofuran (200 ml), allyl bromide (2.53 ml, 29.3 mrnol), and cesium carbonate (4.77 g, 14.63 mmol). Heated in a 60°C oil bath for 18 hours. HPLC/MS o.k. Cooled. Evaporated. Added 2.5% potassium bisulfate. Extracted three times with ethyl acetate, washed combined organic portions with brine, dried with anhydrous magnesium sulfate, filtered and rotary evaporated filtrate. Subjected to flash column chromatography eluting with 70 hexane / 30 ethyl acetate. Evaporation of fractions containing product gave the title compound as a pale oil. (4.56g, 12.05 mmol, 82 % yield). LCMS (ES+) m/z 379.3 (M+H)⁺.

Step 6: 1-tert-Butγl 2-methyϊ (2-?,4/gV4-rf l-aIlyl-6-oxo-l ,6-dihydropvridin-2- ypoxyjp yrrolidme- 1 ,2-di carboxylate

An oven-dried 10OmL Kjeldahl Flask under nitrogen was charged with the product of Step 5 (2.82g, 7.45 mmol) and toluene (15 ml). Degassed 5 minutes with nitrogen. Added tetrakis(triphenylphosphme)palladium(0) (0.431 g, 0.373 mmol). Heated in a 60°C oil bath for 1 hour. HPLC/MS shows product. HPLC/MS looks good after 2 hours. Cooled. Evaporated. Subjected to flash column chromatography eluting with 99 methylene chloride / 1 methanol. Evaporation of fractions containing product gave the title compound as an oil / foam. (1.9 Ig, 5.05 mmol, 67.7 % yield). LCMS (ES+) m/z 379.3 (M+H)⁺.

Step 7: (25^r,4i?)-4-[(l -Allyl-6-oxo-l ,6-dihydropyridin-2-vDoχγ1-2-(methoxycarbonyl)- pyrrolidinium chloride

HCI

A lOOrnL round bottom flask was charged with the product of Step 6 (1.9 Ig, 5.05 mmol). Added HCl (4.0M in dioxane) (30 mL, 120 mmol). Stirred at room temperature for 1 hour. Evaporated. Added ethanol. Evaporated. Repeated. The title compound remained as a pale foam. (1.59%, 5.05 mmol, 100 % yield). LCMS (ES+) m/z 279.3 (M+H)⁺.

Step 8: Methyl (4JgM-ffl-allyl-6-oxo-L6-dihvdropwidifl-2-γ1)oxv]-l -If 25)-2-[({[f lJg,2S)-2- but-3-en- 1 -ylcyclopentyljoxyl carbonγl)amino]-2-cyclopentylacetyl) -L-prolinate

The title compound was prepared in a similar manner as Example 1 Step 3, utilizing (2,S,4i?)-4- [(l-allyl-6-oxo-l ,6-dihydropyridin-2-yl)oxy]-2-(methoxycarbonyl)-pyrroHdinium chloride and Intermediate B7 in 71% yield. LCMS (ES+) m/z 570.3 (M+H)⁺.

Step 9: Methyl (IRAOE, and 10Z,14>S^FJ8ig,225',255)-22-cyclopentyl-7,20,234rioxo-2J9-dioxa- 8,21.24-triazatetracyclor22.2.1.0³'⁸.0¹⁴'¹⁸lheυtacosa-3,5.10-triene-25-carboxylate O

The title compound was prepared in a similar manner as Example 1 Step 4, utilizing methyl (4i?)-4-[(l-allyl-6-oxo-1,6-dihydropyridin-2-yl)oxy]-1-{(2₁S)-2-[({[(l_JR,25)-2-but-3-en-1- ylcyclopentyl]oxy}carbonyl) amino] -2-cyclopentylacetyl}-L-prolinate in 100% yield. LCMS (ES+) m/z 542.3 (M+H)⁺.

Step 10: (IRΛOE, and 10ZJ4JU8i?22£25^-22-CvclopentyI-7,20.23-trioxo-2.19-dioxa- 8.21.24-triazatetracvclof22.2.1 ,0³'⁸.0¹⁴'^l8]heptacosa-3,5,l O-triene-25-carboxylic acid

The title compound was prepared in a similar manner as Example 1 Step 5, utilizing methyl

(lΛ_ϊ10i5₉ and l0Z,145,18Λ,225,25₁S)-22-cyclopentyl-7,20,23-trioxo-2,19-dioxa-8,21,24- triazatetracyclo[22.2.1.0³'⁸.0^l4>ls]heptacosa-3_;5,10-triene-25-carboxylate in 83% yield. LCMS (ES+) m/z 528.3 (M+H)⁺.

Step 11: (IRΛOE, and 10Z.145,18J?.22.S',2S5)-22-Cvclopentyl-.V-ffl_JR^_JSf)-1-Ufcvclo- propylsulfonyl)aminoicarbonyl} -2-vinylcyclopropyl)-7,20,23-trioxo-2,l 9-dioxa-8,21 ,24- triazatetracvcloF22.2.1.0³'⁸,θ'^4<ι8lheptacosa-3,5JO-triene-25-carboxamide

The title compound was prepared in a similar manner as Example 1 Step 6, utilizing (li?,10£, and l0Z,14_iS',18J?,22₎S,255)-22-Cyclopentyl-7,20,23-trioxo-2,19-dioxa-8,21₃24- triazatetracyclo[22.2.1.0³'⁸.0¹⁴'¹⁸]heptacosa-3,5_f10-triene-25-carboxylic acid to give a 31% yield. LCMS (ES+) m/z 740.3 (M+H)⁺.

Example 82 qj?J4i?J8/;,225,25^-22-cvclopentyl-iV-f(lJ?,2^)-1-

{ [(cyclopropylsulfonyl)amino] carbonyl } -2- vinylcyclopropyl)-7,20,23-trioxo-2 , 19-dioxa-

8,21 ,24-triazatetracvolo[22,2.1.0³'⁸.0¹⁴'¹⁸lheptacosa-3.5-diene-25-carboxamide

Step 1 : Methyl (lJ?.14^,18ig,225,2S.?)-22-cvclopentyl-7.2023-trioxo-2,19-dioxa-8_<2^] .24- triazatetracyclor22.2.1.0³'^S.θ'⁴'^iS1heptacosa--3,5-diene-25-carboxvlate

The title compound was prepared in a similar manner as Example 13 Step 1, utilizing methyl (IR₇IOE, and 10Z,14S,l 8Λ,225^55)-22-cycloρentyl-7,20,23-trioxo-2,19-dioxa-8,21,24- triazatetracyclo[22.2.1.0³'⁸.0¹⁴'¹⁸]heptacosa-3,5,10-triene-25-carboxylate and replacing THF with ethanol as the solvent to give a 94% yield. LCMS (ES+) m/z 544.3 (M+H)⁺.

Step 2: (lJ?J4_JRJ8i?,22^255^r)-22-Cvclopentyl-7,20,23-trioxo-2.19-dioxa-8,21,24- triazatetracvclo[22.2.1.0³'^S.Q'⁴'¹⁸lheptacosa-3,5-diene-25-carboxylic acid

The title compound was prepared in a similar manner as Example 1 Step 5, utilizing methyl (liζl4^18/ζ22S,25^-22-cyclopentyl-7,20,23-trioxo-2,19-dioxa-8,21,24- triazatetracyclo[22.2.1.0³'⁸.0¹⁴'¹⁸]heptacosa-3,5-diene-25-carboxylate to give a 100% yield. LCMS (ES+) m/z 530.3 (M+H)⁺.

Step 3: d^J4^J8^,22^25^-22-cvclopentyl-JV-f(lJ^!?,25^r)-1-{r(cvclopropylsulfonyl)- aminol carbonyl I -2-vinyl cvcloυroυ yl)-7 ,20,23 -trioxo-2 J 9-dioxa- 8.21 ,24- triazatetracvcloi-22.2.1.0³'^s.0^l4'^ls1heptaco$a-3,5-diene-25-carboxamide

The title compound was prepared in a similar manner as Example 1 Step 6, utilizing (lΛ,14Λ,18Λ,225,255)-22-Cyclopentyl-7,20,23-trioxo-2,19-dioxa-8,21₎24- triazatetracyclo[22.2.1,0³'⁸.0¹⁴'¹⁸]heptacosa-3,5-diene-25-carboxylic acid to give a 61% yield. LCMS (ES+) m/z 742.3 (M+H)⁺.

Example 83 : (IRΛ4RΛSR22S25,SV 6- bromo-22 -cyclopent yl-iV-Cf 1 R2S)- 1 - ([(cyclopropylsulfonyl)aminolcarbonyl}-2-vinylcyclopropyl')-7,20,23-trioxc)-2,19-dioxa-

8.21 ,24-triazatetracvclor22.2.1.0³'⁸.0¹⁴'¹⁸lheptacosa-3,5-diene-25-carboxamide

Step 1: Methyl fli?J4i?J8^,225,255^r)-6-bromo-22-cvclopentyl-7.20.23-trioxo-2J9-dioxa- 8,2L24-triazatetracvclor22.2.1.Q³'⁸,0¹⁴'^ls]heptacosa-3.5-diene-25-carboxylate

A 25rnL round bottom flask under nitrogen was charged with the product of Example 82 Step 1

(238 mg, 0.438 mmol), NBS (39.0 mg, 0.219 mmol), and acetonitrile (15 ml). Stirred at room temperature 10 minutes. HPLC/MS shows mainly desired; little starting material. Added an additional lOmg NBS. Still a little starting material remaining. Added another lOmgNBS. HPLC/MS looks good. Cooled. Evaporated. Subjected to ISCO chromatography eluting with a hexane / ethyl acetate gradient. Evaporated fractions containing product to give the title compound as a white solid . (224 mg, 0.360 mmol, 82 % yield). LCMS (ES+) τaJz 623.3 (M+H)⁺.

Step 2: flJ?,14j?J8i;,225',25y>-6-bromo-22-cvclopentyl-7,20,23-trioxo-2.19-dioxa-8,21.24- triazatetracvclo[22.2.1.0³'⁸,0¹⁴'¹⁸lheptacosa-3,5-dieae-25-carboxvlic acid

The title compound was prepared in a similar manner as Example 1 Step 5, utilizing methyl (lΛ,14Λ,18Λ,225,25^-6-bromo-22-cyclopentyl-7.20,23-trioxo-2,19-dioxa-8,21,24- tria2atetracyclo[22.2.1.0^3<8,0^14<I8]heptacosa-3,5-diene-25-carboxylate to give a 100% yield. LCMS (ES+) m/z 609.8 (M+H)⁺.

Step 3: (l^a4^J 8^22^25_i$^f)-6-bromo-22-cyclopentvl-N-rπJ?,25)-1- {[fcyclopropylsijlfonyl)amino]carbonyl)-2-vinylcvcloρroρyl)-7,20,23-trioxo-2,19-dioxa- 8,21,24-triazatetracvclof 22,2, 1.0³'⁸.0¹⁴'^lslheptacosa-3,5-diene-25-carboxamide

The title compound was prepared in a similar manner as Example 1 Step 6, utilizing

(\R,l4R,l 8i?,22S,255)-6-bromo-22-cyclopentyl-7_?20_f23-trioxo-2,l 9-dioxa-8,21 ,24- triazatetracyclo[22.2.1,0 '^S.O¹⁴' ]heptacosa-3,5-diene-25-carboxylic acid to give a 49% yield. LCMS (ES+) m/z 821.3 (M+H)⁺.

Example 84: aRΛ4RΛ^R22S25S)~22-cvc\θOentγ\-N-((lR2S)-\~

{ [f cyclopropylsulfon yl)amino] carbonyl I -2 - vinylcyclopropyl)-7 ,20,23 -trioxo-6-phenyl -2,19- dioxa-8,21 ,24-triazatetracyclor22.2, 1.0³'⁸.0¹⁴'ⁱ⁸]heptacosa-3.5-diene-25-carboxamide

Step 1: Methyl flJ?,147?J8J?.22^.25.y>-22-cyclopentyl-7.2Q,23-trioxo-6-t>henvl-2J9-dioxa- 8,21 ,24-triazatetracyclor22.2.1.Q³'⁸.0^14>ls1heptacosa-3 ,5-diene-25-carboxylate

An oven-dried 10OmL Kjeldahl Flask under nitrogen was charged with the product of Example 83 Step 1 (100 mg, 0.161 mmol), dioxane (5 ml), phenylboronic acid (24.48 mg, 0.201 mmol), cesium carbonate (65.4 mg, 0.201 mmol), tricyclohexylphosphine (13.51 mg, 0.048 mmol), and tris(dibenzylideneacetone)dipalladium(0) (17.65 mg, 0.019 mmol). Heated in an 80°C oil bath for 20 hours. HPLC/MS looks good. Cooled. Poured into 2.5% potassium bisulfate. Extracted three times with ethyl acetate, washed combined organic portions with brine, dried with anhydrous magnesium sulfate, filtered and rotary evaporated filtrate. Subjected to ISCO chromatography eluting with a hexane / ethyl acetate gradient. Evaporation of fractions containing product gave the title compound as a white solid remained, (75 rng, 0.121 mmol, 75 % yield). LCMS (ES+) m/z 620.3 (M+H)⁺.

Step 2: πJ?J4J?J8/?.22^25^-22-cvclopentyl-7,20,23-trioxo-6-phenyl-2J9-dioxa-8,2L24- triazatetracvclof22.2,1.0³'^s.0^ϊ4'^ls1heptacosa-3.5-diene-25-carbox:ylic acid

The title compound was prepared in a similar manner as Example 1 Step 5, utilizing methyl (lΛ,14J?,18i?,225,25_J5)-22-cyclopentyl-7,20,23-trioxo-6-phenyl-2_;19-dioxa-8,21,24- triazatetracyclo[22.2.1.0^3>8.0^l4'¹⁸]heptacosa-3,5-diene-25-carboxylate to give a 100% yield. LCMS (ES+) m/z 606.3 (M+H)⁺.

Step 3: (lJΪ,14J?.18i;,22_J?,25^)-22-cvdopentyl-^-f (1^.25)-!-

{[(cyclopropylsulfonvπamino]carbonyl|-2-vinylcvclopropyl)-7_h20,23-trioxo-6-phenyl-2,19- dioxa-8,2L24-triazatetracvcIor22.2.1.0³'⁸.0ⁱ⁴'¹⁸1heτ>tacosa-3.5-diene-25-carboxamide

The title compound was prepared in a similar manner as Example 1 Step 6, utilizing (l_JR,14i?,18i?,22_iS',25₁S^!)-22-cyclopentyl-7,20,23-trioxo-6-phenyl-2,19-dioxa-8,21,24- triazatetracyclo[22,2.1.0³'^S.0^!4>183heptacosa-3,5-diene-25-carboxylic acid to give a 50% yield. LCMS (ES+) m/z 818.3 (M+H)⁺.

Example 87: NS3 protease Ib Rl 55K Ki Activity

The HCV NS3 protease inhibitory activity was measured using the protease time- resolved fluorescence (TRF) assay as described below and in International Patent Application Publication WO 2006/102087. The assay was performed with HCV genotype Ib (BK) NS3 modified with a R 155K mutation .

The assay was performed in a final volume of 100 μl in assay buffer containing 50 mM HEPES, pH 7.5, 150 mM NaCl, 15 % glycerol, 0.15 % TRITON X-100, 10 mM DTT, and 0.1 % PEG 8000. NS3 and NS4A protease is pre-incubated with various concentrations of inhibitors in DMSO for 30 minutes. The reaction is initiated by adding the TRF peptide substrate (final concentration 100 nM). NS3 mediated hydrolysis of the substrate is quenched after 1 hour at room temperature with 100 μl of 500 mM MES₃ pH 5.5. Product fluorescence is detected using either a VICTOR V2 or FUSION fluorophotometer (Perkin Elmer Life and Analytical Sciences) with excitation at 340 nm and emission at 615 nm with a 400 μs delay. Testing concentrations of the enzyme containing the R155K mutation were selected to result in a signal to background ratio (S/B) of 10-30. IC₅₀ values are derived using a standard four- parameter fit to the data. K; values are derived from IC₅₀ values using the following formula,

IC₅₀ = K₁ (1 + [S] / KM), Eqn (I), where [S] is the concentration of substrate peptide in the reaction and K_M is the Michaelis constant. See P. Gallinari et al., 38 BiOCHEM. 5620-32(1999); P. Gallinari et al, 72 J. ViROL. 6758-69 (1998); M. Taliani et al, 240 ANAL. BIOCHEM. 60-67 (1996).

The activity table provided below illustrates the observed activity, where compound activity fell within the following ranges: A: Ki 1 nM to 5 nM B: K, 0.5 nM to 1 nM C: K, 0.1 nM to 0.5 nM D: K₁ < 0.1 nM Activity Table

Other embodiments are within the following claims. While several embodiments have been shown and described, various modifications may be made without departing from the spirit and scope of the present invention.

Claims

WHAT IS CLAIMED IS:

1. A compound of formula (I), or a pharmaceutical acceptable salt thereof:

(I) wherein:

is selected from the group consisting of:

each L is independently selected from the group consisting of N and CH, provided that the total number of L that are N is from 1 to 4,

R¹ is selected from the group consisting of -CO₂H, and -CONHS0₂(cyclopropyl);

R² is selected from the group consisting of ethyl and ethenyl;

R³ is selected from the group consisting of cyclopentyl, cyclohexyl, and

-C(CHa)₃; each R⁴ is independently selected from the group consisting of halogen atoms,

C₁-C₄ alkoxy, C₁-C₇ aϊkyl, C₂-C₇ alkenyl, -CN, -CF₃, -OCF₃, SCH₃, -SO₂(CH₃), C₃-C₈ cycloalkyl, C₃-C₈ cycloalkoxy, C₁-C₆ haloalkyl, phenyl, naphthyl and heteroaryl groups, wherien each said R⁴ heteroaryl is selected from the group consisting of 5- and

6-membered aromatic rings having 1, 2 or 3 heteroatoms independently selected from N, O and S, and said R⁴ heteroaryl is attached through a ring atom selected from C or N, each said R⁴ phenyl, naphthyl and heteroaryl groups are substituted with 0 to 4 substituents independently selected from the group consisting of halogen atoms, -OR , -SR⁵, -N(R⁵)₂, -N(C₁-C₆ alkyl)O(C₁-C₆ alkyl), C₁-C₆ alkyl, C-C₆ haloalkyl, (C₁-C₆ haloalkoxy), -NO₂, -CN₃ -CF₃, -SO₂(C₁-C₆ alkyl), -S(O)(C₁-C₆ alkyl), -NR⁵SO₂R⁶, SO₂N(R⁶)₂, -NHCOOR⁶, -NHCOR⁶, -NHCONHR⁶, -CO₂R^S, -C(O)R⁵ and -CON(R^S)₂, and 2 adjacent substituents of said R⁴ phenyl, naphthyl and heteroaryl groups may be taken together to form a 3- to 6-membered cyclic ring containing O to 3 heteroatoms independently selected from N, O and S;

Y is selected from the group consisting of -C(O)-, -C(O)O- and -C(O)NH-; each R⁵ is independently H or C₁-C₆ alkyl; each R⁶ is independently H, C₁-C₆ alkyl, C₃-C₆ cycloalkyl, C₃-C₆ cycloalkyl(C₁-C₅)alkyl, aryl, aryl(C₁-C₄)alkyl, heteroaryl, heteroaryl(C₁-C₄ alkyl), heterocyclyl, or heterocyclyl(C₁-Cs alkyl);

M is selected from the group consisting of C₄-C₇ alkylene and C₄-C₇ alkenylene, wherein said M is substituted with O to 3 substituents independently selected C₁-C₈ alkyl, provided that two adjacent substituents can together form a 3 to 6 membered ring.

2. The compound according to claim 1 , wherein

is selected from the group consisting of:

3. The compound according to claim 2, wherein O or 1 R⁴ is present and, if present, is selected from the group consisting of -Br, -Cl, -CN, phenyl, -O-phenyl, -OCF3, -OCH₃, -C(O)OH, -CH₃ and -C(O)CH₃.

4. The compound according to any one of claims 1-3, wherein R¹ is -C(O)OH.

5. The compound according to any one of claims 1-3, wherein R is -C(O)NHSO₂cyclopropyl.

6. The compound according to any one of claims 1-5, wherein R is -CH₂CH₃.

7. The compound according to any one of claims 1-5, wherein R is

-CH=CH₂.

8. The compound according to any one of claims 1-7, wherein R is cyclopentyl.

9. The compound according to any one of claims 1-7, wherein R is cyclohexyl.

10. The compound according to any one of claims 1 -7, wherein R³ is

-C(CH₃)_J.

11. The compound according to any one of claims 1-10, wherein M is selected from the group consisting of

, where Z is

12. The compound according to any one of claims 1-11, wherein said compound has the following structure or a pharmaceutical acceptable salt thereof:

wherein "a" is an optionally present bond and R⁴ if present is selected from the group consisting of -Br, -Cl, -CN, phenyl, -O-phenyl, -OCF₃, -OCH₃, -C(O)OH, -CH₃ and -C(O)CH₃.

13. The compound according to claim 1, wherein said compound has the following structure or a pharmaceutically acceptable salt thereof:

wherein "a" is an optionally present bond and R⁴ if present is selected from the group consisting of -Br, -Cl, -CN, phenyl, -O-phenyl, -OCF₃, -OCH₃, -C(O)OH, -CH₃ and -C(O)CH₃.

14. A compound selected from the group consisting of the compound of

Examples 1-86 or a pharmaceutical acceptable salt thereof.

15 A pharmaceutical composition comprising an effective amount of the compound according to any one of claims 1-14, and a pharmaceutically acceptable carrier.

16. The pharmaceutical composition according to claim 15, further comprising a second therapeutic agent selected from the group consisting of HCV antiviral agents, immunomodulators, and anti-infective agents.

17. The pharmaceutical composition according to claim 15, further comprising a second therapeutic agent selected from the group consisting of HCV protease inhibitors and HCV NS5B polymerase inhibitors.

18. A use of the compound according to anyone of claims 1-14 in the preparation of a medicament for inhibiting HCV NS3 protease activity in a subject in mod thereof.

19. A use of the compound according to anyone of claims 1-14 in the preparation of a medicament for preventing or treating infection by HCV in a subject in need thereof.

20. A method of treating a patient infected with HCV comprising the step of administering to said patient a therapeutically effective amount of the compound according to any one of claims 1-14.