WO2020219808A1

WO2020219808A1 - Dicarbamate inhibitors of ns5a for treatment of hepatitis c virus infections and related diseases

Info

Publication number: WO2020219808A1
Application number: PCT/US2020/029717
Authority: WO
Inventors: Michael John COSTANZO; Michael Alan GREEN; Jirong Peng; Don Zhang
Original assignee: Beta Pharma, Inc.
Priority date: 2019-04-25
Filing date: 2020-04-24
Publication date: 2020-10-29
Also published as: TW202106677A

Abstract

Dicarbamate compounds as inhibitors of NS5A, and therapeutic uses and methods of preparation thereof, are disclosed. These compounds, and pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, and prodmgs, and pharmaceutical compositions thereof, are useful in treating diseases and disorders caused by the hepatitis C virus in humans, including cirrhosis and liver cancer.

Description

DICARBAMATE INHIBITORS OF NS5A FOR TREATMENT OF HEPATITIS C VIRUS INFECTIONS AND RELATED DISEASES

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Application Serial No. 62/838,925, filed on April 25, 2019, the disclosure of which is incorporated herein by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to novel dicarbamate compounds, compositions and methods for the treatment or prevention of a disease, disorder, or medical condition caused by the hepatitis C virus in humans, including cirrhosis and liver cancer.

BACKGROUND OF THE INVENTION

Hepatitis C is a liver disease that is caused by the hepatitis C virus (HCV). HCV can cause both acute and chronic hepatitis, ranging in severity from a mild infection lasting only a few weeks to a serious, lifelong illness. In 2018, it was estimated by the WHO that 71 million people globally were living with chronic hepatitis C. A significant number of these chronically infected individuals will eventually develop cirrhosis and/or liver cancer, resulting in approximately 399,000 deaths worldwide each year.

The hepatitis C virus (HCV) is a member of the Flaviviridae family of enveloped, positive-strand RNA viruses for which there are no clinically proven vaccines. For decades, the antiviral treatment of chronic hepatitis C was based upon the use of interferon (IFN)-a combined with ribavirin and was associated with suboptimal response rates and/or high incidence of side effects (Bassetti, M. et al., World J. Gastroenterol. 2015, 27(38), 10760- 10775). Complicating the discovery of new HCV therapies is the highly complex and incompletely understood nature of the viral life cycle. The HCV genome consists of a single strand of RNA of about 9,600 nucleotides encoding a polypeptide precursor of about 3,000 amino acids. Co- and posttranslational proteolytic cleavage of this precursor by cellular and viral enzymes yields the nonstructural (NS) proteins NS2, NS3, NS4A, NS4B, NS5A, and NS5B, which are required for membrane-associated RNA replication (Kim, C. W., Chang, K.-M., Clin. Mol. Hepatol. 2013, 19, 17-25).

The nonstructural protein NS5A has been identified as a promising drug target for antiviral therapeutic intervention because it exerts functionally essential effects in regulation of HCV replication, assembly and egress (Pawlotsky, J.-M., J. Hepatol. 2013, 59, 375-382). It is a proline-rich and hydrophilic phosphoprotein of 447 residues with three domains. While no clear enzymatic functions have been assigned to NS5A, it appears to function through interactions with other HCV proteins and host cell factors. Inhibition of NS5A at picomolar concentrations has been associated with significant reductions in HCV RNA levels in cell culture-based models. In the clinic, small molecule inhibitors of NS5A have been shown to be effective when administered in combination therapy (Schinazi, R. F., Infect. Drug Resist. 2014, 7, 41-56).

Daclatasvir (W02008021927A2) and ledipasvir (W02010132601A1) are representative examples of dicarbamate-based NS5A inhibitors that are currently in the clinic as approved drugs. Other clinically successful examples include elbasvir

( W 02012040923 A 1 ) , odalasvir (WO2012166716A2), vepadasvir (W02013075029A1) and ruzasvir (W02016004899A1).

daclatasvir ledipasvir

However, despite their clinical successes, NS5A inhibitors have certain limitations. Although many of the marketed NS5A inhibitors display potent activity against the HCV genotypes la and lb, their relative antiviral effectiveness against HCV genotypes 2-6 can vary dramatically from one structural motif to another (Gao, M., Curr. Opin. Virol. 2013, 3, 514-520). In addition, the emergence of antiviral resistance in the form of resistance-associated variants (RAVs) continues to be a problem and can render existing clinical NS5A inhibitors ineffective (Chayama, K., Biochem. Biophys. Res. Commun. 2018, 500, 152-157). Clearly a need still exists to develop new small molecule NS5A inhibitors that have pan-genotypic activity and/or are potent against RAVs.

SUMMARY OF THE INVENTION

The present invention relates to dicarbamate compounds of the following general formula I, wherein the dash represents a covalent bond.

In one aspect, the present invention provides a compound of formula I:

B¹ -A¹ -Q¹ -P¹ -M-P²-Q²-A²-B²

I or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, wherein:

M is selected from the formulas II, III, and IV, where R¹ at each occurrence is independently C1-C6 alkyl:

II III IV

P¹ and P² are the same or different and are independently absent (a direct bond) or selected from the formulas:

wherein X is CH or N, L¹ is a direct bond to Q 1¹ or Q -_>2 ², and L 2² is a direct bond to M;

Q¹ and Q -_>2 ² are independently absent (a direct bond),

wherein L¹ is a direct bond to A¹ or A², and L² is a direct bond to P¹ or P²; provided, however, that P¹ and Q¹ are not both absent, and P² and Q² are not both absent; and that P¹

and P² are not

when M is II;

A¹ and A² are the same or different and are independently selected from formulas:

wherein L³ at each occurrence is independently a direct bond to B¹ or B²; L⁴ at each occurrence is independently a direct bond to Q¹ or Q²; and R² and R³ at each occurrence are the same or different and are independently selected from H and C1-C6 alkyl optionally substituted, or taken together along with the carbon atom to which they are attached form a C₃-C₆ cycloalkyl optionally substituted;

B¹ and B² are the same or different and are independently selected from formula:

wherein R⁴ is C₁-C₆ alkyl or aryl-(Ci-C₃) alkyl, each optionally substituted; and R⁵ is C₁-C₆ alkyl, aryl, heteroaryl, substituted C₁-C₆ alkyl, aryl-(Ci-C₃) alkyl, substituted aryl-(Ci-C₃) alkyl, heteroaryl-(Ci-C₃) alkyl, substituted heteroaryl-(Ci-C₃) alkyl, substituted aryl, or substituted heteroaryl; and L⁵ is a direct bond to A¹ or A².

Another aspect of the present invention is directed to a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, and one or more pharmaceutically acceptable excipients, such as adjuvants, diluents, and/or carriers.

Another aspect of the invention is directed to a method of inhibiting the function of an HCV NS5A protein comprising contacting a biological sample containing the HCV NS5A protein with a compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug,.

Another aspect of the invention is directed to a method for the treatment or prevention of a disease, disorder or medical condition caused by HCV in a patient, comprising administering to the patient a therapeutically effective amount of the compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate or prodrug thereof.

In one embodiment, the diseases, disorders or medical conditions caused by HCV may include cirrhosis, liver cancer or hepatitis C infection.

In another aspect, the present invention provides use of a compound of formula (I) according to any embodiments described herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, prodrug, or composition thereof, in the manufacture of a medicament for treatment of a disease or disorder associated caused by HCV.

The compounds according to this invention can generally be prepared by a series of steps according to the synthetic methods described herein, in combination with the knowledge known to a person skilled in synthetic organic chemistry. Other aspects or advantages of the present invention will be apparent to those skilled in the art in view of the following detailed description and claims in combination with the knowledge and skills generally known in the field.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides novel dicarbamate compounds useful as inhibitors of

NS5A for the treatment of hepatitis C virus (HCV) infections and diseases or conditions associated with HCV infections.

In one aspect, the present invention provides a compound of formula I:

B¹ -A¹ -Q¹ -P¹ -M-P²-Q²-A²-B²

I

or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, wherein:

wherein X is CH or N, L¹ is a direct bond to Q¹ or Q², and L² is a direct bond to M;

_ -N

I 'AL²

Q¹ and Q² are independently absent (a direct bond),

, or L- H wherein L¹ is a direct bond to A¹ or A², and L² is a direct bond to P¹ or P²; provided, however, that P¹ and Q¹ are not both absent, and P² and Q² are not both absent; and that P¹

and P² are not

when M is II;

wherein L³ at each occurrence is independently a direct bond to B¹ or B²; L⁴ at each occurrence is independently a direct bond to Q¹ or Q²; and R² and R³ at each occurrence are the same or different and are independently selected from H and Ci-G, alkyl optionally substituted, or taken together along with the carbon atom to which they are attached form a C₃-C₆ cycloalkyl optionally substituted;

wherein R⁴ is C₁-C₆ alkyl or aryl-(Ci-C₃) alkyl, each optionally substituted; and R⁵ is C₁-C₆ alkyl, aryl, heteroaryl, substituted C1-C6 alkyl, aryl-(Ci-C3) alkyl, substituted aryl-(Ci-C3) alkyl, heteroaryl-(Ci-C3) alkyl, substituted heteroaryl-(Ci-C3) alkyl, substituted aryl, or substituted heteroaryl; and L⁵ is a direct bond to A¹ or A².

In one embodiment of this aspect, in the compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, M is II;

P¹ and P² are each

; Q¹ and Q² are absent; A¹ and A² are each

R⁴ is C₁-C₆ alkyl optionally substituted; and R⁵ is C₁-C₆ alkyl optionally substituted.

In one embodiment of this aspect, in the compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, each X is CH; R² and R³ are independently hydrogen, methyl, ethyl, or taken together along with the carbon atom to which they are attached form a C₃-C₆ cycloalkyl optionally substituted. In one embodiment of this aspect, in the compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, R² and R³ are independently hydrogen or methyl; R⁴ is methyl or ethyl; and R⁵ is isopropyl.

In one embodiment of this aspect, in the compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, M is

substituted.

I_1^ JTVN L²

III; P¹ and P² are each absent; Q¹ and Q² are each H ; A¹ and A² are each

optionally substituted; and R⁵ is C1-C6 alkyl optionally substituted.

I _A ^{’ L2}

II; P¹ is absent; Q¹ is ^L H ; P² is X ; Q² is absent;

optionally substituted; and R⁵ is C1-C6 alkyl optionally substituted.

In one embodiment of this aspect, in the compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, wherein each X is CH; R² and R³ are independently hydrogen, methyl, ethyl, or taken together along with the carbon atom to which they are attached form a C3-C6 cycloalkyl optionally substituted.

In one embodiment of this aspect, in the compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, R² and R³ are independently hydrogen or methyl; R⁴ is methyl or ethyl, each optionally substituted; and R⁵ is isopropyl optionally substituted.

IV; P and P are each

, wherein X is CH; Q¹ and Q² are absent; A¹ and A

are each

optionally substituted; and R⁵ is Ci-G, alkyl or Ce- Cio aryl, each optionally substituted.

IV; P¹ and P² are each

, wherein X is N; Q¹ and Q ,2² are absen ft; A D ΐ¹ and A i 2² are

each

optionally substituted; and R⁵ is Ci-G, alkyl or aryl, each optionally substituted.

In one embodiment of this aspect, in the compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, R⁵ is isopropyl or phenyl, each optionally substituted.

In one embodiment of this aspect, in the compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, wherein

M is IV; P¹ and P² are each

, wherein X is CH or N; Q¹ and Q² are absent; A¹ and A² are each

optionally substituted; and R⁵ is C₆-C₁₀ aryl, each optionally substituted.

optionally substituted; and R⁵ is C₆-C₁₀ aryl optionally substituted.

substituted.

In one embodiment of this aspect, in the compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, R⁵ is phenyl optionally substituted.

In one embodiment of this aspect, in the compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, R² and R³ are independently hydrogen, methyl, ethyl, or taken together along with the carbon atom to which they are attached form a C₃-C₆ cycloalkyl optionally substituted; and R⁴ is C₁-C₄ alkyl optionally substituted. In one embodiment of this aspect, in the compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, R² and R³ are independently hydrogen or methyl; R⁴ is methyl or ethyl.

In other embodiments, the present invention encompasses any and all possible combinations of the embodiments disclosed herein.

In one embodiment of this aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, selected from the group consisting of:

In another embodiment, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, selected from the group consisting of:

4, 12-bis-|/V-(methoxycarbonyl )-L-valyl-L-prolyl-/V-(3-aminophenyl ) |-| 2.2 Iparacyclophane;

methyl ((5)-l-((5)-2-(5-(9,9-difluoro-7-(3-((5)-l-((methoxycarbonyl)-L-valyl)pyrrolidine-2- carboxamido)phenyl)-9//-fluoren-2-yl)- l//-imidazol-2-yl)pyrrolidin- 1 -yl)-3 -methyl- 1 - oxobutan-2-yl)carbamate;

dimethyl ((2S,2'S)-((2S,2'S)-(((( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-.v-indacene-4,8- diyl)bis(3 , 1 -phenylene))bis(azanediyl))bis(carbonyl))bis(pyrrolidine-2, 1 -diyl))bis(3 -methyl- 1 -oxobutane- 1 ,2-diyl))dicarbamate;

dimethyl (( 1 /?, 1 'R)-((2S,2'S)-(((( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-.v-indacene-4,8- diyl)bis(3,l-phenylene))bis(azanediyl))bis(carbonyl))bis(pyrrolidine-2,l-diyl))bis(2-oxo-l- phenylethane-2,l-diyl))dicarbamate;

dimethyl (( 1 /?, 1 'R)-((2S,2'S)-(((( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-.v-indacene-4,8- diyl)bis(pyridine-5,3-diyl))bis(azanediyl))bis(carbonyl))bis(pyrrolidine-2,l-diyl))bis(2-oxo-l- phenylethane-2,l-diyl))dicarbamate;

diyl)bis(3, 1 -phenylene))bis( 1 //-imidazole-5,2-diyl ))bis( pyrrolidine-2, 1 -diyl ))bis(2-oxo- 1 - phenylethane-2, l-diyl))dicarbamate; and

diyl)bis(4, 1 -phenylene))his( 1 //-imidazole-5,2-diyl ))bis( pyrrolidine-2, 1 -diyl ))bis(2-oxo- 1 - phenylethane-2,l-diyl))dicarbamate.

In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I) according to any of the embodiments disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, and one or more pharmaceutically acceptable excipients, such as adjuvants, diluents, and/or carriers.

In one embodiment of this aspect, the pharmaceutical composition further comprises a second agent having anti-HCV activity.

In one embodiment of this aspect, in the pharmaceutical composition, the second agent having anti-HCV activity is selected from the group consisting of a recombinant Human Interferon Alfa, a nucleoside analog, a direct acting antiviral, a NS3/4A protease inhibitor, a nucleotide NS5B polymerase inhibitor, a NS5A inhibitors, a non-nucleoside NS5B polymerase inhibitors, and combinations thereof.

In one embodiment of this aspect, in the pharmaceutical composition, the second agent is selected from the group consisting of peginterferon, ribavirin, daclatasvir, boceprevir, telapravir, simeprevir, sofosbuvir, dasabuvir, ombitasvir, velpatasvir, ledipasvir, paritaprevir, ritonavir, elbasvir, grazoprevir, asunaprevir, beclabuvir, and combinations thereof.

In another aspect, the present invention provides a method of inhibiting the function of the HCV NS5A protein, comprising contacting a biological sample containing the HCV NS5A protein with a compound according to any embodiment disclosed herein, or a salt, stereoisomer, tautomer, solvate, or prodrug thereof.

In another aspect, the present invention provides a method of treating or preventing a disease, disorder, or medical condition associated with an HCV activity in a subject, comprising administering to the subject a therapeutically effective amount of the compound of according to any embodiment disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof.

In one embodiment of this aspect, the method is further in conjunction with administration of a second agent having anti-HCV activity to the subject.

In one embodiment of this aspect, the second agent is selected from the group consisting of a recombinant Human Interferon Alfa, a nucleoside analog, a direct acting antiviral, a NS3/4A protease inhibitor, a nucleotide NS5B polymerase inhibitor, a NS5A inhibitors, a non-nucleoside NS5B polymerase inhibitors, including, but limited to, peginterferon, ribavirin, daclatasvir, boceprevir, telapravir, simeprevir, sofosbuvir, dasabuvir, ombitasvir, velpatasvir, ledipasvir, paritaprevir, ritonavir, elbasvir, grazoprevir, asunaprevir, beclabuvir, and combinations thereof.

In another embodiment, the method for the treatment or prevention of a disease, disorder, or medical condition associated with HCV activity in a subject comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition according to any embodiment disclosed herein.

The disease, disorder, or medical condition associated with HCV activity includes, but is not limited to, hepatitis C infection, chronic hepatitis, cirrhosis, hepatocellular carcinoma (HCC), liver cancer, mixed cryoglobulinemia, porphyria cutanea tarda, leukocytoclastic vasculitis, lichen planus (LP), sicca syndrome, urticaria, pruritus, thrombocytopenic purpura, and psoriasis.

In one embodiment, the diseases, disorders or medical conditions caused by HCV may include cirrhosis, liver cancer and hepatitis C infection.

In one embodiment, the disease is cirrhosis.

In one embodiment, the disease is liver cancer.

In one embodiment, the disease is hepatitis C infection.

Unless otherwise indicated, the term“alkyl,” as used herein, is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups containing 1 to 8 carbons, preferably 1 to 6, more preferably 1 to 4, carbons. The term encompasses, but is not limited to, methyl, ethyl, propyl, isopropyl, butyl, t-butyl, isobutyl, pentyl, hexyl, or the like.

Unless otherwise indicated, the term“alkylene,” as used herein, refers to a bivalent saturated aliphatic radical derived from an alkane by removal of two hydrogen atoms. Examples include, but are not limited to, methylene (— Ctk— ), ethylene (— CH₂CH₂— ), propylene (— CH₂CH₂CH₂— ), or the like.

Unless otherwise indicated, the term“cycloalkyl”, as used herein alone or as a part of another group, includes saturated cyclic hydrocarbon radical having 3 to 8 carbons forming the ring. Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

Unless otherwise indicated, the term“aryl”, as used herein alone or as part of another group, refers to monocyclic or bicyclic aromatic radical containing 6 to 10 carbons in the ring portion (such as phenyl and naphthyl, including 1 -naphthyl and 2-naphthyl).

The term "heteroaryl," as used herein, refers to 5- to 14-membered monocyclic, bicyclic, or tricyclic, sometimes preferably 5- to 10-membered monocyclic or bicyclic, aromatic radical comprising one or more, preferably one to four, sometimes preferably one to three, heteroatoms independently selected from nitrogen (N), oxygen (O), and sulfur (S) in the aromatic ring(s). As is well known to those skilled in the art, heteroaryl rings have less aromatic character than their all-carbon counterparts. Thus, for the purposes of the invention, a heteroaryl group need only have some degree of aromatic character.

“Halo” or“halogen” as used herein, refers to fluoro (F), chloro (Cl), bromo (Br), and iodo (I).

When any group, for example, alkyl, alkenyl,“cycloalkyl,”“aryl,”“heterocyclyl,” or “heteroaryl”, is said to be“optionally substituted,” unless specifically defined, it means that the group is or is not substituted by from one to five, sometimes preferably one to three, substituents independently selected from halogen, C₁-C₆ alkyl, C₁-C₆ alkoxy, Ci-Ce haloalkyl, C₁-C₆ haloalkoxy, hydroxy, oxo, C₁-C₆ acyl, cyano, nitro, and NR^aR^b (R^a and R^b are each independently H or C₁-C₄ alkyl), or the like, provided that such substitution would not violate the conventional bonding principles known to a person of skill in the art. When the phrase“optionally substituted” is used before a list of groups, it means that each one of the groups listed may be optionally substituted.

As a person of skill in the art would understand, when an aryl, heteroaryl, cycloalkyl, heterocyclyl, or the like, is between two or more groups, it should be interpreted as a divalent group with a proper name“arylene,”“heteroarylene,”“cycloalkylene,”“heterocycylene,” or the like. Although sometimes no such distinction is made, for example,“aryl” should be interpreted as“arylene”, as a person of skill in the art would understand.

The term“optionally substituted” means the substitution may or may not occur and includes instances where said substitution occurs and instances in which it does not. One of ordinary skill in the art would understand that with respect to any molecule described as containing one or more substituents, only sterically practical and/or synthetically feasible compounds are meant to be included. Unless otherwise specified in this specification, when a variable is said to optionally substituted or substituted with a substituent(s), this is to be understood that this substitution occurs by replacing a hydrogen that is covalently bound to the variable with one these substituent(s).

As used herein, the term“pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. See, e.g., S. M. Berge et a , J. Pharm. Sci, 1977, 66, 1-19, which is incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, lumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2- naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like.

Stereochemical definitions and conventions used herein generally follow McGraw- Hill Dictionary of Chemical Terms, S. P. Parker, Ed., McGraw-Hill Book Company, New York (1984) and Stereochemistry of Organic Compounds, Eliel, E. and Wilen, S., John Wiley & Sons, Inc., New York (1994). Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule about its chiral center(s). The prefixes d and 1 or (+) and (-) are employed to designate the sign of rotation of plane-polarized light by the compound, with (-) or 1 meaning that the compound is levorotatory and (+) or d, meaning the compound, is dextrorotatory. For a given chemical structure, these compounds, called stereoisomers, are identical except that they are mirror images of one another. A specific stereoisomer of a mirror image pair may also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture. With reference to the instances where ( R ) or (S) is used, it is to designate the absolute configuration of a substituent in context to the whole compound and not in context to the substituent alone.

The term“tautomer” refers to a molecule in which a proton may shift from one atom to another atom in the same molecule, for example, in amide and carbamide bonds or heterocycles such as imidazole. The compounds presented herein may exist as tautomers. In circumstances where tautomerization is possible, a chemical equilibrium of the tautomers will exist, and the exact ratio of the tautomers may depend on several factors, including physical state, temperature, solvent, and pH, etc. For illustration purpose, some examples of tautomeric equilibrium (not exhaustive) may include:

Tautomerization of Compound 9

Tautomerization of Compound 17 The terms“racemic mixture” and“racemate” refer to an equimolar mixture of two enantiomeric species, devoid of optical activity.

The terms “Enantiopure” or “enantomerically pure” means a pure stereoisomer uncontaminated by its enantiomer.“Enantiomerically enriched” means a compound in which one of two (or more) enantiomers is found in greater concentrations in a given sample than another enantiomer.

As used herein, the term“substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. Illustrative substituents include, for example, those described herein. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valencies of the heteroatoms. This invention is not intended to be limited in any manner by the permissible substituents of organic compounds.

The term“solvate,” as used herein, means a physical association of a compound of this invention with a stoichiometric or non- stoichiometric amount of solvent molecules. For example, one molecule of the compound associates with one or more, preferably one to three, solvent molecules. It is also possible that multiple (e.g., 1.5 or 2) molecules of the compound share one solvent molecule. This physical association may include hydrogen bonding. In certain instances the solvates will be capable of isolation as crystalline solid. The solvent molecules in the solvate may be present in a regular arrangement and/or a non-ordered arrangement. Exemplary solvates include, but are not limited to, hydrates, ethanolates, methanolates, and isoprop anolates. Methods of solvation are generally known in the art.

The term“prodrug” as used herein means derivatives of the compounds of the invention which have chemically or metabolically cleavable groups and become, by solvolysis or under physiological conditions, the compounds of the invention which are pharmaceutically active in vivo. A prodrug of a compound may be formed in a conventional manner with a functional group of the compounds such as with an amino, hydroxy or carboxy group when present. The prodrug derivative form often offers advantages of solubility, tissue compatibility, or delayed release in a mammalian organism (see, Bundgard, H., Design of Prodrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985). Prodrugs include acid derivatives well known to practitioners of the art, such as, for example, esters prepared by reaction of the parent acidic compound with a suitable alcohol, or amides prepared by reaction of the parent acid compound with a suitable amine.

When it is possible that, for use in therapy, therapeutically effective amounts of a compound of the present invention, or pharmaceutically acceptable salts or solvates thereof, may be administered as the raw chemical, it is possible to present the active ingredient as a pharmaceutical composition. Accordingly, the disclosure further provides pharmaceutical compositions, which include any compounds of the present invention, or pharmaceutically acceptable salts or solvates thereof, and one or more, preferably one to three, pharmaceutically acceptable carriers, diluents, or other excipients. The carrier(s), diluent(s), or other excipient(s) must be acceptable in the sense of being compatible with the other ingredients of the formulation and not deleterious to the subject being treated.

The compositions of the present invention may be formulated in a conventional manner using one or more pharmaceutically acceptable carriers. Pharmaceutically acceptable carriers that may be used in these pharmaceutical compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as prolamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose- based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.

The term“pharmaceutically acceptable,” as used herein, refers to the property of those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of patients without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit/risk ratio, and are effective for their intended use.

The compositions of the present invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. The term “parenteral” as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques.

Pharmaceutical formulations adapted for oral administration may be presented as discrete units such as capsules or tablets; powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; edible foams or whips; or oil-in-water liquid emulsions or water-in-oil emulsions.

For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Powders are prepared by comminuting the compound to a suitable fine size and mixing with a similarly comminuted pharmaceutical carrier such as an edible carbohydrate, as, for example, starch or mannitol. Flavoring, preservative, dispersing, and coloring agent can also be present.

Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents, and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, and the like. Lubricants used in these dosage forms include sodium oleate, sodium chloride, and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, betonite, xanthan gum, and the like. Tablets are formulated, for example, by preparing a powder mixture, granulating or slugging, adding a lubricant and disintegrant, and pressing into tablets. A powder mixture is prepared by mixing the compound, suitable comminuted, with a diluent or base as described above, and optionally, with a binder such as carboxymethylcellulose, an aliginate, gelating, or polyvinyl pyrrolidone, a solution retardant such as paraffin, a resorption accelerator such as a quaternary salt and/or and absorption agent such as betonite, kaolin, or dicalcium phosphate. The powder mixture can be granulated by wetting with a binder such as syrup, starch paste, acadia mucilage, or solutions of cellulosic or polymeric materials and forcing through a screen. As an alternative to granulating, the powder mixture can be run through the tablet machine and the result is imperfectly formed slugs broken into granules. The granules can be lubricated to prevent sticking to the tablet forming dies by means of the addition of stearic acid, a stearate salt, talc, or mineral oil. The lubricated mixture is then compressed into tablets. The compounds of the present disclosure can also be combined with a free flowing inert carrier and compressed into tablets directly without going through the granulating or slugging steps. A clear or opaque protective coating consisting of a sealing coat of shellac, a coating of sugar or polymeric material, and a polish coating of wax can be provided. Dyestuffs can be added to these coatings to distinguish different unit dosages.

Oral fluids such as solution, syrups, and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of the compound. Syrups can be prepared by dissolving the compound in a suitably flavored aqueous solution, while elixirs are prepared through the use of a non-toxic vehicle. Solubilizers and emulsifiers such as ethoxylated isostearyl alcohols and polyoxyethylene sorbitol ethers, preservatives, flavor additive such as peppermint oil or natural sweeteners, or saccharin or other artificial sweeteners, and the like can also be added.

Where appropriate, dosage unit formulations for oral administration can be microencapsulated. The formulation can also be prepared to prolong or sustain the release, for example, by coating or embedding particulate material in polymers, wax, or the like.

It should be understood that in addition to the ingredients particularly mentioned above, the formulations may include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavoring agents.

The term“subject” or“patient” includes both humans and other mammalian animals, including but not limited to horses, cats, dogs, monkeys, and cows, and preferably humans.

The term "treatment" includes partial or total inhibition of a disease, disorder, or medical condition caused by the heptatitis C virus in humans, including cirrhosis and liver cancer. The term "prevention" includes either preventing the onset of hepatitis C infection altogether or preventing the onset of hepatitis C infection in individuals at risk.

Other aspects or advantages of the present invention will be apparent to those skilled in the art in view of the following detailed description and claims in combination with the knowledge and skills generally known in the field.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

It is noted here that, as used in this specification and the appended claims, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly dictates otherwise. The terms“including,”“comprising,”“containing,” or“having” and variations thereof are meant to encompass the items listed thereafter and equivalents thereof as well as additional subject matter unless otherwise noted.

Chemical Synthesis

The compounds described herein, and/or the pharmaceutically acceptable salts and solvates thereof, can be synthesized from commercially available starting materials by methods well known to those skilled in the art. The following general synthetic schemes illustrate methods for most of compound preparation. In each of these schemes, G¹ and G² are leaving groups that are the same or different and are exemplified but not limited to halogen, mesylate, tosylate or triflate. In addition, the reagents, solvents, temperatures, catalysts and ligands are not limited to what is depicted for illustrative purposes. Certain abbreviations and acronyms well known to those trained in the art that are used in the schemes are listed below for clarity.

Abbreviations and Acronyms

The following abbreviations and acronyms may be used in this application:

anhyd. = anhydrous

aq. = aqueous;

br = broad

brine = saturated aqueous NaCl;

Ebpiro = bis(pinacolato)diboron;

/7-BU3P = tri-n-butylphosphine;

CAS# = Chemical Abstracts Service Registry Number;

Compd = compound;

coned. = concentrated

d = day(s);

DCM = dichloromethane;

DIEA = DIPEA = /V,/V-diisopropylethylamine;

DMF = N, /V-di meth y 1 Form am i de ;

DMSO = dimethylsulfoxide;

DMA = /V,/V-dimethylacetamide;

dppf = l,r-bis(diphenylphosphino)ferrocene;

EtOAc = ethyl acetate;

Ex = Example;

FCC = flash column chromatography using silica;

h = hour(s);

HATU = 1 -|bis(dimethylamino)methylene|- 1 H- 1 ,2,3-triazolo|4,5-/; Ipyridinium 3- oxide hexafluorophosphate;

IBCF = isobutylchloroformate;

KOAc = potassium acetate;

LDA = lithium diisopropylamide;

LiHMDS = lithium bis(trimethylsilyl)amide [LiN(SiMe3)2];

MeOH = methanol; min. = minutes;

NMM = N- methyl morpho line;

Pd2(dba)3 = tris(dibenzylideneaeetone)dipalladium(0);

Pd(PPh3)2Cl2 = bis(triphenylphosphine)palladium(II) dichloride;

Pd(dppf)Cl2 = [1 ,1 '-bis(diphenylphosphino)ferrocene]dichloropalladium(II);

Pd(dppf)Cl2· CH2CI2 = [1 ,1 '-bis(diphenylphosphino)ferrocene]dichloropalladium(II), complex with dichloromethane

r.t. = room temperature;

satd. = saturated;

T3P = 2,4,6-tripropyl- 1 ,3 ,5,2,4,6-trioxatriphosphorinane-2,4,6-trioxide;

TEA = triethylamine;

TFA = trifluoroacetic acid;

THF = tetrahydrofuran;

wt. = weight;

wt. % = weight percent;

w/v = weight to volume ratio;

Xantphos = 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene;

XtalFluor-E = (diethylamino)difluorosulfonium tetrafluoroborate.

General Synthetic Schemes

Scheme 1 shows a general synthesis of the compounds of the formula I of the invention when the compounds have a symmetric structure of formula 5, wherein a transition metal-mediated cross-coupling reaction is used to construct the P'-M and M-P² bonds. For illustrative purposes, the Suzuki reaction is employed to couple an intermediate G'-M-G² 1, where G¹ and G² are the same or different, with boronic acids or esters 2 using a palladium catalyst such as Pd(dppf)2Cl2. Alternatively, boronic acids or esters 3 can be coupled under similar conditions with 4 to furnish compound 5 of the formula I. Other transition metal- mediated cross-coupling reactions that enable P*-M and M-P² bond formation but employ alternative coupling partners and reagents include but are not limited to the Negishi, Kumada, Sonagasira and Stille reactions.

Scheme 1: Representative synthesis of compounds of the formula I of a symmetric structure

3 4

Scheme 2 shows a synthesis of the compounds of the formula I of the invention that contain a carboxamide linkage. Coupling of carboxylic acids 6 with amines 7 using peptide coupling reagents such as HATU affords carboxamides 8 of the formula I. Alternatively, the reaction of amines 7 with the corresponding carboxylic acid halide, activated ester or mixed anhydride derivatives of 6 can also provide carboxamides 8 of the formula I. Scheme 2

6 7 8

Other general schemes for synthesis of the compounds of formula I include, but are not limited to, Schemes 3 to 8, below.

Scheme 3

X-M-X + B¹-A¹-Q¹-P¹-Y

B¹-A¹-Q¹-P¹-X + Y-M-Y

Scheme 4

Scheme 5

G²-Q¹-X²

X¹-M-X¹ + G¹-P¹-Y G¹-P¹-M-P¹-G¹ - ► G²-Q¹-P¹-M-P¹-Q¹-G²

B¹-A¹-X³

- B¹-A¹-Q¹-P¹-M-P¹-Q¹-A¹-B¹ Scheme 6

G²-A¹-Q¹-X²

X¹-M-X¹ + G¹-P¹-Y G¹-P¹-M-P¹-G¹ - »-

B¹-X³

G²-A¹-Q¹-P¹-M-P¹-Q¹-A¹-G² B¹-A¹-Q¹-P¹-M-P¹-Q¹-A¹-B¹

Scheme 7

B¹-A¹-Q¹-P¹-G¹ +

B¹-A¹-Q¹-pi-B(OR)₂

Scheme 8

G²-A¹ -Q¹ -Y²

X¹-M-X² + G¹-P¹-Y¹ G¹-P¹-M-X² - ^► G²-A¹-Q¹-P¹-M-X²

B¹-G³ G²-P²-Y²

B¹-A¹-Q¹-P¹-M-X² - B¹-A¹-Q¹-P¹-M-P²-G²

B¹-A¹-Q¹-P¹-M-P²-Q²-A²-B²

These general schemes are provided for illustration purpose. In any of the synthetic schemes provided, any of the X, X¹, X², X³, Y, Y¹, Y², Y³, G, G¹, G², and G³ may be selected, as needed, from various coupling, leaving, or protecting groups, preferably with good selectivities. Where needed, protection and deprotection of certain functional groups, for example, hydroxyl (-OH), amino (-NFb), and carboxyl group (-CO2H) may be implemented.

The compounds thus obtained can be further modified at their peripheral positions to provide the desired compounds. Synthetic chemistry transformations are described, for example, in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T.W. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley and Sons (1999); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995) and subsequent editions thereof. The compounds of the formula I and/or their pharmaceutically acceptable salts and solvates described herein can be purified by column chromatography, high performance liquid chromatography, crystallization, or other suitable methods.

EXAMPLES The following non- limiting Examples further illustrate certain aspects of the present invention. These compounds are prepared according to the general synthetic schemes described above.

Example 1

(±)-4,12-Bis-[W-(methoxycarbonyl)-L-valyl-L-prolyl-W-(3-aminophenyl)]- [2.2]paracyclophane (9)

Compound 9 was prepared according to Scheme 9. Scheme 9

(±)-4,12-Bis-(3-aminophenyl)-[2.2]paracyclophane (12). (+)-4,12-Dibromo[2.2]- paracyclophane (10) (100 mg, 0.2732 mmol; CAS#23927-40-4), 3-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)benzenamine (11) (239 mg, 1.09 mmol; CAS#210907-84-9) and K3PO4 (476 mg, 2.24 mmol) were combined with 10.6 mL of 10:1 mixture of 1,4-dioxane and water (v/v) in a microwave reaction tube. The resulting white slurry was degassed via N2 bubbling for 30 min., treated with Pd(dppf)Cl2· CH2CI2 (22 mg, 0.0273 mmol) and heated on a microwave reactor at 80 °C with vigorous stirring for 2 h. The reaction mixture was cooled to room temperature and partitioned between water and CPhCk/MeOH (19:1). The aqueous layer was extracted 2 more times with CPhCb/MeOH (19:1) and the combined organic extracts were washed twice with brine, dried (Na2SC>4), filtered and concentrated in vacuo. The residue was purified via chromatography on silica gel eluting with a gradient of 5-30% EtOAc in hexanes to give 22 mg (21%) of (±)-4,12-bis-(3-aminophenyl)-[2.2]paracyclophane (12) as a white foam: MS (m/z) MH⁺ 391.

(±)-4,12-Bis-[/V-(fer/-butoxycarbonyl)- -prolyl-/V-(3-aminophenyl)]-[2.2]paracyclophane

(14). N-Uert-Bu to x y c ar b o n y 1 ) - L - p ro line (13) (27 mg, 0.126 mmol) and DIEA (30 pL, 0.1713 mmol) were dissolved in DMF (1.0 mL) and treated with HATU (65 mg, 0.1713 mmol). The reaction mixture was placed under a nitrogen atmosphere, stirred at r.t. for 30 min. and then treated with a solution of (±)-4,12-bis-(3-aminophenyl)-[2.2]paracyclophane (12) (22 mg, 0.0571 mmol) in DMF (1.5 mL). After 18 h, the reaction mixture was quenched by the addition of water (5 mL), diluted with brine (5 mL) and extracted 3 times with 5 mL of EtOAc. The combined EtOAc layers were washed twice with 5 mL of a 1:1 mixture of satd. aq. NaHCCL and brine (v/v), then twice with 5 mL of 10% aq. citric acid, dried (Na2S04), filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with a gradient of 10-50% EtOAc in hexanes to afford 37 mg (83%) of (±)-4,12-bis- I AA/cw-butoxycarbonyl J-L-prolyl-AA 3-ami nophenyl ) |-| 2.2 Iparacyclophane (14) as a beige solid: MS (m/z) MH⁺ 785 (weak).

(±)-4,12-Bis-[ -prolyl-/V-(3-aminophenyl)]-[2.2]paracyclophane (15). (±)-4,12-Bis-[A-

( /cw-butoxycarbonyO-L-prolyl- V-i 3 -aminophenyl ) |- 12.2 Iparacyclophane (14) (37 mg, 0.0471 mmol) was dissolved in CH2CI2 (1.0 mL) and treated with trifluoroacetic acid (1.0 mL) while stirring at r.t. under a nitrogen atmosphere. After 25 h, the reaction mixture was concentrated in vacuo and the residue was partitioned between IN aq. NaOH (10 mL) and CH2CI2 (5 mL). The basic aq. layer was extracted 2 more times with CH2CI2 (5 mL) and the combined organic extracts were washed twice with brine, dried (Na2S04), filtered and concentrated in vacuo. The residue was purified via chromatography on silica gel eluting with a gradient of 5-30% MeOH/NELOH (49:1) in CH2CI2 to give 11.4 mg (41%) of (±)-4,12-bis-[L-prolyl-A- (3-aminophenyl)]-[2.2]paracyclophane (15) as a white solid: MS (m/z) MH⁺ 585.

(±)-4,12-Bis-[/V-(methoxycarbonyl)- -valyl- -prolyl-/V-(3-aminophenyl)]- [2.2]paracyclophane (9). N- ( m etho x ycarb o n y 1 ) - L- va 1 i n e (16) (8.5 mg (0.0487 mmol) and DIEA (10 pL, 0.0585 mmol) were dissolved in DMF (2.0 mL) and treated with HATU (18.5 mg, 0.0487 mmol) while stirring at r.t. under a nitrogen atmosphere. After 30 min., (±)-4,12- b i s - 1 L- p ro 1 y 1 - - ( 3 - a m i no p hen y 1 ) | - 12.21 p arac yc 1 o p ha ne (15) (11.4 mg, 0.0195 mmol) was added and the resulting slurry was stirred at r.t. for 16 h. The reaction was quenched by the addition of water (5 mL), diluted with brine (5 mL) and extracted 3 times with CH2CI2 (5 mL). The combined organic layers were extracted twice with 10% aq. citric acid (5 mL), twice with brine (5 mL), dried (NaaSCU), filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with a gradient of 0-5 % MeOH in CH2CI2 to furnish 13.8 mg (78%) of (±)-4,12-bis-[A-(methoxycarbonyl)-L-valyl-L-prolyl-A-(3- aminophenyl)]-[2.2]paracyclophane (9) as a white solid: MS (m/z) MH⁺ 899; ¹ H NMR (300 MHz, CD3OD): d 7.98 (br s, 0.2H), 7.95 (br s, 0.8H), 7.90 (br s, 0.2H), 7.88 (br s, 0.8H), 7.60-7.49 (m, 2H), 7.40 (dd, J = 7.8, 7.9 Hz, 2H), 7.27-7.17 (m, 2H), 6.81-6.57 (m, 6H), 4.66-4.56 (m, 2H), 4.28-4.18 (m, 2H), 4.06-3.92 (m, 2H), 3.98-3.71 (m, 2H), 3.65 (s, 6H), 3.60-3.39 (m, 2H), 3.11-2.91 (m, 2H), 2.91-2.76 (m, 2H), 2.76-2.61 (m, 1.6H), 2.61-2.47 (m, 0.4H), 2.43-2.26 (m, 2H), 2.26-1.91 (m, 8H), 1.12-0.81 (m, 12H).

Example 2 Methyl ((5)-l-((5)-2-(5-(9,9-difluoro-7-(3-((5)-l-((methoxycarbonyl)-L-valyl)pyrrolidine-2- carboxamido)phenyl)-977-fluoren-2-yl)- 177-imidazol-2-yl)pyrrolidin- 1 -yl)-3 -methyl- 1 - oxobutan-2-yl)carbamate (17)

Compound 17 was prepared according to Scheme 10. Scheme 10

to7-Butyl (S)-2-(5-(7-(3-aminophenyl)-9,9-difluoro-9//-fluoren-2-yl)-l/7-imidazol-2- yl)pyrrolidine-l-carboxylate (20). /_<?/7- Butyl (5)-2-(5-(7-bromo-9,9-difluoro-9//-fluoren-2- yl J- 17/-imida/ol-2-yl)pyrrolidine- 1 -carboxylate (18) (100 mg, 0.1936 mmol; CAS #1256388- 03-0; J. Med. Chem. 2014, 57(5), 2033-2046), 3-aminophenylboronic acid pinacol ester (19) (85 mg, 0,3873 mmol), K3PO4 (123 mg, 0.5808 mmol), water (0.25 mL) and 1,4-dioxane (2.5 mL) were combined in a microwave reaction tube. The resulting mixture was degassed via bubbling with N₂ while stirring for 60 min., then Pd(dppf)Cl2 (14.2 mg, 0.0194 mmol) was added. The resulting mixture was heated on a CEM Discover SP microwave reactor in PowerMax mode at 80 °C with vigorous stirring for 2 h. The reaction was cooled to r.t., diluted with EtOAc (15 mL), extracted twice with 15 mL of a 1:1 mixture of satd. aq. NaHCCb and brine (v/v), washed twice with 15 mL of brine, dried (Na₂SC>₄), filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with a gradient of 10-40% EtOAc in CH2CI2 to afford 100 mg (98%) of ieri-butyl (S)-2-(5-(7- ( 3-ami nophenyl )-9,9-dinuoro-97/-lluoren-2-yl)- 17/-imidazol-2-yl)pyrrolidine- 1 -carboxyl ate (20) as a light orange solid: MS (m/z) MH⁺ 529. to7-Butyl (S)-2-(5-(9,9-difluoro-7-(3-((S)-l-((methoxycarbonyl)- -valyl)pyrrolidine-2- carboxamido)phenyl)-9/7-fluoren-2-yl)-l/7-imidazol-2-yl)pyrrolidine-l -carboxylate (22).

A solution of (methoxycarbonyl)-L-valyl-L-proline (21) (66 mg, 0.2435 mmol; CAS #181827-47-4) and DIEA (98 pL, 0.5619 mmol) in DMF (2.0 mL) was treated with HATU (93 mg, 0.2435 mmol) while stirring at r.t under a nitrogen atmosphere. After 30 min., a solution of ieri-butyl (5)-2-(5-(7-(3-aminophenyl)-9,9-difluoro-977-fluoren-2-yl)-177- imidazol-2-yl)pyrrolidine-l-carboxylate (20) (99 mg, 0.1873 mmol) in DMF (1.0 mL) was added. After 15 h, the reaction was diluted with EtOAc (15 mL), extracted twice with 15 mL of a 1:1 mixture of satd. aq. NaHC0₃ and brine (v/v), washed twice with 15 mL of brine, dried (Na₂S0₄), filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with a gradient of 10-60% EtOAc in CH2CI2 to yield 97 mg, (66%) of ieri-butyl (5)-2-(5-(9,9-difluoro-7-(3-((5)-l-((methoxycarbonyl)-L- valyl)pyrrolidine-2-carboxamido)phenyl)-977-fluoren-2-yl)-177-imidazol-2-yl)pyrrolidine-l- carboxylate (22) as a light orange solid: MS (m/z) MH⁺ 783.

Methyl ((.S)-l-((.S)-2-((3-(9,9-difluoro-7-(2-((.S)-pyrrolidin-2-yl)-l//-imidazol-5-yl)-9//- fluoren-2-yl)phenyl)carbamoyl)pyrrolidin-l-yl)-3-methyl-l-oxobutan-2-yl)carbamate trifluoroacetate (1:2) (23). A solution of ieri-butyl (S)-2-(5-(9,9-difluoro-7-(3-((S)-l- ((methoxycarbonyl)-L-valyl)pyrrolidine-2-carboxamido)phenyl)-977-fluoren-2-yl)-177- imidazol-2-yl)pyrrolidine-l -carboxylate (22) (94 mg, 0.1196 mmol) in CH2CI2 (3.0 mL) was treated with trifluoroacetic acid (1.0 mL) while stirring at r.t. under a nitrogen atmosphere. After 2 h, the reaction mixture was concentrated in vacuo, triturated twice with Et₂0 and the resulting solid was dried in vacuo to afford 101 mg (92%) of methyl ((S)-l-((S)-2-((3-(9,9- difluoro-7-(2-((S)-pyrrolidin-2-yl)-l//-imidazol-5-yl)-9//-fluoren-2- yl)phenyl)carbamoyl)pyrrolidin- 1 -yl)- 3 -methyl- 1 -oxobutan-2-yl)carbamate trifluoroacetate

(1:2) (23) as a beige solid: MS (m/z) MH⁺ 683.

Methyl ((S)-l-((S)-2-(5-(9,9-difluoro-7-(3-((S)-l-((methoxycarbonyl)-L-valyl)- pyrrolidine-2-carboxamido)phenyl)-9//-fluoren-2-yl)-l//-imidazol-2-yl)pyrrolidin-l-yl)- 3-methyl-l-oxobutan-2-yl)carbamate (17). A solution of N-( methoxycarbonyl )- - valine (16) (29 mg, 0.1630 mmol) and DIEA (76 pL, 0.4348 mmol) were dissolved in DMF (2.0 mL) and treated with HATU (62 mg, 0.1630 mmol) while stirring at r.t. under a nitrogen atmosphere. After 30 min., a solution of ieri-butyl (.V)-2-(5-(7-(3-ami nophenyl )-9, 9-dill uoro- 977-fluoren-2-yl)-177-imidazol-2-yl)pyrrolidine-l-carboxylate (20) (99 mg, 0.1873 mmol) in DMF (1.0 mL) was added. After 18 h, the reaction mixture was diluted with EtOAc (15 mL), extracted twice with 15 mL of a 1:1 mixture of satd. aq. NaHCCb and brine (v/v), washed twice with brine (15 mL), dried (Na2SC>4), filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with a gradient of 0-5% MeOH in CH2CI2 to yield 67 mg (73%) of methyl ((S)-l-((S)-2-(5-(9,9-difluoro-7-(3-((S)-l- ((methoxycarbonyl)-L-valyl)-pyrrolidine-2-carboxamido)phenyl)-977-fluoren-2-yl)-177- imidazol-2-yl)pyrrolidin-l-yl)-3-methyl-l-oxobutan-2-yl)carbamate (17) as a beige solid: MS (m/z) MH⁺ 840; ¾ NMR (300 MHz, CD3OD): d 8.11-7.23 (m, 11H), 5.19-5.17 (m, 1H), 4.65-4.51 (m, 1H), 4.27-4.16 (m, 2H), 4.14-3.71 (m, 4H), 3.65 (s, 3H), 3.64 (s, 3H), 2.44- 2.07 (m, 10H), 1.11-0.88 (m, 12H).

Example 3

Dimethyl ((2S,2'S)-((2S,2'S)-(((( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-s-indacene-4,8- diyl)bis(3 , 1 -phenylene))bis(azanediyl))bis(carbonyl))bis(pyrrolidine-2, 1 -diyl))bis(3 -methyl- 1 -oxobutane- 1 ,2-diyl))dicarbamate (24).

Compound 24 was prepared according to Scheme 11.

Scheme 11

4,4'-(2,5-Dibromo-l,4-phenylene)bis(2-methylbut-3-yn-2-ol) (27). Triethylamine (125 mL) was added to a mixture of 1,4-dibro mo-2, 5-diiodobenzene (25) (12.2 g, 25.0 mmol), copper (I) iodide (49.6 mg, 0.260 mmol) and Pd(PPli3)2Cl2 (354 mg, 0.504 mmol). The resulting suspension was placed under a N2 atmosphere and degassed by 5 evacuation/N2 blanketing cycles and 2-methyl-3-butyn-2-ol (26) (6.1 mL, 62.9 mmol) was added. The reaction mixture was degassed again and then heated to reflux while stirring, under N2 for 17 h. The cooled reaction mixture was concentrated in vacuo, triturated several times with ethyl acetate and filtered. The filtrate was washed with brine, dried (CaSCL), filtered and concentrated in vacuo. The residue was recrystallized from boiling heptane to yield 9.3 g of

4,4'-(2,5-dibromo-l,4-phenylene)bis(2-methylbut-3-yn-2-ol) (27) as an off white powder: MS (m/z) (M-H₂0)H⁺ = 382; ¾ NMR (300 MHz, DMSO-d6): d 1.49 (s, 12H), 5.60 (s, 2H), 7.79 (s, 2H); ¹³C NMR (75 MHz, DMSO-d6) d 31.7, 64.3, 78.3, 104.0, 123.9, 126.2, 136.3.

4,4'-(3,3"-Dinitro-[l,l':4',l"-terphenyl]-2',5'-diyl)bis(2-methylbut-3-yn-2-ol) (29). 4,4'- (2,5-dibromo-l,4-phenylene)bis(2-methylbut-3-yn-2-ol) (27) (1.14 g, 2.85 mmol); 3- nitrophenylboronic acid (28) (1.43 g, 8.57 mmol); sodium carbonate (7.55 g, 71.3 mmol); Pd(dppf)Cl2· CH2CI2 (250 mg, 0.31 mmol) and tetrabutylammonium chloride hydrate (84.1 mg, 0.30 mmol) were combined at r.t. in a 1 :1 mixture (v/v) of THF and H2O (58 mL). The resulting suspension was degassed via 5 evacuation/N2 blanketing cycles and stirred at r.t. under N2_. After 2.5 h, the reaction mixture washed with brine and the organic layer was concentrated in vacuo and purified by chromatography on silica gel eluting with a gradient of 0-10% MeOH in CH2CI2 to give 1.61 g of light brown solid. This material was recrystallized from boiling ethyl acetate to provide 1.09 g of 4,4'-(3,3"-dinitro-[l,r:4',l"-terphenyl]-2',5'- diyl)bis(2-methylbut-3-yn-2-ol) (29) as a white powder: MS (m/z) (M-H₂0)H⁺ = 467; ¹ H NMR (300 MHz, DMSO-d6): d 1.34 (s, 12H), 5.40 (s, 2H), 7.69 (s, 2H), 7.80 (dd, J = 8.0, 8.0 Hz, 2H), 8.13 (d, J = 8.3 Hz, 2H), 8.32 (m, 2H), 8.49 (t, J = 1.9 Hz, 2H) ppm; ¹³C NMR (75 MHz, DMSO-d6) d 31.5, 64.1, 79.1, 102.1, 121.7, 123.4, 124.1, 130.4, 134.1, 136.2, 140.0, 140.6, 147.9.

4,4'-(3,3"-Diamino-[l,l':4',l"-terphenyl]-2',5'-diyl)bis(2-methylbutan-2-ol) (30). 10 wt.

% Pd on carbon (311 mg dry basis; 50 wt. % H2O; Degussa type E101) was added under N2 at r.t. to a rapidly stirred suspension of 4,4'-(3,3"-dinitro-[l,r:4',l"-terphenyl]-2',5'-diyl)bis(2- methylbut-3-yn-2-ol) (29) (1.09 g, 2.25 mmol) in EtOAc (200 mL). The suspension was subsequently sparged with hydrogen gas for 5 min. and then stirred under a hydrogen atmosphere for 7 h. The reaction was vented, flushed with N2, filtered through Celite, and the filtrate was concentrated in vacuo. The solid residue was recrystallized from boiling EtOAc/heptane to afford 885 mg of 4,4'-(3,3"-diamino-[l,r:4',l"-terphenyl]-2',5'-diyl)bis(2- methylbutan-2-ol) (30) as a white powder: MS (m/z) (M-H₂0)H⁺ = 415, MH⁺ = 433 (weak), MNa⁺ = 455 (weak): ¾ NMR (300 MHz, DMSO-d6): d 0.98 (s, 12H), 1.54 (m, 2H), 2.56 (m, 2H), 4.09 (s, 2H), 5.10 (s, 4H), 6.46 (d, J = 7.5 Hz, 2H), 6.54 (m, 4H), 6.99 (s, 2H), 7.05 (m, 2H); ¹³C NMR (75 MHz, DMSO-d6) d 27.6, 29.5, 46.4, 69.2, 112.8, 115.1, 117.1, 128.9, 130.7, 137.3, 141.2, 142.3, 148.9.

3,3'-(l,l,5,5-Tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-4,8-diyl)dianiline (31). 4,4'- (3,3"-Diamino-[l,l’:4’,l"-terphenyl]-2’,5’-diyl)bis(2-methylbutan-2-ol), (30) (187 mg, 0.43 mmol) was added in portions as a solid to trifluoroacetic acid (4.3 mL, 56.2 mmol) while stirring at r.t. under N2. The reaction mixture was heated to reflux for 4 d, cooled to r.t., carefully basified with satd. aq. NaHCCb and extracted with EtOAc. The organic extract washed with satd. aq. NaCl (aq.), filtered and concentrated in vacuo. The residue purified by chromatography on silica gel eluting with a gradient of 10-25% EtOAc in hexanes to yield 181 mg of white solid. This solid was recrystallized from EtO Ac/heptane to yield 131 mg of 3,3'-(l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-4,8-diyl)dianiline (31) as a fine white powder: MS (m/z) MH⁺ = 397; ¾ NMR (300 MHz, DMSO-d6): d 0.95 (d, 7 = 0.9 Hz, 6H), 0.99 (d, 7 = 1.4 Hz, 6H), 1.66 (t, 7 = 7.0 Hz, 4H), 2.36 (t, 7 = 7.0, 4H), 5.03 (s, 4H), 6.35 (dd, 7 = 7.4, 1.0 Hz, 2H), 6.41 (d, 7 = 1.7 Hz, 2H), 6.53 (dd, 7 = 8.0, 1.4 Hz, 2H), 7.03 (t, 7 = 7.7 Hz, 2H); ¹³C NMR (75 MHz, DMSO-d6) d 28.8, 28.9, 43.4, 45.5, 112.7, 115.4, 117.4, 128.6, 135.2, 140.7, 140.9, 145.6, 148.5.

Di-to7- butyl 2,2'-((((l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-4,8- diyl)bis(3,l-phenylene))bis(azanediyl))bis(carbonyl))(2S,2'S)-bis(pyrrolidine-l- carboxylate) (32). A mixture of 3,3'-(l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-

4.8-diyl)dianiline (31) (121 mg, 0.31 mmol), (ieri-butoxycarbonyl)-L-proline (13) (139 mg, 0.64 mmol), HATU (248 mg, 0.65 mmol) in THF (20 mL) was treated with DIEA (0.3 mL, 1.72 mmol) while stirring at r.t under N2. After 17 h, the resultant suspension was then partitioned between satd. aq. NaCl and EtOAc. The organic extract appeared as a suspension that was concentrated in vacuo to provide 536 mg of di-ieri-butyl 2,2'-((((l,l,5,5-tetramethyl- l,2,3,5,6,7-hexahydro-s-indacene-4,8-diyl)bis(3,l-phenylene))bis(azanediyl))- bis(carbonyl))(2.V,2A)-bis( pyrrolidine- 1 -carboxyl ate) (32) as a white solid, which was used without further purification: MS (m/z) MH⁺ = 791.

(2S,2'S)-/V^V'-((l,l»5,5-Tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-4,8-diyl)bis(3,l- phenylene))bis(pyrrolidine-2-carboxamide) dihydrochloride (33). Acetyl chloride (2.2 mL, 30.9 mmol) was slowly added via syringe to methanol (200 mL) while stirring at 0 °C. After 10 min, a suspension of di-ieri-butyl 2,2'-((((l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro- s-indacene-4,8-diyl)bis(3,l-phenylene))bis(azanediyl)) bis(carbonyl))(25,2'5)-bis(pyrrolidine- 1-carboxylate) (32) (536 mg, 0.68 mmol) in methanol (50 mL) was added and the resulting suspension was heated at reflux for 2h. The slightly turbid reaction mixture was filtered while hot and the filtrate was reduced in volume and diluted with EtOAc. The resulting precipitated product was isolated by filtration to afford 410 mg of (2S2'S)-N,N'-(( 1 , 1 ,5,5- tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-4,8-diyl)bis(3,l-phenylene))bis(pyrrolidine-2- carboxamide) dihydrochloride (33) as a white powder: MS (m/z) MH⁺ = 591.

Dimethyl ((2S,2'S)-((2S,2'S)-((((l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-

4.8-diyl)bis(3,l-phenylene))bis(azanediyl))bis(carbonyl))bis(pyrrolidine-2,l-diyl))bis(3- methyl-l-oxobutane-l,2-diyl))dicarbamate (24). DIEA (0.6 mL, 3.44 mmol) was added a suspension of (2S,2'S)-N,N'-(( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-s-indacene-4,8- diyl)bis(3,l-phenylene))bis(pyrrolidine-2-carboxamide) dihydrochloride (33) (203 mg, 0.31 mmol) and (methoxycarbonyl)-L- valine (118 mg, 0.67 mmol) in THF (6.2 mL), stirred at r.t. for lh and then treated with a 50 wt. % solution of T3P in EtOAc (0.6 mL, 1.01 mmol). After 17 h at r.t., the reaction mixture was partitioned between 5% aq. NaHCCh (w/v) and EtOAc. The organic extract washed with brine, dried (CaS0₄) and concentrated in vacuo and the residue was purified by chromatography on silica gel eluting with a gradient of 50-100% EtOAc in hexanes. The resulting material was recrystallized from EtOAc/heptane to afford 66.4 mg of dimethyl ((2S,2'S)-((2S,2'S)-(((( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-s- indacene-4,8-diyl)bis(3,l-phenylene))bis(azanediyl))bis(carbonyl))bis(pyrrolidine-2,l- diyl))bis(3 -methyl- 1 -oxobutane- 1 ,2-diyl))dicarbamate (24) as a white powder: MS (m/z) MH⁺ = 905; ¾ NMR (300 MHz, DMSO-d6): d 0.81-1.05 (m, 24 H), 1.68 (t, J = 6.5 Hz, 4H), 1.82-2.06 (m, 8H), 2.09-2.23 (m, 2H), 2.24-2.50 (m, 4H), 3.52 (s, 6H), 3.57-3.69 (m, 2H), 3.75-3.88 (m, 2H), 4.04 (t, J = 8.2 Hz, 2H), 4.41-4.51 (m, 2H), 6.92 (t, J = 5.5 Hz, 2H), 7.32-7.68 (m, 8H), 10.05 (s, 2H); ¹³C NMR (75 MHz, DMSO-d6): d 18.9, 19.1, 19.3, 25.1,

28.8, 29.9, 30.3, 43.4, 45.4, 47.6, 51.9, 58.4, 60.7, 117.7, 120.1, 124.6, 128.7, 134.6, 139.2, 140.7, 141.1, 146.0, 157.3, 170.

Example 4

Dimethyl (( 1 /?, 1 'R)-((2S,2'S)-(((( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-s-indacene-4,8- diyl)bis(3,l-phenylene))bis(azanediyl))bis(carbonyl))bis(pyrrolidine-2,l-diyl))bis(2-oxo-l- phenylethane-2, l-diyl))dicarbamate (34).

Compound 34 was prepared according to Scheme 12. Scheme 12

Dimethyl ((l/?,17?)-((2S,2'S)-((((l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-

4,8-diyl)bis(3,l-phenylene))bis(azanediyl))bis(carbonyl))bis(pyrrolidine-2,l-diyl))bis(2- oxo-l-phenylethane-2,l-diyl))dicarbamate (34). DIEA (0.6 mL, 3.44 mmol) was added at r.t. to a stirred suspension of (2S2'S)-N,N'-(( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-s- indacene-4,8-diyl)bis(3,l-phenylene))bis(pyrrolidine-2-carboxamide) dihydrochloride (33) (207 mg, 0.31 mmol) and (R)-2-((methoxycarbonyl)amino)-2-phenylacetic acid (35) (229 mg, 1.09 mmol; CAS #50890-96-5) in THF (6.2 mL), stirred at r.t for 1 h and then treated with a 50 wt. % solution of T3P in EtOAc (0.6 mL, 1.01 mmol). After 17 h at r.t., the reaction mixture was partitioned between 5% aq. NaHCCb (w/v) and EtOAc. The organic extract washed with brine, dried (CaS0₄) and concentrated in vacuo and the residue was purified by chromatography on silica gel eluting with a gradient of 50-100% EtOAc in hexanes. The resulting material was recrystallized from EtOAc/heptane to afford 74 mg of dimethyl (( 1 /?, 1 'R)-((2S,2'S)-(((( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-s-indacene-4,8- diyl)bis(3,l-phenylene))bis(azanediyl))bis-(carbonyl))bis-(pyrrolidine-2,l-diyl))bis(2-oxo-l- phenylethane-2,l-diyl))dicarbamate (34) as a fine white powder: MS (m/z) MH⁺ = 973; ¹ H NMR (300 MHz, DMSO-d6): d 0.94 (d, J = 3.0 Hz, 6H), 1.00 (d, J = 5.6 Hz, 6H), 1.70 (t, J = 6.9 Hz, 4H), 1.75-1.83 (m, 2H), 1.88-2.10 (m, 6H), 2.26-2.44 (m, 4H), 3.18 (br s, 2H), 3.49-3.55 (m, 6H), 3.82 (br s, 2H), 4.41 (d, J = 6.7 Hz, 2H), 5.34-5.51 (m, 2H), 6.94 (d, J = 7.5 Hz, 2H), 7.04-7.11 (m, 2H), 7.24-7.52 (m, 12H), 7.59-7.80 (m, 4H), 9.99 (s, 2H); ¹³C NMR (75 MHz, DMSO-d6): d 22.6, 24.7, 28.9, 29.7, 43.3, 45.5, 47.4, 52.0, 57.2, 61.2, 118.0, 118.2, 120.6, 124.8, 128.2, 128.5, 128.7, 129.1, 134.6, 137.5, 139.0, 140.7, 141.0, 145.9, 156.6, 168.9, 170.7.

Example 5

Dimethyl (( 1 /?, 1 'R)-((2S,2'S)-(((( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-s-indacene-4,8- diyl)bis(pyridine-5,3-diyl))bis(azanediyl))bis(carbonyl))bis(pyrrolidine-2,l-diyl))bis(2-oxo-l- phenylethane-2, l-diyl))dicarbamate (36).

36

Compound 36 was prepared according to Scheme 13.

Scheme 13

4,4'-(l,4-Phenylene)bis(2-methylbut-3-yn-2-ol) (38). Triethylamine (172 mL) was combined with 1,4-diiodobenzene (37) (11.4 g, 34.4 mmol), copper(I) iodide (329 mg, 1.7 mmol) and Pd(PPh3)2Cl2 (1.2 g, 1.7 mmol). The resulting suspension was degassed through 5 evacuation/N2 blanketing cycles while stirring. 2-Methyl-3-butyn-2-ol (6.1 mL, 62.9 mmol) was added and the reaction mixture was again degassed as above, then heated to reflux while stirring under N2. After 3 h, the reaction mixture was cooled to r.t. and concentrated in vacuo and triturated with hot EtOAc. The combined triturations were filtered through Celite and the filtrate was washed with 0.1 N aq. HC1, brine, dried (CaSC ), heated to boiling and then diluted with heptane to precipitate crude product. The precipitate was isolated by filtration and recrystallized from boiling heptane to yield 7.3 g of 4,4'-(l,4-phenylene)bis(2-methylbut- 3-yn-2-ol) (38) as a tan powder: MS (m/z) (M-H₂0)H⁺ = 225; ¾ NMR (300 MHz, DMSO- d6): 57.36 (s, 4H), 5.51 (s, 2H), 1.47 (s, 12H).

4,4'-(l,4-Phenylene)bis(2-methylbutan-2-ol) (39). 10 wt. % Pd on carbon (619 mg dry basis; 50 wt. % H₂0; Degussa type E101) was added under N2 at r.t. to a rapidly stirred suspension of 4,4'-(l,4-phenylene)bis(2-methylbut-3-yn-2-ol) (38) (4.1 g, 17.0 mmol) in MeOH (200 mL). The suspension was subsequently sparged with hydrogen gas for 5 min. and then stirred under a hydrogen atmosphere for approximately 1 week. The reaction mixture was filtered through Celite and the filtrate was concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with a gradient of 10-40% EtOAc in hexanes and then recrystallized from EtO Ac/hexanes to provide 805 mg of 4,4'-(l,4- phenylene)bis(2-methylbutan-2-ol) (39) as a white powder: ¹ H NMR (300 MHz, DMSO-d6): 5 7.07 (s, 4H), 4.23 (s, 2H), 2.59-2.50 (m, 4H), 1.63-1.57 (m, 4H), 1.13 (s, 12H); ¹³C NMR (75 MHz, DMSO-d6): 5 140.4, 128.5, 69.1, 46.3, 30.3, 29.7. l,l,5,5-Tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene (40). Methanesulfonic acid (10.4 mL, 160 mmol) was added rapidly at r.t. to a stirred solution of 4,4'-(l,4-phenylene)bis(2- methylbutan-2-ol) (39) (804 mg, 3.2 mmol) in CH2CI2 (160 mL) and the solution immediately turns reddish-brown. After 17 h, the reaction mixture was extracted sequentially with water, 5% (w/v) aq. NaHCCb, dried (CaSC ), filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with 100% hexanes to furnish 370 mg of l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene (40) as a white solid: ¹ H NMR (300 MHz, CDCb): 5 6.95 (s, 2H), 2.84 (t, J = 7.1 Hz, 4H), 1.91 (t, J = 7.1 Hz, 4H), 1.24 (s, 12H); ¹³C NMR (75 MHz, CDCb): 5 150.7, 141.0, 117.9, 43.4, 41.9, 29.9, 28.8.

4,8-Dibromo-l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene (41). Neat bromine (32 DL, 0.6 mmol) was added at r.t. to a stirred solution of 1, 1,5,5 -tetramethyl- 1,2, 3 ,5,6,7- hexahydro-s-indacene (40) (51 mg, 0.2 mmol) and iodine (12.3 mg, 0.1 mmol) in anhyd. CH2CI2 (2.4 mL) and stirred for 1 h in the dark. The reaction was diluted with additional CH2CI2, washed with 5 M aq. NaOH and the resulting colorless CH2CI2 extract was concentrated in vacuo. The residue was dissolved in EtOAc, filtered through Celite, and the filtrate was evaporated to yield 65 mg of 4,8-dibromo-l,l,5,5-tetramethyl-l,2,3,5,6,7- hexahydro-s-indacene (41) as a white solid: ¹ H NMR (300 MHz, CDCb): d 2.87 (t, J = 7.5 Hz, 4H), 1.94 (t, J = 7.5 Hz, 4H), 1.41 (s, 12H); ¹³C NMR (75 MHz, CDCb): d 148.3, 145.2, 116.2, 48.2, 41.8, 31.8, 26.9. tert-Butyl (5)-2-((5-bromopyridin-3-yl)carbamoyl)pyrrolidine-l-carboxylate (42).

Isobutyl chloroformate (4.5 mL, 34.7 mmol) was added slowly to a solution of (tert- butoxycarbonyl)-L-proline (13) (6.8 g, 31.6 mmol) and NMM (7.7 mL, 70 mmol) in EtOAc (160 mL) while stirring under N2 at 0 °C. After lh, the resulting suspension was slowly treated at 0 °C with a solution of 5-bromopyridin-3-amine (5.5 g, 31.6 mmol) in EtOAc (20 mL) and then slowly warmed to r.t. overnight. The reaction mixture was extracted with H2O and then with 5% aq. NaHC0₃, which resulted in the precipitation of product in the EtOAc phase. The EtOAc phase containing the precipitated product was concentrated in vacuo and the solid residue was triturated with boiling EtOAc and isolated by filtration to provide 7.6 g of ieri-butyl (.V)-2-((5-bromopyridin-3-yl (carbamoyl (pyrrolidine- 1 -carboxyl ate (42) as a white powder: MS (m/z) MH⁺ = 370.

(S)-(5-(l-(ter/-Butoxycarbonyl)pyrrolidine-2-carboxamido)pyridin-3-yl)boronic acid (43). ieri-Butyl (.V)-2-((5-bromopyridin-3-yl (carbamoyl (pyrrolidine- 1 -carboxyl ate (42) (2.0 g, 5.4 mmol), bis(pinacolato)diboron (1.5 g, 6.0 mmol), potassium acetate (1.6 g, 16.3 mmol) and Pd(dppi)Cb· CH2CI2 (224 mg, 0.3 mmol) were combined in anhydrous DMSO (54 mL) and then degassed. The reaction mixture was heated at 80 °C while stirring under N2 for 17 h, cooled to r.t. and partitioned between H2O water and EtOAc. The aqueous layer was extracted several times with EtOAc and the combined organic extracts were dried (CaS0₄), filtered and concentrated in vacuo to provide 3.5 g of (S)-(5-(l-(tert- butoxycarbonyl)pyrrolidine-2-carboxamido)pyridin-3-yl)boronic acid (43) as a black tar: MS (m/z) MH⁺= 336.

Di-to7- butyl 2,2'-((((l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-4,8- diyl)bis(pyridine-5,3-diyl))bis(azanediyl))bis(carbonyl))(2S,2'S)-bis(pyrrolidine-l- carboxylate) (44). 4,8-Dibromo-l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene (41) (221 mg, 0.6 mmol),

1 -(/_<?/7-butoxycarbonyl (pyrrol idine-2-carboxamido )pyridin-3- yl)boronic acid (43) (892 mg, 2.7 mmol), Na₂C0₃ (1.6 g, 14.7 mmol), Pd(dppf)Cl2· CH2CI2 (49 mg, 0.1 mmol), and tetrabutylammonium chloride hydrate (20 mg, 0.1 mmol) were combined in 20 mL of a 1:1 mixture of THF and H2O (v/v). The resulting suspension was degassed via 5 evacuation/N2 blanketing cycles and then heated at reflux for 3 h while stirring under N2. The reaction mixture was cooled to r.t, diluted with additional THF and extracted with brine. The organic layer was concentrated in vacuo and the residue was purified by chromatography on silica gel eluting with a gradient of 30-100% EtOAc in hexanes to yield 144 mg of di-ieri-butyl 2,2'-((((l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s- indacene-4,8-diyl)bis(pyridine-5,3-diyl))bis(azanediyl))bis(carbonyl))(25,2'5)- bis(pyrrolidine- 1 -carboxylate) (44) as an off white solid: MS (m/z) MH⁺ = 794.

(2S,2'S)-/V^V'-((l,l_»5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-4,8- diyl)bis(pyridine-5,3-diyl))bis(pyrrolidine-2-carboxamide) tetrahydrochloride (1:4) (45).

A stirred suspension of di-ieri-butyl 2,2'-((((l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s- indacene-4,8-diyl)bis(pyridine-5,3-diyl))bis(azanediyl))bis(carbonyl))(25,2'5)- bis(pyrrolidine- 1 -carboxylate) (44) (144 mg, 0.2 mmol) and coned aq. HC1 (2.0 mL) in methanol (90 mL) was heated at reflux. After 2 h, the reaction mixture was cooled to r.t. and concentrated in vacuo provide 160 mg of (2S,2'S)-N,N'-(( 1 , 1 ,5,5-tetramethyl- 1 ,2, 3, 5, 6, 7- hexahydro-s-indacene-4,8-diyl)bis(pyridine-5,3-diyl))bis(pyrrolidine-2-carboxamide) tetrahydrochloride (1 :4) (45) as a light brown solid: MS (m/z) MH⁺ = 593.

Dimethyl (( 1 R,1 'R)-((2S,2'S)-((((1 ,1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-s-indacene- 4,8-diyl)bis(pyridine-5,3-diyl))bis(azanediyl))bis(carbonyl))bis(pyrrolidine-2,l- diyl))bis(2-oxo-l-phenylethane-2,l-diyl))dicarbamate (36). DIEA (0.5 mL, 2.9 mmol) was added at r.t. to a stirred suspension of (2S,2'S)-N,N'-(( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7- hexahydro-s-indacene-4,8-diyl)bis(pyridine-5,3-diyl))bis(pyrrolidine-2-carboxamide) tetrahydrochloride (1:4) (45) (160 mg, 0.2 mmol) (207 mg, 0.31 mmol) and ( R)-2 - ((methoxycarbonyl)amino)-2-phenylacetic acid (35) (202 mg, 1.0 mmol) in EtOAc (85 mL), stirred at r.t for 1 h and then treated with a 50 wt. % solution of T3P in EtOAc (0.6 mL, 1.01 mmol). After 17 h at r.t., the reaction mixture was partitioned between 5% aq. NaHC0₃ (w/v) and EtOAc. The organic extract washed with brine, dried (CaS0₄) and concentrated in vacuo and the residue was purified by chromatography on silica gel eluting with a gradient of 0-10% MeOH in CH2CI2. The resulting material was recrystallized from EtO Ac/heptane to afford 94 mg of dimethyl (( 1 /?, 1 'R)-((2S 'S)-(((( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-s- indacene-4,8-diyl)bis(pyridine-5,3-diyl))bis(azanediyl))bis(carbonyl))bis(pyrrolidine-2,l- diyl))bis(2-oxo-l-phenylethane-2,l-diyl))dicarbamate (36) as a white solid: MS (m/z) MH⁺ = 976; ¾ NMR (300 MHz, DMSO-d6): d 10.18 (s, 2H), 8.83-8.82 (m, 2H), 7.93-7.92(m, 2H), 7.75 (t, 7 = 7.8 Hz, 2H), 7.43-7.32 (m, 12H), 3.87-3.80 (m, 2H), 3.52-3.51 (m, 6H), 3.25-3.15 (m, 2H), 2.41-2.32 (m, 4H), 2.09-1.96 (m, 8H), 1.81-1.73 (m, 8H), 1.01-0.99 (m, 6H), 0.96- 0.93 (m, 6H). Example 6

Dimethyl (( 1 /?, 1 'R)-((2S,2'S)-((( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-s-indacene-4,8- diyl)bis(3, 1 -phenylene))bis( 17/-imidazole-5,2-diyl ))his( pyrrolidine-2, 1 -diyl))bis(2-oxo- 1 - phenylethane-2, l-diyl))dicarbamate (46).

Compound 46 was prepared according to Scheme 14.

Scheme 14

Benzyl ((/?)-2-((methoxycarbonyl)amino)-2-phenylacetyl)- -prolinate (48). A suspension of (/?)-2-((methoxycarbonyl)amino)-2-phenylacetic acid (35) (2.5 g, 12.1 mmol) and benzyl L-prolinate hydrochloride (47) (2.9 g, 12.1 mmol) in EtOAc (120 mL) was treated with a 50 wt. % solution of T3P in EtOAc (7.2 mL, 12.2 mmol) at r.t under a N2 followed by DIEA (6.3 mL, 36.2 mmol). After 1 h, the reaction mixture was partitioned between water and extracted with additional EtOAc. The combined organic extracts were washed with 5% aq. NaHC0₃ (w/v), brine, dried (CaS0₄), filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with a gradient of 10-50% EtOAc to afford 2.2 g of benzyl ( (R)-2-(( methoxycarbonyl )ami no )-2-phenylacetyl )-L-prolinate (48) as a light yellow solid: MS (m/z) MH⁺= 397.

((/?)-2-((Methoxycarbonyl)amino)-2-phenylacetyl)- -proline (49). 10 wt. % Pd on carbon (890 mg dry basis; 50 wt. % H2O; Degussa type E101) was added under N2 at r.t. to a rapidly stirred suspension of ((/?)-2-((methoxycarbonyl)amino)-2-phenylacetyl)-L-prolinate (48) (2.2 g, 5.5 mmol) in EtOAc (175 mL). The suspension was subsequently sparged with hydrogen gas for 5 min. and then stirred under a hydrogen atmosphere for 2.5 h. The reaction mixture was filtered through Celite, and the filtrate was concentrated in vacuo to provide 2.4 g of (( /?)-2-( (methoxycarbonyl)ami no )-2-phenyl acetyl )-L-proline (49) as a colorless oil: MS (m/z) MH⁺= 307.

2-(3-Bromophenyl)-2-oxoethyl ((/?)-2-((methoxycarbonyl)amino)-2-phenylacetyl)- - prolinate (51). A solution of 2-bromo-l-(3-bromophenyl)ethan-l-one (50) (2.2 g, 7.7 mmol), and ((/?)-2-((methoxycarbonyl)amino)-2-phenylacetyl)-L-proline (49) (2.4 g, 7.7 mmol) in acetonitrile (80 mL) was treated dropwise at r.t. with TEA (2.2 mL, 15.8 mmol). After lh, the reaction mixture was partitioned between brine and EtOAc. The organic extract was dried (CaS0₄), filtered and concentrated in vacuo to afford 3.4 g of 2-(3-bromophenyl)- 2-oxoethyl ((/?)-2-((methoxycarbonyl)amino)-2-phenylacetyl)-L-prolinate (51) as a white foam: MS (m/z) MH⁺ = 503.

Methyl ((/f)-2-((S)-2-(5-(3-bromophenyl)-l/7-imidazole-2-earbonyl)pyrrolidin-l-yl)-2- oxo-l-phenylethyl)carbamate (52). A mixture of 2-(3-bromophenyl)-2-oxoethyl ((/?)- 2- ((methoxycarbonyl)amino)-2-phenylacetyl)-L-prolinate (51) (3.3 g, 6.5 mmol) and ammonium acetate (2.5 g, 32.6 mmol) was heated at reflux in toluene (110 mL) while stirring under N2. After 17 h, the reaction mixture was cooled to r.t., partitioned between water and EtOAc and the organic extract was dried (CaS0₄), filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with a gradient of 10-80% EtOAc in hexanes to yield 2.0 g of methyl (( ?)-2-((.V)-2-(5-(3-bromophenyl)- 1 //-imidazole-2- carbonyl)pyrrolidin-l-yl)-2-oxo-l-phenylethyl)carbamate (52) as a light yellow foam: MS (m/z) MH⁺= 483.

Methyl ((/?)-2-oxo-l-phenyl-2-((S)-2-(5-(3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)phenyl)-l/7-imidazole-2-earbonyl)pyrrolidin-l-yl)ethyl)earbamate (53). A stirred mixture of methyl (( ?)-2-((.V)-2-(5-(3-bromophenyl)- 1 //-imidazole-2-carbonyl)pyrrolidin- 1 - yl)-2-oxo-l-phenylethyl)carbamate (52) (2.0 g, 4.1 mmol), bis(pinacolato)diboron (1.3 g, 5.0 mmol), KOAc (970 mg, 9.9 mmol) and Pd(dppf)Cl2· CH2CI2 (338 mg, 0.4 mmol) in anhydrous DMSO (21 mL) was degassed and then heated at 80°C for 1 h under N2. The cooled reaction mixture was partitioned between brine and EtOAc. The aqueous layer was extracted again with EtOAc and the combined extracts were dried (CaS0₄), filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with a gradient of 0-10% methanol in CH2CI2 and then recrystallized from EtOAc/hexanes to furnish 970 mg of methyl ((/?)-2-oxo- 1 -phenyl-2-((.V)-2-(5-(3-(4,4,5,5-tetramethyl- 1 ,3,2- dioxaborolan-2-yl)phenyl)-177-imidazole-2-carbonyl)-pyrrolidin-l-yl)ethyl)carbamate (53) as a tan foam: MS (m/z) MH⁺= 531.

Dimethyl ((l/?,1 ?)-((2S,2'S)-(((l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene- 4,8-diyl)bis(3,l-phenylene))bis(l //-imidazole-5, 2-diyl))bis(pyrrolidine-2,l-diyl))bis(2- oxo-l-phenylethane-2,l-diyl))dicarbamate (46). 4,8-Dibromo-l,l,5,5-tetramethyl- 1,2,3,5,6,7-hexahydro-s-indacene (41) (196 mg, 0.5 mmol), methyl ((R)-2-oxo- 1 -phenyl-2- ((5)-2-(5-(3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)phenyl)-l//-imidazole-2- carbonyl)pyrrolidin-l-yl)ethyl)carbamate (53) (823 mg, 1.6 mmol), NaC0₃ (1.4 g, 13.1 mmol), Pd(dppf)Cl2· CH2CI2 (87 mg, 0.1 mmol), and tetrabutylammonium chloride hydrate (32.4 mg, 0.1 mmol) were combined in 20 mL of a 1 :1 mixture of THF/H2O (v/v). The resulting suspension was degassed via 5 evacuation/N2 blanketing cycles and then heated at reflux for 3 h while stirring under N2. The reaction mixture cooled to r.t, diluted with additional THF and washed with brine. The organic layer was concentrated in vacuo and the residue was purified by chromatography on silica gel eluting with 0-10% MeOH in CH2CI2 and then recrystallized from EtOAc/hexanes to yield 189 mg of dimethyl (( 1 R ’R)-((2S,2'S)- (((l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-4,8-diyl)bis(3,l-phenylene))bis(l/Z- imidazole-5 ,2-diyl))bis(pyrrolidine-2, 1 -diyl))bis(2-oxo- 1 -phenylethane-2, 1 -diyl))dicarbamate (46) as a white powder: MS (m/z) MH⁺ = 1020; ¾ NMR (300 MHz, DMSO-d6): d 11.94- 11.74 (m, 2H), 7.98-6.89 (m, 22H), 5.50-5.06 (m, 4H), 3.88-3.11 (m, 10H), 4.08 (br s, 4H), 2.00 (br s, 4H), 1.73-1.71 (m, 4H), 1.00-0.96 (m, 12H).

Example 7 Dimethyl (( 1 /?, 1 'R)-((2S,2'S)-((( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-s-indacene-4,8- diyl)bis(4, 1 -phenylene))bis( 17/-imidazole-5,2-diyl ))his( pyrrolidine-2, 1 -diyl))bis(2-oxo- 1 - phenylethane-2, l-diyl))dicarbamate (54).

Compound 54 was prepared according to Scheme 15. Scheme 15

2-(2-(4-Bromophenyl)-2-oxoethyl) 1 -(tot-butyl) (S)-pyrrolidine-l,2-dicarboxylate (56).

Et3N (6.0 mL, 43 mmol) was slowly added at r.t. to a stirred, slightly turbid solution of 2- bromo-l-(4-bromophenyl)ethan-l-one (55) (5.7 g, 21 mmol) and (i_<?ri-butoxycarbonyl)-L- proline (13) (5.2 g, 24.2 mmol) in acetonitrile (69 mL). After 2 h, the reaction mixture was partitioned between brine and EtOAc. The aqueous layer was extracted again with EtOAc and the combined organic extracts were dried (CaSCU), filtered and concentrated in vacuo to provide 7.8 g of 2-(2-(4-bromophenyl)-2-oxoethyl) l-(ieri-butyl) (5)-pyrrolidine-l,2- dicarboxylate (56) as a clear yellow oil: MS (m/z) MNa⁺ 434, (M-Boc)H⁺= 312. tert-Butyl (S)-2-(5-(4-bromophenyl)-l/ -imidazol-2-yl)pyrrolidine-l-carboxylate (57). A mixture of ammonium acetate (7.3 g, 94.2 mmol) and 2-(2-(4-bromophenyl)-2-oxoethyl) 1- (ieri-butyl) (5)-pyrrolidine-l,2-dicarboxylate (56) (7.8 g, 18.9 mmol) in toluene (125 mL) was heated at reflux. After 17 h, the reaction mixture was cooled to r.t. and partitioned between brine and EtOAc. The organic extract was dried (CaS0₄), filtered, and concentrated in vacuo. The resulting light yellow foam was crystalized from ethyl acetate/hexanes to yield 3.9 g of ieri-butyl (.V)-2-(5-(4-bromophenyl)- 17/-imidazol-2-yl)pyrrolidine- 1 -carboxyl ate (57) as a white powder: MS (m/z) MH⁺= 392. tert-Butyl (S)-2-(5-(4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)phenyl)-l/ -imidazol- 2-yl)pyrrolidine-l-carboxylate (58). A mixture of ieri-butyl (5)-2-(5-(4-bromophenyl)-177- imidazol-2-yl)pyrrolidine-l-carboxylate (57) (3.8 g, 9.7 mmol), bis(pinacolato)diboron (3.0 g, 11.7 mmol), potassium acetate (2.4 g, 24.3 mmol) and Pd(dppf)Cl2· CH2CI2 (795 mg, 1.0 mmol) in anhydrous DMSO (49 mL) was degassed and heated at 80°C for 17 h while stirring under a N2 atmosphere. The cooled reaction mixture was diluted with brine and extracted several times with EtOAc. The combined organic extracts were dried (CaS0₄), filtered and concentrated in vacuo. The dark brown solid residue was recrystallized from boiling EtOAc/heptane to afford 1.3 g of ieri-butyl (5)-2-(5-(4-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)phenyl)- 17/-imidazol-2-yl)pyrrolidine- 1 -carboxyl ate (58) as a white powder: MS (m/z) MH⁺= 440.

Di-to7- butyl 2,2'-(((l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-4,8- diyl)bis(4,l-phenylene))bis(l //-imidazole-5, 2-diyl))(2S, 2^,S)-bis(pyrrolidine- 1- carboxylate) (59). The title compound (59) was prepared in an analogous manner described for Example 6 (46) from 4,8-dibromo-l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene (41) (437 mg, 1.2 mmol), ieri-butyl (5)-2-(5-(4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)phenyl)-l//-imidazol-2-yl)pyrrolidine-l-carboxylate (58) (2.2 g, 5.1 mmol), sodium carbonate (3.1 g, 29 mmol), Pd(dppf)Cl2· CH2CI2 (192 mg, 0.2 mmol), and tetrabutylammonium chloride hydrate (66 mg, 0.2 mmol) in 1 :1 mixture (v/v) of THF and water (47 mL). The product was purified by chromatography on silica gel eluting with a gradient of 0-10% MeOH in CH2CI2 followed by recrystallization from EtOAc/hexanes to yield 167 mg of di-ieri-butyl 2,2'-((( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-.v-indacene-

4.8-diyl)bis(4, 1 -phenylene))bis( 17/-imidazole-5,2-diyl))(2.V,2'.V)-bis( pyrrolidine- 1 - carboxylate) (59): MS (m/z) MH⁺ = 838.

5,5'-((l,l,5,5-Tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-4,8-diyl)bis(4,l- phenylene))bis(2-((S)-pyrrolidin-2-yl)-l //-imidazole) dihydrochloride (60). A stirred suspension of di-ieri-butyl 2,2'-(((l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-4,8- diyl)bis(4,l-phenylene))bis(l//-imidazole-5,2-diyl))(25,2'5)-bis(pyrrolidine-l -carboxylate) (59) (117 mg, 0.2 mmol) and coned aq. HC1 (1.0 mL) in methanol (4.0 mL) was heated at reflux for 0.5 h, cooled, and the precipitated product isolated by filtration to provide 138 mg of 5,5'-((l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-4,8-diyl)bis(4,l-phenylene)) bis(2-((5)-pyrrolidin-2-yl)- \H- imidazole) dihydrochloride (60) as a white solid: MS (m/z) MEL = 637.

Dimethyl ((l/?,17?)-((2S,2'S)-(((l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-

4.8-diyl)bis(4,l-phenylene))bis(l/ -imidazole-5,2-diyl))bis(pyrrolidine-2,l-diyl))bis(2- oxo-l-phenylethane-2,l-diyl))dicarbamate (54). DIEA (0.2 mL, 1.0 mmol) was added to a suspension of 5,5'-((l,l,5,5-tetramethyl-l,2,3,5,6,7-hexahydro-s-indacene-4,8-diyl)bis(4,l- phenylene))bis(2-((.V)-pyrrolidin-2-yl)- 1 //-imidazole) dihydrochloride (60) (138 mg, 0.2 mmol) and (/?)-2-((methoxycarbonyl)amino)-2-phenylacetic acid (35) in THF (19 mL) while stirring at r.t. under N2. After 0.5 h, the reaction mixture was treated with a 50 wt. % solution of T3P in EtOAc (0.5 mL, 0.8 mmol), stirred for 10 min. and then diluted with brine. The layers were separated and the aqueous layer was extracted with additional THF. The combined THF extracts were washed with satd. aq. NaHCCL, filtered, dried (CaSCL), and concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with a gradient of 0-10% MeOH in CH2CI2 followed by recrystallization from EtOAc/heptane provide 102 mg of dimethyl (( 1 /?, 1 'R)-((2S,2'S)-((( 1 , 1 ,5,5-tetramethyl- l,2,3,5,6,7-hexahydro-s-indacene-4,8-diyl)bis(4,l-phenylene))bis(l//-imidazole-5,2- diyl))bis(pyrrolidine-2,l-diyl))bis(2-oxo-l-phenylethane-2,l-diyl))dicarbamate (54) as a white solid: MS (m/z) MH⁺ = 1020; ¾ NMR (300 MHz, DMSO-d6): d 11.97-11.75 (m, 2H), 8.03-7.00 (m, 22H), 5.51-5.08 (m, 4H), 3.90 (br s, 1H), 3.62-3.54 (m, 6H), 3.16 (br s, 1H), 2.41-2.37 (m, 4H), 2.03 (br s, 4H), 1.99-1.84 (m, 2H), 1.71 (br s, 4H), 0.97 (br s, 12H).

Biological Assays

The compounds were assayed side-by-side with standard (STD) control compounds Daclatasvir and Sofosbuvir against selected HCV genotypes. Both control compounds are FDA approved direct-acting antiviral agents against Hepatitis C Virus (HCV) used for the treatment of chronic HCV infection and have published HCV genotype selectivity and potency values from which the reliability of the assay can be inferred and compared with the test compound assayed profile.

GTlb-Luc (Con-1) Replicon Assay. The HCV GTlb (Con-1) replicon assay (luciferase endpoint) was conducted in Huh Luc/Neo cells by Southern Research, Frederick, Maryland, USA. The HCV GTlb (Con-1) replicon assay evaluated the antiviral activity of compounds at six serial dilutions in triplicate. Sofosbuvir was included in each run as a reference standard. The Huh Luc/Neo replicon cells were plated at 5,000 cells/well into separate 96- well plates for analysis of cell viability or antiviral activity. On the following day, diluted test samples were added to the appropriate wells. Cells were processed 72 hours later when they were still sub-confluent. The HCV replicon levels were quantified by replicon-derived luciferase activity. Curve fitting software was used to generate EC50 values (the concentration inhibiting HCV replicon by 50%).

GTla WT Replicon, GTlb/3a NS5a and GTlb/6a NS5a Chimeric Replicons and Cytotoxicity (CCso) Assays. These assays were conducted in Huh7 cells by WuXi Apptec Co. Ltd. (Shanghai, China) and daclatasvir was used as a reference standard. The chimeric replicons GTlb/3a and GTlb/6a were constructed using the GTlb replicon as a backbone.

The compounds were tested according to the following procedure:

1) Compounds were added to 96- well plates by Echo at 9 concentrations with a serial 3- fold dilution, in duplicate. The final concentration of DMSO in the cell culture medium was 0.5%. 2) The replicon RNAs were in vitro transcribed using the replicon plasmid DNAs.

3) The Huh7 cells were transiently transfected with the replicon RNAs by electroporation and seeded at a density of 10,000 cells/well in 96-well plates. The cells were cultured and treated with the compounds at 37 °C and 5% CO2 for 3 days.

4) The cell viability was determined with the CellTiter- Fluor kit in cells of GTla replicon assay in accordance with the protocol provided by the supplier.

5) The supernatants were removed from the wells. The antiviral activity was determined by monitoring replicon reporter firefly luciferase using Bright-Glo in accordance with the protocol provided by the supplier. The raw data (RLU) were used for calculating the antiviral activity (% inhibition) of the compounds.

Inhibition% = (CPD-ZPE)/(HPE-ZPE)*100%

*CPD: Signal from a well containing a test compound.

ZPE: Average of signals from DMSO control wells.

HPE: Average of signals from medium control wells.

The 50% effective concentrations (EC50) and 50% cytotoxicity concentration (CC50) values were calculated with the GraphPad Prism software. The results for Examples 1-7 are shown in Table 1.

Table 1. Anti-HCV Activity of Examples 1-7

STD = reference standard; WT = wild type; GTlb-Luc (Con 1) assay was conducted in Huh Luc/Neo cells; Gtla-wt, Gtlb/3a NS5a and Gtlb/6a NS5a are transient transfected replicon assays and were conducted in Huh7 cells; NT = not tested; * = Literature values from the Solvaldi package insert (rev. 03/2015) and Gao, M. in Curr. Opin.in Virology 2013, 3, 514-520.

The structures claimed in this application were designed in an effort to develop a selectivity and potency profile improvement over currently approved agents. Favorable comparison of the genotypic selectivity, potency, and cytotoxicity of the claimed structures against that of daclatasvir and dofosbuvir provides a rationale for further development of these compounds.

It will be understood by those of skill in the art that numerous and various modifications can be made to the compounds, compositions, and/or methods of the present invention without departing from the spirit of the invention. Therefore, the various examples and embodiments of the present invention described herein are illustrative only and are not intended to limit the scope of the invention in any way. All references cited herein are incorporated by reference in their entirety.

Claims

CLAIMS What is claimed is:

1. A compound of formula I:

B¹ -A¹ -Q¹ -P¹ -M-P²-Q²-A²-B²

I

M is selected from the formulas II, III, and IV, where R¹ at each occurrence is independently Ci-G, alkyl:

II III IV

, wherein X is CH or N, L¹ is a direct bond to Q¹ or Q², and L² is a direct bond to M;

Q¹ and Q² are independently absent (a direct bond),

and P² are not

when M is II;

occurrence is independently a direct bond to Q¹ or Q²; and R² and R³ at each occurrence are the same or different and are independently selected from H and C₁-C₆ alkyl optionally substituted, or taken together along with the carbon atom to which they are attached form a C₃-C₆ cycloalkyl optionally substituted;

2. The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer,

tautomer, solvate, or prodrug thereof, wherein M is II; P¹ and P² are each

Q¹ and Q² are absent; A¹ and A² are each

optionally substituted; and R⁵ is C₁-C₆ alkyl optionally substituted.

3. The compound of claim 2, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, wherein each X is CH; R² and R³ are independently hydrogen, methyl, ethyl, or taken together along with the carbon atom to which they are attached form a C3-C6 cycloalkyl optionally substituted.

4. The compound of claim 2 or 3, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, wherein R² and R³ are independently hydrogen or methyl; R⁴ is methyl or ethyl; and R⁵ is isopropyl.

5. The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer,

tautomer, solvate, or prodrug thereof, wherein M is III; P¹ and P² are each

Q¹ and Q² are absent; A¹ and A² are each

optionally substituted; and R⁵ is C1-C6 alkyl optionally substituted.

6. The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, wherein M is III; P¹ and P² are each absent; Q¹ and Q²

_L

are each

optionally substituted; and R⁵ is C₁-C₆ alkyl optionally substituted.

7. The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer,

N"

A Y, ^_L2

I_G N

tautomer, solvate, or prodrug thereof, wherein M is III; P¹ is absent; Q¹ is H ; P² is

optionally substituted; and R⁵ is C1-C6 alkyl optionally substituted.

8. The compound of any one of claims 5 to 7, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, wherein each X is CH; R² and R³ are independently hydrogen, methyl, ethyl, or taken together along with the carbon atom to which they are attached form a C₃-C₆ cycloalkyl optionally substituted.

9. The compound of any one of claims 5 to 8, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodmg thereof, wherein R² and R³ are independently hydrogen or methyl; R⁴ is methyl or ethyl, each optionally substituted; and R⁵ is isopropyl optionally substituted.

10. The compound of Claim 1, or a pharmaceutically acceptable salt, stereoisomer,

tautomer, solvate, or prodrug thereof, wherein M is IV; P¹ and P² are each

wherein X is CH; Q¹ and Q² are absent; A¹ and A² are each

optionally substituted; and R⁵ is C1-C6 alkyl or C6-C10 aryl, each optionally substituted.

11. The compound of Claim 1, or a pharmaceutically acceptable salt, stereoisomer,

tautomer, solvate, or prodrug thereof, wherein M is IV; P¹ and P² are each

wherein X is N; Q¹ and Q² are absent; A¹ and A² are each

optionally substituted; and R⁵ is C1-C6 alkyl or aryl, each optionally substituted.

12. The compound of claim 10 or 11, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodmg thereof, wherein R⁵ is isopropyl or phenyl, each optionally substituted.

13. The compound of Claim 1, or a pharmaceutically acceptable salt, stereoisomer,

tautomer, solvate, or prodmg thereof, wherein M is IV; P¹ and P² are each

wherein X is CH or N; Q¹ and Q² are absent; A¹ and A² are each

alkyl optionally substituted; and R⁵ is C6-C10 aryl, each optionally substituted.

14. The compound of Claim 1, or a pharmaceutically acceptable salt, stereoisomer,

tautomer, solvate, or prodrug thereof, wherein M is IV; P¹ and P² are each

; Q¹

_L

and Q² are each ^L H ; A¹ and A² are each

optionally substituted; and R⁵ is C6-C10 aryl optionally substituted.

15. The compound of Claim 1, or a pharmaceutically acceptable salt, stereoisomer,

tautomer, solvate, or prodrug thereof, wherein M is IV; P¹ and P² are each

1

; Q

and Q² are each L¹ H ; A¹ and A² are each L 3^’ ; R⁴ is C1-C6 alkyl optionally substituted; and R³ is C6-C10 aryl optionally substituted.

16. The compound of any one of claims 13 to 15, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, wherein R⁵ is phenyl optionally substituted.

17. The compound of any one of claims 10 to 16, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, wherein R² and R³ are independently hydrogen, methyl, ethyl, or taken together along with the carbon atom to which they are attached form a C₃-C₆ cycloalkyl optionally substituted; and R⁴ is C₁-C₄ alkyl optionally substituted.

18. The compound of any one of claims 10 to 16, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, wherein R² and R³ are independently hydrogen or methyl; R⁴ is methyl or ethyl.

19. The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, selected from the group consisting of:

20. The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, selected from the group consisting of:

4, 12- Bis- 1 V-imethoxycarhonylj-L-valyl-L-prolyl- V-i 3-ami nophenyl ) |- [2.2]paracyclophane;

methyl ((5)-l-((5)-2-(5-(9,9-difluoro-7-(3-((5)-l-((methoxycarbonyl)-L- valyl)pyrrolidine-2-carboxamido)phenyl)-9H-fluoren-2-yl)- l//-imidazol-2-yl)pyrrolidin- 1 - yl)-3-methyl-l-oxobutan-2-yl)carbamate;

dimethyl ((2S,2'S)-((2S,2'S)-((((1 ,1 ,5,5-tetramethyl- 1 ,2,3,5 ,6,7-hexahydro-.v-indacene- 4,8-diyl)bis(3 , 1 -phenylene))bis(azanediyl))bis(carbonyl))bis(pyrrolidine-2 , 1 -diyl))bis(3 - methyl- 1 -oxobutane- 1 ,2-diyl))dicarbamate; dimethyl (( 1 /?, 1 'R)-((2S,2'S)-(((( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-.v-indacene-

4.8-diyl)bis(3,l-phenylene))bis(azanediyl))bis(carbonyl))bis(pyrrolidine-2,l-diyl))bis(2-oxo- 1 -phenylethane-2, 1 -diyl))dicarbamate;

dimethyl (( 1 /?, 1 'R)-((2S,2'S)-(((( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-.v-indacene-

4.8-diyl)bis(pyridine-5,3-diyl))bis(azanediyl))bis(carbonyl))bis(pyrrolidine-2,l-diyl))bis(2- oxo- 1 -phenylethane-2 , 1 -diyl))dicarbamate;

dimethyl (( 1 /?, 1 'R)-((2S,2'S)-((( 1 , 1 ,5,5-tetramethyl- 1 ,2,3,5,6,7-hexahydro-.v-indacene-

4.8-diyl)bis(3,l-phenylene))bis(l//- imidazole-5, 2-diyl))bis(pyrrolidine-2,l-diyl))bis(2-oxo-l- phenylethane-2, l-diyl))dicarbamate; and

4.8-diyl)bis(4, 1 -phenylene))bis( 1 //-imidazole-5, 2-diyl))bis(pyrrolidine-2, 1 -diyl))bis(2-oxo- 1 - phenylethane-2, l-diyl))dicarbamate.

21. A pharmaceutical composition comprising a compound of formula (I) according to any one of claims 1 to 20, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, and one or more pharmaceutically acceptable excipients, such as adjuvants, diluents, and/or carriers.

22. The composition of Claim 21 further comprising a second agent having anti- HCV activity.

23. The composition of Claim 22, wherein the second agent having anti-HCV activity is selected from the group consisting of a recombinant Human Interferon Alfa, a nucleoside analog, a direct acting antiviral, a NS3/4A protease inhibitor, a nucleotide NS5B polymerase inhibitor, a NS5A inhibitors, a non-nucleoside NS5B polymerase inhibitors, and

combinations thereof.

24. The composition of claim 22, wherein the second agent is selected from the group consisting of peginterferon, ribavirin, daclatasvir, boceprevir, telapravir,

simeprevir, sofosbuvir, dasabuvir, ombitasvir, velpatasvir, ledipasvir, paritaprevir, ritonavir, elbasvir, grazoprevir, asunaprevir, beclabuvir, and combinations thereof.

25. A method of inhibiting the function of the HCV NS5A protein comprising contacting a biological sample comprising the HCV NS5A protein with the compound of any one of claims 1 to 20, or a salt, stereoisomer, tautomer, solvate, or prodrug thereof.

26. A method for the treatment or prevention of a disease, disorder, or medical condition associated with HCV activity in a subject, comprising administering to the subject a therapeutically effective amount of the compound of according to any one of claims 1 to 20, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof.

27. The method of claim 26, further in conjunction with administration of a second agent having anti-HCV activity to the subject.

28. The method of claim 27, wherein the second agent is selected from the group consisting of a recombinant Human Interferon Alfa, a nucleoside analog, a direct acting antiviral, a NS3/4A protease inhibitor, a nucleotide NS5B polymerase inhibitor, a NS5A inhibitors, a non-nucleoside NS5B polymerase inhibitors, including, but limited to, peginterferon, ribavirin, daclatasvir, boceprevir, telapravir, simeprevir, sofosbuvir, dasabuvir, ombitasvir, velpatasvir, ledipasvir, paritaprevir, ritonavir, elbasvir, grazoprevir, asunaprevir, beclabuvir, and combinations thereof.

29. A method for the treatment or prevention of a disease, disorder, or medical condition associated with HCV activity in a subject, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition according to any one of claims 21 to 24.

30. The method of any one of claims 26 to 29, wherein the disease, disorder, or condition is selected from the group consisting of hepatitis C infection, chronic hepatitis, cirrhosis, hepatocellular carcinoma (HCC), liver cancer, mixed cryoglobulinemia, porphyria cutanea tarda, leukocytoclastic vasculitis, lichen planus (LP), sicca syndrome, urticaria, pruritus, thrombocytopenic purpura, and psoriasis.

31. The method of any one of claims 26 to 29, wherein the disease or disorder is cirrhosis.

32. The method of any one of claims 26 to 29, wherein the disease or condition is liver cancer.

33. The method of any one of claims 26 to 29, wherein the disease is hepatitis C infection.