WO2011050146A1 - Chemical compounds - Google Patents

Abstract

Disclosed are compounds of Formula (I). Also disclosed are pharmaceutical compositions comprising the compounds, and methods for treating HCV invention by administration of the compounds.

Description

CHEMICAL COMPOUNDS

FIELD OF THE INVENTION

The present disclosure relates to antiviral compounds.

BACKGROUND OF THE INVENTION

Chronic infection with HCV is a major health problem associated with liver cirrhosis, hepatocellular carcinoma and liver failure. An estimated 170 million chronic carriers worldwide are at risk of developing liver disease. See, for example, Szabo, et al., Pathol. Oncol. Res. 2003, 9:215-221 , and Hoofnagle JH, Hepatology 1997 ^', 26:15S-20S. In the United States alone 2.7 million are chronically infected with HCV, and the number of HCV-related deaths in 2000 was estimated between 8,000 and 10,000, a number that is expected to increase significantly over the next years. Infection by HCV is insidious in a high proportion of chronically infected (and infectious) carriers who may not experience clinical symptoms for many years. Liver cirrhosis can ultimately lead to liver failure. Liver failure resulting from chronic HCV infection is now recognized as a leading cause of liver transplantation.

HCV is a member of the Flaviviridae family of RNA viruses that affect animals and humans. The genome is a single ~9.6-kilobase strand of RNA, and consists of one open reading frame that encodes for a polyprotein of -3000 amino acids flanked by untranslated regions at both 5' and 3' ends (5'- and 3'-UTR). The polyprotein serves as the precursor to at least 10 separate viral proteins critical for replication and assembly of progeny viral particles. The organization of structural and non-structural proteins in the HCV polyprotein is as follows: C-E1 -E2-p7-NS2-NS3-NS4a-NS4b-NS5a-NS5b. Because the replicative cycle of HCV does not involve any DNA intermediate and the virus is not integrated into the host genome, HCV infection can theoretically be cured. While the pathology of HCV infection affects mainly the liver, the virus is found in other cell types in the body including peripheral blood lymphocytes. See, for example, Thomson BJ and Finch RG, Clin Microbial Infect. 2005, 1 1 :86-94, and Moriishi K and Matsuura Y, Antivir. Chem. Chemother. 2003, 14:285-297.

At present, the standard treatment for chronic HCV is interferon alpha (IFN-alpha) in combination with ribavirin and this requires at least six (6) months of treatment. IFN-alpha belongs to a family of naturally occurring small proteins with characteristic biological effects such as antiviral, immunoregulatory and antitumoral activities that are produced and secreted by most animal nucleated cells in response to several diseases, in particular viral infections. IFN- alpha is an important regulator of growth and differentiation affecting cellular communication and immunological control. Treatment of HCV with interferon has frequently been associated with adverse side effects such as fatigue, fever, chills, headache, myalgias, arthralgias, mild alopecia, psychiatric effects and associated disorders, autoimmune phenomena and associated disorders and thyroid dysfunction. Ribavirin, an inhibitor of inosine 5'-monophosphate dehydrogenase (IMPDH), enhances the efficacy of IFN-alpha in the treatment of HCV. Despite the introduction of ribavirin, more than 50% of the patients do not eliminate the virus with the current standard therapy of interferon-alpha (IFN) and ribavirin. By now, standard therapy of chronic hepatitis C has been changed to the

combination of pegylated IFN-alpha plus ribavirin. However, a number of patients still have significant side effects, primarily related to ribavirin. Ribavirin causes significant hemolysis in 10-20% of patients treated at currently

recommended doses, and the drug is both teratogenic and embryotoxic. Even with recent improvements, a substantial fraction of patients do not respond with a sustained reduction in viral load and there is a clear need for more effective antiviral therapy of HCV infection. See, for example, Fried, et al. N. Engl. J Med 2002, 347:975-982.

A number of approaches are being pursued to combat the virus. They include, for example, application of antisense oligonucleotides or ribozymes for inhibiting HCV replication. Furthermore, low-molecular weight compounds that directly inhibit HCV proteins and interfere with viral replication are considered as attractive strategies to control HCV infection. Among the viral targets, the NS3/4A protease/helicase and the NS5b RNA-dependent RNA polymerase are considered the most promising viral targets for new drugs. See, for example, Ni, Z. J. and Wagman, A. S. Curr. Opin. Drug Discov. Devel. 2004, 7, 446-459, Beaulieu, P. L. and Tsantrizos, Y. S. Curr. Opin. Investig. Drugs 2004, 5, 838- 850, and Griffith, et al., Ann. Rep. Med. Chem 39, 223-237, 2004.

Besides targeting viral genes and their transcription and translation products, antiviral activity can also be achieved by targeting host cell proteins that are necessary for viral replication. For example, Watashi, et al, Molecular Cell, 19, 1 1 1 -122, 2005, show how antiviral activity can be achieved by inhibiting host cell cyclophilins. Alternatively, a potent TLR7 agonist has been shown to reduce HCV plasma levels in humans. See, Horsmans, et al, Hepatology, 42, 724-731 , 2005.

Compounds said to be useful for treating HCV infection are disclosed, for example, in WO 2008/064218 (Leivers et. al), WO 2008/244380 (Bachand et. al), and US 2009/0068140 (Bachand et. al). These references also disclose methods for preparing the compounds, compositions comprising the compounds, compositions comprising the compounds and additional compounds, methods of treating HCV, and etc.

SUMMARY OF THE INVENTION

Briefly, in one aspect, the present invention discloses compounds of

Formula I;

wherein A is a phenyl, pyridyl, furanyl, or thiofuranyl ring;

each R¹ is independently H, Ci_-6alkyl, Ci_-6cycloalkyl, or phenyl:

each R² is independently H, C_halky!, Ci-3acyl, C3-6cycloacyl, or C(O)O-Ci-3alkyl, or two alkyl R²s may, together with the nitrogen atom to which they are bound, join together to form a 5 or 6-membered saturated heterocycylic ring;

each R³ is independently H, F, d_-3alkyl, or Ci_-3alkoxy, or when two R³s are alkyl and bound to adjacent carbon atoms, they may, together with the two carbon atoms to which they are bound, join to from a 5 or 6-membered cycloalkyl ring,

R³s may optionally be substituted by one or two fluorine atoms;

within each ring, 1 of the 3 Zs are CH and the other 2 are CH or N; and

each R⁴ is independently H, F, or C_halky!. In another aspect, the present invention discloses compounds of Formula II:

II

wherein A, R¹ , R², R³, Z, and R⁴ are as defined above;

each X is independently CRR, O, or S;

each n is independently 2 or 3; and

each R is independently methyl, hydrogen, or deuterium. In another aspect, the present invention discloses compounds of

formula III;

III wherein A, R¹ , R², Z, and R⁴ are as defined above;

each X is independently CRR, O, or S;

each n is independently 2 or 3; and

each R is independently methyl, hydrogen, or deuterium.

In another aspect, the present invention discloses pharmaceutically acceptable salts of the compounds of Formula I, Formula II, or formula III. In another aspect, the present invention discloses pharmaceutical compositions comprising a compound of Formula I, Formula II, or Formula III, or a pharmaceutically acceptable salt thereof.

In another aspect, the present invention discloses a method for treating a viral infection, for example infection with HCV, in a human, comprising administration of a compound of Formula I, Formula II, or Formula III, or a pharmaceutically acceptable salt thereof.

DETAILED DESCRIPTION OF THE INVENTION

Pharmaceutically acceptable salts can be prepared by methods well known in the art. Suitable salts include those described, for example, in P.

Heinrich Stahl, Camille G. Wermuth (eds.), handbook of Pharmaceutical Salts properties, selection, and Use; 2002.

In one aspect of the invention, A is phenyl

When A is either phenyl or pyridyl, the two large groups ending in R², are preferably located para to each other on said phenyl or pyridyl A group.

In one aspect, R¹ is C_halky!, in particular i-propyl.

In one aspect, with respect to each nitrogen to which they are attached, one R² is H. In particular, one R² is C(O))-Ci-3alkyl and the other R² is H.

In one aspect, at least 2 Zs in each ring are CH.

Example

A table of abbreviations used in this Experimental section is set forth below.

DCM Dichloromethane

DMF N,N-dimethylformamide

HATU (O-7-azabenzotriazol-1 -yl)-N,N,N',N'- tetramethyluronium hexafluorophosphate)

ES LC-MS Electrospray Liquid Chromatography Mass

Spectrometry

THF Tetrahydrofuran

DIEA diisopropylethylamine

DMSO Dimethylsulfoxide

DME Dimethoxyethane Triethylamine

Isopropyl alcohol

Dess-Martin periodinane

High Resolution Mass Spectroscopy

1 ,1 '-Bis(diphenylphosphino)ferrocene

4-Dimethylaminopyridine

1 -Ethyl-3-(3-dimethylanninopropyl)carbodiinnide

Boc anhydride

N-hydroxybenzotriazole

Ethyl acetate

Room temperature

hour

methanol

Reverse Phase High Performance Liquid

Chromatography

1 ,2-Dimethoxyethane

The Synthetic schemes l-VII below are intended to illustrate synthetic methods which can be used to prepare compounds of the present invention. When specific compounds are shown, they are meant to be illustrative of the general method and not restricted to the specific compound shown. It is intended that these chemistries are not limited to compounds in Schemes l-VI and are generally applicable to compounds of Formula I, II, III and R, R1 -R4 claimed herein.

Scheme I cNH cNH

Scheme II

Scheme

Scheme IV

X,X=H,H Example 7

Spirocyclic intermediates 60 and 61 can be synthesized according to Scheme VII.

Scheme VII

Intermediate 6

L-valine (49.09 g, 419 mmol), sodium hydroxide (16.76 g, 419 mmol) (89 mL as a 1 M solution) and sodium carbonate (22.21 g, 210 mmol) were combined and methyl chloroformate (35.7 ml, 461 mmol) added dropwise over 10 min. After stirring at RT for 16 hours, ether (200 mL) was added and the reaction stirred for 20 min. The organic layer was removed and the aqueous layer pH was adjusted to ~ 3 with 1 M HCI. Following extraction with DCM (4 x 400 mL), the combined organ ics were dried over MgSO₄, filtered and concentrated to afford 68 g of N- [(methyloxy)carbonyl]-L-valine 6 as a white solid. Intermediate 56: Methyl (4R)-4-hydroxy-L-prolinate 55 (50 g, 275 mmol), N- [(methyloxy)carbonyl]-L-valine (50.6 g, 289 mmol) 6 and HOBT (52.7 g, 344 mmol) were suspended in Dichloromethane (DCM) (500 mL) and cooled to 0 ⁰ C. DIEA (99 mL, 564 mmol) was added and when the mixture was added EDC (66.0 g, 344 mmol), stirred at RT for 2hrs. The clear solution was extracted with water (300mL), washed with saturated sodium carbonate (3x100mL), brine (100mL), dried with sodium sulfate and evaporated to give 56 (70.6g, 234 mmol, 85 % yield) as a sticky oil.

Intermediate 57: In a 5L 4-neck round bottom flask equipped with an overhead stirrer and a bleach bubbler trap was added dichloromethane (1300 mL) and oxalyl chloride (58.3 mL, 667 mmol). The mixture was cooled to -72°C (IPA/dry ice) and DMSO (63.1 mL, 889 mmol) added over 40 min, keeping the temp below -62°C, during which gas evolution was observed. A pale yellow solution of methyl N-[(methyloxy)carbonyl]-L-valyl-(4R)-4-hydroxy-L-prolinate 56 (134.34 g, 444 mmol) in DCM (1000 mL) was added over 1 .5 h, keeping the temp below -64°C. The resulting opaque pale yellow mixture was stirred for 1 h. TEA (217 mL, 1555 mmol) was added over 40 min below -65°C, subsequently IPA/CO₂ bath was removed and the mixture was allowed to warm up to 0°C. Saturated NaHCO3 solution (500 mL) was then added and layers were separated, the organic layer was washed with a 1 N HCI solution (500 mL) and a saturated NaCI solution (500 mL). After drying over Na₂SO₄ overnight, the slurry was filtered and solution concentrated yielding 127.61 g of 57. Intermediate 58: A mixture of 80g (1290 mmol) of ethylene glycol, 77.46g, (258 mmol) of methyl N-[(methyloxy)carbonyl]-Lvalyl-4-oxo-L-prolinate 57 and 4.91 g (25.8 mmol) of TsOH in 775 mL toluene was heated to reflux in a flask fitted with a Dean Stark trap and condenser. After 1 .5 hours the reaction mixture was allowed to cool down and then washed with saturated aqueous NaHCO3. The aqueous layer was back extracted with EtOAc and the combined organic layers were washed with brine and dried with Na₂SO₄, filtered and concentrated. The crude material was dissolved in 50 mL of 1 :1 hexane : DCM and

chromatographed on silica with 10-100% gradient if EtOAc in hexanes. Fractions containing pure product were combined to yield 32.54g (36.6% yield) of 58. Intermediate 60:

(8S)-7-{N-[(methyloxy)carbonyl]-L-valyl}-1,4-dioxa-7-azaspiro[4.4]nonane-8- carboxylic acid

Methyl (8S)-7-{N-[(methyloxy)carbonyl]-L-valyl}-1 ,4-dioxa-7-azaspiro[4.4]nonane- 8-carboxylate 58 (38.41 g, 112 mmol) was dissolved in a cold solvent mixture of tetrahydrofuran (THF) (295 mL), t-butanol (73.8 mL) and water (73.8 mL). LiOH (5.62 g, 134 mmol) was added as solid and the mixture was stirred at RT for 1 hr. The mixture was quenched with HCI (139 mL, 139 mmol), diluted with ethyl acetate (590 mL) and brine (200mL). The organic phase was washed twice with brine (100mL), dried (Na₂SO₄) and evaporated to give a dark solid which was sonicated with 200 mL ether and filtered to yield (8S)-7-{N-[(methyloxy)carbonyl]- L-valyl}-1 ,4-dioxa-7-azaspiro[4.4]nonane-8-carboxylic acid 60 (30.004g, 91 mmol, 81 % yield)

¹ H NMR (400 MHz, DMSO-c/₆) δ ppm 12.56 (br. s., 1 H) 7.40 (d, J=8.4 Hz, 1 H) 4.33 (dd, J=8.8, 7.02 Hz, 1 H) 3.81 - 4.10 (m, 5 H) 3.41 - 3.66 (m, 5 H) 2.28 - 2.43 (m, 1 H) 1 .94 - 2.11 (m, 1 H) 1 .74 - 1 .94 (m, 1 H) 0.67 - 1 .05 (m, 6 H). ES LC-MS m/z =331 .6 (M+H)⁺.

Intermediate 61 ; {3S)-2-{N-[(methyloxy)carbonyl]-L-valyl}-6, 10-dioxa-2- azaspiro[4.5]decane-3-carboxylic acid

Prepared using process outlined in preparation of intermediate 60 using propylene glycol in place of ethylene glycol.

¹ H NMR (400 MHz, DMSO-c/₆) δ ppm 12.56 (br. s., 1 H) 7.43 (d, J=8 Hz, 1 H) 4.28 - 4.39 (m, 1 H) 4.24 (t, J=8 Hz, 1 H) 3.77 - 3.94 (m, 5 H) 3.45 - 3.63 (m, 5 H) 2.05 (dd, J=13, 8 Hz, 1 H) 1 .91 (br. s., 1 H) 1 .54 - 1 .76 (m, 2 H) 0.83 - 0.98 (m, 6 H)

General HATU coupling

Schemes I step 6, Scheme II step 4, Scheme VI step 1 , Scheme VI step 2

To a solution of 1 .1 eq. (to 1 .0 eq. of amino component to be coupled) acid, 1 .1 eq HATU in DMF (volume to make a 0.2 mol/L solution) 2.2 eq. DIEA is added drop-wise. The mixture is stirred at RT for 5 minutes then 1 eq. of the amino component is added (followed by an additional 1 eq. DIEA if the amine is a salt). The mixture is stirred at RT until completion (monitored by LCMS). The DMF is evaporated; the residue is taken up in a saturated NaHCO3 - EtOAc mixture, the organic phase is washed with NaHCO3 (1 x), brine (1x), dried (is^SO^ and evaporated. The crude product is purified on RP-HPLC.

Intermediate 45a

1, 1-Dimethylethyl(2S)-2-(5-bromo-1H-benzimidazol-2-yl)-1-pyrrolid

Scheme V step 1 , 2

A solution of 1 -{[(1 ,1 -dimethylethyl)oxy]carbonyl}-L-proline 20 in DMF was stirred as Hunig's base was added followed by HATU. After ~5 minutes 4-bromo-1 ,2- benzenediamine 1 was added and the resulting dark solution was allowed to stir for 2.5 hours. The solution was partitioned between EtOAc and NaHCO3 solution. The organic layer was washed with brine, dried with MgSO^ filtered and concentrated. The residue was chromatographed on a 330 g silica gel column eluted with 0-70% EtOAc in hexanes. Fractions containing the product were combined and concentrated to yield 23.46 g of a brown solid. A solution of this brown oil in acetic acid was heated in an 80°C oil bath for 3.5 hours. The solution was then concentrated and the residue dissolved in EtOAc. The organic layer was washed with NaHCO3 solution then brine, dried with MgSO^ filtered and concentrated. The residue was chromatographed on a 330 g silica gel column eluted with 0-2.5% MeOH in DCM. Fractions containing the product were combined and concentrated to yield brown oil. Mixed fractions containing the product were combined and concentrated. Both samples were slurried in hot EtOAc. A pale solid formed. The EtOAc was decanted off and the residual solids were dissolved in MeOH combined and concentrated to yield 1 ,1 -dimethylethyl (2S)-2-(5-bromo-1 H-benzimidazol-2-yl)-1 -pyrrolidinecarboxylate (8.59 g, 34% yield) as an orange foam.

¹ H NMR (400 MHz, DMSO-c/₆) δ ppm 12.50 (br. s., 2 H) 7.74 (d, J=1 .5 Hz, 1 H) 7.61 (br. s., 1 H) 7.51 (d, J=8.5 Hz, 1 H) 7.42 (d, J=8.4 Hz, 1 H) 7.18 - 7.34 (m, 2 H) 4.68 - 5.1 1 (m, 2 H) 4.1 1 (q, J=5.2 Hz, 1 H) 3.59 (br. s., 2 H) 3.37 - 3.52 (m, 2 H) 2.33 (br. s., 2 H) 1 .79 - 2.13 (m, 6 H) 0.87 - 1 .56 (m, 18 H)

In rmediate 45b

1, 1 -dimethylethyl (2S)-2-(5-bromo-6-fluoro- 1 H-benzimidazol-2-yl)-1 - pyrrolidinecarboxylate

Prepared analogously to intermediate 45a (Scheme V step 1 , 2)

4-Bromo-5-fluoro-1 ,2-diaminobenzene lb (0.487 g, 2.375 mmol) and 1 -{[(1 ,1 - dimethylethyl)oxy]carbonyl}-L-proline 20 (0.51 1 g, 2.375 mmol) were combined in Dichloromethane (DCM) (50 ml_). DIEA (0.830 ml_, 4.75 mmol) was added and the reaction stirred for 10 min followed by addition of HATU (0.903 g, 2.375 mmol). After stirring at RT overnight, water (50 ml_) was added and the reaction stirred for 30 min. The solution was filtered through a hydrophobic frit and the organics were concentrated. The residue was taken up in acetic acid (20 mL) and heated to 70°C for 1 h and then concentrated. The crude material was purified on silica using 0-100% EtOAc gradient in hexanes to afford 0.635g (1 .65 mmol, 70%) of the product as a light brown foam. ¹H NMR (400 MHz, CHLOROFORM- d) δ 7.65 - 7.84 (m, 1 H), 7.29 - 7.43 (m, 1 H), 5.10 (dd, J=7.78, 2.51 Hz, 1 H), 3.46 (d, J=7.78 Hz, 2 H), 3.00 (d, J=5.52 Hz, 1 H), 2.1 1 - 2.28 (m, 2 H), 1 .97 - 2.07 (m, 2 H), 1 .53 (s, 9 H)

LCMS (M + H) 384, 386. Intermediate 47

Methyl ((1 S)-1-{[(2S)-2-(5-bromo-6-fluoro-1H-benzimidazol-2-yl)-1- pyrrolidinyl]carbonyl}-2-methylpropyl)carbamate

Scheme V, step 6, 7

1 ,1 -dimethylethyl (2S)-2-(5-bromo-6-fluoro-1 H-benzimidazol-2-yl)-1 - pyrrol id inecarboxylate 45b (obtained along the lines of Scheme V, step 1 , 2) (0.635 g, 1 .653 mmol) was dissolved in 1 ,4-dioxane (5.00 mL) and treated with 4M HCI in dioxane (1 mL). After 3hrs the reaction was concentrated. The residue was then suspended in DCM (5 mL) and DIEA (0.866 mL, 4.96 mmol) was added, reaction stirred for 5 minutes and N-[(methyloxy)carbonyl]-L-valine (0.290 g, 1 .653 mmol) was added followed by HATU (0.628 g, 1 .653 mmol). The reaction was stirred overnight at RT, water (50 mL) added and reaction stirred for 5 minutes, the solution filtered through a hydrophobic frit and the organics concentrated. The crude material was purified on silica using 0-100% EtOAc gradient in hexanes to afford 0.579g (1 .31 mmol, 79%) of the product as a light brown oil. ¹ H NMR (400 MHz, CHLOROFORM-c/) δ 0.88 (dd, J=6.5, 3.26 Hz, 6 H) 1 .92 - 2.02 (m, 1 H) 2.12 - 2.43 (m, 4 H) 2.93 - 3.03 (m, 1 H) 3.72(s, 4 H) 3.82 - 3.95 (m, 1 H) 4.28 - 4.39 (m, 1 H) 5.36 (br. s., 2 H) 7.29 - 7.40 (m, 1 H) 7.68 - 7.78 (m, 1 H).

Intermediate 30

methyl ((1S)-1 -{[(2S)-2-(5-bromo-7-fluoro-1H-benzimidazol-2-yl)-1- pyrrolidinyl]carbonyl}-2-methylpropyl)carbamate

Scheme III step 1 , 2

N-[(Methyloxy)carbonyl]-L-valyl-L-proline 9 (0.4124 g, 1 .515 mmol) and 5-bromo- 2,3-diaminofluorobenzene 25 (0.31 1 g, 1 .515 mmol) were combined in DCM (50 mL). DIEA (0.529 mL, 3.03 mmol) was added and the reaction stirred for 10 minutes, following the addition of HATU (0.576 g, 1 .515 mmol). The reaction was stirred at RT overnight. Water (50 mL) was added and the reaction stirred for 20 minutes, the reaction filtered through hydrophobic frit and the organics were concentrated. The residue was dissolved in acetic acid (20.0 mL) and heated to 70 °C for I hour, reaction concentrated and purified on silica using 0-100% EtOAc gradient in hexanes to afford 0.604g (1 .37 mmol, 90%) of the desired product as a brown oil. ¹H NMR (400 MHz, DMSO-c/6) δ 0.90 (dd, J=16.0, 6.6 Hz, 6 H) 1 .90 (d, J=3.9 Hz, 5 H) 2.03 - 2.25 (m, 1 H) 3.51 (s, 3 H) 3.71 - 3.86 (m,1 H) 4.00 (s, 1 H) 4.17 - 4.30 (m, 1 H) 7.32 - 7.40 (m, 1 H). LCMS (M + H) 440, 442

Intermediate 22

bis(1, 1-dimethylethyl) (2S,2'S)-2,2'-[benzene-1 ,4-diylbis(1H-benzimidazole-5,2- diyl)]di(1-pyrrolidinecarboxylate)

Scheme V, step 3

1 ,1 -dimethylethyl(2S)-2-(5-bromo-1 H-benzimidazol-2-yl)-1 -pyrrolidinecarboxylate (intermediate 45a) (5.00g, 12.76 mmol), benzene-1 ,4-diyldiboronic acid 3 (1 .057 g, 6.38 mmol), PdCI₂(dppf)-CH₂CI₂ adduct (0.512 g, 0.638 mmol), potassium carbonate (2.76 g, 20 mmol) in 1 ,2-dimethoxyethane (30 mL) and water (10 mL) were heated in an oil bath at 100°C for 2 hours. The sample was filtered through celite and washed with DCM. The sample was concentrated and the residue partitioned between DCM and water, the organic layer dried with MgSO^ filtered and concentrated. The residue was chromatographed on a silica using 0-5% (2N NH3 in MeOH) in DCM. Fractions were concentrated to yield bis(1, 1- dimethylethyl) (2S,2'S)-2,2'-[benzene-1 -diylbis(1H-benzimidazole-5,2-diyl)]di pyrrolidinecarboxylate) (2.50 g, yield 55%) as an orange solid. ¹H NMR (400 MHz, DMSO-c/e) δ ppm 12.03 (br. s., 2 H) 7.83 - 7.89 (m, 1 H) 7.75 (s, 5 H) 7.61 (s, 1 H) 7.52 (br. s., 3 H) 4.99 (br. s., 2 H) 3.79 (td, J=3.4, 1 .6 Hz, 1 H) 3.61 (br. s., 2 H) 3.48 (dt, J=10.2, 7.1 Hz, 2 H) 3.20 (d, J=5.3 Hz, 2 H) 2.34 (br. s., 2 H) 2.05 (d, J=5.9 Hz, 4 H) 1 .93 (d, J=7.1 Hz, 2 H) 1 .28 (br. s., 18 H). ES LC-MS m/z =672.2 (M+H).

Intermediate 5

1, 1':4', 1"-Terphenyl-3,3",4,4"-tetramine tetrahydrochloride salt

Scheme I step 2, 3

4-bromo-N,N'-diBoc-1 ,2-benzenediamine 2 (4.61 g, 1 1 .90 mmol), benzene-1 ,4- diyldiboronic acid 3 (0.988 g 5.96 mmol), potassium carbonate (2.396 g, 17.34 mmol) and PdCI₂(dppf)-CH₂CI₂ adduct (0.487 g, 0.596 mmol) in 1 ,2- dimethoxyethane (26 mL) and water (8.67 mL) were heated in a oil bath at 100°C for 18 hours. The sample was filtered through celite, concentrated and the residue partitioned between brine and EtOAc. The organic layer was dried with MgSO4 filtered, concentrated and the product chromatographed on a silica column eluted with 0-40% EtOAc in hexanes. The product was concentrated to yield tetrakis(1 ,1 -dimethylethyl) 1 ,1 ':4',1 "-terphenyl-3,3",4,4"- tetrayltetrakiscarbamate (3.20 g, 73%) as a tan solid.

¹ H NMR (400 MHz, DMSO-c/₆) δ ppm 8.63 (d, J=10.0 Hz, 4 H) 7.84 (br. s., 2 H) 7.70 (s, 4 H) 7.61 (d, J=8.4 Hz, 2 H) 7.44 (dd, J=8.4, 2.0 Hz, 2 H) 1 .50 (s, 36 H). To tetrakis(1 ,1 -dimethylethyl) 1 ,1 ':4',1 "-terphenyl-3,3",4,4"- tetrayltetrakiscarbamate (1 .0 g, 1 .45 mmol) in dioxane (10 mL) was added 4N HCI in dioxane (26.1 mL, 104 mmol), stirred for 3 hours, the mixture diluted with ether (150 mL), filtered, solid washed with ether and dried at 45°C in vacuum to yield 1, 1':4', 1"-Terphenyl-3,3",4,4"-tetramine tetrahydrochloride salt (630 mg 100%) as a tan solid.

¹ H NMR (400 MHz, DMSO-c/₆) δ ppm 7.63 (s, 4 H), 7.38 (br s, 2 H), 7.26 (br s, 2 H), 7.10 (d, J=8 Hz, 2 H).

Intermediate 5b

4,4'-(2,5-pyridinediyl)di(1,2-benzenediamine) pentahydrochloride

A mixture of 2,5-dibromopyridine (50 mg, 0.21 1 mmol), bis(1 ,1 -dimethylethyl) [4- (4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)benzene-1 ,2-diyl]biscarbamate (193 mg, 0.443 mmol), aqueous Na₂CO₃ solution(1 M, 1 .266 mL, 1 .266 mmol) and PdCl2(dppf)-CH₂Cl2 adduct (34.5 mg, 0.042 mmol) in dioxane (5 mL) was heated in a sealed tube at 90 °C overnight. After cooling to room temperature, the product was purified on silica with 10-100% EtOAc/hexane yielding tetrakis(1 ,1 - dimethylethyl) (2,5-pyridinediyldibenzene-4,1 ,2-triyl)tetrakiscarbamate (91 mg, 62%) as a white solid:

¹ H NMR (400 MHz, DMSO-c/6) δ ppm 8.91 (d, J=1 .8, 1 H) 8.64 (d, J=5.9, 4H) 8.31 (s, 1 H) 8.08 (dd, J=8.3, 2.4, 1 H) 7.96 (d, J=8.4 Hz, 1 H) 7.84 (dd, J=8.6, 2.0 Hz, 2H) 7.60 - 7.73 (m, 2H) 7.51 (dd, J=8.4, 2.2 Hz, 1 H) 1 .31 - 1 .66 (m, 36 H); ES-LCMS m/z 692 (M+H)

Tetrakis(1 ,1 -dimethylethyl) (2,5-pyridinediyldibenzene-4,1 ,2- triyl)tetrakiscarbamate (88 mg, 0.127 mmol) was taken up into a solution of HCI in 1 ,4-dioxane (4N, 5 mL). After 2 h the reaction mixture was concentrated to afford 4,4'-(2,5-pyridinediyl)di(1,2-benzenediamine)pentahydrochloride (56 mg, 93%) as a white solid.

ES-LCMS MH⁺ 292.4.

Intermediate 23

5, 5 '-Benzene- 1, 4-diylbis{2-[ ( 2S)-2-pyrrolidinyl]- 1 H-benzimidazole} hydrochloride Scheme V, step 4

Bis(1 ,1 -dimethylethyl) (2S,2'S)-2,2'-[benzene-1 ,4-diylbis(1 H-benzimidazole-5,2- diyl)]di(1 -pyrrolidinecarboxylate) (intermediate 22) (2.49 g , 3.84 mmol) was dissolved in 20 ml_ of dioxane and concentrated. A solution of 4N HCI in dioxane (45 ml_, 180 mmol) was added, the mixture sonicated and allowed to stir for 20 minutes, diluted with ether and filtered to yield 5,5'-benzene-1,4-diylbis{2-[(2S)-2- pyrrolidinyl]-1H-benzimidazole} hydrochloride as a tan solid, (1 .86 g, 82%).

1 H NMR (400 MHz, DMSO-c/₆) δ ppm 10.39 (br. s., 2 H) 9.56 (br. s., 2 H) 7.96 (s, 2 H) 7.85 (s, 4 H) 7.75 - 7.81 (m, 2 H) 7.67 - 7.73 (m, 2 H) 5.05 (br. s., 2 H), 3.39 (m, 2 H) 2.23 - 2.39 (m, 2 H) 1 .95 - 2.23 (m, 2 H).

Intermediate 23b

5,5'-benzene-1,4-diylbis{2-[(2S)-4,4-difluoro-2-pyrrolidinyl]-1H- benzimidazolejtetrahydrochloride

Scheme II step 1 , 2, 3

A solution of 1 -{[(1 ,1 -dimethylethyl)oxy]carbonyl}-4,4-difluoro-L-proline (0.559g, 2.225 mmol ) in DMF (8.56 ml_) and DIEA (1 .399 ml_, 8.01 mmol) was added HATU (0.846 g, 2.225 mmol). After ~5 minutes (1 ,1 ':4',1 "-terphenyl-3,3",4,4"- tetramine 5 (0.50 g, 1 .1 13 mmol ) was added and the resulting dark solution was allowed to stir for 2.5 hours. The solution was then partitioned between EtOAc and NaHCO3, the organic layer washed with brine, dried with MgSO^ filtered and concentrated. The residue was chromatographed on silica using a gradient of 0-10% MeOH in DCM. The concentrated product was dissolved in acetic acid (14 mL) and heated in at 80°C for 2 hours, concentrated and partitioned between EtOAc and NaHCO3, the organic layer washed with brine, dried with MgSO^ filtered and concentrated. The residue was chromatographed on silica using a gradient of 0-10% MeOH in DCM or 0-100% EtOAc in hexanes yielding bis(1 ,1 - dimethylethyl) (2S,2'S)-2,2'-[benzene-1 ,4-diylbis(1 H-benzimidazole-5,2- diyl)]bis(4,4-difluoro-1 -pyrrolidinecarboxylate) (365 mg, 45%) as a yellow glass. ¹ H NMR (400 MHz, DMSO-c/₆) δ ppm 12.54 (br. s., 2 H) 7.91 (br. s., 1 H) 7.74 - 7.84 (m, 5 H) 7.67 (d, J=8 Hz, 1 H) 7.45 - 7.62 (m, 3 H) 5.23 (br. s., 2 H) 3.92 - 4.10 (m, 4 H) 3.09 (br. s., 2 H) 2.68 (br. s., 2 H) 1 .44 (br. s., 8 H) 1 .18 ((br. s.10 H).

Next, solution of 4N HCI in dioxane (5.9 mL, 23.6 mmol) was added to a flask containing bis(1 ,1 -dimethylethyl) (2S,2'S)-2,2'-[benzene-1 ,4-diylbis(1 H- benzimidazole-5,2-diyl)]bis(4,4-difluoro-1 -pyrrolidinecarboxylate) (361 mg, 0.50 mmol). The mixture was sonicated and then allowed to stir for 2 hours. The mixture was then diluted with ether and filtered to yield 5,5'-benzene-1,4- diylbis{2-[(2S)-4,4-difluoro-2-pyrrolidinyl]-1H-benzimidazole} tetrahydrochloride (316 mg) as a tan solid. ¹H NMR (400 MHz, DMSO-c/₆) δ ppm 7.98 (s, 2 H) 7.86 (s, 4 H) 7.77 - 7.82 (m, 2 H) 7.68 - 7.76 (m, 2 H) 5.35 (t, J=8 Hz, 2 H) 3.85 (t, J=12 Hz, 2 H) 2.88 - 3.31 (m, 2 H).

Intermediate 23c

5,5'-benzene-1,4-diylbis{2-[(2S,4R)-4-fluoro-2-pyrrolidinyl]-1H- benzimidazolejtetrahydrochloride

Synthesized analogously to intermediate 23b (Scheme II step 1 , 2, 3)

5,5'-benzene-1 -diylbis{2-[(2S R)-4-fluoro-2^yrrolidinyl]-1H-benzi

was synthesized from intermediate 5 and (4R)-1 -{[(1 ,1 - dimethylethyl)oxy]carbonyl}-4-fluoro-L-proline using general HATU coupling procedure and cyclization to yield the desired product in 63% yield.

1 H NMR (400 MHz, DMSO-c/₆) δ ppm 12.48 (d, J=9 Hz, 2 H) 7.88 (br. s., 1 H) 7.71 - 7.80 (m, 5 H) 7.64 (d, J=8 Hz, 1 H) 7.46 - 7.58 (m, 3 H) 5.29 - 5.56 (m, 2 H) 4.93 - 5.14 (m, 2 H) 3.84 (br. s., 4 H) 2.68 (d, J=6 Hz, 2 H) 2.13 - 2.43 (m, 2 H) 1 .39 & 1 .06 (each s, 18 H combined).

The product was deprotected under acidic conditions yielding 80% of 5,5'- Benzene-1 ,4-diylbis{2-[(2S,4R)-4-fluoro-2-pyrrolidinyl]-1 H-benzimidazole}.

1 H NMR (400 MHz, DMSO-c/₆) δ ppm 10.69 (br. s., 1 H) 9.82 (br. s., 1 H) 7.94 (s, 1 H) 7.83 (s, 3 H) 7.71 - 7.79 (m, 1 H) 7.66 (dd, J=8.5, 1 .3 Hz, 1 H) 5.50 - 5.83 (m, 2 H) 5.19 (br. s., 2 H) 3.60 - 3.90 (m, 10 H) 3.48 (dd, J=1 1 .6, 4.3 Hz, 2 H) 2.88 (br. s., 1 H) 2.55 - 2.73 (m, 2 H).

Intermediate 23d

5,5'-Benzene-1 ,4-diylbis{7-methyl-2-[(2S)-2-pyrrolidinyl]-1H-benzimidazole}

Scheme V step 1 . 2. 3. 4

1 ,1 -Dimethylethyl (2S)-2-(5-bromo-7-methyl-1 H-benzimidazol-2-yl)-1 - pyrrol id inecarboxylate was synthesized according to the general procedure for intermediate 45a using 5-bromo-3-methyl-1 ,2-benzenediamine instead of 4- bromo-1 ,2-benzenediamine 1 to yield the desired product in 71 % yield.

1 H NMR (400 MHz, DMSO-c/₆) δ ppm 1 1 .96 - 12.66 (m, 1 H) 7.55 and 7.42 (s, 1 H) 7.1 1 (d, J=7 Hz, 1 H) 4.82 - 5.00 (m, 1 H) 3.60 (br. s., 1 H) 3.42 (dd, J=7, 3 Hz, 1 H) 2.48 (d, J=2 Hz, 3 H) 2.34 (br. s., 1 H) 1 .83 - 2.09 (m, 3 H) 1 .00 - 1 .46(m, 9 H).

Bis(1 ,1 -dimethylethyl) (2S,2'S)-2,2'-[benzene-1 ,4-diylbis(4-methyl-1 H- benzimidazole-6,2-diyl)]di(1 -pyrrolidinecarboxylate) was synthesized according to the general procedure for bis(1 ,1 -dimethylethyl) (2S,2'S)-2,2'-[benzene-1 ,4- diylbis(1 H-benzimidazole-5,2-diyl)]di(1 -pyrrolidinecarboxylate) 22 using 1 ,1 - dimethylethyl (2S)-2-(5-bromo-7-methyl-1 H-benzimidazol-2-yl)-1 - pyrrol id inecarboxylate in place of 1 ,1 -dimethylethyl (2S)-2-(5-bromo-1 /-/- benzimidazol-2-yl)-1 -pyrrolidinecarboxylate to yield 57% of the desired product. ¹ H NMR (400 MHz, DMSO-c/₆) δ ppm 12.28 (s, 2 H) 7.75 (s, 4 H) 7.52 (s, 2 H) 7.33 (d, J=8 Hz, 2 H) 4.94 (br. s., 2 H) 3.63 (br. s., 2 H) 3.40 - 3.55 (m, 2 H) 2.57 (d, J=3 Hz, 6 H) 2.34 (br. s., 2 H) 1 .83 - 2.14 (m, 6 H) 1 .05 - 1 .5 (m, 18 H) 5,5'-Benzene-1 ,4-diylbis{7-methyl-2-[(2S)-2-pyrrolidinyl]-1 /-/-benzimidazole} hydrochloride was synthesized according to the general procedure for 23. The product was then deprotection with acids resulting in 23d in 76% yield.

1 H NMR (400 MHz, DMSO-c/₆) δ ppm 10.42 (br. s., 2 H) 9.60 (br. s., 2 H) 7.83 (s, 4 H) 7.79 (s, 2 H) 7.56 (s, 2 H) 5.06 (br. s., 4 H) 3.47 (m., 2 H) 2.67 (s, 6 H) 2.53 - 2.62 (m, 2 H) 2.37 (dd, J=13, 8 Hz, 2 H) 1 .97 - 2.27 (m, 4 H).

Intermediate 23e

4 HCI

5, 5'- θηζθηθ-1 -ΰίγΙ 8{2-[(28 5)-4-Αυο -2 γποΙί ίηγΙ]-1Η- θηζί

tetrahydrochloride

Scheme II step 1 , 2, 3

A mixture of 1 ,1 ':4',1 "-terphenyl-3,3",4,4"-tetramine hydrochloride salt

(intermediate 5) (0.45 g, 1 .03 mmol) in DMF (5 mL) and DIEA (0.720 mL, 4.13 mmol) was added to a flask containing (4S)-1 -{[(1 ,1 -dimethylethyl)oxy]carbonyl}- 4-fluoro-L-proline (0.481 g, 2.063 mmol), HATU (0.785 g, 2.063 mmol) and DIEA (0.36 mL, 2.06 mmol) in DMF (5 mL), which were premixed for 15 minutes. After stirring for one hour the solution was concentrated and residue partitioned between EtOAc and water, the organic layer separated, dried over sodium sulfate, filtered, concentrated and chromatographed on silica with 0-2.5% MeOH in DCM. The product was dissolved in acetic acid (20 mL) and the solution heated at 80° for one hour. Solvents were removed providing bis(1 ,1 - dimethylethyl) (2S,4S,2'S,4'S)-2,2'-[benzene-1 ,4-diylbis(1 H-benzimidazole-5,2- diyl)]bis(4-fluoro-1 -pyrrolidinecarboxylate) (652 mg, greater than theoretical yield). ¹ H NMR (400 MHz, DMSO-c/₆) δ ppm 1 1 .98 (br. s., 7 H) 7.72 - 8.01 (m, 6 H) 7.49 - 7.73 (m, 4 H) 5.09 - 5.68 (m, 2 H) 3.49 - 4.09 (m, 6 H) 3.05 - 3.22 (m, 2 H) 2.3 - 2.96 (m overlapping DMSO, 2 H) 1 .12 - 1 .54 (m, 18 H)

ES-LCMS m/z 685 (M+H)

Next, bis(1 ,1 -dimethylethyl) (2S,4S,2'S,4'S)-2,2'-[benzene-1 ,4-diylbis(1 H- benzimidazole-5,2-diyl)]bis(4-fluoro-1 -pyrrolidinecarboxylate) (0.149 g, 0.218 mmol) in THF (4 mL) was treated with 4N HCI in dioxane (2 mL, 8 mmol), allowed to stir for 30 minutes and concentrated to yield quantitative amount of 5,5'- benzene-1,4-diylbis{2-[(2S,4S)-4-fluoro-2-pyrrolidinyl]-1H-benzimidazole} tetrahydrochloride

Intermediate 41 b

methyl ((1S)-1 -{[(2S)-2-(5-bromo- 1 H-benzimidazol-2-yl)-1 -pyrrolidinyl]carbonyl}-2- methylpropyl)carbamate (non-preferred name)

Scheme III, step 1 . 2

4-bromo-1 ,2-benzenediamine 1 (0.8g, 4.28 mmol) A/-[(methyloxy)carbonyl]-L- valyl-proline 9 (1 .1 g, 4.28 mmol), HATU (2.439 g, 6.42 mmol), and DIEA (1 .121 ml, 6.42 mmol) were dissolved in Ν,Ν-dimethylformamide (20ml) and stirred at RT for 12h. The reaction mixture was concentrated and partitioned between ethyl acetate (50) mL and NaHCO3 (20) mL. The organic phase was separated and dried over sodium sulphate, solvents removed vacuo and the crude product dissolved in acetic acid (4.90 ml, 86 mmol) and heated at 60°C for 12h.

Saturated NaHCO3 was added and the product was extracted with

dicholoromethane, following by purification on silica column with 5% MeOH/DCM to yield the product as a tan foam.

1 H NMR (400 MHz, CDCI₃) δ ppm

7.64 - 7.73 (m, 1 H), 7.39 - 7.46 (m, 1 H), 7.34 (s, 1 H), 5.34 - 5.50 (m, 2 H), 4.27 - 4.39 (m, 1 H), 3.88 (br. s., 1 H), 3.71 (s, 3 H), 3.17 (br. s., 1 H), 2.97 (s, 2 H), 2.32 - 2.43 (m, 1 H), 2.20 - 2.32 (m, 1 H), 2.13 - 2.20 (m, 1 H), 1 .96 (br. s., 1 H), 1 .45 (dd, J=15.3, 6.3 Hz, 6 H)

ES LC-MS m/z = 425.0 (M+H)⁺ 1, 1-dimethylethyl 6-bromo-2-((2S)-1-{N-[(methyloxy)carbonyl]-L-valyl}-2- pyrrolidinyl)- 1 H-benzimidazole- 1 -carboxylate

Scheme IV, step 1

To a mixture of ((1 S)-1 -{[(2S)-2-(5-bromo-1 H-benzimidazol-2-yl)-1 - pyrrolidinyl]carbonyl}-2-methylpropyl)carbamate 30b (0.5g, 1 .181 mmol) and triethylamine (0.823 mL, 5.91 mmol) was added to a solution of B0C2O (0.823 mL, 3.54 mmol) in DCM (3mL), catalytic amount of DMAP and the reaction mixture was allowed to stir for 12 h. The reaction was added water and DCM (2 X 10mL), layers were separated, washed with brine, dried with MgSO4, and the concentrated crude product was purified on silica gel column using 5% MeOH/DCM to give 41 b in 60% yield.

1 H NMR (400 MHz, CDCI₃) δ ppm

8.08 (d, J=1 .7 Hz, 1 H), 7.76 (d, J=8.8 Hz, 1 H), 7.71 (d, J=1 .8 Hz, 1 H), ), 7.40 - 7.43 (m, 1 H), 5.85 - 5.93 (m, 1 H), 5.35 (d, J=9.3 Hz, 1 H), 4.42 (dd, J=9.2, 5.9 Hz, 1 H), 3.95 - 4.03 (m, 1 H), 3.90 (br. s., 1 H), 3.67 (s, 3 H), 2.35 - 2.46 (m, 1 H), 2.17 (dd, J=13.0, 6.7 Hz, 1 H), 2.01 - 2.1 1 (m, 1 H), ), 1 .76 (d, J=3.8 Hz, 1 H), 1 .71 (d, J=3.8 Hz, 6 H), 1 .48 (s, 9 H).

ES LC-MS m/z = 525.2 (M+H)⁺

Intermediate 10b

Dimethyl (2,5-pyridinediylbis{(2-aminobenzene-4,1 - diyl)imino(oxomethanediyl)(2S)-2,1 -pyrrolidinediyl[(2S)-3-methyl-1 -oxo-1 ,2- butanediyl]})biscarbamate

Scheme I step 6

4,4'-(2,5-pyridinediyl)di(1 ,2-benzenediamine) pentahydrochloride 5b (56 mg, 0.1 18 mmol), N-[(methyloxy)carbonyl]-L-valyl-L-proline 9 (67.6 mg, 0.248 mmol) and DIEA (0.206 mL, 1 .182 mmol) in DMF (5 mL) were cooled to 0 °C. HATU (94 mg, 0.248 mmol) was added and the reaction mixture was allowed to reach the RT and proceed for 5 hrs. The product was partitioned between EtOAc (100 mL) and 1 N HCI solution (100 mL), layers separated, the aqueous layer neutralized with solid Na₂CO3 and extracted with EtOAc (100 mL). The product was purified on silica gel column using 0-20%MeOH/EtOAc to yield dimethyl (2,5- pyridinediylbis{(2-aminobenzene-4,1 -diyl)imino(oxomethanediyl)(2S)-2,1 - pyrrolidinediyl[(2S)-3-methyl-1 -oxo-1 ,2-butanediyl]})biscarbamate (44 mg, 47%) as yellow solid

ES-LCMS MH⁺ 800.5.

Example 1

Scheme I step 7

Dimethyl (2, 5-pyridinediylbis{1H-benzimidazole-5, 2-diyl(2S)-2, 1- pyrrolidinediyl[(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

A bright orange solution of dimethyl (2,5-pyridinediylbis{(2-aminobenzene-4,1 - diyl)imino(oxomethanediyl)(2S)-2,1 -pyrrolidinediyl[(2S)-3-methyl-1 -oxo-1 ,2- butanediyl]})biscarbamate (intermediate 10b) (42 mg, 0.053 mmol) in acetic acid (4 mL) was heated at 80 °C for 1 .5 h. The mixture was cooled to room

temperature and purified on RP-HPLC using 10-80% CH₃CN /H₂O (both containing 0.2%NH₃) to afford dimethyl (2,5-pyridinediylbis{1 H-benzimidazole- 5,2-diyl(2S)-2,1 -pyrrolidinediyl[(2S)-3-methyl-1 -oxo-1 ,2-butanediyl]})biscarbamate (8 mg, 20%) as a white solid.

1 H NMR (400 MHz, DMSO-d6) δ ppm 12.28 (br. s., 2 H) 8.97 (br. s., 1 H) 7.72 - 8.36 (m, 5 H) 7.57 (br. s., 3 H) 7.33 (s, 2 H) 5.21 (br. s., 2 H) 4.09 (s, 2 H) 3.86 (br. s., 3 H) 3.54 (s, 6 H) 3.16 - 3.28 (m,1 H) 1 .79 - 2.39 (m, 10 H) 0.76-0.98 (m, 12 H). ES-LCMS m/z 764 M+H), purity 100% Example 2

Diethyl (benzene- 1 ,4-diylbis{1 H-benzimidazole-5, 2-diyl(2S)-2, 1 - pyrrolidinediyl[(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

5,5'-benzene-1 ,4-diylbis{2-[(2S)-2-pyrrolidinyl]-1 H-benzimidazole} (intermediate 23) and N-[(ethyloxy)carbonyl]-L-valine were reacted following the general HATU coupling procedure to give the title compound in 52% yield.

¹ H NMR (400 MHz, DMSO-d₆) δ 12.07 - 12.68 (m, 2H), 7.44 - 7.90 (m, 1 1 H), 7.25 (d, J = 8.4 Hz, 2H), 5.12 - 5.29 (m, 2H), 4.10 (t, J = 8.3 Hz, 2H), 3.95 - 4.05 (m, 4H), 3.86 (d, J = 4.9 Hz, 3H), 2.16 - 2.37 (m, 4H), 1 .84 - 2.16 (m, 6H), 1 .18 (t, J = 7.1 Hz, 6H), 0.86 (m,12H)

HRMS for C₄₄H₅₄N₈O₆ (M + H)⁺ calc: 791 .4245, found: 791 .4244.

Purity (LC/MS) 97%

Example 3

Bis(1, 1-dimethylethyl) (benzene-1 ,4-diylbis{1H-benzimidazole-5,2-diyl(2S)-2, 1- pyrrolidinedi l[(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

5,5'-benzene-1 ,4-diylbis{2-[(2S)-2-pyrrolidinyl]-1 H-benzimidazole} (intermediate 23) and N-{[(1 ,1 -dimethylethyl)oxy]carbonyl}-L-valine were reacted following the general HATU coupling procedure to give the title compound in 47% yield.

1 H NMR (400 MHz, DMSO-d₆) δ 12.05 - 12.74 (m, 2H), 7.43 - 7.93 (m, 10H), 6.83 (d, J = 8.3 Hz, 2H), 5.12 - 5.33 (m, 2H), 4.08 (t, J = 8.1 Hz, 2H), 3.79 - 3.91 (m, 3H), 2.25 (d, J = 6.4 Hz, 4H), 1 .85 - 2.15 (m, 5H), 1 .40 (s, 18H), 1 .18 (d, J = 7.6 Hz, 2H), 0.77 - 0.93 (m, 12H); HRMS for C48H62N8O6 (M +H )+ calc: 847.4871 , found: 847.4868; Purity (LC/MS) 98%

Example 4 (Benzene-1 ,4-diylbis{1H-benzimidazole-5,2-diyl(2S)-2, 1 -pyrrolidinediyl[(2S)-3- methyl-1 -oxo-1 ,2-butanediyl]})diformamide

5,5'-benzene-1 ,4-diylbis{2-[(2S)-2-pyrrolidinyl]-1 H-benzimidazole} (intermediate 23) and /V-formyl-L-valine were reacted following the general HATU coupling procedure to give the title compound in 36% yield.

¹ H NMR (400MHz, DMSO-d₆) δ 12.97 - 1 1 .95 (m, 2 H), 8.64 - 7.38 (m, 12 H), 5.19 (d, J = 7.0 Hz, 1 H), 4.71 - 4.18 (m, 2 H), 4.10 - 3.50 (m, 9 H), 2.43 - 1 .78 (m, 8 H), 1 .28 - 0.44 (m, 12 H)

HRMS for C₄oH₄₆N₈O₄ (M + H)⁺ calc: 703.3720, found: 703.3719

Purity (LC/MS) 100%

Example 5

N-{(1S)-1-[((2S)-2-{5-[4-(2-{(2S)-1-[(2S)-2-(acetylamino)^^^

pyrrolidinyl}-1H-benzimidazol-6-yl)phenyl]- 1 H-benzimidazol-2-yl}- 1 - pyrrolidinyl)carbon l]-2-methylpropyl}acetamide

5,5'-benzene-1 ,4-diylbis{2-[(2S)-2-pyrrolidinyl]-1 H-benzimidazole} (intermediate 23) and

were reacted following the general HATU coupling procedure to give the title compound in 38% yield.

¹ H NMR (400MHz, DMSO-d₆) δ 12.91 - 1 1 .80 (m, 2 H), 8.35 - 7.33 (m, 12 H), 5.31 - 5.10 (m, 1 H), 4.61 - 4.24 (m, 2 H), 4.08 - 3.62 (m, 6 H), 2.36 - 1 .68 (m, 15 H), 1 .31 - 0.57 (m, 12 H),

LCMS for C₄₂H₅oN₈O₄ (M + H)⁺ calc: 731 .4, found: 731 .4

Purity (LC/MS) 100% (sum of diastereomers) Example 6

(1R, 1 'R)-2,2'-{benzene-1 -diylbis[1H-benzimidazole-5,2^ 1- pyrrolidinediyl]}bis N,N-dimethyl-2-oxo-1-phenylethanamine)

5,5'-benzene-1 ,4-diylbis{2-[(2S)-2-pyrrolidinyl]-1 H-benzimidazole} (intermediate 23) and (2R) (dimethylamino)(phenyl)ethanoic acid were reacted following the general HATU coupling procedure to give the title compound in 28% yield.

1 H NMR (400MHz, DMSO-d₆) δ 12.29 (br. s., 2 H), 8.13 - 6.59 (m, 20 H), 5.13 (d, J = 7.3 Hz, 2 H), 4.36 - 3.87 (m, 8 H), 2.37 - 1 .60 (m, 18 H)

HRMS for C₄₈H₅oN₈O2 (M + H)⁺ calc: 771 .4135, found: 771 .4537

Purity (LC/MS) 100%

Example 7

[(1S)-1-({(2S)-2-[5-(2,5-difluoro-4-{2-[(2S)-1-((2S)-3-m^^

{[(methyloxy)carbonyl]amino}butanoyl)-2^yrrolidinyl]-^

yl}phenyl)-1H-benzi idazol-2-yl]-1-pyrrolidinyl}carbonyl)-2- acid)

Scheme IV step 2

The mixture of 1 ,1 -dimethylethyl 6-bromo-2-((2S)-1 -{/V-[(methyloxy)carbonyl]-L- valyl}-2-pyrrolidinyl)-1 H-benzimidazole-1 -carboxylate 41b (129 mg, 0.246 mmol), 2,2'-(2,5-difluorobenzene-1 ,4-diyl)bis(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolane) 3b (30 mg, 0.082 mmol) and Na₂CO₃ (1 M soln in H₂O, 0.246 mol) in dioxane (2ml_) and Pd(dppf)CI₂ (6mg, 8.2mmol) was purged with nitrogen and microwaved at 140° for 30 minutes. After cooling down, the mixture was filtered, diluted with EtOAc, and washed with aqueous NaHCO3, brine, dried over Na₂SO₄ and concentrated. The crude product was purified on silica gel (MeOH/DCM 0-20%) or by RP-HPLC using 10-80% acetonithle/H₂O (both containing 0.2%NH₃) to afford the title compound as yellow solid (20mg, Yield: 30.5%)

¹ H NMR (400 MHz, DMSO-d6) δ 12.36 (br. s., 2 H), 7.72 (br. s., 2 H), 7.58 (br. s.,

1 H),7.52 (t, J=9.0 Hz, 2 H),7.39 (d, J=8.3 Hz, 2 H), 7.32 (s, 1 H), 5.08 - 5.27 (m,

2 H), 3.94 - 4.20 (m, 2 H), 3.80 - 3.93 (m, 4 H), 3.54 (s, 6 H), 3.25 (br. s., 2 H), 2.24 (d, J=6.2 Hz, 4 H), 1 .99 (s, 6 H), 0.73 - 0.98 (m, 12 H)

ES LC-MS m/z = 799.4 (M+H)⁺

Example 8

[(1S)-1-({(2S)-2-[5-(2,5-dimethyl-4-{2-[(2S)-1-((2S)-3-me^

{[(methyloxy)carbonyl]amino}butanoyl)-2-pyrrolidinyl]-1H-benzimidazol-5- yl}phenyl)-1H-benzimidazol-2-yl]-1-pyrrolidinyl}carbonyl)-2-methylpropyl]carbam acid non-preferred name)

Prepared similar to procedure described in example 7, yield: 13.5%

1 H NMR (400 MHz, DMSO-d6) δ

12.1 1 - 12.34 (m, 2 H), 7.58 - 7.69 (m, 2 H), 7.53 - 7.59 (m, 2 H), 7.45 - 7.52 (m, 2 H), 7.37 - 7.43 (m, 1 H), 7.29 - 7.36 (m, 2 H), 7.07 - 7.21 (m,3 H), 5.1 1 - 5.26 (m, 2 H), 4.00 - 4.20 (m, 2 H), 3.78 - 3.93 (m, 4 H), 3.54 (s, 6 H), 2.24 (br. s., 5 H), 2.13 - 2.19 (m, 1 H), 1 .79 - 2.1 1 (m, 8 H), 0.86 (m, 12 H);

ES LC-MS m/z = 791 .5 (M+H)⁺

Example 9

dimethyl (2, 5-furandiylbis{1H-benzimidazole-5, 2-diyl(2S)-2, 1-pyrrolidinediyl[(2S)- 3-methyl-1 -oxo-1 ,2-butanediyl]})biscarbamate

Prepared similar to procedure described in example 7 in 12.8% yield.

1 H NMR (400 MHz, DMSO-d6) δ 12.27 (br. s., 2 H), 7.82 - 7.98 (m, 1 H), 7.60 (br s., 2 H), 7.49 - 7.57 (m, 2 H), 7.39 - 7.48 (m, 1 H), 7.25 - 7.38 (m,2 H), 6.97 (s, 2 H), 5.10 - 5.26 (m, 2 H), 4.01 - 4.22 (m, 2 H), 3.77 - 3.90 (m, 4 H), 3.54 (s, 6 H), 2.14 - 2.32 (m, 4H), 1 .79 - 2.12 (m, 6 H), 0.85 (m, 12 H)

ES LC-MS m/z = 753.4 (M+H)⁺

Example 10

dimethyl (2, 5-thienediylbis{1H-benzimidazole-5, 2-diyl(2S)-2, 1-pyrrolidinediyl[(2S)- 3-meth l-1 -oxo-1 ,2-butanediyl]})biscarbamate

Prepared similar to procedure described in example 7, yield: 14.3%.

1 H NMR (400 MHz, DMSO-d6 ) δ 12.03 - 12.49 (m, 2 H), 7.58 - 7.89 (m, 3 H), 7.42 - 7.53 (m, 5 H), 7.33 (d, J=8.5 Hz, 2H), 5.12 - 5.22 (m, 2 H), 4.03 - 4.12 (m, 2 H), 3.85 (br. s., 4 H), 3.54 (s, 6 H), 2.15 -2.29 (m, 4 H), 1 .98 - 2.1 1 (m, 4 H), 1 .87 - 1 .97 (m, 2 H), 0.73 - 0.93 (m, 12 H);

ES LC-MS m/z = 769.6 (M+H)⁺

Example 11

Dimethyl (benzene-1,4-diylbis{1H-benzimidazole-5,2-diyl(2S)-2, 1- pyrrolidinediyl[(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

To a solution of N-[(methyloxy)carbonyl]-L-valine 6 (1 18 mg, 0.673 mmol), DIEA (0.364 ml_) and HATU (256 mg, 0.673 mmol) in DMF (2.6 ml_) was added 5,5'- benzene-1 ,4-diylbis{2-[(2S)-2-pyrrolidinyl]-1 H-benzimidazole} intermediate 23 (200 mg, 0.336 mmol) and reaction allowed to proceed at RT for 1 hour. Following purification by RP-HPLC dimethyl (benzene-1 ,4-diylbis{1 H- benzimidazole-5,2-diyl(2S)-2, 1-pyrrolidinediyl[(2S)-3-methyl-1-oxo-1,2- butanediyl]})biscarbamate (138 mg, 54%) was obtained as a white solid.

1 H NMR (400 MHz, METHANOL-c/₄) δ ppm 7.67 - 7.87 (m, 6 H) 7.56 (br. s., 4 H) 5.27 (dd, J=8.0, 5.1 Hz, 2 H) 4.25 (d, J=7.4 Hz, 2 H) 3.98 - 4.13 (m, 2 H) 3.93 (br. s., 2 H) 3.64 (s, 6 H) 1 .92 - 2.54 (m, 10 H), 0.69 - 1 .03 (m, 12 H).

ES-LCMS m/z 763 (M+H), Purity 100%

Example 12

Dimethyl (benzene-1 ,4-diylbis{1H-benzimidazole-5,2-diyl(2S)-2, 1- pyrrolidinediyl[(2S)-3,3-dimethyl-1 -oxo-1, 2-butanediyl]})biscarbamate

5,5'-benzene-1 ,4-diylbis{2-[(2S)-2-pyrrolidinyl]-1 H-benzimidazole} intermediate 23 and 3-methyl-N-[(methyloxy)carbonyl]-L-valine were reacted following the general HATU coupling procedure to yield dimethyl (benzene-1 ,4-diylbis{1 H- benzimidazole-5,2-diyl(2S)-2,1 -pyrrol idinediyl[(2S)-3,3-dimethyl-1 -oxo-1 ,2- butanediyl]})biscarbamate in 52% yield.

¹ H NMR (400 MHz, METHANOL-c/₄) δ ppm 7.66 - 7.84 (m, 6 H) 7.43 - 7.66 (m, 4 H) 6.90 and 6.56 (d, J=9 Hz, 2 H) 5.51 and 5.28(m, 2 H), 4.36 and 4.17 (d, J=9.0 Hz, 2 H), 3.83 - 4.1 1 (m, 4 H) 3.65 (s, 6 H) 2.21 - 2.63 (m, 5 H) 2.12 (dd, J=1 1 , 6 Hz, 1 H) 0.88 - 1 .03 (m, 18 H).

HRMS for C₄₄H₅5N₈O6 (M + H)⁺ calc: 791 .4245, found: 791 .4245.

ES-LCMS m/z 791 (M+H), Purity 100%

Example 13

Dimethyl (benzene-1,4-diylbis{1H-benzimidazole-5,2-diyl(2S)-2, 1- rrolidinediyl[(2S)-1 -oxo-1, 2-propanediyl]})biscarbamate

5,5'-benzene-1 ,4-diylbis{2-[(2S)-2-pyrrolidinyl]-1 H-benzimidazole} intermediate 23 and N-[(methyloxy)carbonyl]-L-alanine were reacted according to the general HATU coupling procedure to yield dimethyl (benzene-1 ,4-diylbis{1 H- benzimidazole-5,2-diyl(2S)-2,1 -pyrrol idinediyl[(2S)-1 -oxo-1 ,2- propanediyl]})biscarbamate (50% yield).

1 H NMR (400 MHz, METHANOL-c/₄) δ ppm 7.66 - 7.87 (m, 6 H), 7.50 - 7.65 (m, 4 H), 5.29 (dd, J=8, 4 Hz, 2 H), 4.51 (q, J=7 Hz, 2 H), 3.92 (m, 4 H), 3.63 (s, 6 H), 2.33 - 2.58 (m, 2 H), 1 .84 - 2.33 (m, 6 H), 1 .34 (d, J=7 Hz, 6 H).

ES-LCMS m/z 707 (M+H), Purity 100%

HRMS for C38H₄₃N₈O6 (M + H)⁺ calc: 707.3306, found: 707.3306.

Example 14

Dimethyl (benzene-1,4-diylbis{1H-benzimidazole-5,2-diyl(2S)-2, 1- pyrrolidinediyl[(1S)-1-cyclohexyl-2-oxo-2, 1-ethanediyl]})biscarbamate

5,5'-benzene-1 ,4-diylbis{2-[(2S)-2-pyrrolidinyl]-1 H-benzimidazole} intermediate 23 and (2S)-cyclohexyl{[(methyloxy)carbonyl]annino}ethanoic acid were reacted following the general HATU coupling procedure to yield dimethyl (benzene-1 ,4- diylbis{1 /-/-benzimidazole-5,2-diyl(2S)-2,1 -pyrrolidinediyl[(1 S)-1 -cyclohexyl-2-oxo- 2,1 -ethanediyl]})biscarbamate in 40% yield.

¹ H NMR (400 MHz, METHANOL-c/₄) δ ppm 7.66 - 7.89 (m, 6 H), 7.50 - 7.65 (m, 4 H), 5.16 - 5.60 (m, 2 H), 4.25 (d, J=8 Hz, 2 H), 3.98 -4.17 (m, 2 H), 3.82 - 3.92 (m, 2H), 3.64 (s, 6 H), 1 .89 - 2.66 (m, 8 H) 1 .55 - 1 .75 (m, 12 H), 0.75 - 1 .45 (m, 10 H).

ES-LCMS m/z 843 (M+H), purity 100%

HRMS for C₄₈H59N₈O6 (M + H)⁺ calc: 843.4558, found: 843.4565.

Example 15

Dimethyl (benzene-1 ,4-diylbis{1 H-benzimidazole-5,2-diyl(2S)-2, 1- rrolidinediyl[(1S)-2-oxo-1-phenyl-2, 1-ethanediyl]})biscarbamate

5,5'-benzene-1 ,4-diylbis{2-[(2S)-2-pyrrolidinyl]-1 H-benzimidazole} intermediate 23 and (2S)-{[(methyloxy)carbonyl]amino}(phenyl)ethanoic acid were reacted following the general HATU coupling to yield dimethyl (benzene-1 ,4-diylbis{1 H- benzimidazole-5,2-diyl(2S)-2,1 -pyrrolidinediyl [(1 S)-2-oxo-1 -phenyl-2,1 - ethanediyl]})biscarbamate (21 % yield). ¹ H NMR (400 MHz, METHANOL-c/₄) δ ppm 7.68 - 7.93 (m, 6 H), 7.57 (br. s., 4 H), 7.22 - 7.51 (m, 10 H), 5.46 - 5.64 (m, 2 H), 5.23 - 5.42 (m, 2 H), 3.79 - 4.23 (m, 2 H), 3.64 (m, 6 H), 3.47 (m, 2 H), 1 .81 - 2.66 (m, 8 H).

ES-LCMS m/z 831 (M+H), Purity 100%

HRMS for C^H^NsOe (M + H)⁺ calc: 831 .3619, found: 831 .3617.

Example 16

Dimethyl (benzene-1, 4-diylbis{1 H-benzimidazole-5, 2-diyl(2S)-2, 1 - rrolidinediyl[(2S)-4-methyl-1 -oxo-1, 2-pentanediyl]})biscarbamate

5,5'-benzene-1 ,4-diylbis{2-[(2S)-2-pyrrolidinyl]-1 H-benzimidazole} intermediate 23 and N-[(methyloxy)carbonyl]-L-leucine were reacted following the general HATU coupling procedure to yield dimethyl (benzene-1 ,4-diylbis{1 H- benzimidazole-5,2-diyl(2S)-2,1 -pyrrol idinediyl[(2S)-4-methyl-1 -oxo-1 ,2- pentanediyl]})biscarbamate (19% yield).

¹ H NMR (400 MHz, METHANOL-c/₄) δ ppm 7.67 - 7.90 (m, 6 H), 7.47 - 7.67 (m, 4 H), 5.28 (m, 2 H), 4.49 (dd, J=1 1 , 4 Hz, 2 H), 3.78 - 4.05 (m, 4 H), 3.64 (s, 6 H), 1 .83 - 2.53 (m, 8 H), 1 .39 - 1 .82 (m, 6 H), 0.76 - 1 .12 (m, 12 H).

ES-LCMS m/z 791 (M+H), Purity 100%

Example 17

Dimethyl (benzene-1, 4-diylbis{1 H-benzimidazole-5, 2-diyl[(2S)-4,4-difluoro-2, 1- pyrrolidinediyl][(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

To a solution of N-[(methyloxy)carbonyl]-L-valine (35 mg, 0.198 mmol), DIEA (0.14 mL) and HATU (75 mg, 0.198 mmol) in DMF (0.8 mL) stirred at room temperature was added solid intermediate 23b 5,5'-benzene-1 ,4-diylbis{2-[(2S)- 4,4-difluoro-2-pyrrolidinyl]-1 H-benzimidazole}. The reaction was allowed to stir at RT for 1 hour. The product was purified by RP-HPLC to yield dimethyl (benzene- 1 ,4-diylbis{1 H-benzimidazole-5,2-diyl[(2S)-4,4-difluoro-2,1 -pyrrol id inediyl] [(2S)-3- methyl-1 -oxo-1 , 2-butanediyl]})biscarbamate (39 mg, 47%) as a white solid.

1 H NMR (400 MHz, DMSO-c/₆) δ ppm 12.44 (br. s., 2 H) 7.71- 7.87 (m, 6 H) 7.43 - 7.69 (m, 6 H) 5.44 (t, J=8 Hz, 2 H) 4.58 (br. s., 2 H) 4.25 (br. s., 2 H) 3.98 (t, J=8 Hz, 2 H) 3.57 (s, 6 H) 3.04 (br. s., 2 H) 2.84 (br. s., 2 H) 1 .91 (m, 2 H) 0.73 - 1 .02 (m, 12 H)

HRMS for C₄₂H₄₇N₈O₆F₄ (M + H)⁺ calc: 835.3555, found: 835.3553.

ES-LCMS m/z 835 (M+H), 100%

Example 18

Dimethyl (benzene-1 ,4-diylbis{(4-methyl-1 H-benzimidazole-6,2-diyl)(2S)-2, 1- pyrrolidinediyl[(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

Dimethyl (benzene-1 ,4-diylbis{(4-methyl-1 H-benzimidazole-6,2-diyl)(2S)-2,1 - pyrrolidinediyl[(2S)-3-methyl-1 -oxo-1 ,2-butanediyl]})biscarbamate was synthesized from 5,5'-benzene-1 ,4-diylbis{7-methyl-2-[(2S)-2-pyrrolidinyl]-1 H- benzimidazole} 23d and 6 using general HATU coupling procedure to yield the desired product in 58% yield.

¹ H NMR (400 MHz, DMSO-c/₆) δ ppm 12.03 - 12.42 (m, 2 H), 7.73 (d, J=5 Hz, 4 H,) 7.64 (br. s., 1 H), 7.53 (s, 1 H), 7.13 - 7.41 (m, 4 H), 5.12 - 5.27 (m, 2 H), 4.04 - 4.18 (m, 2 H), 3.89 (br. s., 4 H), 3.55 (s, 6 H), 2.54 (d, J = 5 Hz, 6 H), 2.27 (br. s., 4 H), 2.02 (br. s., 6 H), 0.64 - 1 .1 1 (m, 12 H).

HRMS for C₄₄H₅5N₈O6 (M + H)⁺ calc: 791 .4245, found: 791 .4241 .

ES-LCMS m/z 791 (M+H), Purity 100% Example 19

Dimethyl (benzene- 1, 4-diylbis{1H-benzimidazole-5, 2-diyl( 8S)-1 ,4-dioxa-7- azaspiro[4.4]nonane-8, 7-diyl[(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate Scheme I step 6, 7

A solution of (8S)-7-{N-[(methyloxy)carbonyl]-L-valyl}-1 ,4-dioxa-7- azaspiro[4.4]nonane-8-carboxylic acid intermediate 60 was coupled to 1 ,1 ':4',1 "- terphenyl-3,3",4,4"-tetramine tetrahydrochloride 5 using the general HATU procedure. The product was partitioned between EtOAc and NaHCO3 solution and the organic layer washed with brine, dried with MgSO^ filtered and purified on silica gel column with 0-10% MeOH in DCM. The product was dissolved in acetic acid and heated at 70°C for 2.5 hours. The product was partitioned between EtOAc and NaHCO3, organic layer dried and the residue purified by RP- HPLC to yield dimethyl (benzene-1 ,4-diylbis{1 H-benzimidazole-5,2-diyl(8S)-1 ,4- dioxa-7-azaspiro[4.4]nonane-8,7-diyl[(2S)-3-methyl-1 -oxo-1 ,2- butanediyl]})biscarbamate (17 mg, 7%) as a white solid.

¹ H NMR (400 MHz, DMSO-c/₆ 140°C) δ ppm 1 1 .72 (br. s., 2 H), 7.73 (s, 6 H), 7.53 - 7.63 (m, 2 H), 7.50 (s, 2 H), 6.27 (br. s., 2 H), 5.27 - 5.51 (m, 2 H), 4.14 (dd, J=8, 7 Hz, 2 H), 3.86 - 4.09 (m, 10 H), 3.80 (br. s., 2 H), 3.53 (br. s., 6 H), 2.58 (d, J=7 Hz, 4 H), 1 .92 - 2.13 (m, 2 H), 0.75 - 0.95 (m, 12 H)

HRMS for C₄₆H₅₅N₈Oio (M + H)⁺ calc: 879.4041 , found: 879.4041 .

ES-LCMS m/z 879 (M+H), Purity 100%

Example 20

Dimethyl (benzene-1 ,4-diylbis{1 H-benzimidazole-5,2-diyl(3S)-6, 10-dioxa-2- azaspiro[4.5]decane-3,2-diyl[(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate Synthesized as example 19, except that intermediate 61 was used instead of 60.

¹ H NMR (400 MHz, DMSO-c/₆) δ ppm 1 1 .72 (br. s., 2 H), 7.73 (s, 6 H), 7.53 - 7.63 (m, 2 H), 7.50 (s, 2 H), 6.27 (br. s., 2 H), 5.27 - 5.51 (m, 2 H), 4.14 (dd, J=8, 7 Hz, 2 H), 3.86 - 4.09 (m, 10 H), 3.80 (br. s., 2 H), 3.53 (br. s., 6 H), 2.58 (d, J=7 Hz, 4 H), 1 .92 - 2.13 (m, 2 H), 0.75 - 0.95 (m, 12 H)

ES LC-MS m/z =907 (M+H). purity 96%

Example 21

Methyl [(1 S)-2-methyl-1 -({(2S)-2-[5-(4-{2-[(8S)-7 '-((2S)-3-methyl-2-

{[(methyloxy)carbonyl]amino}butanoyl)-1,4-dioxa-7-azaspiro[4.4]non-8-yl]-1H- benzimidazol-5-yl}phenyl)-1 H-benzimidazol-2-yl]- 1 - pyrrolidinyl}carbonyl)propyl]carbamate

Scheme VI step1 -3

N-[(methyloxy)carbonyl]-L-valyl-L-proline 9 was reacted with 1 ,1 ':4',1 "-terphenyl- 3,3",4,4"-tetramine 5 using general HATU coupling conditions. After 2.5 hours, the reaction was terminated by partitioning between EtOAc and NaHCO3, the organic layer was concentrated and residue was chromatographed on a silica gel column with 0-10% (2N NH3 in MeOH) / DCM. Product 50 dissolved in 0.2 mL DMF was coupled to (8S)-7-{N-[(Methyloxy)carbonyl]-L-valyl}-1 ,4-dioxa-7- azaspiro[4.4]nonane-8-carboxylic acid 60 (6.5 mg, 0.020 mmol) in presence of DIEA (4.33 μΙ_, 0.025 mmol) and HATU (7.54 mg, 0.020 mmol) for 2.5 hours. Following EtOAc and NaHCO3 workup, the organic layer was concentrated to yield 14mg of 51 , dissolved in acetic acid (0.43 mL) and heated at 70°C for 1 .5 hours. RP-HPLC purification (10-70% MeCN/water with 0.2% NH₃) yielded methyl [(1S)-2-methyl-1-({(2S)-2-[5-(4-{2-[(8S)-7-((2S)-3-methyl-2- {[(methyloxy)carbonyl]amino}butanoyl)-1,4-dioxa-7-azaspiro[4.4]non-8-yl]-1H- benzimidazol-5-yl}phenyl)-1H-benzimidazol-2-yl]-1- pyrrolidinyl}carbonyl)propyl]carbamate (2.3 mg, 1 %) as a white solid

1 H NMR (400 MHz, DMSO-d6 140°C) δ ppm 1 1 .72 (br. s., 2 H), 7.73 (s, 6 H) 7.53 - 7.63 (m, 2 H), 7.50 (s, 2 H), 6.27 (br. s., 2 H), 5.27 - 5.51 (m, 2 H), 4.14 (dd, J=8, 7 Hz, 2 H), 3.86 - 4.09 (m, 10 H), 3.80 (br. s., 2 H), 3.53 (br. s., 6 H), 2.58 (d, J=7 Hz, 4 H), 1 .92 - 2.13 (m, 2 H), 0.75 - 0.95 (m, 12 H)

HRMS for C44H53N8O8 (M + H)+ calc: 821 .3986, found: 821 .3992.

ES-LCMS m/z 821 (M+H), purity 86%

Example 22

Dimethyl (benzene-1,4-diylbis{1H-benzimidazole-5,2-diyl[(2S,4R)-4-fluoro-2, 1- pyrrolidinediyl][(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

dihydrochloride

Scheme II, step 4

To a solution of N-[(methyloxy)carbonyl]-L-valine 6, (151 mg, 0.86 mmol), DIEA (0.60 ml_) and HATU (327 mg, 0.86 mmol) in Ν,Ν-Dimethylformamide (DMF) (3.44 ml_) stirred at room temperature was added solid 5,5'-benzene-1 ,4- diylbis{2-[(2S,4R)-4-fluoro-2-pyrrolidinyl]-1 H-benzimidazole} 23c (271 mg, 0.43 mmol) and the reaction was allowed to proceed at RT for 1 hour. The reaction mixture was concentrated and the product was partitioned between EtOAc and NaHCO3 solution. The organic layer was washed with brine, dried with MgSO₄ filtered, concentrated and chromatographed on silica gel with a gradient of 0-7.5 % MeOH in DCM yielding dimethyl (benzene-1 ,4-diylbis{1 H-benzimidazole-5,2- diyl[(2S,4R)-4-fluoro-2,1 -pyrrolidinediyl][(2S)-3-methyl-1 -oxo-1 ,2- butanediyl]})biscarbamate (207 mg, 60%) as a tan solid

¹ H NMR (400 MHz, DMSO-d6) δ ppm 12.46 (br. s., 2 H) 7.66 - 8.01 (m, 6 H) 7.44 - 7.71 (m, 4 H) 7.37 (d, J=9 Hz, 2 H) 5.37 - 5.69 (m, 2 H) 5.22 (t, J=8 Hz, 2 H) 4.35 (br. s., 2 H) 4.03 (m, 4 H) 3.54 (s, 6 H) 2.53 - 2.77 (m, 4 H) 1 .80 - 1 .96 (m, 2 H) 0.69 - 0.96 (m, 12 H).

HRMS for C₄₂H₄9N₈O6F2 (M + H)+ calc: 799.3743, found: 799.3745.

ES-LCMS m/z 799 (M+H), Purity 97% Example 23

Dimethyl (benzene-1 ,4-diylbis{1 H-benzimidazole-5,2-diyl[(2S,4S)-4-fluoro-2, 1- pyrrolidinediyl][(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

Scheme II, step 4

5,5'-benzene-1,4-diylbis{2-[(2S,4S)-4-fluoro-2-pyrrolidinyl]-1H-benzimidazole} tetrahydrochloride ) 23e (0.218 mmol) in DMF (2 mL) was added HATU (0.165 g, 0.435 mmol), DIEA (0.152 mL, 0.870 mmol) and N-[(methyloxy)carbonyl]-L-valine 6 (0.076 g, 0.435 mmol) and the reaction allowed to proceed overnight. RP-HPLC purification yielded dimethyl (benzene-1 ,4-diylbis{1 H-benzimidazole-5,2- diyl[(2S, 4S)-4-fluoro-2, 1 -pyrrolidinediyl][(2S)-3-methyl- 1 -oxo-1,2- butanediyl]})biscarbamate (45 mg, 26% over 4 steps) as a white solid.

1 H NMR (400 MHz, methanol-c/₄) δ ppm 7.66 - 7.92 (m, 6 H), 7.57 (m, 4 H), 5.14 - 5.72 (m, 4H), 3.79 - 4.41 (m, 6 H), 3.66 (s, 6 H), 2.40 - 2.94 (m, 4 H), 1 .87 - 2.29 (m, 2 H), 0.77 - 1 .13 (m, 12 H).

ES-LCMS m/z 798 (M+H). Purity 92%

Example 24

dimethyl (benzene-1, 4-diylbis{(4-fluoro-1H-benzimidazole-6,2-diyl)(2S)-2, 1- pyrrolidinediyl[(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

Scheme III ste 3

A mixture of benzene-1 ,4-diyldiboronic acid 3 (0.1 13 g, 0.684 mmol), methyl ((1 S)-1 -{[(2S)-2-(5-bromo-7-fluoro-1 H-benzimidazol-2-yl)-1 -pyrrol idinyl]carbonyl}- 2-methylpropyl)carbamate 30 (0.604 g, 1 .369 mmol), PdCI₂(dppf)-CH₂CI₂ adduct (0.1 12 g, 0.137 mmol) and potassium carbonate (0.284 g, 2.053 mmol) in DME (6 mL) and Water (1 .00 mL) was heated at 100 °C for 4h. The product was purified on a silica column using 0-20% MeOH (2M NH₃) / CH₂CI₂ gradient to afford 0.368g (0.461 mmol, 34%) of the product as a dark yellow brown solid.

1 H NMR (400 MHz, methanol-d4) δ 7.72 - 7.80 (m, 4H), 7.44 - 7.71 (m, 4H), 7.24 - 7.39 (m, 2H), 5.20 - 5.35 (m, 2H), 4.29 (d, J = 7.28 Hz, 2H), 3.88 - 4.12 (m, 4H), 3.68 (s, 8H), 1 .98 - 2.56 (m, 10H), 0.82 - 1 .06 (m, 12H)

HRMS for C44H54N8O6 (M + H)+ calc: 799.3743, found: 799.370. Example 25

dimethyl (benzene-1,4-diylbis{(5-fluoro-1H-benzimidazole-6,2-diyl)(2S)-2, 1- rrolidinediyl[(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})

Scheme III step 3

Benzene-1 ,4-diyldiboronic acid 3 (0.109 g, 0.656 mmol), methyl ((1 S)-1 -{[(2S)-2- (5-bromo-6-fluoro-1 H-benzimidazol-2-yl)-1 -pyrrolidinyl]carbonyl}-2- methylpropyl)carbamate 30c (0.579 g, 1 .312 mmol), PdCI₂(dppf)-CH₂CI₂ adduct (0.107 g, 0.131 mmol) and potassium carbonate (0.272 g, 1 .968 mmol) in DME (6 ml_)/ water (1 .00 ml_) were heated at 100 °C for 4h. RP-HPLC purification afforded 14 mg (0.018 mmol, 1 .3%) of the product as a white solid. 1 H NMR (400 MHz), methanol-d4) δ 7.70-7.60 (m, 6H), 7.32-7.40 (m 2H), 5.20-5.36 (m, 2H), 4.21 -4.35 (m, 2H), 4.01 -4.15 (m, 2H), 3.87 - 4.01 (m, 2H), 3.63 -3.72 (m, 8H), 2.30 - 2.52 (m, 4H), 2.24 - 2.29 (m, 4H), 2.01 -2.21 (m, 4H), 0.79 - 1 .04 (m, 12H).

HRMS for C44H54N8O6 (M + H)+ calc: 799.3743, found: 799.3743.

Example 26

dimethyl (benzene-1,4-diylbis{1H-imidazo[4,5-b]pyridine-6,2-diyl(2S)-2, 1- rrolidinediyl[(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

Scheme V, step 8

Synthesized as in example 25, except that methyl ((1 S)-1 -{[(2S)-2-(6-bromo-1 H- imidazo[4,5-i ]pyridin-2-yl)-1 -pyrrol was used instead of methyl ((1 S)-1 -{[(2S)-2-(5-bromo-6-fluoro-1 H-benzimidazol-2-yl)- 1 -pyrrolidinyl]carbonyl}-2-methylpropyl)carbamate.

LC/MS purity 100%

Scheme VIII

Synthesis of Intermediate C

[2-amino-4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]amine (5.00 g, 21 .36 mmol), 1 -bromo-4-iodobenzene (5.49 g, 19.42 mmol), bis(1 ,1 - dimethylethyl) [4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)benzene-1 ,2- diyl]biscarbamate (Intermediate A, 8.43 g, 19.42 mmol), PdCI2(dppf)-CH₂CI₂ adduct (2.378 g, 2.91 mmol) in 1 ,4-Dioxane (20 ml_) was purged with nitrogen and reaction mixture stirred at 90°C for 2 hr. The volatiles were removed and the residue was column chromatographed to give bis(1 ,1 -dimethylethyl) (3",4"- diamino-1 ,1 ':4',1 "-terphenyl-3,4-diyl)biscarbamate (Intermediate C, 4.1 g, 43%). MS (ESI): [M+H]⁺.

Synthesis of Intermediate D

bis(1 ,1 -dimethylethyl) (3",4"-diamino-1 ,1 ':4',1 "-terphenyl-3,4-diyl)biscarbamate (Intermediate C, 1 .033 g, 2.1 1 mmol), (8S)-7-{A/-[(methyloxy)carbonyl]-L-valyl}- 1 ,4-dioxa-7-azaspiro[4.4]nonane-8-carboxylic acid (Intermediate 60, 0.730 g, 2.21 mmol), HATU (0.881 g, 2.316 mmol) in Λ/,/V-Dimethylformamide (DMF) (10 mL) was added DIEA (1 .103 mL, 6.32 mmol) and stirred overnight at RT. After solvent removal, acetic acid (12.05 mL, 21 1 mmol) was added and the mixture heated at 70°C for 4 hours. The volatiles were removed and the residue column chromatographed to give methyl [(1 S)-1 -({(8S)-8-[5-(3',4'-diamino-4-biphenylyl)- 1 /-/-benzimidazol-2-yl]-1 ,4-dioxa-7-azaspiro[4.4]non-7-yl}carbonyl)-2- methylpropyl]carbamate (Intermediate D, 0.83 g, 68%). MS (ESI): 585 [M+H]⁺.

Intermediate E can be synthesized as described for compound 8.

1 H NMR (400MHz, CDCI₃): δ 0.78-1 .50 (12H, m), 1 .62-1 .81 (1 H, m), 1 .93-2.10 (2H, m), 3.34 (0.5H, d), 3.62-3.82 (4.5H, m), 4.16 (0.5H, m), 4.28 (0.8H, d), 4.55 (t, 0.7H), 5.58 (1 H, m).

Intermediate F

Intermediate F-1 Intermediate F-2 Intermediate F-3 Intermediate F-4

Intermediate F-5 Intermediate F

Intermediate F-2 can be obtained by treating F-1 in dichloromethane/methanol (1 :1 ) with 2.2 eqivalents of TMS-CHN₂ (2M in hexane). F-2 (4g, 16.58 mmol) in toluene (30 mL) was then added 50.5 mL of 1 .1 M diethylzinc in toluene (6.14 g, 49.7 mmol) over 10 minutes at -22 °C, followed by dropwise addition of chloroiodomethane (17.5g, 7.22 mL, 99 mmol) over 5 min. The mixture was stirred at -22 °C for 6 hrs and purified on silica, yielding 1 .35g of the cyclopropyl product F-3. Deprotection of F-3 with excess 4N HCI in dioxane yielded F-4. Intermediate F-5 can be obtained by coupling F-4 and intermediate 6 using the method described for intermediate 8. The final intermediate F was obtained by treating 0.25g (0.8 mmol) of F-5 dissolved in 2 mL water and 2 mL dioxane with 0.023g (0.960 mmol) of lithium hydroxide monohydrate for 2 hrs at RT. After the reaction was complete, solvent were removed, the residue acidified with 1 M aqueous citric acid and extracted with ethyl acetate. The combined organic fractions were dried, yielding intermediate F. MS found 298 (calc 298).

Intermediate G

G-4 Intermediate G

G-2

CF₂Br₂ (21 .56 g, 103 mmol) and HMPT (18.4 g, 103 mmol) were added into a solution of (S)-1 -tert-butyl 2-methyl 4-oxopyrrolidine-1 ,2-dicarboxylate (5.0 g, 20.6 mmol) in THF (250 mL) at 0 °C. The resulting mixture was warmed to 0 °C and Zn (8.0 g, 123 mmol) was added in. The mixture was refluxed for 3.5 hours, cooled, diluted with water (100 mL) and Et₂O (150 mL). The aqueous phase was separated and extracted with Et₂O (150 mL x 2). The combined organic layer was washed with water (100 mL), brine (100 mL), dried over Na₂SO , filtered and concentrated to dryness. The residue was purified on silica gel chromatography (EtOAc:petroleum ether = 1 :20) to give G-2 (1 .12 g, 20%) as a colorless oil. ¹H- NMR (300 MHz, CDCI₃) δ 4.56-4.52 (m, 1 H), 4.13-4.08 (m, 2H), 3.74 (s, 3H), 2.88 (m, 1 H), 2.63 (m, 1 H), 1 .47 and 1 .42 (s, 9H). LCMS (m/z) ES+ 278 (m+1 ), purity 90%.

G-3

A solution of G-2 (300 mg, 1 .08 mmol) in dioxane (1 .5 mL) was cooled to 0 °C and 5 N HCI/dioxane (3.0 mL) was added in. The mixture was stirred at RT for 2 hours. The solvents were removed, and water (5 mL) was added, the aqueous phase extracted with EtOAc (10 mL x 2), pH was adjusted to 1 -2 with 1 N HCI, and the aqueous phase was extracted with DCM (10 mL x 2). The combined organic layers were dried over Na₂SO₄, filtered and concentrated to dry to give G- 3 (230 mg, crude) as dark brown gum which was used in the next step without further purification. LCMS (m/z) ES+ 178 (m+1 ). Purity 80%. G-4

A solution of G-3 (230 mg, 1 .08 mmol), (S)-2-(methoxycarbonylamino)-3-methyl butanoic acid (213 mg, 1 .22 mmol), EDCI (195 mg, 1 .22 mmol), HOBt (164 mg, 1 .22 mmol), DIPEA (296 mg, 2.3 mmol) in DMF (5 mL) was stirred at RT overnight. The reaction solution was diluted with EtOAc (10 mL), and washed with 5% citric acid (10 mL), saturated NaHCO₃ (10 mL), brine (10 mL), dried over Na₂SO₄, filtered and concentrated to dry to give a residue which was purified on silica gel column (EtOAc:petroleum ether = 1 : 5) to give G-4 (144 mg, 40%) as a colorless oil. LCMS (m/z) ES+ 335 (m+1 ), purity 90%. Intermediate G

Intermediate G-4 (522 mg, 1 .56 mmol) was treated with a solution of LiOH (95 mg, 4.68 mmol) in THF/H₂O (8 mL/8mL) at r.t. overnight. The THF was removed in vacuo, and aqueous phase was extracted with EtOAc (10 mL x 2). The pH of the solution was adjusted to 1 -2 with 1 N HCI, the resulting solution was extracted with EtOAc (10 mL x 2). The combined organic layer was dried over Na₂SO₄, filtered and concentrated to dry to give compound G (291 mg, 58%) as colorless oil. ¹H-NMR (300 MHz, CDCI₃) δ 5.36 (m, 1 H), 4.85 (m, 1 H), 4.57 (m, 1 H), 4.31 - 4.21 (m, 2H), 3.67 (s, 3H), 2.94 (m, 2H), 2.34 (m, 1 H), 1 .03-1 .90 (m, 6H). LCMS (m/z) ES+ 321 (m+1 ), purity 90%.

Intermediate H

OH OH OH O

$ [J Μ^βΟΗ I N _C|H DCM ¾ DCM j

DOC DOC

H-2 H-3 H-4

Intermediate H

H-7 H-8 Intermediate H

H-2

Thionyl chloride SOCI₂ (26.0 mL, 495.3 mmol) was added dropwise to (2S,4R)-4- hydroxypyrrolidine-2-carboxylic acid (50.0 g, 381 mmol) in MeOH (290 mL). The mixture was stirred at RT for 2 hours, concentrated to dryness to give a crude H- 2 (65.0 g) as pale yellow solid which was used in the next step without further purification.

H-3

Boc₂O (94.0 g, 428 mmol) was added into a solution of H-2 (65.0 g, 357.0 mmol) and Et₃N (124.0 mL, 892 mmol) in DCM (300 mL) at 0 °C. The resulting mixture was stirred at r.t. for 4 hours before the reaction solution was washed with 1 N HCI (300 mL), brine (300 mL) and dried over Na₂SO₄, filtered and concentrated to dryness to give H-3 (79.0 g, 90%) as a white solid.

H- 4

PCC (87.9 g, 408 mmol) was added to solution of H-3 (50.0 g, 204 mmol) in DCM (1 L) and the resulting mixture was stirred at RT overnight. The solution was filtered through celite pad, solvents evaporated and the resulting residue purified on silica (EtOAc:petroleum ether = 1 :2) to give H-4 (32.0 g, 65%) as a colorless oil. LC/MS (m/z) ES+ 244 (m+1 ). Purity 90%.

H- 5

NaHMDS (24.0 g, 65.8 mmol) was added into a solution of H-4 (32.0 g, 131 mmol) in THF (150 mL) at -78 °C under N₂ and the resulting mixture was stirred at -78 °C for 30 minutes before adding Tf₂NPh (48.7 g, 137.0 mmol) in THF (150 mL). The mixture was stirred at -78 °C for 2 hours, and then continued overnight at RT. The reaction was quenched with the addition of saturated NH CI (aq. 300 mL) and the aqueous phase was extracted with EtOAc (300 mL x 3). Combined organic layers were washed with brine, dried over Na₂SO₄, filtered and

concentrated to dryness to give a residue which was purified on silica gel column (EtOAc:petroleum ether = 1 :30) to give H-5 (18.0 g, 37%) as colorless oil. ¹ H- NMR (300 MHz, CDCI₃) δ 5.72 (m, 1 H), 5.03 (m, 1 H), 4.38-4.28 (m, 2H), 3.77 and 3.75 (s, 3H), 1 .48 and 1 .43 (s, 9H). LCMS (m/z) ES+ 376 (m+1 ). Purity 90%. H-6

A suspension of H-5 (18.0 g, 47.9 mmol), CH₃B(OH)₂ (7.18 g, 120 mmol), NaBr (4.93 g, 47.9 mmol), KF (9.18 g, 158.0 mmol) and Pd(PPh₃) (2.76 g, 2.4 mmol) in toluene (100 mL) was stirred at reflux for 5 hours. The reaction mixture was filtered through celite pad. The filtration was concentrated to dry and the residue was purified by chromatography (EtOAc:petroleum ether = 1 :5) to give H-6 (5.0 g, 50%) as a yellow oil. ¹H-NMR (300 MHz, CDCI₃) δ 5.35 (m, 1 H), 4.90 (m, 1 H), 4.16-4.04 (m, 2H), 3.72 (s, 3H), 1 .79 (s, 3H), 1 .47 and 1 .42 (s, 9H). LCMS (m/z) ES+ 242 (m+1 ). Purity 95%.

H-7

Intermediate H-6 (1 .0 g, 2.14 mmol) was treated with HCI/dioxane (1 1 .0 mL) at RT for 4 hours before the solvent was evaporated out in vacuo to give a crude H- 7 (0.6 g) as yellow oil which was used in the next step without further purification. H-8

To a solution of H-7 (0.58 g, 4.12 mmol) and DIPEA (1 .86 g, 14.42 mmol) in DCM (10 mL) was added (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (0.76 g, 4.34 mmol), HOBt (0.70 g, 5.13 mmol) and EDCI (0.99 g, 5.16 mmol). The resulting mixture was stirred at r.t. overnight, washed with water (10mL), saturated NaHCO₃ (10 mL x 2) and brine (10 mL x 1 ). The combined organic layer was dried over Na₂SO₄, filtered and concentrated to dryness. The residue was purified by chromatography (EtOAc:petroleum ether = 1 :5) to give H-8 (0.90, 73%) as a colorless oil. ¹ H-NMR (300 MHz, CDCI₃) δ 5.44 (m, 1 H), 5.32 (m, 1 H), 5.16 (m, 1 H), 4.93-4.22 (m, 3H), 3.71 (s, 3H), 3.65 (s, 3H), 2.06 (m, 1 H), 1 .81 (s, 3H), 1 .06 (d, 3H), 0.98 (d, 3H). LCMS (m/z) ES+ 299 (m+1 ). Purity 90%.

Intermediate H:_(S)-1 -((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-4- methyl- 2,5-dihydro-1 H-pyrrole-2-carboxylic acid

To a solution of H-8 (0.90 g, 2.98 mmol) in THF (10 mL) was added a solution of LiOH (0.15 g, 3.58 mmol) in water (2 mL). The mixture was stirred at r.t.

overnight. The mixture was concentrated and water (15 mL) was added in. The aqueous phase was extracted with EtOAc (15 mL x 2), acidified with 1 N HCI to pH 2-3, extracted with DCM (15 mL x 2). The combined organics were washed with water (15 mL), dried over Na₂SO₄, filtered and concentrated to dryness to give H (600 mg, 70%) as a white foam. ¹H-NMR (300 MHz, DMSO-c/₆) δ 12.58 (br s, 1 H), 5.51 (m, 1 H), 4.87 (m, 1 H), 4.50 (m, 1 H), 4.34 (m, 1 H), 4.04-3.90 (m, 2H), 3.51 (s, 3H), 1 .99 (m, 1 H), 1 .77 (s, 3H), 0.96-0.84 (m, 6H). LCMs (m/z) ES+ 285 (m+1 ). Purity 84%.

Scheme for Intermediate I

Intermediate la

Intermediate lb Intermediate I

Intermediate lb.

Trifluoroacetic acid TFA (20 ml_, 260 mmol) was added at RT to 1 -(1 ,1 - dimethylethyl) 2-methyl 4-methylidene-1 ,2-pyrrolidinedicarboxylate (4 g, 16.58 mmol) (intermediate la) in dichloromethane (160 ml_) and stirred with for 1 hour.

The volatiles were removed under reduced pressure to give methyl 4- methylideneprolinate (intermediate lb, 1 .7 g, 73%). ¹ H NMR (CHLOROFORM- d) δ 4.90 (d, J = 12.7 Hz, 2H), 3.84 (t, J = 6.9 Hz, 1 H), 3.66 - 3.76 (m, 3H), 3.63 (br. s., 1 H), 3.38 - 3.57 (m, 1 H), 2.76 (dd, J = 15.9, 7.9 Hz, 1 H), 2.53 (dd, J =

16.2, 3.9 Hz, 1 H), 2.34 (br. s., 1 H).

Intermediate I

N-[(methyloxy)carbonyl]-L-valine (1 .97 g, 1 1 .22 mmol), HATU (4.27 g, 1 1 .22 mmol) and DIEA (3.96 g, 30.6 mmol) were added to methyl 4- methylideneprolinate intermediate lb (1 .44 g, 10.20 mmol) in DMF. The mixture was stirred at room temperature overnight. After the work up, the residue was dissolved in THF and treated with lithium hydroxide (1 .22 g, 51 .0 mmol) in water for 3 hours, giving after the work up A/-[(methyloxy)carbonyl]-L-valyl-4- methylideneproline (Intermediate I), 1 .5 g, 52%. ¹H NMR (CHLOROFORM-d) δ: 1 1 .03 (br. s., 1 H), 5.87 (d, J = 9.2 Hz, 1 H), 5.09 (br. s., 1 H), 4.75 (dd, J = 9.5, 3.4 Hz, 1 H), 4.53 (d, J = 13.7 Hz, 1 H), 4.17 - 4.35 (m, 2H), 3.65 (s, 2H), 2.89 - 3.02 (m, 1 H), 2.73 - 2.84 (m, 1 H), 2.68 (s, 2H), 1 .97 - 2.1 1 (m, 1 H), 0.80 - 1 .08 (m, 6H). Example 27

methyl [(1S)-1-({(2S)-4 -dimethyl-2-[5-(4-{2-[(8S)-7-(^

{[(methyloxy)carbonyl]amino}butanoyl)-1,4-dioxa-7-azaspiro[4.4]non-8-yl]-1H- benzimidazol-5-yl}phenyl)-1H-benzimidazol-2-yl]-1-pyrrolidinyl}carbon

methylpropyl]carbamate

Synthesized from intermediates D and E using the method described in Example 21 .

HR MS for C^Hs_/NsOs (M+H)⁺ calc = 849.4299 (found 849.4300) Example 28

methyl [(1S)-1-({(2S)-4-(difluoromethylidene)-2-[5-(4-{2-[(8S^

{[(methyloxy)carbonyl]amino}butanoyl)-1,4-dioxa-7-azaspiro[4.4]non-8-yl]-1H- benzimidazol-5-yl}phenyl)-1H-benzimidazol-2-yl]-1-pyrrolidinyl}carbonyl)-2- methylpropyl]carbamate

Synthesized from intermediates D and G using the method described in Exampk 21 .

HR MS for C₄₅H5i N₈O8F2 (M+H)⁺ calc = 869.3798 (found 869.3800) Example 29

methyl [(1 S)-2-methyl-1-({(8S)-8-[5-(4-{2-[(2S)-4-methyl-1-((2S)-3-methy^ {[(methyloxy)carbonyl]amino}butanoyl)-2,5-dihydro-1H-pyrrol-2-yl]-1H- benzimidazol-5-yl}phenyl)-1H-benzimidazol-2-yl]-1,4-dioxa-7-azaspiro[4.4]non-7- yl}carbonyl)propyl]carbamate

Synthesized from intermediates D and H using the method described in Exampl 21 .

¹ H NMR (500 MHz, CD3OD) δ ppm: 7.80-7.60 (m, 10H), 5.93 (m, 1 H), 5.61 (m, 1 H), 5.31 (m, 1 H), 4.73 (m, 1 H), 4.63 (m, 1 H), 4.18 (m, 1 H), 4.17 (m, 1 H), 4.14 (m, 1 H), 4.07-3.95 (m, 4H), 3.94 (m, 1 H), 3.65 (s+s, 6H), 2.58 (m, 1 H), 2.54 (m, 1 H), 2.00 (m, 2H), 1 .95 (s, 3H), 0.90 (m, 3H), 0.88 (m, 6H), 0.84 (m, 3H).

HR MS for C₄₅H53N₈O8 (M+H)⁺ calc = 833.3986 (found 833.3984) Example 30

methyl [(1 S)-2-methyl-1 -({(8S)-8-[5-(4-{2-[(6S)-5-((2S)-3-methyl-2- {[(methyloxy)carbonyl]amino}butanoyl)-5-azaspiro[2.4]hept-6-yl]-1H- benzimidazol-5-yl}phenyl)-1H-benzimidazol-2-yl]-1,4-dioxa-7-azaspiro[4.4]non-7-

Synthesized from intermediates D and F using the method described in Example 21 .

HR MS for C^HssNsOs (M+H)⁺ calc = 847.4143 (found 847.4147) Example 31

methyl [(1S)-2-methyl-1-({(2S)-4-methylidene-2-[5-(4-{2-[(8S)^

{[(methyloxy)carbonyl]amino}butanoyl)-1,4-dioxa-7-azaspiro[4.4]non-8-yl]-1H- benzimidazol-5-yl}phenyl)-1 H-benzimidazol-2-yl]- 1 -

Synthesized from intermediates D and I using the method described in Example 21 .

¹ H NMR (500 MHz, CD3OD) δ ppm: 7.79-7.54 (m, 10H), 5.51 (m, 1 H), 5.31 (m, 1 H), 5.23 (m, 1 H), 5.18 (m, 1 H), 4.73 (m, 1 H), 4.59 (m, 1 H), 4.20 (m, 1 H), 4.06- 3.97 (m, 4H), 4.14 (m, 1 H), 3.93 (m, 1 H), 4.13 (m, 1 H), 3.65 (s+s, 6H), 3.18 (m, 1 H), 2.90 (m, 1 H), 2.57 (m, 1 H), 2.54 (m, 1 H), 2.05 (m, 1 H), 2.03 (m, 1 H), 0.90- 0.84 (m, 12H).

HR MS for C₄₅H53N₈O8 (M+H)⁺ calc = 833.3986 (found 833.3986) Example 32

Dimethyl (benzene-1,4-diylbis{1H-benzimidazole-5,2-diyl(2S,3aS, 7aS)octahydro- 1 H-indole-2, 1 -diyl[(2S)-3-methyl-1 -oxo-1 ,2-butanediyl]})biscarbamate

To a solution of (2S,3aS,7aS)-1 -{N-[(methyloxy)carbonyl]-L-valyl}octahydro-1 H- indole-2-carboxylic acid, (144 mg, 0.44 mmol) DIEA (0.315 ml_) and HATU (176 mg, 0.462 mmol) in Ν,Ν-Dimethylformamide (DMF) (0.88 ml_) stirred at room temp was added solid 1 ,1 ':4',1 "-terphenyl-3,3",4,4"-tetramine tetra hydrochloride intermediate 5 (96 mg, 0.22 mmol). The reaction was allowed to stir at room temperature for ~ 2.5 hours. The reaction mixture was partitioned between

EtOAc and NaHCO3 solution. The organic layer was washed with brine, dried with MgSO4, filtered and concentrated. The residue was chromatographed on a

40 g silica gel column eluted with 0-7% MeOH in DCM. Fractions containing the product were combined and concentrated to yield a brown oil. The residue was dissolved in acetic acid (1 .92 ml_) and heated in a 70°C oil bath for 1 .5 hours. Reaction mixture concentrated and the residue was purified by reverse phase HPLC. Fractions containing the product were freeze-dried to yield the dimethyl (benzene-1 ,4-diylbis{1 H-benzimidazole-5,2-diyl(2S,3aS,7aS)octahydro-1 H- indole-2,1 -diyl[(2S)-3-methyl-1 -oxo-1 ,2-butanediyl]})biscarbamate (42 mg, 22%) as a white solid .

¹ H NMR (400 MHz, DMSO-c/₆) δ ppm 1 1 .98 - 12.44 (m, 2 H) 7.87 (d, J=4 Hz, 1 H) 7.69 - 7.82 (m, 5 H) 7.61 (br. s., 1 H) 7.41 - 7.58 (m, 5 H), 4.93 - 5.27 (m, 2 H) 4.44 (br. s., 2 H) 3.84 (t, J=9 Hz, 2 H) 3.55 (s, 6 H) 2.41 (br. s., 4 H) 2.1 1 - 2.27 (m, 2 H) 1 .59 - 2.07 (m, 12 H) 1 .08 - 1 .56(m, 6 H) 0.54 - 1 .00 (m, 12 H).

ES-LCMS m/z 871 (M+H), Purity 100%

Example 33

Dimethyl (benzene- 1, 4-diylbis{1 H-benzimidazole-5, 2-diyl( 8S)-1,4-dithia-7- azaspiro[4.4]nonane-8, 7-diyl[(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

Methyl (8S)-7-{/V-[(methyloxy)carbonyl]-L-valyl}-1 ,4-dithia-7-azaspiro[4.4]nonane- 8-carboxylate

To a stirred solution of methyl N-[(methyloxy)carbonyl]-L-valyl-4-oxo-L-prolinate (1 .0g, 3.33 mmol) in anhydrous DCM (68 mL) was added 1 ,2-ethanedithiol (0.56 mL, 6.66 mmol) and followed by addition of born trifluoride diethyl etherate (0.164 mL) . The resulting mixture was stirred overnight at room temperature before quenched with a saturated aqueous solution of NaHCO3. The layers were separated and the organic phase was dried, filtered and evaporated. The residue was chromatographed on silica gel column eluted with 0 to 70 %

EtOAc/Hexanes. Fractions containing the product were combined and

concentrated to yield methyl (8S)-7-{N-[(methyloxy)carbonyl]-L-valyl}-1 ,4-dithia-7- azaspiro[4.4]nonane-8-carboxylate (0.637 g, 51 % yield) as an oil.

1 H NMR (400 MHz, DMSO-c/6) δ ppm 7.40 (d, J=9 Hz, 1 H) 4.38 (t, J=8 Hz, 1 H) 4.29 (d, J=1 1 Hz, 1 H) 3.85 - 4.07 (m, 2 H) 3.62 (s, 3 H), 3.52 (s, 3 H) 3.38 (d, J=4 Hz, 4 H) 2.65 - 2.76 (m, 1 H) 2.31 - 2.43 (m, 1 H) 1 .88 - 1 .98 (m, 1 H) 0.77- 0 98 (m, 6 H) (8S)-7-{/V-[(methyloxy)carbonyl]-L-valyl}-1 ,4-dithia-7-azaspiro[4.4]nonane-8- carboxylic acid

Methyl (8S)-7-{N-[(methyloxy)carbonyl]-L-valyl}-1 ,4-dithia-7-azaspiro[4.4]nonane-

8-carboxylate was dissolved in a mixture of tetrahydrofuran (4.5 ml_), tert- butanol (1 .125 mL) and water (1 .125 ml_). The solution was cooled in an ice/water bath and allowed to cool before the addition of lithium hydroxide hydrate (0.106 g, 2.52 mmol). The mixture was stirred at room temperature for 2 hours. Quenched with 1 N HCI (2.6 mL), then partitioned between EtOAc and brine. The organic phase was washed two more times with brine, dried (Na2SO4) filtered and concentrated to yield (8S)-7-{N-[(methyloxy)carbonyl]-L-valyl}-1 ,4- dithia-7-azaspiro[4.4]nonane-8-carboxylic acid (585 mg, 86% yield) as a tan foam.

¹ H NMR (400 MHz, DMSO-c/6) δ ppm 12.65 (br. s., 1 H) 7.38 (d, J=8 Hz, 1 H) 4.28 (d, J=8 Hz, 2 H) 3.93 - 4.1 1 (m, 1 H) 3.88 (d, J=1 1 Hz, 1 H) 3.52 (s, 3 H) 3.35 - 3.45 (m, 4 H) 2.69 (dd, J=13, 8 Hz, 1 H) 2.34 (dd, J=13, 8 Hz, 1 H) 1 .87 - 1 .97 (m, 1 H) 0.80 - 0.96 (m, 6 H)

ES-LCMS m/z 363 (M+H), Purity 99%

Dimethyl (benzene- 1, 4-diylbis{1 H-benzimidazole-5, 2-diyl( 8S)-1,4-dithia-7- azaspiro[4.4]nonane-8, 7-diyl[(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

This compound was made according to the general procedure described for Example 32.

¹ H NMR (400 MHz, DMSO-c/6) δ ppm 12.35 (br. s., 2 H) 7.84 (br s, 1 H) 7.70 - 7.80 (m, 5 H) 7.61 (d, J = 8 Hz, 1 H) 7.45 - 7.58 (m, 3 H) 7.33 (d, J=8 Hz, 2 H) 5.17 (t, J = 8 Hz, 2 H) 4.44 (d, J = 12 Hz, 2 H) 3.99 - 4.15 (m, 4 H) 3.54 (s, 6 H) 3.36 - 3.50 (m, 8 H) 2.14 - 2.29 (m, 4 H) 1 .76 - 2.02 (m, 2 H) 0.74 - 0.88 (m, 12 H)

HR MS for C₄₆H55N₈O6S₄ (M + H)⁺ calc: 943.3127, found: 943.3127

ES-LCMS m/z 943 (M+H), Purity 100% Example 34

Dimethyl (benzene-1 ,4-diylbis{1 H-benzimidazole-5, 2-diyl[(2S, 4R)-4-hydroxy-2, 1 - pyrrolidinediyl][(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

/V-[(methyloxy)carbonyl]-L-valyl-(4R)-4-hyclroxy-L-proline

/V-[(methyloxy)carbonyl]-L-valyl-(4R)-4-hyclroxy-L-proline was synthesized according to the general procedure used for (8S)-7-{/V-[(methyloxy)carbonyl]-L- valyl}-1 ,4-dithia-7-azaspiro[4.4]nonane-8-carboxylic acid using methyl N- [(methyloxy)carbonyl]-L-valyl-(4R)-4-hydroxy-L-prolinate in place of methyl (8S)- 7-{N-[(methyloxy)carbonyl]-L-valyl}-1 ,4-dithia-7-azaspiro[4.4]nonane-8- carboxylate to yield Methyl (8S)-7-{N-[(methyloxy)carbonyl]-L-valyl}-1 ,4-dithia-7- azaspiro[4.4]nonane-8-carboxylate (1 .586 g, 75% yield as 0.2 EtOAc)

1 H NMR (400 MHz, DMSO-c/6) δ ppm 12.43 (br. s., 1 H) 7.29 (d, J=9 Hz, 1 H) 5.17 (d, J=4 Hz, 1 H) 4.33 (d, J = 3, 1 H) 4.24 (t, J = 8 Hz, 1 H), 3.98 - 4.07 (m, 1 H) 3.64 (d, J = 3, 2H) 3.52 (s, 3H), 2.03 - 2.15 (m, 1 H) 1 .82 - 1 .97 (m, 2H) 0.72 - 0.98 (m, 6 H)

ES-LCMS m/z 289 (M+H), Purity 96%

Dimethyl (benzene-1,4-diylbis{1H-benzimidazole-5,2-diyl[(2S,4R)-4-hydroxy-2, 1- pyrrolidinediyl][(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

This compound was made according to the general procedure described for Example 32. ¹ H NMR (400 MHz, DMSO-c/6) δ ppm 12.35 (br. s., 2 H) 8.34 (s, 1 H) 7.75 (s, 5 H) 7.49 (br. s., 4 H) 7.22 (d, J=9 Hz, 2 H) 5.00 - 5.55 (m, 4 H) 4.56 (br. s., 2 H) 4.01 - 4.23 (m, 2 H) 3.84 - 4.02 (m, 2 H) 3.64 - 3.82 (m, 2 H) 3.55 (s, 6 H) 2.14 - 2.29 (m, 4 H) 1 .76 - 2.02 (m, 2 H) 0.56 - 1 .05 (m,12 H)

HR MS for C₄₂H5i N₈O8 (M + H)⁺ calc: 795.3830, found: 795.3830

Example 35

Dimethyl (benzene-1 ,4-diylbis{(4-fluoro-1 H-benzimidazole-6, 2-diyl)(8S)-1, 4- dioxa-7-azaspiro[4.4]nonane-8, 7-diyl[(2S)-3-methyl-1 -oxo-1 ,2- butanediyl]})biscarbamate

Methyl ((1 S)-1 -{[(8S)-8-(5-bromo-7-fluoro-1 H-benzimidazol-2-yl)-1 ,4-d

azaspiro[4.4]non-7-yl]carbonyl}-2-methylpropyl)carbannate

(8S)-7-{N-[(methyloxy)carbonyl]-L-valyl}-1 ,4-dioxa-7-azaspiro[4.4]nonane-8- carboxylic acid (403 mg, 1 .22 mmol) and 5-bromo-2,3-diaminofluorobenzene (250 mg, 1 .219 mmol) were combined in dichloromethane (40 mL). DIEA (0.426 mL, 2.439 mmol) was added and the reaction stirred for 10 min. HATU (464 mg, 1 .219 mmol) was then added. The reaction was stirred at room temperature overnight. Water (50 mL) was added and the reaction stirred for 20 minutes. The solution was filtered through a hydrophobic frit. Organic layer was concentrated. The residue dissolved in acetic acid (20.0 mL) heated to 70 °C, stirred for 1 hour then concentrated. The residue then was partitioned between EtOAc and saturated aqueous NaHCO3 solution. The organic layer was dried with MgSO^ filtered and concentrated. The residue was chromatographed on 40g silica gel column (0-100% EtOAc in Hex) to afford methyl ((1 S)-1 -{[(8S)-8-(5-bromo-7- fluoro-1 H-benzimidazol-2-yl)-1 ,4-dioxa-7-azaspiro[4.4]non-7-yl]carbonyl}-2- methylpropyl)carbamate ( 457 mg, 70%) as a tan solid.

¹ H NMR (400 MHz, DMSO-c/6) δ ppm 12.71 (br. s., 1 H) 7.52 (br. s., 1 H) 7.33 (d, J=8 Hz, 1 H) 7.21 (d, J=10 Hz, 1 H) 5.15 (t, J=8 Hz, 1 H) 3.84 - 4.25 (m, 6 H) 3.78 (d, J=10 Hz, 1 H) 3.54 (s, 3 H) 3.20 (s, 1 H) 2.22 - 2.44 (m, 1 H) 1 .81 - 1 .97 (m, 1 H) 0.71 - 0.94 (m, 6 H)

ES-LCMS m/z 500 (M+H), Purity 93% Dimethyl (benzene-1 ,4-diylbis{(4-fluoro-1 H-benzimidazole-6,2-diyl)(8S)-1 ,4- dioxa-7-azaspiro[4.4]nonane-8, 7-diyl[(2S)-3-methyl-1 -oxo-1 ,2- butanediyl]})biscarbamate

A mixture of benzene-1 ,4-diyldiboronic acid (75 mg, 0.45 mmol), methyl ((1 S)-1 - {[(8S)-8-(5-bromo-7-fluoro-1 H-benzimidazol-2-yl)-1 ,4-dioxa-7-azaspiro[4.4]non-7- yl]carbonyl}-2-methylpropyl)carbamate (453 mg, 0.91 mmol), and potassium carbonate (188 mg, 1 .36 mmol) in DME and water was degassed with nitrogen before the addition of PdCI₂(dppf)-CH₂CI₂ adduct (74 mg, 0.09 mmol). The solution was then heated at 100°C for three hours. The solution was then concentrated and the residue dissolved in EtOAc and filtered through celite. The filtrate was concentrated. The residue was purified first by chromatography on silica gel eluted with 0 - 10 % MeOH in DCM. A portion of the sample was then purified by reverse phase chromatography to yield dimethyl (benzene-1 , 4- diylbis{(4-fluoro-1 H-benzimidazole-6,2-diyl)(8S)-1 ,4-dioxa-7-azaspiro[4.4]nonane- 8,7-diyl[(2S)-3-methyl-1 -oxo-1 ,2-butanediyl]})biscarbamate (24 mg, 6% yield).

¹ H NMR (400 MHz, DMSO-c/6) δ ppm 12.63 -12.86 m., 2 H) 8.13 (br. s., 1 H) 7.79 (m, 5 H) 7.60 (s, 2 H) 7.28 - 7.43 (m, 4 H) 5.17 (s, 2 H) 3.73 - 4.19 (m, 14 H) 3.54 (s, 6 H) 2.37 - 2.56 (m, 4 H overlapping DMSO) 1 .81 - 2.03 (m, 2 H) 0.66 - 1 .00 (m, 12 H)

ES-LCMS m/z 915 (M+H), Purity 100% HR MS for C₄₆H53N₈Oio (M + H)⁺ calc: 915.3853, found: 915.3845

Example 36

Dimethyl (benzene-1 ,4-diylbis{1H-benzimidazole-5,2-diyl(3S)-8-oxa-2- azaspiro[4.5]decane-3,2-diyl[(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

This compound was made according to the procedure used in Example 32 using

(3S)-2-{N-[(methyloxy)carbonyl]-L-valyl}-8-oxa-2-azaspiro[4.5]decane-3- carboxylic.

1 H NMR (400 MHz, DMSO-c/₆) δ ppm 12.25 - 12.39 (m, 2 H) 7.82 (br. s., 1 H) 7.70 - 7.77 (m, 5 H) 7.59 (d, J=8 Hz, 1 H) 7.43 - 7.56 (m, 3 H) 7.35 (d, J=8 Hz, 2 H) 5.1 1 (t, J=8 Hz, 2 H) 4.21 (d, J=10 Hz, 2 H) 4.06 (t, J=8 Hz, 2 H) 3.68 (br. s., 4 H) 3.47 - 3.65 (m, 12 H) 2.28 - 2.43 (m, 2 H) 2.02 (dd, J=12, 10 Hz, 2 H) 1 .88 (d, J=6 Hz, 2 H) 1 .65 (br. s., 4 H) 1 .50 (br. s., 4 H) 0.84 (t, J=6 Hz, 12 H)

ES-LCMS m/z 903 (M+H), Purity 100%

HR MS for CsoHesNsOs (M + H)⁺ calc: 903.4769, found: 903.4772

Example 37

Bis(l-methylethyl) (benzene-1, 4-diylbis{1H-benzimidazole-5,2-diyl(2S)-2, 1- pyrrolidinediyl[(2S)-3-methyl-1 -oxo-1, 2-butanediyl]})biscarbamate

To a solution of N-{[(1 -methylethyl)oxy]carbonyl}-L-valine (83 mg, 0.336 mmol), DIEA (0.182 mL, 1 .043) and HATU (128 mg, 0.336 mmol) in DMF (1 .3 mL) was added 5,5'-benzene-1 ,4-diylbis{2-[(2S)-2-pyrrolidinyl]-1 H-benzimidazole} intermediate 23 (100 mg, 0.168 mmol) and reaction allowed to proceed at RT for 3 hours. Following purification by RP-HPLC bis(l -methylethyl) (benzene-1 ,4- diylbis{1 H-benzimidazole-5,2-diyl(2S)-2,1 -pyrrolidinediyl[(2S)-3-methyl-1 -oxo-1 ,2- butanediyl]})biscarbamate (64 mg, 46%) was obtained as a white solid.

¹ H NMR (400 MHz, DMSO-c/₆) δ ppm 12.09 - 12.60 (m, 2 H) 7.83 (br. s., 1 H)

7.70 - 7.80 (m, 5 H) 7.43 - 7.66 (m, 4 H) 7.13 (d, J=8 Hz, 2 H) 5.1 1 - 5.29 (m, 2 H)

4.71 - 4.83 (m, 2 H) 4.09 (t, J=8 Hz, 2 H) 3.79 - 3.92 (m, 4 H) 2.17 - 2.35 (m, 4 H) 1 .85 - 2.16 (m, 6 H) 1 .18 (d, J=6 Hz, 12 H) 0.80 - 0.94 (m, 12 H).

HR MS for C₄₆H59N₈O6 (M + H)⁺ calc: 819.4558, found: 819.4555

Scheme IX

bis( -dimethylethyl) (4-bromobenzene-1 ,2-diyl)biscarbamate (IX-1 )

A solution of 4-bromo-1 ,2-benzenediamine (64 g, 342 mmol) and bis(1 ,1 - dimethylethyl) dicarbonate (318 mL, 1369 mmol) in CHCI₃ (1000 mL) was stirred at room temperature for 6 days. The resulting mixture was washed with a sat. NaHCO₃ solution (300 mL), a sat. NaCI solution (500 mL), dried (Na₂SO₄) and concentrated. The residue was triturated with hexane to obtain bis(1 ,1 - dimethylethyl) (4-bromobenzene-1 ,2-diyl)biscarbamate (1 13 g, 85%) as a white solid: ¹H NMR (400 MHz, DMSO-c/₆) δ ppm 8.66 (br. s., 1 H) 8.62 (br. s., 1 H) 7.73 (s, 1 H) 7.44 (d, J=8.6 Hz, 1 H) 7.23 (dd, J=8.8, 2.3 Hz, 1 H) 1 .47 (s, 9 H) 1 .46 (s, 9 H); ES LC-MS m/z =387.1 (Br⁷⁹, M+H)⁺; ES LC-MS m/z =389.1 (Br⁸¹, M+H)⁺. bis(1 ,1 -dimethylethyl) [4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)ben

1 ,2- iyl]biscarbamate (IX-2)

A degassed slurry of bis(1 ,1 -dimethylethyl) (4-bromobenzene-1 ,2- diyl)biscarbamate (30.27 g, 78 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi-1 ,3,2- dioxaborolane (21 .83 g, 86 mmol), PdCI₂(dppf)-CH₂CI₂ adduct (1 .277 g, 1 .563 mmol) and potassium acetate (19.18 g, 195 mmol) in anh. 1 ,4-dioxane (500 mL) was heated at the reflux temperature for 2 h. The resulting mixture was allowed to cool to room temperature. Na₂SO₄ was added and the mixture was filtered through a pad of Celite with the aid of EtOAc. The filtrate was concentrated, the residue dissolved in a minimal amount of CH₂CI₂ and filtered through a plug of silica gel with the aid of 40% EtOAc in hexane. The filtrate was concentrated to afford bis(1 ,1 -dimethylethyl) [4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)benzene-1 ,2-diyl]biscarbamate (41 .1g, 80% pure, 97% yield) as a white foamy solid: ¹H NMR (400 MHz, DMSO-c/₆) δ ppm 8.60 (s, 1 H) 8.54 (br. s., 1 H) 7.77 (s, 1 H) 7.59 (d, J=8.0 Hz, 1 H) 7.36 (d, J=8.6 Hz, 1 H) 1 .47 (s, 18 H) 1 .28 (s, 12 H) ; ES LC-MS m/z =435.2 (M+H)⁺. bis( -dimethylethyl) (4'-bromo-3,4-biphenyldiyl)biscarbamate (IX-3)

A degassed mixture of 1 -bromo-4-iodobenzene (5.58 g, 19.71 mmol), bis(1 ,1 - dimethylethyl) [4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)benzene-1 ,2- diyl]biscarbamate (5.35 g, 9.85 mmol) tetrakis(triphenylphosphine)palladium(0) (0.569 g, 0.493 mmol) and sodium carbonate (20.89 g, 197 mmol) in water (300 mL) and DME (300 mL) was heated at 60 °C for 22 h, then at the reflux

temperature for 1 h. The resulting mixture was allowed to cool to room

temperature and diluted with CH₂CI₂ (500 mL) and water (500 mL). The organic layer was washed with a sat. NaCI solution (300 mL), dried (Na₂SO₄) and concentrated. The residue was dissolved in CH₂CI₂, concentrated onto Celite and purified by column chromatography (silica gel, 0 to 20% EtOAc in hexane) to obtain bis(1 ,1 -dimethylethyl) (4'-bromo-3,4-biphenyldiyl)biscarbamate (3.51 g, 77%) as a white foamy solid: ¹ H NMR (400 MHz, DMSO-c/₆) δ ppm 8.63 (br. s., 2 H) 7.78 (s, 1 H) 7.62 - 7.68 (m, 2 H) 7.50 - 7.62 (m, 3 H) 7.38 (dd, J=8.6, 2.1 Hz, 1 H) 1 .49 (s, 18 H); ES LC-MS m/z =463.1 (Br⁷⁹, M+H)⁺; ES LC-MS m/z =465.1 (Br⁸¹ , M+H)⁺.

bis(phenylmethyl) (4-bromobenzene-1 ,2-diyl)biscarbamate (X-1 )

A deep brown slurry of 4-bromo-1 ,2-benzenediamine (18.27 g, 98 mmol) and K2CO3 (33.8 g, 244 mmol) was placed in an ice-bath for a few minutes. Benzyl chloroformate (30.2 mL, 215 mmol) was added and the ice-bath was removed. After 1 h more benzyl chloroformate (3 mL, 0.2 eq.) was added. After 1 h more K₂CO₃ (3.2 g, 0.25 eq.) was added. After 30 min the slurry was filtered through a pad of Celite. The filtrate was taken up into EtOAc (200 mL), washed with a 1 N HCI solution (200 mL), a sat NaHCO₃ solution (2x200 mL), a sat. NaCI solution (100 mL), dried (Na₂SO₄) and concentrated to a thick dark oil. CH2CI2 and hexane were added and the mixture was concentrated to a wet solid. Hexane (200 mL) was added and the resulting slurry was filtered, solid washed with hexane to afford bis(phenylmethyl) (4-bromobenzene-1 ,2-diyl)biscarbamate (23.46 g, 53%) as a light brown solid: ¹H NMR (400 MHz, DMSO-c/₆) δ ppm 9.13 (br. s., 1 H) 9.04 (br. s., 1 H) 7.79 (s, 1 H) 7.50 (d, J=8.6 Hz, 1 H) 7.24 - 7.46 (m, 1 1 H) 5.15 (s, 2 H) 5.13 (s, 2 H); ES LC-MS m/z =455.7 (Br⁷⁹, M+H)⁺; ES LC-MS m/z =457.1 (Br⁸¹ , M+H)⁺. bis(phenylmethyl) [4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)benzene-1 ,2- diyl]biscarbamate (X-2)

A degassed slurry of bis(phenylmethyl) (4-bromobenzene-1 ,2-diyl)biscarbamate (18.19 g, 40.0 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi-1 ,3,2-dioxaborolane (1 1 .16 g, 43.9 mmol), PdCI₂(dppf)-CH₂CI₂ adduct (0.653 g, 0.799 mmol) and potassium acetate (9.80 g, 100 mmol) in anh. 1 ,4-dioxane (400 mL) was heated at the reflux temperature for 1 .5 h. The resulting mixture was allowed to cool to rt. Solid Na₂SO₄ was added and the mixture was filtered through a pad of Celite with the aid of EtOAc. The filtrate was concentrated, the residue was reevaporated twice from toluene. The residue was dissolved in CH₂CI₂/hexane, concentrated onto Celite and purified by column chromatography (silica gel, 0 to 30% EtOAc in hexane) to obtain bis(phenylmethyl) [4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)benzene-1 ,2-diyl]biscarbamate (12.4 g, 62%) as a white solid: ¹H NMR (400 MHz, DMSO-c/e) δ ppm 9.07 (s, 1 H) 8.96 (br. s., 1 H) 7.81 (s, 1 H) 7.67 (d, J=8.0 Hz, 1 H) 7.28 - 7.48 (m, 1 1 H) 5.15 (d, J=4.3 Hz, 4 H) 1 .29 (s, 12 H); ES LC-MS m/z =503.2 (M+H)⁺.

Scheme XI

bis(1 ,1 -dimethylethyl) bis(phenylmethyl) 1 ,1 ':4',1 "-terphenyl-3,3",4,4"- tetrayltetrakiscarbamate (XI-1 )

A degassed slurry of bis(phenylmethyl) [4-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)benzene-1 ,2-diyl]biscarbamate (3.81 g, 7.58 mmol), bis(1 ,1 - dimethylethyl) (4'-bromo-3,4-biphenyldiyl)biscarbamate (3.51 g, 7.58 mmol), PdCI₂(dppf)-CH₂Cl₂ adduct (0.124 g, 0.152 mmol) and sodium carbonate (16.06 g, 152 mmol) in DME (100 ml_) and water (100 ml_) was heated at 60 °C. After 20 min the resulting thick brown slurry was filtered hot through paper filter. The solid was washed with water and MeOH and dried in vacuum to afford bis(1 ,1 - dimethylethyl) bis(phenylmethyl) 1 ,1 ':4',1 "-terphenyl-3,3",4,4"- tetrayltetrakiscarbamate (4.90 g, 85%) as a grey solid: ¹H NMR (400 MHz, DMSO-c/e) δ ppm 9.06 (br. s., 2 H) 8.63 (d, J=10.5 Hz, 2 H) 7.77 - 7.98 (m, 2 H) 7.55 - 7.78 (m, 6 H) 7.26 - 7.55 (m, 1 1 H) 5.17 (s, 4 H) 1 .49 (s, 18 H). bis(1 ,1 -dimethylethyl) (3",4"-diamino-1 ,1 ':4',1 "-terphenyl-3,4-diyl)biscarbamate (XI-2

10% palladium on carbon (200 mg, 1 .879 mmol) was placed under N₂

atmosphere, THF (100 ml_) was added, followed by bis(1 ,1 -dimethylethyl) bis(phenylmethyl) 1 ,1 ':4',1 "-terphenyl-3,3",4,4"-tetrayltetrakiscarbamate (2 g, 2.64 mmol). The resulting solution was diluted with MeOH (50 ml_). Placed under 60 psi H₂ pressure and heated at 80 °C for 3 h. A slurry of 50 mg catalyst in MeOH (2 ml_) was then added and the reaction mixture was placed under 60 psi H₂ pressure and heated at 80 °C for 5 h. The resulting mixture was filtered through a pad of Celite and. filtrate was concentrated to obtain bis(1 ,1 -dimethylethyl) (3",4^M- diamino-1 ,1 ':4',1 "-terphenyl-3,4-diyl)biscarbamate (1 .29 g, 100%) as a white solid: ¹H NMR (400 MHz, DMSO-c/₆) δ ppm 8.60 (br. s., 1 H) 7.80 (br. s., 1 H) 7.51 - 7.64 (m, 4 H) 7.39 (dd, J=8.5, 1 .9 Hz, 1 H) 6.85 - 6.93 (m, 1 H) 6.78 (dd, J=8.0, 1 .8 Hz, 1 H) 6.58 (d, J=8.0 Hz, 1 H) 4.65 (s, 1 H) 4.57 (s, 1 H) 1 .55-1 .43 (m, 18 H, apparent 2 singlets); ES LC-MS m/z =491 .3 (M+H)⁺.

Scheme XII

Example 38

methyl ('ίS -2-me^ /- ('{('2S,3aS,7aS -2- 5- 4-{2- ('8S -7-('('2Sj-3-me^ /-2- {[(methyloxy)carbonyl]amino}butanoyl)-1,4-dioxa-7-azaspiro[4.4]non-8-yl]-1H- benzimidazol-5-yl}phenyl)-1 H-benzimidazol-2-yl]octahydro- 1 H-indol- 1 - yl}carbonyl)propyl]carbamate

A mixture of bis(1 ,1 -dimethylethyl) (3",4"-diamino-1 ,1 ':4',1 "-terphenyl-3,4- diyl)biscarbamate (XI-2) (1310 mg, 2.67 mmol), (8S)-7-{N-[(methyloxy)carbonyl]- L-valyl}-1 ,4-dioxa-7-azaspiro[4.4]nonane-8-carboxylic acid (926 mg, 2.80 mmol) and DIEA (0.933 mL, 5.34 mmol) in DMF (30 mL) was treated with HATU (1 1 17 mg, 2.94 mmol) at 0 °C. The reaction mixture was allowed to warm slowly to room temperature. After 2.5 h water (100 mL) was added. The resulting slurry was filtered. The solids were washed with water and hexane, then taken up into CH2CI2 (50 mL). The layers were separated, the organic layer was dried

(Na2SO₄) and concentrated down to about 15 mL. The resulting mixture was purified by column chromatography (silica gel, 0-100% EtOAc/hexane). The resulting regioisomeric mixture (XII-1 ) was taken up into a 4N HCI solution in dioxane (12 mL). The resulting orange-brown solution was allowed to sit. After 10 min Et₂O (100 mL) was added and the mixture was sonicated and filtered. The resulting regioisomeric mixture (XII-2) was dissolved in in DMF (2 mL), cooled to 0 °C and treated with (2S,3aS,7aS)-1 -{N-[(methyloxy)carbonyl]-L-valyl}octahydro- 1 H-indole-2-carboxylic acid (60.5 mg, 0.185 mmol), DIEA (0.441 mL, 2.53 mmol) and HATU (77 mg, 0.202 mmol). After 3 h water (50 mL) was added and the resulting slurry was filtered. The resulting solid (XII-3) was dissolved in AcOH (2 mL) and heated at 80 °C for 1 .5 h. The reaction mixture was allowed to cool to room temperature and concentrated. The residue was purified by HPLC (5-70% H₂O in CH₃CN, both containing 0.1 % formic acid) to obtain methyl [(1 S)-2-methyl- 1 -({(2S,3aS,7aS)-2-[5-(4-{2-[(8S)-7-((2S)-3-methyl-2- {[(methyloxy)carbonyl]amino}butanoyl)-1 ,4-dioxa-7-azaspiro[4.4]non-8-yl]-1 H- benzimidazol-5-yl}phenyl)-1 H-benzimidazol-2-yl]octahydro-1 H-indol-1 - yl}carbonyl)propyl]carbamate (39 mg) as a white solid: ¹H NMR (400 MHz, DMSO-c/e) δ ppm 12.14 - 12.30 (m, 2 H, apparent 2 broad singlets) 7.68 - 8.35 (m, 6 H) 7.53 (d, J=8.4 Hz, 5 H) 7.33 (d, 1 H) 5.18 (t, J=8.3 Hz, 1 H) 5.10 (t, J=8.1 Hz, 1 H) 4.44 (br. s., 1 H) 3.74 - 4.17 (m, 7 H) 3.06 - 3.67 (m, 9 H) 1 .06 - 2.55 (m, 13 H) 0.64 - 0.90 (m, 12 H); HRMS for

(M + H)⁺ calc: 875.4456, found: 875.4453; 100% pure

Example 39

methyl [(1S)-1-({(2S)-4A-difluoro-2-[5-(4-{2-[(8S)-7-((2S)-3-meth

{[(methyloxy)carbonyl]amino}butanoyl)-1,4-dioxa-7-azaspiro[4.4]non-8-yl]-1H- benzimidazol-5-yl}phenyl)-1H-benzimidazol-2-yl]-1^yrrolidi

methylpropyl]carbamate

A mixture of methyl {(1 S)-2-methyl-1 -[((8S)-8-{[(3,3",4"-triamino-1 ,1 ':4',1 "- terphenyl-4-yl)amino]carbonyl}-1 ,4-dioxa-7-azaspiro[4.4]non-7- yl)carbonyl]propyl}carbamate XII-2 (120 mg, 0.169 mmol), N- [(methyloxy)carbonyl]-L-valyl-4,4-difluoro-L-proline (57.1 mg, 0.185 mmol) and DIEA (0.441 mL, 2.53 mmol) in DMF (2 mL) was treated with HATU (77 mg, 0.202 mmol) at 0 °C. After 3 h water (50 mL) was added. The resulting slurry was filtered. The solids were dissolved in AcOH (2 mL) and heated at 80 °C for 1 .5 h. The resulting mixture was concentrated and purified by HPLC (5-70% H2O/CH3CN, both containing 0.1 % formic acid) to obtain methyl [(1 S)-1 -({(2S)- 4,4-difluoro-2-[5-(4-{2-[(8S)-7-((2S)-3-methyl-2-

{[(methyloxy)carbonyl]amino}butanoyl)-1 ,4-dioxa-7-azaspiro[4.4]non-8-yl]-1 H- benzimidazol-5-yl}phenyl)-1 H-benzimidazol-2-yl]-1 -pyrrolidinyl}carbonyl)-2- methylpropyl]carbamate (29 mg, 21 % for 2 steps) as a white solid: ¹H NMR (400 MHz, DMSO-c/e) δ ppm 7.63 - 8.22 (m, 10 H) 7.53 (d, J=8.0 Hz, 1 H) 7.32 - 7.46 (m, 1 H) 5.46 (t, J=8.1 Hz, 1 H) 5.28 (t, J=8.5 Hz, 1 H) 4.50 - 4.66 (m, 1 H) 4.28 - 4.50 (m, 1 H) 3.89 - 4.22 (m, 7 H) 3.18 - 3.84 (m, 9 H) 3.02 - 3.16 (m, 1 H) 2.82 - 3.02 (m, 1 H) 2.53 - 2.70 (m, 2 H) 1 .82 - 2.06 (m, 2 H) 0.62 - 0.91 (m, 12 H);

HRMS for C₄₄H5i F₂N₈O8 (M + H)⁺ calc: 857.3800, found: 857.3798; 100% pure.

Protocol for testing and data analysis of compounds in the HCV replicon assay Compounds were assayed for activity against HCV using the genotype 1 a and 1 b subgenomic replicon model systems. Stable cell lines bearing the genotype 1 a and 1 b replicons were used for screening of compounds. Both replicons are bicistronic and contain the firefly luciferase gene. The ET cell line is stably transfected with RNA transcripts harboring a l₃₈₉luc-ubi-neo/NS3-37ET replicon with firefly luciferase-ubiquitin-neomycin phosphotransferase fusion protein and EMCV-IRES driven NS3-5B polyprotein containing the cell culture adaptive mutations (E1202G; T1280I; K1846T) (Krieger at al, 2001 and unpublished). The genotype 1 a replicon is a stable cell line licensed from Apath LLC, modified to contain the firefly luciferase gene. The cells were grown in DMEM, supplemented with 10% fetal calf serum, 2 mM Glutamine, Penicillin (100 IU/mL)/Streptomycin (100 g/mL), 1 x nonessential amino acids, and 250-500 g/mL G418 ("Geneticin"). They were all available through Life Technologies (Bethesda, Md.). The cells were plated at 0.5 x 10⁴ cells/well in 384 well plates containing compounds. The final concentration of compounds ranged between 0.03 pM to 50 μηη and the final DMSO concentration of 0.5-1 %.

Luciferase activity was measured 48 hours later by adding a Steady glo (Promega, Madison, Wis.). Percent inhibition of replication data was plotted relative to no compound control. Under the same condition, cytotoxicity of the compounds was determined using cell titer glo (Promega, Madison, Wis). EC50s were determined from a 10 point dose response curve using 3-4-fold serial dilution for each compound, which spans a concentration range > 1000 fold. BioAssay determines the level of inhibition for each compound by normalizing cross-talk corrected plate values against the negative (low or background, cells with no compound present) and positive (high DMSO, no cells) controls to determine Percent Inhibition:

100 ^* (1 -(Cross-talk corrected value - Compound Positive Control Mean))

DMSO Negative Control Mean - Compound Positive Control Mean

These normalized values are exported to EC50 where they are plotted against the molar compound concentrations using the standard four parameter logistic equation:

B-A

Where:

A = minimum y D= slope factor

B = maximum y x = logio compound concentration [M]

C = logi₀EC₅o pECso = -C

As shown below, the tested compounds tested were found to inhibit the activity of the replicon with pECso >5.

Claims

What is claimed is:

Claim 1 . A compound of Formula I;

wherein A is a phenyl, pyridyl, furanyl, or thiofuranyl ring;

each R¹ is independently H, Ci_-6alkyl, Ci_-6cycloalkyl, or phenyl:

each R² is independently H, C_halky!, Ci-3acyl, C3-6cycloacyl, or C(O)O-Ci-3alkyl, or two alkyl R²s may, together with the nitrogen atom to which they are bound, join together to form a 5 or 6-membered saturated heterocycylic ring;

each R³ is independently H, F, d_-3alkyl, or Ci_-3alkoxy, or when two R³s are alkyl and bound to adjacent carbon atoms, they may, together with the two carbon atoms to which they are bound, join to from a 5 or 6-membered cycloalkyi ring,

R³s may optionally be substituted by one or two fluorine atoms;

within each ring, one of the 3 Zs is CH and the other two are CH or N; and each R⁴ is independently H, F, or C_halky!.

A compound of Formula II

II wherein A is a phenyl, pyridyl, furanyl, or thiofuranyl ring;

each R¹ is independently H, C_halky!, Ci-₆cycloalkyl, or phenyl: each R² is independently H, C_halky!, Ci-3acyl, C3-6cycloacyl, or C(O)O-Ci-3alkyl, or two alkyl R²s may, together with the nitrogen atom to which they are bound, join together to form a 5 or 6-membered saturated heterocycylic ring;

each R³ is independently H, F, C_halky!, or

or when two R³s are alkyl and bound to adjacent carbon atoms, they may, together with the two carbon atoms to which they are bound, join to from a 5 or 6-membered cycloalkyi ring,

R³s may optionally be substituted by one or two fluorine atoms;

within each ring, one of the 3 Zs is CH and the other two are CH or N;

each R⁴ is independently H, F, or C_halky!;

each X is independently CRR, O, or S;

each n is independently 2 or 3; and

each R is independently methyl, hydrogen, or deuterium.

Claim 3. A compound of Formula III;

III

wherein A is a phenyl, pyridyl, furanyl, or thiofuranyl ring;

each R¹ is independently H, C_halky!, Ci-₆cycloalkyl, or phenyl:

each R² is independently H, C_halky!, Ci-3acyl, C3-6cycloacyl, or C(O)O-Ci-3alkyl, or two alkyl R²s may, together with the nitrogen atom to which they are bound, join together to form a 5 or 6-membered saturated heterocycylic ring;

within each ring, one of the 3 Zs is CH and the other two are CH or N;

each R⁴ is independently H, F, or C_halky!;

each X is independently CRR, O, or S;

each n is independently 2 or 3; and

each R is independently methyl, hydrogen, or deuterium. Claim 4. A compound according to any preceding claim wherein A is phenyl or pyridyl.

Claim 5. A compound according to Claim 4 wherein the groups ending in R² are located para to each other on said phenyl or said pyridyl.

Claim 6. A compound according to any preceding claim wherein A is phenyl.

Claim 7. A compound according to any preceding claim wherein, with respect to each N to which they are bound, one R² is C(O)O-Ci-3alkyl and the other R² is H .

Claim 8. A compound according to any preceding claim wherein each R¹ is i- propyl.

Claim 9. A compound according to any preceding claim wherein within each ring, two of the 3 Zs are CH and the other is CH or N.

Claim 10. A pharmaceutically acceptable salts of a compound according to any preceding claim.

Claim 1 1 . A pharmaceutical compositions comprising a compound or salt according to any of claims 1 -10.

Claim 12. A method for treating a viral infection in a human comprising

administration of a compound or salt according to any of claims 1 -10.

Claim 13. The method of Claim 12 wherein said viral infection is an HCV infection.

Claim 14. A compound or salt as defined in any of claims 1 -10 For use in the treatment of a viral infection in a human.

Claim 15. The use of a compound or salt as defined in any of claims 1 -10 In the manufacture of a medicament for use in the treatment of a viral infection in a human.