WO2019046364A1 - Inhibitors of rsv replication and applications thereof - Google Patents

Abstract

The invention provides a compound of formula (A) or a salt thereof for use in the treatment of RSV infections. The invention also relates to a method of treating viral infections with RSV using compounds of formula (A), optionally in combination with one or more other antiviral agents, as well as pharmaceutical compositions containing a compound of formula ( A) and optionally one or more other antiviral agents.

Description

INHIBITORS OF RSV REPLICATION AND APPLICATIONS THEREOF

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No. 62/551,777 filed August 29, 2017. The entirety of this application is hereby incorporated by reference for all purposes.

TECHNICAL FIELD

The present disclosure relates to benzimidazole derivatives having antiviral activity, and in particular having an inhibitory activity on the replication of Respiratory Syncytial Virus (RSV). It further relates to methods for preparing these compounds, pharmaceutical compositions comprising these compounds and uses thereof for the treatment and prevention of RSV.

BACKGROUND

Respiratory Syncytial Virus (RSV) is a single stranded, negative sense RNA orthopneumovirus first discovered in 1956. It causes upper and lower respiratory tract infections and infects 60% of infants in their first viral season. It will infect nearly all children by 2-3 years of age. Of those infected by RSV, approximately 3% will develop an infection severe enough to warrant hospitalization. However, RSV infection is not only a problem for the young; severe infections in elderly patients have been increasing in frequency.

Symptoms of RSV are similar to the common cold, and spread easily from person to person contact. While RSV may occur in any population segment, infants, the elderly and immunocompromised are especially vulnerable.

There are some drugs available for use against RSV infection. One such drug is Virazole® an aerosol form of ribavirin, a nucleoside analogue, which is used for the treatment of serious RSV infection in hospitalized children. The aerosol route of administration, the toxicity (risk of teratogenicity), the cost and the highly variable efficacy, however, limit its use. Two other drugs, RespiGam® (RSV-IG) and Synagis ® (palivizumab) are polyclonal and monoclonal antibody immunostimulants which are intended as prophylactic agents for prevention of infection. However, both are expensive thus limiting their use to high risk infants in the developed world. Furthermore, palivizumab is not effective in the treatment of established RSV infection. A newer version monoclonal antibody, motavizumab, designed as a potential replacement of palivizumab, failed to show additional benefits in recent Phase III clinical trials.

Yu et al. (Respiratory syncytial virus fusion inhibitors. Part 4: Optimization for oral bioavailability, Bioorganic and Medicinal Chemistry Letters, 17, 2007, 895-901) disclose a series of benzimidazole-based inhibitors of RSV fusion and identified BMS-433771 as a potent inhibitor of RSV in vitro that is orally bioavailable in 4 species and demonstrates antiviral activity in both the BALB/c mouse and cotton rat models of RSV infection following oral administration. The antiviral activity was determined as the reduction of the cytopathic effect induced by the Long (A) strain of virus replicating in HEp-2 human lung epithelial carcinoma cells.

Combrink et al. (Respiratory syncytial virus fusion inhibitors. Part 6: An examination of the effect of structural variation of the benzimidazol-2-one heterocycle moiety, Bioorganic and Medicinal Chemistry Letters, 17, 2007, 4784-4790) reported on the effect of structural variation of the RSV fusion inhibitors related to BMS-433771, with the introduction of an amino-methyl substituent at the 5 position of the benzimidazole moiety. They found that the introduction of a more polar nitrile-containing side chain reduced potency and may reflect an observed trend in which the combination of polar moieties at two sites of the pharmacophore lead to reduced potency.

PCT application no. WO 2001/095910 discloses imidazopyridine and imidazopyrimidine antiviral agents with antiviral activity showing a wide range of antiviral activity from 0.001 μΜ to as high as 50 μΜ, which is not a particularly desired biological activity. Another PCT publication no. WO 2013/068769 discloses pharmaceutical compounds with an isopentyl side chain, having an EC50 of 80 μΜ or lower, without mentioning any specific values or particularly active compounds. PCT publication no. WO 2012/080446 discloses halogenated derivatives of benzimidazole compounds. Also see PCT publication nos. WO 2014/184163 and WO 2015/022263.

There is thus a major unmet medical need for safe and effective therapeutics capable of both treating active RSV infections and being administered prophylactically to prevent infection.

SUMMARY

According to one embodiment of the present disclosure there is provided a compound according to Formula A or salt thereof,

wherein R¹ is an optionally substituted alkyl and when X is N then R² is a heterocyclyl, and R³ is absent; and when X is C, then R² and R³ are linked to form a carbocyclyl and R⁴ is selected from H, alkyl, alkoxy, halogen, amino, or nitro.

Further features provide for R¹ to be an optionally substituted C₃ to C₆ alkyl, preferably an optionally substituted C₄ alkyl, and for the substituents to be selected from≡N, -S0₂(alkyl) and - OH; and for R² to be a C₃ to C₆ heterocyclyl, preferably a C₃ or C₄ heterocyclyl, and for the heteroatoms of the heterocyclyl to be selected from O, N, S or S0₂ or for R² and R³ to be linked to form a C₃-C₆ carbocyclyl, preferably a C₃ carbocyclyl and for R⁴ to be H or halogen.

Yet further features of this embodiment provide for the compound of Formula A to be selected from Formula Al or Formula A2, or salts thereof

Formula Al Formula A2 wherein n is 1, 2, 3, or 4; Y is selected from N, O, S, or SO2; and R⁵ is OH , S0₂(alkyl) or

In a preferred embodiment, the compounds of Formula A are selected from:

According to a further embodiment there is provided compounds of Formula A or salts thereof for use in the treatment of RSV infections,

In another embodiment there is provided a method of treating viral RSV infections in a subject in need thereof, the method comprising administering an effective amount of the compound of Formula A or salts thereof to the subject.

In certain embodiments, the disclosure contemplates derivatives of compounds disclosed herein such as those containing one or more, the same or different, substituents.

In still another embodiment there is provided a pharmaceutical composition comprising a compound of Formula A or salts thereof and a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition is in the form of a tablet, capsule, pill, gel, granules, aerosol, or aqueous buffer, such as a saline or phosphate buffer, or a nanoparticle formulation, emulsion, liposome, etc. The pharmaceutical composition may also include one or more further active antiviral agents, or may be administered in combination with one or more such active agent. In a yet further embodiment there is provided methods for preparing the compounds of Formula A or salts thereof comprising mixing one or more starting materials with reagents under conditions such that the products are formed. BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 shows a crystal structure of l '-((l-(4-hydroxybutyl)-lH-benzo[d]imidazol-2- yl)methyl)spiro[cyclopropane-l,3'-pyrrolo[2,3-c]pyridin]-2'( iI)-one (designated herein as ZD-2- 160);

Figure 2 shows a crystal structure of 4-(2-((2-oxo-l-(tetrahydro-2H-pyran-4-yl)-l,2- dihydro-3H-imidazo[4,5-c]pyridin-3-yl)methyl)-lH-benzo[d]imidazol-l-yl)butanenitrile

(designated herein as RJW-2-01 1);

Figure 3 shows a crystal structure of l-(l, l-dioxidothietan-3-yl)-3-((l-(4-hydroxybutyl)- lH-benzo[d]imidazol-2-yl)methyl)-l,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one (designated herein as EJ-2143);

Figure 4 is a graph showing CCso data for some exemplary compounds of the disclosure; and

Figure 5 is a graph showing the ICso data for exemplary compounds of this disclosure against the wild type and RSV-F489 mutant virus. DETAILED DESCRIPTION

Terms

Before the present disclosure is described in greater detail, it is to be understood that this disclosure is not limited to particular embodiments described, and as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference and are incorporated herein by reference to disclose and describe the methods and/or materials in connection with which the publications are cited. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present disclosure is not entitled to antedate such publication by virtue of prior disclosure. Further, the dates of publication provided could be different from the actual publication dates that may need to be independently confirmed.

As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present disclosure. Any recited method can be carried out in the order of events recited or in any other order that is logically possible.

Embodiments of the present disclosure will employ, unless otherwise indicated, techniques of medicine, organic chemistry, biochemistry, molecular biology, pharmacology, and the like, which are within the skill of the art. Such techniques are explained fully in the literature.

To the extent that any chemical formulas reported herein contain one or more chiral centers, the formulas are intended to encompass all stable stereoisomers, enantiomers, and diastereomers. It is also understood that formulas encompass all tautomeric forms.

It must be noted that, as used in the specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise.

"Subject" refers any animal, preferably a human patient, livestock, mouse model or domestic pet.

As used herein, the terms "prevent" and "preventing" include the prevention of the recurrence, spread or onset. It is not intended that the present disclosure be limited to complete prevention. In some embodiments, the onset is delayed, or the severity of the disease is reduced.

As used herein, the terms "treat" and "treating" are not limited to the case where the subject (e.g. patient) is cured and the disease is eradicated. Rather, embodiments, of the present disclosure also contemplate treatment that merely reduces symptoms, and/or delays disease progression. As used herein, the term "combination with" when used to describe administration with an additional treatment means that the agent may be administered prior to, together with, or after the additional treatment, or a combination thereof.

As used herein, "alkyl" means a noncyclic straight chain or branched, unsaturated or saturated hydrocarbon such as those containing from 1 to 20 carbon atoms. A "higher alkyl" refers to unsaturated or saturated hydrocarbon having 6 or more carbon atoms. A "Cs-Cis" refers to an alkyl containing 8 to 18 carbon atoms. Likewise, a "C6-C22" refers to an alkyl containing 6 to 22 carbon atoms. Representative saturated straight chain alkyls include methyl, ethyl, n-propyl, n- butyl, n-pentyl, n-hexyl, n-septyl, n-octyl, n-nonyl, hexadecyl, dodecyl, tetradecyl, izosonyl, octadecyl, and the like; while saturated branched alkyls include isopropyl, sec-butyl, isobutyl, tert- butyl, isopentyl, and the like. Unsaturated alkyls contain at least one double or triple bond between adjacent carbon atoms (referred to as an "alkenyl" or "alkynyl", respectively). Representative straight chain and branched alkenyls include ethylenyl, propylenyl, 1-butenyl, 2-butenyl, isobutylenyl, 1-pentenyl, 2-pentenyl, 3 -methyl- 1-butenyl, 2-methyl-2-butenyl, 2,3- dimethyl-2- butenyl, and the like; while representative straight chain and branched alkynyls include acetylenyl, propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, 3- methyl- 1-butynyl, and the like.

Non-aromatic mono or polycyclic alkyls are referred to herein as "carbocycles" or "carbocyclyl" groups. Representative saturated carbocycles include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like; while unsaturated carbocycles include cyclopentenyl and cyclohexenyl, and the like. Carbocyclyls include cycloalkyls and cycloalkenyls.

"Heterocarbocycles" or "heterocarbocyclyl" groups are carbocycles which contain from 1 to 4 heteroatoms independently selected from nitrogen, phosphorous, oxygen and sulphur which may be saturated or unsaturated (but not aromatic), monocyclic or polycyclic, and wherein the nitrogen and sulphur heteroatoms may be optionally oxidized (e.g. -S(O)-, -SO2-, -N(O)-), and the nitrogen heteroatom may be optionally quaternized. Heterocarbocycles include morpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyridinyl, tetrahydroprimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like.

As used herein, "heterocycle" or "heterocyclyl" refers to mono- and polycyclic ring systems having 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur, and containing at least 1 carbon atom. The mono- and polycyclic ring systems may be aromatic, non-aromatic or mixtures of aromatic and non-aromatic rings. Heterocycle includes heterocarbocycles, heteroaryls, and the like.

The terms "cycloalkyl" and "cycloalkenyl" refer to mono-, bi-, or tri homocyclic ring groups of 3 to 15 carbon atoms which are, respectively, fully saturated and partially unsaturated.

The term "substituted" refers to a molecule wherein at least one hydrogen atom is replaced with a substituent. When substituted, one or more of the groups are "substituents". The molecule may be multiply substituted. In the case of an oxo substituent (=0), two hydrogen atoms are replaced. Example substituents within this context may include halogen, hydroxy, alkyl, alkoxy, nitro, cyano, oxo, carbocyclyl, carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, - RaRb, - RaC(=0)Rb, - RaC(=0) Ra Rb, - RaC(=0)ORb, - RaSOzRb, -C(=0)Ra, -C(=0)ORa, -C(=0) RaRb, -OC(=0) RaRb, -ORa, -SRa, -SORa, - S(=0)₂Ra, -OS(=0)₂Ra and -S(=0)₂ORa. Ra and Rb in this context may be the same or different and independently hydrogen, halogen hydroxyl, alkyl, alkoxy, alkyl, amino, alkylamino, dialkylamino, carbocyclyl, carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl.

The term "optionally substituted", as used herein, means that substitution is optional and therefore it is possible for the designated atom to be unsubstituted.

As used herein, the term "derivative" refers to a structurally similar compound that retains sufficient functional attributes of the identified analogue. The derivative may be structurally similar because it is lacking one or more atoms, substituted, a salt, in different hydration/oxidation states, or because one or more atoms within the molecule are switched, such as, but not limited to, adding a hydroxyl group, replacing an oxygen atom with a sulfur atom, or replacing an amino group with a hydroxyl group, oxidizing a hydroxyl group to a carbonyl group, reducing a carbonyl group to a hydroxyl group, and reducing a carbon-to-carbon double bond to an alkyl group or oxidizing a carbon-to-carbon single bond to a double bond. A derivative optionally has one or more, the same or different, substitutions. Derivatives may be prepared by any variety of synthetic methods or appropriate adaptations presented in synthetic or organic chemistry text books, such as those provided in "March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure", Wiley, 6th Edition (2007) Michael B. Smith or "Domino Reactions in Organic Synthesis", Wiley (2006) Lutz F. Tietze, hereby incorporated by reference. The present disclosure relates to com ounds of the general Formula A or salt thereof,

Formula A

wherein R¹ is an optionally substituted alkyl and X may be either N or C. In the case where X is N, then R² will be a heterocyclyl, and R³ will be absent; whereas when X is C, then R² and R³ will be linked to form a carbocyclyl.

The substituent R¹ can be an optionally substituted C₃ to C₆ alkyl such as a propyl, butyl, pentyl or hexyl chain, which may be straight, branched, saturated or unsaturated. In a preferred embodiment, R¹ is an optionally substituted C₄ alkyl, i.e. a butyl chain and is terminally substituted with a nitrile or alcohol group (i.e.≡N or -OH).

When X is N, the substituent R² can be a C₃ to C₆ heterocyclyl and in a preferred embodiment is a C₃ or C₄ heterocyclyl and the heteroatoms are selected from O, N, S or SO2. The heterocycles are preferably selected from tetrahydropyran and thietane- 1, 1 -dioxide.

When X is C, the substituents R² and R³ are linked to form a C₃-C₆ carbocycle. In a preferred embodiment it is a C₃ carbocyclyl (i.e. cyclopropyl).

The compound of Formula A may be further exemplified as compounds of Formula Al or Formula A2, or salts thereof,

Formula Al Formula A2 wherein n is 1, 2, 3, or 4; Y is selected from N, O, S, or SO2; and R⁵ iis OH or N. In a preferred embodiment, the compounds of Formula A are selected from:

In present disclosure also contemplates derivatives of the compounds disclosed herein such as those containing one or more, the same or different, substituents, or where single bonds are replaced with double or triple bonds.

Methods of Use

In certain embodiments, the disclosure relates to methods of treating or preventing a viral infection comprising administering in effective amount of a compound of Formual A or salts thereof disclosed herein to a subject in need thereof. In some embodiments, the subject is at risk of, exhibiting symptoms of, suffering from, or diagnosed with a viral infection such as RSV infections.

In some embodiments, the subject is at risk of, exhibiting symptoms of, or diagnosed with Respiratory syncytial virus (RSV), influenza A virus including subtype H1N1, influenza B virus, influenza C virus, rotavirus A, rotavirus B, rotavirus C, rotavirus D, rotavirus E, SARS coronavirus, human adenovirus types (HAdV-1 to 55), human papillomavirus (UPV) Types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, parvovirus B 19, molluscum contagiosum virus, JC virus (JCV), BK vims, Merkel cell polyomavirus, coxsackie A vims, norovirus, Rubella vims, lymphocytic choriomeningitis vims (LCMV), yellow fever vims, measles vims, mumps vims, rinderpest vims, California encephalitis vims, hantavims, rabies vims, ebola vims, marburg vims, herpes simplex virus- 1 (HSV-1), herpes simplex virus-2 (HSV-2), varicella zoster vims (VZV), Epstein-Barr vims (EBV), cytomegalovims (CMV), herpes lymphotropic vims, roseolovims, Kaposi's sarcoma-associated herpesvims, hepatitis A (HAV), hepatitis B (HBV), hepatitis C (HCV), hepatitis D (HDV), hepatitis E (HEV), human immunodeficiency vims (HIV), The Human T-lymphotropic vims Type I (HTLV-1), Friend spleen focus-forming vims (SFFV) or Xenotropic MuLV-Related Vims (XMRV); or is at risk of, exhibiting symptoms of, or diagnosed with RSV in addition to a co-infection with one of the above listed diseases.

In certain embodiments, methods disclosed herein are contemplated to be administered in combination with other the antiviral agent(s) such as abacavir, acyclovir, adefovir, amantadine, amprenavir, ampligen, arbidol, atazanavir, atripla, boceprevir, cidofovir, combivir, complera, damnavir, delavirdine, didanosine, docosanol, dolutegravir, edoxudine, efavirenz, emtricitabine, enfuvirtide, entecavir, famciclovir, fomivirsen, fosamprenavir, foscarnet, fosfonet, ganciclovir, ibacitabine, imunovir, idoxuridine, imiquimod, indinavir, inosine, interferon type III, interferon type II, interferon type I, lamivudine, lopinavir, loviride, maraviroc, moroxydine, methisazone, nelfinavir, nevirapine, nexavir, oseltamivir, peginterferon alfa-2a, penciclovir, peramivir, pleconaril, podophyllotoxin , raltegravir, ribavirin, rimantadine, ritonavir, pyramidine, saquinavir, stavudine, stribild, tenofovir, tenofovir disoproxil, tenofovir alafenamide fumarate, tipranavir, trifluridine, trizivir, tromantadine, tmvada, valaciclovir, valganciclovir, vicriviroc, vidarabine, viramidine, zalcitabine, zanamivir, or zidovudine, and combinations thereof. The methods of treatment and the pharmaceutical compounds of the present disclosure may also be administered with heretofore undisclosed antiviral agents acting of different components of RSV.

In certain embodiments, the disclosure contemplates the treatment or prevention of a viral infection using compounds disclosed herein, wherein viral infection is Respiratory syncytial vims (RSV).

Formulations

The present disclosure includes pharmaceutical compositions comprising a compound of

Formula A or salt thereof in combination with pharmaceutically acceptable excipients. Pharmaceutical compositions disclosed herein may be in the form of pharmaceutically acceptable salts, as generally described below. Some preferred, but non-limiting examples of suitable pharmaceutically acceptable organic and/or inorganic acids are hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, acetic acid and citric acid, as well as other pharmaceutically acceptable acids known per se (for which reference is made to the references referred to below).

When the compounds of the disclosure contain an acidic group as well as a basic group, the compounds of the disclosure may also form internal salts, and such compounds are within the scope of the disclosure. When a compound contains a hydrogen-donating heteroatom (e.g. H), salts are contemplated to covers isomers formed by transfer of said hydrogen atom to a basic group or atom within the molecule.

Pharmaceutically acceptable salts of the compounds include the acid addition and base salts thereof. Suitable acid addition salts are formed from acids which form non-toxic salts. Examples include the acetate, adipate, aspartate, benzoate, besylate, bicarbonate/carbonate, bisulphate/sulphate, borate, camsylate, citrate, cyclamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methyl sulphate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate and xinofoate salts. Suitable base salts are formed from bases which form non-toxic salts. Examples include the aluminium, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine and zinc salts. Hemisalts of acids and bases may also be formed, for example, hemisulphate and hemicalcium salts. For a review on suitable salts, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use by Stahl and Wermuth (Wiley - VCH, 2002), incorporated herein by reference.

Pharmaceutical compositions for use in the present disclosure typically comprise an effective amount of a compound and a suitable pharmaceutically acceptable carrier. The preparations may be prepared in any manner known per se, which usually involves mixing the at least one compound according to the disclosure with the one or more pharmaceutically acceptable carriers, and, if desired, in combination with other pharmaceutical active compounds, when necessary under aseptic conditions. Reference is made to U.S. Pat. Nos. 6,372,778; 6,369,086; 6,369,087 and 6,372,733 and the further references mentioned above, as well as to the standard handbooks, such as the latest edition of Remington's Pharmaceutical Sciences.

Generally, for pharmaceutical use, the compounds may be formulated as a pharmaceutical preparation comprising at least one compound of the present disclosure and at least one pharmaceutically acceptable carrier, diluent or excipient and/or adjuvant, and optionally one or more further pharmaceutically active compounds.

In certain embodiments, the pharmaceutical composition is in the form of a tablet, capsule, pill, gel, granules, aerosol, or aqueous buffer, such as a saline or phosphate buffer, or a nanoparticle formulation, emulsion, liposome, etc. The pharmaceutical preparations of the disclosure are preferably in a unit dosage form, and may be suitably packaged, for example in a box, blister, vial, bottle, sachet, ampoule or in any other suitable single-dose or multi-dose holder or container (which may be properly labeled); optionally with one or more leaflets containing product information and/or instructions for use. Generally, such unit dosages will contain between 1 and 1000 mg, and usually between 5 and 500 mg, of the at least one compound of the disclosure, e.g. about 10, 25, 50, 100, 200, 300, 400 or 500 mg per unit dosage.

The compounds can be administered by a variety of routes including the oral, ocular, rectal, transdermal, subcutaneous, intravenous, intramuscular or intranasal routes, depending mainly on the specific preparation used. In certain embodiments, the compound is administered by inhalation through the lungs.

The compound will generally be administered in an "effective amount", by which is meant any amount of a compound that, upon suitable administration, is sufficient to achieve the desired therapeutic or prophylactic effect in the subject to which it is administered. Usually, depending on the condition to be prevented or treated and the route of administration, such an effective amount will usually be between 0.01 to 1000 mg per kilogram body weight of the patient per day, more often between 0.1 and 500 mg, such as between 1 and 250 mg, for example about 5, 10, 20, 50, 100, 150, 200 or 250 mg, per kilogram body weight of the patient per day, which may be administered as a single daily dose, divided over one or more daily doses. The amount(s) to be administered, the route of administration and the further treatment regimen may be determined by the treating clinician, depending on factors such as the age, gender and general condition of the patient and the nature and severity of the disease/symptoms to be treated. Reference is again made to U.S. Pat. Nos. 6,372,778; 6,369,086; 6,369,087 and 6,372,733 and the further references mentioned above, as well as to the standard handbooks, such as the latest edition of Remington's Pharmaceutical Sciences.

Depending upon the manner of introduction, the compounds described herein may be formulated in a variety of ways. Formulations containing one or more compounds can be prepared in various pharmaceutical forms, such as granules, tablets, capsules, suppositories, powders, controlled release formulations, suspensions, emulsions, creams, gels, ointments, salves, lotions, nanoparticles, aerosols and the like. Preferably, these formulations are employed in solid dosage forms suitable for simple, and preferably oral, administration of precise dosages. Solid dosage forms for oral administration include, but are not limited to, tablets, soft or hard gelatin or non- gelatin capsules, and caplets. However, liquid dosage forms, such as solutions, syrups, suspension, shakes, etc. can also be utilized. In another embodiment, the formulation is administered topically. Suitable topical formulations include, but are not limited to, lotions, ointments, creams, and gels. In a preferred embodiment, the topical formulation is a gel. In another embodiment, the formulation is administered intranasally.

In certain embodiments, the pharmaceutical composition comprises a compound disclosed herein and a propellant. In certain embodiments, an aerosolizing propellant is compressed air, ethanol, nitrogen, carbon dioxide, nitrous oxide, hydrofluoroalkanes (HFAs), 1,1,1,2,- tetrafluoroethane, 1,1, 1,2,3,3,3-heptafluoropropane or combinations thereof.

In certain embodiments, the disclosure contemplates a pressurized or unpressurized container comprising a compound herein. In certain embodiments, the container is a manual pump spray, inhaler, meter-dosed inhaler, dry powder inhaler, nebulizer, vibrating mesh nebulizer, jet nebulizer, or ultrasonic wave nebulizer.

Formulations containing one or more of the compounds described herein may be prepared using a pharmaceutically acceptable carrier composed of materials that are considered safe and effective and may be administered to an individual without causing undesirable biological side effects or unwanted interactions. The carrier is all components present in the pharmaceutical formulation other than the active ingredient or ingredients. As generally used herein "carrier" includes, but is not limited to, diluents, binders, lubricants, disintegrators, fillers, pH modifying agents, preservatives, antioxidants, solubility enhancers, and coating compositions. Carrier also includes all components of the coating composition which may include plasticizers, pigments, colorants, stabilizing agents, and glidants. Delayed release, extended release, and/or pulsatile release dosage formulations may be prepared as described in standard references such as "Pharmaceutical dosage form tablets", eds. Liberman et al. (New York, Marcel Dekker, Inc., 1989), "Remington - The science and practice of pharmacy", 20th ed., Lippincott Williams & Wilkins, Baltimore, MD, 2000, and "Pharmaceutical dosage forms and drug delivery systems", 6th Edition, Ansel et al., (Media, PA: Williams and Wilkins, 1995). These references provide information on carriers, materials, equipment and processes for preparing tablets and capsules and delayed release dosage forms of tablets, capsules, and granules.

Examples of suitable coating materials include, but are not limited to, cellulose polymers such as cellulose acetate phthalate, hydroxypropyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate and hydroxypropyl methylcellulose acetate succinate; polyvinyl acetate phthalate, acrylic acid polymers and copolymers, and methacrylic resins that are commercially available under the trade name Eudragit® (Roth Pharma, Westerstadt, Germany), zein, shellac, and polysaccharides.

Additionally, the coating material may contain conventional carriers such as plasticizers, pigments, colorants, glidants, stabilization agents, pore formers and surfactants.

Optional pharmaceutically acceptable excipients present in the drug-containing tablets, beads, granules or particles include, but are not limited to, diluents, binders, lubricants, disintegrants, colorants, stabilizers, and surfactants. Diluents, also referred to as "fillers", are typically necessary to increase the bulk of a solid dosage form so that a practical size is provided for compression of tablets or formation of beads and granules. Suitable diluents include, but are not limited to, dicalcium phosphate dihydrate, calcium sulfate, lactose, sucrose, mannitol, sorbitol, cellulose, microcrystalline cellulose, kaolin, sodium chloride, dry starch, hydrolyzed starches, pregelatinized starch, silicone dioxide, titanium oxide, magnesium aluminum silicate and powdered sugar.

Binders are used to impart cohesive qualities to a solid dosage formulation, and thus ensure that a tablet or bead or granule remains intact after the formation of the dosage forms. Suitable binder materials include, but are not limited to, starch, pregelatinized starch, gelatin, sugars (including sucrose, glucose, dextrose, lactose and sorbitol), polyethylene glycol, waxes, natural and synthetic gums such as acacia, tragacanth, sodium alginate, cellulose, including hydroxypropylmethylcellulose, hydroxypropylcellulose, ethylcellulose, and veegum, and synthetic polymers such as acrylic acid and methacrylic acid copolymers, methacrylic acid copolymers, methyl methacrylate copolymers, aminoalkyl methacrylate copolymers, polyacrylic acid/polymethacrylic acid and polyvinylpyrrolidone.

Lubricants are used to facilitate tablet manufacture. Examples of suitable lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, glycerol behenate, polyethylene glycol, talc, and mineral oil.

Disintegrants are used to facilitate dosage form disintegration or "breakup" after administration, and generally include, but are not limited to, starch, sodium starch glycolate, sodium carboxymethyl starch, sodium carboxymethylcellulose, hydroxypropyl cellulose, pregelatinized starch, clays, cellulose, alginine, gums or cross linked polymers, such as cross- linked PVP (Polyplasdone XL from GAF Chemical Corp).

Stabilizers are used to inhibit or retard drug decomposition reactions which include, by way of example, oxidative reactions.

Surfactants may be anionic, cationic, amphoteric or nonionic surface active agents.

Suitable anionic surfactants include, but are not limited to, those containing carboxylate, sulfonate and sulfate ions. Examples of anionic surfactants include sodium, potassium, ammonium of long chain alkyl sulfonates and alkyl aryl sulfonates such as sodium dodecylbenzene sulfonate; dialkyl sodium sulfosuccinates, such as sodium dodecylbenzene sulfonate; dialkyl sodium sulfosuccinates, such as sodium bis-(2-ethylthioxyl)-sulfosuccinate; and alkyl sulfates such as sodium lauryl sulfate. Cationic surfactants include, but are not limited to, quaternary ammonium compounds such as benzalkonium chloride, benzethonium chloride, cetrimonium bromide, stearyl dimethylbenzyl ammonium chloride, polyoxyethylene and coconut amine. Examples of nonionic surfactants include ethylene glycol monostearate, propylene glycol myristate, glyceryl monostearate, glyceryl stearate, polyglyceryl-4-oleate, sorbitan acylate, sucrose acylate, PEG- 150 laurate, PEG-400 monolaurate, polyoxyethylene monolaurate, polysorbates, polyoxyethylene octylphenylether, PEG- 1000 cetyl ether, polyoxyethylene tridecyl ether, polypropylene glycol butyl ether, Poloxamer® 401, stearoyl monoisopropanolamide, and polyoxyethylene hydrogenated tallow amide. Examples of amphoteric surfactants include sodium N-dodecyl-β- alanine, sodium N-lauryl-P-iminodipropionate, myristoamphoacetate, lauryl betaine and lauryl sulfobetaine. If desired, the tablets, beads, granules, or particles may also contain minor amount of nontoxic auxiliary substances such as wetting or emulsifying agents, dyes, pH buffering agents, or preservatives.

The concentration of the compound to carrier and/or other substances may vary from about 0.5 to about 100 wt % (weight percent). For oral use, the pharmaceutical formulation will generally contain from about 5 to about 100% by weight of the active material. For other uses, the pharmaceutical formulation will generally have from about 0.5 to about 50 wt. % of the active material.

The compositions described herein can be formulated for modified or controlled release. Examples of controlled release dosage forms include extended release dosage forms, delayed release dosage forms, pulsatile release dosage forms, and combinations thereof.

The extended release formulations are generally prepared as diffusion or osmotic systems, for example, as described in "Remington - The science and practice of pharmacy" (20th ed., Lippincott Williams & Wilkins, Baltimore, MD, 2000). A diffusion system typically consists of two types of devices, a reservoir and a matrix, and is well known and described in the art. The matrix devices are generally prepared by compressing the drug with a slowly dissolving polymer carrier into a tablet form. The three major types of materials used in the preparation of matrix devices are insoluble plastics, hydrophilic polymers, and fatty compounds. Plastic matrices include, but are not limited to, methyl acrylate-methyl methacrylate, polyvinyl chloride, and polyethylene. Hydrophilic polymers include, but are not limited to, cellulosic polymers such as methyl and ethyl cellulose, hydroxyalkylcelluloses such as hydroxypropyl-cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and carbopol Carbopol® 934, polyethylene oxides and mixtures thereof. Fatty compounds include, but are not limited to, various waxes such as carnauba wax and glyceryl tristearate and wax-type substances including hydrogenated castor oil or hydrogenated vegetable oil, or mixtures thereof.

In certain preferred embodiments, the plastic material is a pharmaceutically acceptable acrylic polymer, including but not limited to, acrylic acid and methacrylic acid copolymers, methyl methacrylate, methyl methacrylate copolymers, ethoxyethyl methacrylates, cyanoethyl methacrylate, aminoalkyl methacrylate copolymer, poly(acrylic acid), poly(methacrylic acid), methacrylic acid alkylamine copolymer poly(methyl methacrylate), poly(methacrylic acid)(anhydride), polymethacrylate, polyacrylamide, poly(methacrylic acid anhydride), and glycidyl methacrylate copolymers.

In certain preferred embodiments, the acrylic polymer is comprised of one or more ammonio methacrylate copolymers. Ammonio methacrylate copolymers are well known in the art, and are described in F XVII as fully polymerized copolymers of acrylic and methacrylic acid esters with a low content of quaternary ammonium groups.

In one preferred embodiment, the acrylic polymer is an acrylic resin lacquer such as that which is commercially available from Rohm Pharma under the Tradename tradename Eudragit®. In further preferred embodiments, the acrylic polymer comprises a mixture of two acrylic resin lacquers commercially available from Rohm Pharma under the Tradenames tradenames Eudragit® RL30D and Eudragit ® RS30D, respectively. Eudragit® RL30D and Eudragit® RS30D are copolymers of acrylic and methacrylic esters with a low content of quaternary ammonium groups, the molar ratio of ammonium groups to the remaining neutral (meth)acrylic esters being 1 :20 in Eudragit® RL30D and 1 :40 in Eudragit® RS30D. The mean molecular weight is about 150,000. Edragit® S-100 and Eudragit® L-100 are also preferred. The code designations RL (high permeability) and RS (low permeability) refer to the permeability properties of these agents. Eudragit® RL/RS mixtures are insoluble in water and in digestive fluids. However, multiparticulate systems formed to include the same are swellable and permeable in aqueous solutions and digestive fluids.

The polymers described above such as Eudragit®.RTM. RL/RS may be mixed together in any desired ratio in order to ultimately obtain a sustained-release formulation having a desirable dissolution profile. Desirable sustained-release multiparticulate systems may be obtained, for instance, from 100% Eudragit® RL, 50% Eudragit® RL and 50% Eudragit® RS, and 10% Eudragit® RL and 90% :Eudragit® 90% RS. One skilled in the art will recognize that other acrylic polymers may also be used, such as, for example, Eudragit® L.

Alternatively, extended release formulations can be prepared using osmotic systems or by applying a semi-permeable coating to the dosage form. In the latter case, the desired drug release profile can be achieved by combining low permeable and high permeable coating materials in suitable proportion. The devices with different drug release mechanisms described above can be combined in a final dosage form comprising single or multiple units. Examples of multiple units include, but are not limited to, multilayer tablets and, capsules containing tablets, beads, or granules, etc.

An immediate release portion can be added to the extended release system by means of either applying an immediate release layer on top of the extended release core using a coating or compression process or in a multiple unit system such as a capsule containing extended and immediate release beads.

Extended release tablets containing hydrophilic polymers are prepared by techniques commonly known in the art such as direct compression, wet granulation, or dry granulation processes. Their formulations usually incorporate polymers, diluents, binders, and lubricants as well as the active pharmaceutical ingredient. The usual diluents include inert powdered substances such as starches, powdered cellulose, especially crystalline and microcrystalline cellulose, sugars such as fructose, mannitol and sucrose, grain flours and similar edible powders. Typical diluents include, for example, various types of starch, lactose, mannitol, kaolin, calcium phosphate or sulfate, inorganic salts such as sodium chloride and powdered sugar. Powdered cellulose derivatives are also useful. Typical tablet binders include substances such as starch, gelatin and sugars such as lactose, fructose, and glucose. Natural and synthetic gums, including acacia, alginates, methylcellulose, and polyvinylpyrrolidone can also be used. Polyethylene glycol, hydrophilic polymers, ethylcellulose and waxes can also serve as binders. A lubricant is necessary in a tablet formulation to prevent the tablet and punches from sticking in the die. The lubricant is chosen from such slippery solids as talc, magnesium and calcium stearate, stearic acid and hydrogenated vegetable oils.

Extended release tablets containing wax materials are generally prepared using methods known in the art such as a direct blend method, a congealing method, and an aqueous dispersion method. In the congealing method, the drug is mixed with a wax material and either spray- congealed or congealed and screened and processed.

Delayed release formulations are created by coating a solid dosage form with a polymer film, which is insoluble in the acidic environment of the stomach, and soluble in the neutral environment of the small intestine.

The delayed release dosage units can be prepared, for example, by coating a drug or a drug- containing composition with a selected coating material. The drug-containing composition may be, e.g., a tablet for incorporation into a capsule, a tablet for use as an inner core in a "coated core" dosage form, or a plurality of drug-containing beads, particles or granules, for incorporation into either a tablet or capsule. Preferred coating materials include bioerodible, gradually hydrolyzable, gradually water-soluble, and/or enzymatically degradable polymers, and may be conventional "enteric" polymers. Enteric polymers, as will be appreciated by those skilled in the art, become soluble in the higher pH environment of the lower gastrointestinal tract or slowly erode as the dosage form passes through the gastrointestinal tract, while enzymatically degradable polymers are degraded by bacterial enzymes present in the lower gastrointestinal tract, particularly in the colon. Suitable coating materials for effecting delayed release include, but are not limited to, cellulosic polymers such as hydroxypropyl cellulose, hydroxy ethyl cellulose, hydroxymethyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl methyl cellulose acetate succinate, hydroxypropylmethyl cellulose phthalate, methylcellulose, ethyl cellulose, cellulose acetate, cellulose acetate phthalate, cellulose acetate trimellitate and carboxymethylcellulose sodium; acrylic acid polymers and copolymers, preferably formed from acrylic acid, methacrylic acid, methyl acrylate, ethyl acrylate, methyl methaciylate and/or ethyl methaciylate, and other methacrylic resins that are commercially available under the tradename Eudragit®. (Rohm Pharma; Westerstadt, Germany), including Eudragit® L30D-55 and LI 00-55 (soluble at pH 5.5 and above), Eudragit® L-100 (soluble at pH 6.0 and above), Eudragit® S (soluble at pH 7.0 and above, as a result of a higher degree of esterification), and Eudragits® E, RL and RS (water- insoluble polymers having different degrees of permeability and expandability); vinyl polymers and copolymers such as polyvinyl pyrrolidone, vinyl acetate, vinylacetate phthalate, vinylacetate crotonic acid copolymer, and ethylene-vinyl acetate copolymer; enzymatically degradable polymers such as azo polymers, pectin, chitosan, amylose and guar gum; zein and shellac. Combinations of different coating materials may also be used. Multi-layer coatings using different polymers may also be applied.

The preferred coating weights for particular coating materials may be readily determined by those skilled in the art by evaluating individual release profiles for tablets, beads and granules prepared with different quantities of various coating materials. It is the combination of materials, method and form of application that produce the desired release characteristics, which one can determine only from the clinical studies. The coating composition may include conventional additives, such as plasticizers, pigments, colorants, stabilizing agents, glidants, etc. A plasticizer is normally present to reduce the fragility of the coating, and will generally represent about 10 wt. % to 50 wt. % relative to the dry weight of the polymer. Examples of typical plasticizers include polyethylene glycol, propylene glycol, triacetin, dimethyl phthalate, diethyl phthalate, dibutyl phthalate, dibutyl sebacate, triethyl citrate, tributyl citrate, triethyl acetyl citrate, castor oil and acetylated monoglycerides. A stabilizing agent is preferably used to stabilize particles in the dispersion. Typical stabilizing agents are nonionic emulsifiers such as sorbitan esters, polysorbates and polyvinylpyrrolidone. Glidants are recommended to reduce sticking effects during film formation and drying, and will generally represent approximately 25 wt. % to 100 wt. % of the polymer weight in the coating solution. One effective glidant is talc. Other glidants such as magnesium stearate and glycerol monostearates may also be used. Pigments such as titanium dioxide may also be used. Small quantities of an anti-foaming agent, such as a silicone (e.g., simethicone), may also be added to the coating composition.

The formulation can provide pulsatile delivery of the one or more compounds. By

"pulsatile" is meant that a plurality of drug doses is released at spaced apart intervals of time. Generally, upon ingestion of the dosage form, release of the initial dose is substantially immediate, i.e., the first drug release "pulse" occurs within about one hour of ingestion. This initial pulse is followed by a first time interval (lag time) during which very little or no drug is released from the dosage form, after which a second dose is then released. Similarly, a second nearly drug release- free interval between the second and third drug release pulses may be designed. The duration of the nearly drug release-free time interval will vary depending upon the dosage form design e.g., a twice daily dosing profile, a three times daily dosing profile, etc. For dosage forms providing a twice daily dosage profile, the nearly drug release-free interval has a duration of approximately 3 hours to 14 hours between the first and second dose. For dosage forms providing a three times daily profile, the nearly drug release-free interval has a duration of approximately 2 hours to 8 hours between each of the three doses.

In one embodiment, the pulsatile release profile is achieved with dosage forms that are closed and preferably sealed capsules housing at least two drug-containing "dosage units" wherein each dosage unit within the capsule provides a different drug release profile. Control of the delayed release dosage unit(s) is accomplished by a controlled release polymer coating on the dosage unit, or by incorporation of the active agent in a controlled release polymer matrix. Each dosage unit may comprise a compressed or molded tablet, wherein each tablet within the capsule provides a different drug release profile. For dosage forms mimicking a twice a day dosing profile, a first tablet releases drug substantially immediately following ingestion of the dosage form, while a second tablet releases drug approximately 3 hours to less than 14 hours following ingestion of the dosage form. For dosage forms mimicking a three times daily dosing profile, a first tablet releases drug substantially immediately following ingestion of the dosage form, a second tablet releases drug approximately 3 hours to less than 10 hours following ingestion of the dosage form, and the third tablet releases drug at least 5 hours to approximately 18 hours following ingestion of the dosage form. It is possible that the dosage form includes more than three tablets. While the dosage form will not generally include more than a third tablet, dosage forms housing more than three tablets can be utilized.

Alternatively, each dosage unit in the capsule may comprise a plurality of drug-containing beads, granules or particles. As is known in the art, drug-containing "beads" refer to beads made with drug and one or more excipients or polymers. Drug-containing beads can be produced by applying drug to an inert support, e.g., inert sugar beads coated with drug or by creating a "core" comprising both drug and one or more excipients. As is also known, drug-containing "granules" and "particles" comprise drug particles that may or may not include one or more additional excipients or polymers. In contrast to drug-containing beads, granules and particles do not contain an inert support. Granules generally comprise drug particles and require further processing. Generally, particles are smaller than granules, and are not further processed. Although beads, granules and particles may be formulated to provide immediate release, beads and granules are generally employed to provide delayed release.

In one embodiment, the compound is formulated for topical administration. Suitable topical dosage forms include lotions, creams, ointments, and gels. A "gel" is a semisolid system containing a dispersion of the active agent, i.e., compound, in a liquid vehicle that is rendered semisolid by the action of a thickening agent or polymeric material dissolved or suspended in the liquid vehicle. The liquid may include a lipophilic component, an aqueous component or both. Some emulsions may be gels or otherwise include a gel component. Some gels, however, are not emulsions because they do not contain a homogenized blend of immiscible components. Methods for preparing lotions, creams, ointments, and gels are well known in the art. The compound described herein can be administered adjunctively with other active compounds. These compounds include but are not limited to analgesics, anti-inflammatory drugs, antipyretics, antidepressants, antiepileptics, antihistamines, antimigraine drugs, antimuscarinics, anxioltyics, sedatives, hypnotics, antipsychotics, bronchodilators, anti-asthma drugs, cardiovascular drugs, corticosteroids, dopaminergics, electrolytes, gastro-intestinal drugs, muscle relaxants, nutritional agents, vitamins, parasympathomimetics, stimulants, anorectics and anti- narcoleptics. "Adjunctive administration", as used herein, means the compound can be administered in the same dosage form or in separate dosage forms with one or more other active agents.

Specific examples of compounds that can be adjunctively administered with the compounds include, but are not limited to, aceclofenac, acetaminophen, adomexetine, almotriptan, alprazolam, amantadine, amcinonide, aminocyclopropane, amitriptyline, amolodipine, amoxapine, amphetamine, aripiprazole, aspirin, atomoxetine, azasetron, azatadine, beclomethasone, benactyzine, benoxaprofen, bermoprofen, betamethasone, bicifadine, bromocriptine, budesonide, buprenorphine, bupropion, buspirone, butorphanol, butriptyline, caffeine, carbamazepine, carbidopa, carisoprodol, celecoxib, chlordiazepoxide, chlorpromazine, choline salicylate, citalopram, clomipramine, clonazepam, clonidine, clonitazene, clorazepate, clotiazepam, cloxazolam, clozapine, codeine, corticosterone, cortisone, cyclobenzaprine, cyproheptadine, demexiptiline, desipramine, desomorphine, dexamethasone, dexanabinol, dextroamphetamine sulfate, dextromoramide, dextropropoxyphene, dezocine, diazepam, dibenzepin, diclofenac sodium, diflunisal, dihydrocodeine, dihydroergotamine, dihydromorphine, dimetacrine, divalproxex, dizatriptan, dolasetron, donepezil, dothiepin, doxepin, duloxetine, ergotamine, escitalopram, estazolam, ethosuximide, etodolac, femoxetine, fenamates, fenoprofen, fentanyl, fludiazepam, fluoxetine, fluphenazine, flurazepam, flurbiprofen, flutazolam, fluvoxamine, frovatriptan, gabapentin, galantamine, gepirone, ginko bilboa, granisetron, haloperidol, huperzine A, hydrocodone, hydrocortisone, hydromorphone, hydroxyzine, ibuprofen, imipramine, indiplon, indomethacin, indoprofen, iprindole, ipsapirone, ketaserin, ketoprofen, ketorolac, lesopitron, levodopa, lipase, lofepramine, lorazepam, loxapine, maprotiline, mazindol, mefenamic acid, melatonin, melitracen, memantine, meperidine, meprobamate, mesalamine, metapramine, metaxalone, methadone, methadone, methamphetamine, methocarbamol, methyldopa, methylphenidate, methylsalicylate, methysergid(e), metoclopramide, mianserin, mifepristone, milnacipran, minaprine, mirtazapine, moclobemide, modafinil (an anti-narcoleptic), molindone, morphine, morphine hydrochloride, nabumetone, nadolol, naproxen, naratriptan, nefazodone, neurontin, nomifensine, nortriptyline, olanzapine, olsalazine, ondansetron, opipramol, orphenadrine, oxaflozane, oxaprazin, oxazepam, oxitriptan, oxycodone, oxymorphone, pancrelipase, parecoxib, paroxetine, pemoline, pentazocine, pepsin, perphenazine, phenacetin, phendimetrazine, phenmetrazine, phenylbutazone, phenytoin, phosphatidyl serine, pimozide, pirlindole, piroxicam, pizotifen, pizotyline, pramipexole, prednisolone, prednisone, pregabalin, propanolol, propizepine, propoxyphene, protriptyline, quazepam, quinupramine, reboxitine, reserpine, risperidone, ritanserin, rivastigmine, rizatriptan, rofecoxib, ropinirole, rotigotine, salsalate, sertraline, sibutramine, sildenafil, sulfasalazine, sulindac, sumatriptan, tacrine, temazepam, tetrabenozine, thiazides, thioridazine, thiothixene, tiapride, tiasipirone, tizanidine, tofenacin, tolmetin, toloxatone, topiramate, tramadol, trazodone, triazolam, trifluoperazine, trimethobenzamide, trimipramine, tropisetron, valdecoxib, valproic acid, venlafaxine, viloxazine, vitamin E, zimeldine, ziprasidone, zolmitriptan, Zolpidem, zopiclone and isomers, salts, and combinations thereof.

The additional active agent(s) can be formulated for immediate release, controlled release, or combinations thereof.

EXAMPLES

The present disclosure will now be described in more detail with reference to the following non-limiting examples.

It should be noted that the particular assays used in the examples section are designed to provide an indication of antiviral activity. There are many other assays available to determine the activity of given compounds against RSV and a result in any one particular assay is therefore not determinative.

Example 1: Synthetic experimental procedure

General procedures

All chemicals used in the following experiments were purchased from Sigma Aldrich or Apollo Scientific. All anhydrous solvents used for reaction purposes were obtained from anhydrous septum-sealed DriSolv® bottles. Thin layer chromatography was performed using Merck silica gel 60 F254 coated on aluminium sheets. Visualization was performed with a UV lamp or using common stains such as p-anisaldehyde, ninhydrin (NIN) or a potassium permanganate (ΚΜη0₄) solution followed by gentle heating. Column chromatography was performed using a Teledyne Isco Combiflash® Rf+ automated column machine with Redisep® silica gel-packed columns with one of or combinations of hexane, EtOAc, DCM or MeOH as the mobile phase.

MR spectra (¾, ¹³C) were recorded on a 300 MHz Varian VNMRS (75 MHz for ¹³C), a 400 MHz Varian INOVA (101 MHz for ¹³C), a 400 MHz Varian VNMRS, a 500 MHz Varian INOVA (126 MHz for ¹³C) or a 600 MHz Varian INOVA (150 MHz for ¹³C). Chemical shifts (δ) are reported in ppm and J - values are given in Hz. Multiplicities are reported as a singlet (s), doublet (d), triplet (t), quartet (q), multiplet (m) or doublet of doublet (dd). Chemical shifts were recorded using the residual solvent peak or external reference. All spectra were obtained at 25 °C unless otherwise reported. Spectroscopic data were processed using MestReNova v6.0.2. Mass spectrometry was performed by the Emory University Mass Spectroscopy Center.

All reactions requiring inert conditions were carried out under a positive atmosphere of argon. All glassware was flame-dried while under vacuum or oven dried overnight before purging with argon. Standard Schlenk techniques were employed when necessary. Solvents were removed using a rotary evaporator followed by the removal of trace amounts of solvent using a high vacuum pump at ca. 0.08 mm Hg.

4-(2-(Hydrox methyl)-lH-benzo[d]imidazol-l-yl)butanenitrile

Into a 100 ml three neck RB flask containing Ar(g) was placed DMF (40 ml), followed by (lH-benzo[d]imidazol-2-yl)methanol (2.20 g, 14.9 mmol), thus forming a pale orange solution. This was cooled to 0 °C and 60% sodium hydride (0.772 g, 19.3 mmol) was added in one portion. After stirring for 10 min under Ar(g) at 0 °C, 4-bromobutanenitrile (2.64 g, 17.8 mmol) was added in one portion. The reaction was allowed to warm to rt and left to proceed for 18 h. The reaction mixture was diluted with DCM and then poured onto a brine solution. The organic layer was separated and the aqueous phase was extracted three times with DCM. The combined organic fractions were dried over anhydrous sodium sulfate and then concentrated in vacuo. Purification by column chromatography (100% DCM to 3% MeOH/DCM) afforded the desired product as a viscous oil which solidified to a white solid over a period of hours (2.99 g, 13.9 mmol, 94%).

¾ NMR (400 MHz, DMSO-d6) δ 7.60-7.53 (m, 2H), 7.26 - 7.19 (m, 1H), 7.19 - 7.13 (m, 1H), 5.65 (t, J = 5.8 Hz, 1H), 4.70 (d, J = 5.8 Hz, 2H), 4.39 - 4.25 (m, 2H), 2.56 (t, J = 7.3 Hz, 2H), 2.15 - 2.01 (m, 2H). 4-(2-(Chloromethyl)-lH-benzo d]imidazol-l-yl)butanenitrile

Into a three neck RB flask containing Ar(g) was placed DCM (10 ml), followed by 4-(2- (hydroxymethyl)-lH-benzo[d]imidazol-l-yl)butanenitrile (500 mg, 2.32 mmol), thus forming a clear solution. This was cooled to 0 °C using an ice bath and then SOCh (359 mg, 3.02 mmol) was added in one portion against a flow of Ar(g), followed by triethylamine. The ice bath was removed after 5 min and the reaction was left to proceed at rt for 3 h. The reaction mixture was then concentrated in vacuo to afford a white solid. This was then taken up into DCM and washed with a saturated NaHCCb solution. After separation, the aqueous phase was extracted three times with DCM. The combined organic fractions were concentrated in vacuo. Purification by column chromatography (3% MeOH/DCM) afforded a clear viscous oil, which solidified to a white solid under vacuum for several hours (436 mg, 1.87 mmol, 80%).

¾ MR (400 MHz, CDCh) δ 7.75 - 7.70 (m, 1H), 7.38 - 7.23 (m, 3H), 4.81 (s, 2H), 4.34 (t, J = 7.4 Hz, 2H), 2.40 (t, J = 6.9 Hz, 2H), 2.29 - 2.19 (m, 2H). (l-(4-((Tert-butyl imethylsilyl)oxy)butyl)-lH-benzo[d]imidazol-2-yl)methanol

Into a three neck RB flask containing Ar(g) was placed (lH-benzo[d]imidazol-2- yl)methanol (2.00 g, 13.5 mmol), followed by DMF (40 ml), thus forming a pale yellow solution. To this was added potassium carbonate (3.73 g, 27.0 mmol) and then tert-butyl(4-iodobutoxy)- dimethylsilane (5.09 g, 16.2 mmol). The reaction was left to stir under Ar(g) at rt for 18 h before being diluted with water and then EtOAc. After separation of the organic phase, the aqueous phase was extracted three times with EtOAc and the combined organic fractions were dried over anhydrous sodium sulfate. After evaporation of the solvent in vacuo, the crude material was purified by column chromatography (EtO Ac/Hex) to afford the desired product as a viscous oil (2.99 g, 8.94 mmol, 66%).

¹H MR (400 MHz, CDCh) δ 7.68 - 7.53 (m, 1H), 7.35 - 7.13 (m, 3H), 4.83 (s, 2H), 4.22 (t, J = 7.6 Hz, 2H), 3.59 (t, J = 6.0 Hz, 2H), 1.97 - 1.79 (m, 2H), 1.64 - 1.45 (m, 2H), 0.85 (s, 9H), 0.01 (s, 6H). l-(4-((Tert-butyldimeth lsilyl)oxy)butyl)-2-(chloromethyl)-lH-benzo[d]imidazole

Into a three neck RB flask containing Ar(g) was placed DCM (20 ml), followed by (l-(4- ((tert-butyldimethylsilyl)oxy)butyl)-lH-benzo[d]imidazol-2-yl)methanol (1.70 g, 5.08 mmol), thus forming a clear solution. This was cooled to 0 °C using an ice bath and then SOCh (0.786 g, 6.61 mmol) was added in one portion against a flow of Ar(g), followed by triethylamine (1.03 g, 10.2 mmol). The ice bath was removed after 5 min and the reaction was left to proceed at rt for 3 h. The reaction mixture was then concentrated in vacuo to afford a white solid. This was then taken up into DCM and washed with saturated NaHCCb solution. The organic phase was separated and the aqueous phase was extracted three times with DCM. Purification by column chromatography (EtO Ac/Hex) afforded a clear viscous oil (0.917 mg, 2.60 mmol, 51%) which was used directly in the next reaction without further characterization. 3-((l-(4-((Tert-butyldimethylsilyl)oxy)butyl)-lH-benzo[d]imidazol-2-yl)methyl)-l- cyclopro l-l,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one

Into a three neck RB flask containing Ar(g) was placed DMF (10 ml) followed by l-(4- ((tert-butyldimethylsilyl)oxy)butyl)-2-(chloromethyl)-lH-benzo[d]imidazole and CS2CO3 (1.08 g, 3.31 mmol). To this was added l-cyclopropyl-lH-imidazo[4,5-c]pyridin-2(3H)-one in one portion (0.536 g, 3.06 mmol) and the reaction was heated to 50 °C for 18 h. After this the reaction was diluted with water and EtOAc. The organic phase was separated and the aqueous phase was extracted three times with EtOAc. The combined organic fractions were dried over anhydrous sodium sulfate and filtered. After concentration in vacuo, the crude product was purified by column chromatography (EtOAc/Hex) to afford the desired product as a white solid (0.438 g, 0.890 mmol, 35%).

¾ NMR (400 MHz, CDCh) δ 8.66 (s, 1H), 8.28 (d, J = 5.3 Hz, 1H), 7.78 - 7.69 (m, 1H), 7.35 - 7.18 (m, 3H), 7.08 (d, J = 5.3 Hz, 1H), 5.34 (s, 2H), 4.31 (t, J = 7.5 Hz, 2H), 3.54 (t, J = 6.1 Hz, 2H), 2.89 (tt, J = 7.0, 3.6 Hz, 1H), 1.75 - 1.62 (m, 2H), 1.56 - 1.44 (m, 2H), 1.18 - 1.09 (m, 2H), 1.02 - 0.93 (m, 2H), 0.82 (s, 9H), -0.03 (t, J = 1.0 Hz, 6H). l-Cyclopropyl-3-((l-(4-hydroxybutyl)-lH-benzo[d]imidazol-2-yl)methyl)-l,3-dihyd imidazo [ -c] pyridin-2-one

Into a 100 ml three neck RB flask containing anhydrous THF and Ar(g) was placed 3-((l- (4-((tert-butyldimethylsilyl)oxy)butyl)-lH-benzo[d]imidazol-2-yl)methyl)-l-cyclopropyl-lH- imidazo[4,5-c]pyridin-2(3H)-one (400 mg, 0.814 mmol) thus forming a clear solution. To this was added a TBAF solution (0.90 ml, 0.90 mmol) and the reaction was left to proceed under Ar(g) at rt for 18 h. The reaction mixture was then diluted with water and EtOAc and the organic fraction was separated. The aqueous phase was extracted three times with EtOAc and then the combined organic fractions were dried over anhydrous sodium sulphate. After concentrating in vacuo, the crude product was purified by column chromatography to afford the desired product as a white solid (181 mg, 0.480 mmol, 59%).

¾ NMR (400 MHz, DMSO-d6) δ 8.39 (s, 1H), 8.24 (d, J = 5.2 Hz, 1H), 7.57 (t, J = 8.6 Hz, 2H), 7.32 - 7.20 (m, 2H), 7.17 (t, J = 7.6 Hz, 1H), 5.38 (s, 2H), 4.49 (t, J = 5.0 Hz, 1H), 4.33 (t, J = 7.4 Hz, 2H), 3.06 - 2.92 (m, 1H), 2.50 (s, 3H), 1.70 - 1.57 (m, 2H), 1.50 - 1.37 (m, 2H), 1.12 - 1.02 (m, 2H), 0.98 - 0.87 (m, 2H). Note, 2H m hidden under water signal. ¹³C NMR (101 MHz, DMSO-d6) δ 153.28, 149.20, 143.15, 142.27, 136.57, 135.63, 129.85, 126.63, 122.91, 122.14, 119.50, 110.96, 104.55, 60.68, 43.61, 38.16, 29.94, 26.75, 23.07, 6.05.

4-(2-((2'-oxospiro[cyclopropane-l,3'-pyrrolo[2,3-c]pyridin]-l'(2'H)-yl)methyl)-lH- benzo[d]imidazol-l-yl)butanenitrile (TK3-118)

To a solution of spiro[cyclopropane-l,3'-pyrrolo[2,3-c]pyridin]-2'( H)-one (100 mg, 0.624 mmol) and 4-(2-(chloromethyl)-lH-benzo[d]imidazol-l-yl)butanenitrile (0.146 g, 0.624 mmol) in CH3CN (4.2 ml) was added CS2CO3 at rt. The reaction was stirred at rt and followed by TLC (18 h to completion). The reaction mixture was filtered and washed with MeOH (-50 ml). The filtrate was concentrated in vacuo. The residue was purified by combiflash (0 to 10% MeOH in DCM) to give the desired compound as a beige solid (0.167 g, 75% yield).

¾ MR (400 MHz, CDCh) δ 8.82 (s, 1H), 8.34 (d, J = 4.8 Hz, 1H), 7.81-7.76 (m, 1H),

7.38-7.27 (m, 3H), 6.82 (d, J = 5.2 Hz, 1H), 5.33 (s, 2H), 4.41 (t, J = 7.6 Hz, 2H), 2.43 (t, J = 7.2 Hz, 2H), 2.06-1.99 (m, 2H), 1.94-1.91 (m, 2H), 1.74-1.71 (m, 2H). ¹³C NMR (100 MHz, CDCh) δ 175.6, 147.6, 144.8, 142.3, 139.5, 135.0, 131.2, 123.8, 122.9, 120.4, 118.5, 113.5, 109.5, 42.4, 38.1, 27.2, 6.0, 20.8, 14.7. HRMS (EI-) m/z calculated for C21H20N5O [M+H⁺]⁺: 358.16624, found: 358.16575.

(l-(4-((tert-butyldimethylsilyl)oxy)butyl)-lH-benzo[d]imidazol-2-yl)methanol (ZD-2-125)

An oven-dried 100 ml three neck RB flask was charged with Ar(g), (lH-benzo[d]imidazol-

2-yl)methanol (300 mg, 2.025 mmol), and anhydrous DMF (10 ml) to give a pale orange solution. To this was added potassium carbonate (420 mg, 3.04 mmol) in one portion, followed by tert- butyl(4-iodobutoxy)dimethylsilane (0.629 ml, 2.430 mmol). The reaction was left to stir under Ar(g) at rt for -18 h. The reaction was diluted with DCM and washed with brine solution before being extracted with DCM (2x50 ml). The organic extract was dried over sodium sulfate, filtered and concentrated to -0.865 g of lightly yellow oil, which was loaded onto a 12g RediSep CombiFlash silica gel column for purification. Product was eluted with a gradient of 0-10% MeOH in DCM over -23 minutes, and desired product was isolated as well as unreacted starting material (0.515 g, 76% yield).

¾ MR (500 MHz, CDCh) δ 7.65 (d, J = 8.0 Hz, 1H), 7.30 (d, J = 7.0 Hz, 1H), 7.24-7.19

(m, 2H), 4.86 (s, 2H), 4.25 (t, J = 7.5 Hz, 2H), 3.62 (t, J = 6.0 Hz, 2H), 1.92 (p, J = 7.5 Hz, 2H), 1.57 (p, J = 6.0 Hz, 2H), 0.88 (s, 9H), 0.03 (s, 6H) ppm. ¹³C NMR (125 MHz, CDCh) δ 154.0, 141.6, 135.1, 122.9, 122.2, 119.3, 110.0, 62.5, 56.6, 43.9, 30.0, 26.7, 26.0, 18.4, -5.22, -5.25, ppm. HRMS (ESI+) m/z: [M + H]⁺ Calcd for C18H31N2O2S1 335.2149; Found 335.2144. l-(4-((tert-butyldimethylsilyl)oxy)butyl)-2-(chloromethyl)-lH-benzo[d]imidazole (ZD-2- 158)

An oven-dried three neck 100 ml RB flask was charged with Ar(g), (l-(4-((tert- butyldimethylsilyl)oxy)butyl)-lH-benzo[d]imidazol-2-yl)methanol (0.718 g, 2.146 mmol), and benzene (10.73 ml) to give a light green solution. Triethylamine (0.449 ml, 3.22 mmol) was then added, which turned the solution completely colourless, and the solution was stirred briefly before thionyl chloride (0.282 ml, 3.86 mmol) was added dropwise under Ar(g), immediately releasing a gas and turning the solution light red. The reaction was allowed to stir at rt for -3.5 h when monitoring by TLC showed spot-to-spot conversion of starting material to the desired less polar spot. The reaction was diluted with DCM before being quenched with saturated sodium bicarbonate solution, and the product was extracted with DCM (2x50 ml). The organic extract was dried over sodium sulfate, filtered and concentrated to give the desired product as a red oil (0.702 g, 93% yield).

¹H MR (600 MHz, CDCh) δ 7.78-7.75 (m, 1H), 7.39 (dd, J = 6.6, 0.6 Hz, 1H), 7.33-7.27 (m, 2H), 4.86 (s, 2H), 4.30 (t, J = 7.8 Hz, 2H), 3.68 (t, J = 6.0 Hz, 2H), 2.02-1.97 (m, 2H), 1.65- 1.60 (m, 2H), 0.89 (s, 9H), 0.05 (s, 6H) ppm. ¹³C MR (150 MHz, CDCh) δ 148.8, 135.5, 128.5, 123.7, 122.8, 120.5, 110.2, 62.5, 44.5, 36.9, 30.1, 26.8, 26.1, 18.4, -5.2 ppm. HRMS (ESI+) m/z: [M + H]⁺ Calcd for CisftoClNiOSi 353.1810; Found 353.1811. l'-((l-(4-((tert-butyldimethylsilyl)oxy)butyl)-lH-benzo[d]imidazol-2-yl)methyl)spiro

[cyclopropane-l,3'-pyrrolo[2,3-c] -2-159)

An oven-dried 50 ml RB flask was charged with Ar(g), spiro[cyclopropane-l,3'- pyrrolo[2,3-c]pyridin]-2'(l'H)-one (0.1 g, 0.624 mmol) and anhydrous acetonitrile (2.51 ml, Ratio: 2) to give a brown suspension; l-(4-((tert-butyldimethylsilyl)oxy)butyl)-2-(chloromethyl)-lH- benzo[d]imidazole (1.4 ml, 0.560 mmol) was then added as a -0.45 M solution in anhydrous acetonitrile (1.257 ml, Ratio: 1.000), followed by cesium carbonate (0.199 g, 0.610 mmol). The reaction stirred at 45 °C for -2.5 h when TLC (95:5 DCM:MeOH) showed complete conversion of the starting chloride to a more polar fluorescent spot with some pyrrolidinone remaining. The reaction mixture was filtered and rinsed with MeOH before the filtrate was adsorbed onto -3 g of Celite to dry load a 12g RediSep CombiFlash silica gel column for purification. Product was eluted with 100% DCM for -4 minutes followed by a slow gradient of 0-10% MeOH in DCM over -35 minutes. The desired product eluted at -26 minutes with -8% MeOH (0.103 g, 39% yield).

¾ MR (600 MHz, CDCh) δ 8.73 (s, 1H), 8.28 (d, J = 4.8 Hz, 1H), 7.76-7.73 (m, 1H),

7.33-7.30 (m, 1H), 7.25-7.22 (m, 2H), 6.75 (dd, J = 4.8, 0.6 Hz, 1H), 5.31, (s, 2H), 4.26 (t, J = 7.8 Hz, 2H), 3.56 (t, J = 6.6 Hz), 1.88 (dd, J = 7.8, 4.2 Hz, 2H), 1.72-1.67 (m, 2H), 1.65 (dd, J = 7.8, 4.2 Hz, 2H), 1.54-1.49 (m, 2H), 0.84 (s, 9H), -0.02 (s, 6H) ppm; ¹³C NMR (150 MHz, CDCh) δ 175.4, 147.8, 144.7, 142.5, 139.6, 139.5, 135.5, 131.4, 123.3, 122.4, 120.3, 113.4, 1 10.1, 62.5, 44.0, 38.3, 29.9, 27.2, 26.7, 26.0, 20.7, 18.4, -5.2 ppm; HRMS (ESI+) m/z: [M + Na]⁺ Calcd for C27H3₆N40₂SiNa 499.2500; Found 499.2497. l'-((l-(4-hydroxybutyl)-lH-benzo[d]imidazol-2-yl)methyl)spiro[cyclopropane-l,3'- pyrrolo[2,3-c]pyridin]-2'(l'H)-one -2-160)

To a stirring solution of l'-((l-(4-((tert-butyldimethylsilyl)oxy)butyl)-lH- benzo[d]imidazol-2-yl)methyl)spiro[cyclopropane-l,3' -pyrrolo[2,3-c]pyridin]-2'(l'H)-one (0.15 g, 0.315 mmol) in anhydrous THF (1.573 ml) was dropwise added tetra-n-butylammonium fluoride (0.503 ml, 0.503 mmol) as a ~1.0 M solution in THF under Ar(g) , and the reaction stirred at rt overnight. The following morning, the Ar(g) inlet had evaporated the solvent, leaving a beige solid. The solid was dissolved in MeOH and adsorbed onto ~1.7 g Celite to dry load a 12g RediSep CombiFlash silica gel column for purification. Product was eluted in with 100% DCM for 2 minutes followed by a gradient of 0-20% MeOH in DCM over 20 minutes, with the major product eluting at -18 minutes with -12% MeOH. The relevant fractions were combined and concentrated to -0.1 g of white solid which was subsequently washed with deionized H2O, collected and dried in vacuo to afford the desired final compound (96 mg, 84% yield).

¾ MR (500 MHz, de-DMSO) δ 8.42 (s, 1H), 8.25 (d, J = 5.0 Hz, 1H), 7.60 (d, J = 8.0

Hz, 1H), 7.56 (d, J = 8.0 Hz, 1H), 7.26-7.22 (m, 1H), 7.19-7.16 (m, 2H), 5.35 (s, 2H), 4.47 (t, J = 5.0 Hz, 1H), 4.31 (t, J = 8.0 Hz, 2H), 3.39 (dd, J = 11.5, 6.0 Hz, 2H) 1.85 (dd, J = 8.5, 4.5 Hz, 2H), 1.72 (dd, J = 7.5, 3.0 Hz, 2H), 1.68-1.62 (m, 2H), 1.48-1.42 (m, 2H) ppm; ¹³C NMR (125 MHz, de-DMSO) δ 174.8, 148.6, 143.7, 141.9, 139.4, 139.1, 135.2, 130.0, 122.4, 121.7, 119.1, 114.5,110.5, 60.3, 43.2, 37.1, 29.5, 26.8, 26.3, 20.0 (x2) ppm. HRMS (ESI+) m/z: [M + Na]⁺ Calcd for

386.1698; Found 386.1692.

The following procedures have been adapted from PCT Pub No. WO2016/022644. 3-Nitro-N-(tetrahydro-2H-pyran-4-yl)pyridin-4-amine (RJW-2-002)

To a 75 ml sealed flask with teflon screw cap was added 4-methoxy-3-nitropyridine (6.86 g, 44.5 mmol), tetrahydro-2H-pyran-4-amine (3.07 ml, 29.7 mmol), absolute EtOH (49.4 ml) and Hunig's Base (7.77 ml, 44.5 mmol) then the vessel was tightly sealed and heated in a 90 °C oil bath for 18 h. The reaction was cooled to 0 °C and the yellow precipitate was collected and washed with hexanes. The solid was dried in vacuo (2.0 g, 8.96 mmol, 30.2% yield).

¹H NMR (600 MHz, DMSO-de) δ 9.03 (s, 1H), 8.27 (dd, J = 6.2, 0.8 Hz, 1H), 8.02 (d, J = 7.9 Hz, 1H), 7.15 (d, J = 6.3 Hz, 1H), 3.96 - 3.90 (m, 1H), 3.87 (ddd, J = 12.1, 4.2, 2.7 Hz, 2H), 3.47 (td, J = 11.6, 2.2 Hz, 2H), 1.89 (ddd, J = 12.6, 4.5, 2.2 Hz, 2H), 1.71 - 1.59 (m, 2H).

N4-(tetrahydro-2H-pyran-4-yl)pyridine-3,4-diamine (RJW-2-005)

3-Nitro-N-(tetrahydro-2H-pyran-4-yl)pyridin-4-amine (1.14 g, 5.11 mmol) was hydrogenated in anhydrous MeOH (100 ml) in the presence of 10% palladium on carbon (0.8 g, 7.52 mmol) at 40 psi for 40 minutes. The mixture was filtered through celite and washed with MeOH then concentrated to afford N4-(tetrahydro-2H-pyran-4-yl)pyridine-3,4-diamine as a light orange solid. The light orange solid was used in the next step without purification.

¾ MR (400 MHz, Methanol-d₄) δ 7.68 (s, 1H), 7.65 (d, J = 5.6 Hz, 1H), 6.55 (d, J = 5.7 Hz, 1H), 4.01 - 3.92 (m, 2H), 3.63 (tt, J = 10.6, 4.1 Hz, 1H), 3.55 (td, J = 11.7, 2.1 Hz, 2H), 2.00 (ddd, J = 12.8, 4.4, 2.2 Hz, 2H), 1.63 - 1.48 (m, 2H). l-(Tetrahydro-2H- -2-010)

To a 50 ml RB flask was added N4-(tetrahydro-2H-pyran-4-yl)pyridine-3,4-diamine (0.595 g, 3.08 mmol), MeCN (20 ml) and CDI (0.549 g, 3.39 mmol) at rt. The reaction was heated to reflux for 3 h, cooled to rt and concentrated in vacuo. The residue was dissolved in DCM, washed with water, dried with Na₂S0₄, filtered and concentrated. The residue was purified via combiflash (5:95 to 30:70 MeOH:EtOAc) to afford l-(tetrahydro-2H-pyran-4-yl)-lH- imidazo[4,5-c]pyridin-2(3H)-one (0.140 g, 0.639 mmol, 21% yield).

¾ NMR (400 MHz, DMSO-de) δ 8.19 (s, 1H), 8.14 (d, J = 5.3 Hz, 1H), 7.32 (d, J = 5.3 Hz, 1H), 4.41 (tt, J = 12.3, 4.2 Hz, 1H), 3.98 (dd, J = 11.5, 4.5 Hz, 2H), 3.46 (td, J = 12.0, 1.9 Hz, 2H), 2.32 (qd, J = 12.4, 4.6 Hz, 2H), 1.70 - 1.60 (m, 2H). LC/MS 10-95% MeOH in H₂0 over 6 minutes, r_t= 0.757 at 254 nM, MS (+) 220.2.

4-(2-((2-oxo-l-(tetrahydro-2H-pyran-4-yl)-l,2-dihydro-3H-imidazo[4,5-c]pyridin-3- yl)meth l)-lH-benzo[d]imidazol-l-yl)butanenitrile (RJW-2-011)

To a 15 ml tube was added cesium carbonate (0.250 g, 0.766 mmol), l-(tetrahydro-2H- pyran-4-yl)-lH-imidazo[4,5-c]pyridin-2(3H)-one (0.140 g, 0.639 mmol) and 4-(2- (chloromethyl)-lH-benzo[d]imidazol-l-yl)butanenitrile (0.157 g, 0.670 mmol) dissolved in MeCN (8 ml) then the vessel was sealed with a teflon screw cap and heated in a 70 °C oil bath overnight. The reaction was cooled to rt, filtered over celite with MeOH and concentrated in vacuo. The residue was purified by column (0-10% MeOH in DCM). The fractions were concentrated in vacuo, triturated with CHCh then PhMe and dried in vacuo overnight. The solid (168 mg) was recrystallized using MeOH (7 ml) to obtain white needles (88 mg) co-crystallized with MeOH (X- Ray).

¾ MR (600 MHz, CDCh): δ = 8.83 (s, 1H), 8.31 (d, J= 5.4 Hz, 1H), 7.79 (dd, J= 7.0,

1.6 Hz, 1H), 7.36 (dd, J= 6.8, 1.4 Hz, 1H), 7.31 (m, 2H), 7.13 (dd, J= 5.4, 0.6 Hz, 1H), 5.39 (s, 2H),4.59 (m, 1H), 4.48 (dd, J= 7.5, 7.6 Hz, 2H), 4.15 (dd, J= 11.5, 4.5 Hz, 2H), 3.56 (dt, J= 12.2,

1.7 Hz, 2H), 3.48 (d, J= 5.2, 2H, MeOH), 2.47-2.40 (m, 4H), 2.06 (p, J= 6.9 Hz, 2H), 1.78 (dd, J= 12.4, 2.6 Hz, 2H), 1.08 (q, J= 5.5 Hz, 0.66H, MeOH); ¹³C NMR (150 MHz, CDCh): δ = 152.8, 147.6, 143.7, 142.5, 135.1, 134.3, 131.4, 126.4, 124.0, 123.0, 120.7, 118.4, 109.5, 104.5, 67.4, 50.9, 50.8, 42.5, 38.7, 30.1, 26.2, 14.8; LC/MS 10-95% MeOH in H₂0 over 6 minutes, r_t= 3.523 at 254 nM, MS (+) 417.2, 20-95% MeOH in H2O over 8 minutes, r_t = 2.459 at 254 nM, MS (+) 417.0.

Tert-butyl thietan-3-ylcarbamate

As in N. K. Thong et al. Bioorg. Med. Chem. Lett., 2009, 19, 3832-5 and M. Muehle-Bach et al. WO 2007/080131; tert-butyl thietan-3-ylcarbamate (10.49 g, white solid) was synthesized from 2-aminopropane-l,3-diol in three steps and 48 % overall yield.

¾ NMR (400 MHz, CDCh): 5.08 (s, 1H), 5.01 - 4.90 (m, 1H), 3.31 (d, J = 8.5 Hz, 4H), 1.42 (s, 9H).¹³C NMR (100 MHz, CDCh): 153.89, 79.88, 48.48, 36.38, 28.27.

Tert-butyl (l,l-dioxidothietan-3-yl)c

As in J. A. Burkhard et al. Org. Lett., 2010, 12, 1944-1947; a 1 L RB flask equipped with a magnetic stir bar and rubber septum was charged with 10.36 g of tert-butyl thietan-3-yl carbamate (54.7 mmol, 1 equiv.) and 456 ml of DCM. Then 16.0 ml of Ti(OiPr)₄ was added dropwise, followed by 22.4 ml of H2O2 (219 mmol, 4 equiv.) at 0 °C. After stirring at 0 °C for 15 min, the reaction was allowed to warm to rt and the stirring was continued for 1 h. The reaction mixture was quenched by addition of water and the product was extracted with DCM (3x) and dried over Na₂S0₄. The organics were concentrated and the crude product was purified on silica gel column (40 g) using 0 to 20% EtOAc as eluent affording 2.17 g (still some impurity) and 8.00 g (10.17 g, 84%) of tert-butyl (l, l-dioxidothietan-3-yl)carbamate as a white solid.

¾ MR (400 MHz, CDCh): 5.19 (s, 1H), 4.53 - 4.43 (m, 3H), 4.02 - 3.92 (m, 2H), 1.45 (s, 9H). LC-MS (ESI- API, 254 nm) 95 % MeOH in H₂0 (0.1% HCO2H), 3 min, 1.00 ml/min, C18 (Agilent Zorbax XDB-18, 50 mm x 4.6 mm, 3.5 μιη), m/z = 244.0 (M + Na), t = 0.546 min (MS signal).

3-Aminothietane 1,1-dioxide hydrochl ride

A 500 ml RB flask equipped with a stirrer bar was charged with 8.00 g of tert-butyl (1, 1- dioxidothietan-3-yl)carbamate (36.2 mmol, 1 equiv.), 12.1 ml of 12 M HCl (145 mmol, 4 equiv.) and 181 ml of dioxane. After stirring at rt for 12 h, the suspension was concentrated affording 5.63 g (99%) of 3-aminothietane 1, 1-dioxide hydrochloride as a white solid.

¾ MR (400 MHz, DMSO-de) δ 8.85 (s, 3H), 4.56 - 4.48 (m, 2H), 4.44 - 4.36 (m, 2H), 4.09 (tt, J = 9.0, 6.3 Hz, 1H). LC-MS (ESI-API, 254 nm) 75-95 % MeOH in H2O (0.1% HCO2H), 3 min, 1.00 ml/min, C18 (Agilent Zorbax XDB-18, 50 mm x 4.6 mm, 3.5 μιη), m/z = 122.0 (M + H), t = 0.499 min (MS signal).

3-((3-Nitropyridin-4-yl)amino)thieta -dioxide

A 100 ml Schlenk tube equipped with a magnetic stirrer bar and cold finger condenser was charged with 2.67 g of 3-aminothietane 1, 1-dioxide hydrochloride (16.9 mmol, 1.1 equiv.), 5.65 ml of DIPEA (32.3 mmol, 2.1 equiv.) and 20 ml of abs. EtOH. The suspension was stirred at rt until the amine was dissolved. Then 2.44 g of 4-chloro-3-nitropyridine (15.4 mmol, 1 equiv.) was added and 5.0 ml of EtOH was used to wash in 4-chloro-3-nitropyridine. After heating at refluxing temperature for 5 h, 1.75 g of NaOH (32.3 mmol, 2.1 equiv.) dissolved in 5 ml of MeOH was added, stirred for 12 h and the product was filtered and washed with water. The product was dispensed in water and filtered again affording 2.29 g (61%) of 3-((3-nitropyridin-4- yl)amino)thietane 1,1 -dioxide as a bright yellow solid.

¾ MR (400 MHz, DMSO-de) δ 9.07 (s, 1H), 8.45 (d, J = 5.9 Hz, 1H), 8.37 (d, J = 6.1 Hz, 1H), 7.00 (d, J = 6.1 Hz, 1H), 4.74 - 4.67 (m, 2H), 4.65 - 4.56 (m, 1H), 4.55 - 4.49 (m, 2H). LC-MS (ESI-API, 254 nm) 75-95 % MeOH in H₂0 (0.1% HCO2H), 3 min, 1.00 ml/min, C18 (Agilent Zorbax XDB-18, 50 mm x 4.6 mm, 3.5 μιη), m/z = 244.0 (M + H), 266.0 (M + Na), t = 0.557 min.

3-((3-Aminopyridin-4-yl)amino)thiet -dioxide

A 500 ml Parr shaker bomb was charged with 2.24 g of 3-((3-nitropyridin-4- yl)amino)thietane 1,1-dioxide (9.21 mmol, 1 equiv.), 0.098 g of 10 w% palladium on carbon (0.092 mmol, 0.01 equiv.) and 23 ml of MeOH and the bottle was installed on the Parr shaker. After shaking at rt and 40 psi H2 pressure for 3 h, white precipitate was formed. More MeOH was added to dissolve the product and the suspension was filtered through a celite plug. The plug was slowly washed with MeOH, water and ether. The organics were concentrated and the product was dissolved in dioxane and toluene. After concentration of the organics, the product was dried under high vacuum for 12 h affording 1.97 g (100 %) of 3-((3-aminopyridin-4-yl)amino)thietane 1, 1- dioxide as a beige solid.

¾ NMR (400 MHz, DMSO-de) δ 7.71 (s, 1H), 7.62 (d, J = 5.2 Hz, 1H), 6.29 (d, J = 5.3 Hz, 1H), 5.99 (d, J = 5.5 Hz, 1H), 4.74 - 4.66 (m, 4H), 4.24 (td, J = 8.4, 4.3 Hz, 1H), 4.11 - 4.04 (m, 2H). LC-MS (ESI-API, 254 nm) 75-95 % MeOH in H2O (0.1% HCO2H), 3 min, 1.00 ml/min, C18 (Agilent Zorbax XDB-18, 50 mm x 4.6 mm, 3.5 μιη), m/z = 214.0 (M + H), t = 0.490 min. l-(l,l-Dioxidothietan-3-yl)-l,3-dihyd -2H-imidazo[4,5-c]pyridin-2-one

A 100 ml Schlenk tube equipped with a magnetic stirrer bar and cold finger condenser was charged with 400 mg of 3-((3-aminopyridin-4-yl)amino)thietane 1,1-dioxide (1.88 mmol, 1 equiv.) and 18.8 ml of acetonitrile and heated to 80 °C to dissolve the starting material. The heating was removed and 608 mg of CDI (3.75 mmol, 2 equiv.) dissolved in 18.8 ml of acetonitrile was added. After heating at 80 °C for 12 h, the organics were concentrated, filtered and washed with DCM affording 345 mg (77%) of l-(l, l-dioxidothietan-3-yl)-l,3-dihydro-2H-imidazo[4,5- c]pyridin-2-one as a brownish orange solid.

¾ MR (400 MHz, DMSO-de) 611.41 (s, 1H), 8.24 (d, J = 5.0 Hz, 1H), 8.23 (s, 1H), 7.43 (d, J = 5.4 Hz, 1H), 5.35 (tt, J = 9.8, 7.4 Hz, 1H), 5.02 - 4.94 (m, 2H), 4.67 - 4.58 (m, 2H). LC- MS (ESI-API, 254 nm) 75-95 % MeOH in H₂0 (0.1% HCO2H), 3 min, 1.00 ml/min, C18 (Agilent Zorbax XDB-18, 50 mm x 4.6 mm, 3.5 μιη), m/z = 240.0 (M + H), 262.0 (M + Na), t = 0.495 min.

4-(2-((l-(l,l-dioxidothietan-3-yl)-2-oxo-l,2-dihydro-3H-imidazo[4,5-c]pyridin-3-yl)n lH-benzo[d]imidazol-l-yl)butanenitrile

A 50 ml Schlenk tube equipped with a magnetic stirrer bar and cold finger condenser was charged with 160 mg of l-(l,l-dioxidothietan-3-yl)-l,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one (0.669 mmol, 1 equiv.), 327 mg of CS2CO3 (1.00 mmol, 1.5 equiv.), 11.0 mg of KI (0.067 mmol, 0.1 equiv.), 188 mg of 4-(2-(chloromethyl)-lH-benzo[d]imidazol-l-yl)butanenitrile (0.803 mmol, 1.2 equiv.) and 6.7 ml of DMF. After heating at 80 °C for 8 h, the reaction mixture was quenched by addition of water, extracted with DCM (3x) and dried over Na₂S04. The crude product was purified on silica gel column (120 g) using 0 to 10% MeOH in DCM as eluent affording 89 mg (31%) of 4-(2-((l-(l, l-dioxidothietan-3-yl)-2-oxo-l,2-dihydro-3H-imidazo[4,5-c]pyridin-3- yl)methyl)-lH-benzo[d]imidazol-l-yl)butanenitrile. The product was recrystallized from EtOAc and MeOH affording slightly brown crystals which were submitted for X-ray crystal structure determination.

¾ MR (400 MHz, CDCh) δ 8.87 (d, J = 0.8 Hz, 1H), 8.45 (d, J = 5.4 Hz, 1H), 7.80 - 7.75 (m, 1H), 7.41 (dd, J = 5.4, 0.8 Hz, 1H), 7.39 - 7.28 (m, 3H), 5.50 - 5.37 (m, 1H), 5.40 (s, 2H), 4.93 - 4.83 (m, 2H), 4.69 - 4.58 (m, 2H), 4.49 (dd, J = 8.1, 6.9 Hz, 1H), 3.49 (d, J = 5.4 Hz, 1.3H, MeOH), 2.45 (t, J = 7.0 Hz, 2H), 2.15 (p, J = 7.1 Hz, 2H), 0.97 (q, J = 5.5 Hz,0.43H, MeOH). ¹³C NMR (100 MHz, CDCh) δ 152.36, 146.91, 144.46, 142.36, 134.86, 132.91, 131.80, 126.27, 124.04, 123.09, 120.60, 118.35, 109.52, 103.72, 68.73, 42.38, 38.70, 33.47, 25.88, 14.67. HRMS (ESI+) calcd for C21H21N6O3S ([M+H]⁺): 437.1390. Found: 437.1386, error 0.5 ppm; calcd for C2iH₂oN₆03SNa ([M+Na]⁺): 4591210. Found: 459.1216, error 0.6 pp; LC-MS (ESI-API, 254 nm) 50-95 % MeOH in H2O (0.1% HCO2H), 6 min, 1.00 ml/min, C18 (Agilent Zorbax XDB-18, 50 mm x 4.6 mm, 3.5 μιη), m/z = 437.0 (M + H), 459.0 (M + Na), t = 0.569 min.

3-((l-(4-((Tert-butyldimethylsilyl)oxy)butyl)-lH-benzo[d]imidazol-2-yl)methyl)-l-(l,l- dioxidothietan-3-yl)-l,3-dihydro-2H-imi zo[4,5-c]pyridin-2-one

A 100 ml Schlenk tube equipped with a magnetic stirrer bar and cold finger condenser was charged with 150 mg of l-(l,l-dioxidothietan-3-yl)-l,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one (0.627 mmol, 1 equiv.), 306 mg of CS2CO3 (0.940 mmol, 1.5 equiv.) and 8.0 ml of acetonitrile and warmed up to 80 °C to dissolve the starting material. The heating was removed and 266 mg of 1- (4-((tert-butyldimethylsilyl)oxy)butyl)-2-(chloromethyl)-lH-benzo[d]imidazole (0.752 mmol, 1.2 equiv.) dissolved in 4.5 ml of acetonitrile was added. After heating at 85 °C for 12 h, MeOH and DCM was added and the suspension was filtered and concentrated. The crude product was purified on silica gel column (80 g) using 5 to 30% MeOH in DCM affording 150 mg (43%) of 3- ((1 -(4-((tert-butyldimethylsilyl)oxy)butyl)- lH-benzo[d]imidazol-2-yl)methyl)- 1 -( 1 , 1 - dioxidothietan-3-yl)-l,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one as a beige solid.

¾ MR (400 MHz, CDCh) δ 8.81 (d, J = 0.8 Hz, 1H), 8.42 (d, J = 5.4 Hz, 1H), 7.77 - 7.72 (m, 1H), 7.39 - 7.34 (m, 2H), 7.31 - 7.24 (m, 2H), 5.45 - 5.37 (m, 1H), 5.38 (s, 2H), 4.95 - 4.88 (m, 2H), 4.64 - 4.56 (m, 2H), 4.34 (t, J = 7.5 Hz, 2H), 3.61 (t, J = 6.0 Hz, 2H), 1.86 - 1.76 (m, 2H), 1.58 - 1.49 (m, 2H), 0.86 (s, 9H), 0.02 (s, 6H). LC-MS (ESI-API, 254 nm) 75-95 % MeOH in H₂0 (0.1% HCO2H), 3 min, 1.00 ml/min, C18 (Agilent Zorbax XDB-18, 50 mm x 4.6 mm, 3.5 μιη), m/z = 556.2 (M + H), 278.6 (M/2 + H), t = 1.523 min, 93 % purity. l-(l,l-Dioxidothietan-3-yl)-3-((l-(4-hydroxybutyl)-lH-benzo[d]imidazol-2-yl)methyl)-l,3- dihydro-2H-imidazo [4,5-c] pyridin- -one

A 20 ml vial equipped with magnetic stir bar was charged with 110 mg of 3-((l-(4-((tert- butyldimethylsilyl)oxy)butyl)-lH-benzo[d]imidazol-2-yl)methyl)-l-(l,l-dioxidothietan-3-yl)- l,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one (0.198 mmol, 1 equiv.) dissolved in 2.0 ml of THF. Then 0.59 ml of 1 M TBAF in THF (0.594 mmol, 3 equiv.) and the mixture was stirred at rt for 32 h. Then reaction mixture was quenched by addition of water and the product was extracted with chloroform (4x), washed with water and dried over Na₂S04. The organics were concentrated and the crude product (105 mg) was recrystallized to afford 57 mg (65%) of l-(l,l-dioxidothietan-3- yl)-3-((l-(4-hydroxybutyl)-lH-benzo[d]imidazol-2-yl)methyl)-l,3-dihydro-2H midazo c]pyridin-2-one as light brown crystals.

¾ NMR (400 MHz, DMSO-de) δ 8.50 (s, 1H), 8.30 (dd, J = 5.4, 0.8 Hz, 1H), 7.58 (d, J = 8.6 Hz, 2H), 7.53 (d, J = 5.5 Hz, 1H), 7.29 - 7.20 (m, 1H), 7.22 - 7.12 (m, 1H), 5.51 - 5.39 (m, 1H), 5.44 (s, 2H), 5.00 (dd, J = 14.9, 7.1 Hz, 2H), 4.68 (dd, J = 14.3, 10.5 Hz, 2H), 4.48 (t, J = 5.2 Hz, 1H), 4.36 (t, J = 7.5 Hz, 2H), 3.40 (q, J = 6.4 Hz, 1H), 1.70 (p, J = 7.7 Hz, 2H), 1.45 (p, J = 6.4 Hz, 2H). ¹³C MR (100 MHz, DMSO-de) δ 152.01, 148.54, 142.85, 141.86, 135.24, 134.19, 130.07, 126.43, 122.48, 121.71, 119.08, 110.54, 104.39, 68.25, 60.24, 43.23, 37.79, 33.60, 29.55, 26.36. HRMS (ESI+) calcd. for C21H24N5O4S ([M+H]⁺): 442.1544. Found: 442.1537, error 0.7 ppm. LC-MS (ESI- API, 254 nm) 50-95 % MeOH in H2O (0.1% HCO2H), 6 min, 1.00 ml/min, C18 (Agilent Zorbax XDB- 18, 50 mm x 4.6 mm, 3.5 μιη), m/z = 442.0 (M + H), t = 0.579 min.

General procedure for the N-alk lation of 4-bromo-2-nitroaniline (Procedure A)

In a three-necked 100 mL round-bottomed flask, sodium hydride (60% dispersion in mineral oil, 1.5 eq.) was added to a solution of 4-bromo-2-nitroaniline (1.0 eq.) in DMF at 0°C. After approximately 30 minutes at 0 °C, the appropriate alkyl halide was added to the reaction mixture dropwise and the reaction was allowed to warm to room temperature overnight. For the workup, the reaction was quenched with a solution of saturated ammonium chloride and then extracted three times with EtOAc. The organic phases were subsequently combined, dried over MgSC"4 and concentrated in vacuo. The resulting crude material was then purified by column chromatography.

4-(4-Bromo-2-nitro-anilino)butanenitrile NP-RSV-003)

NP-RSV-003 was synthesized according to general procedure A as described above. The following quantities were employed: 4-bromo-2-nitroaniline (1.0 g, 4.6 mmol), 60% sodium hydride (277 mg, 6.91 mmol) and 4-bromobutyronitrile (1.02 g, 6.91 mmol) in DCM (20 mL). This procedure yielded NP-RSV-003 (399 mg, 1.40 mmol, 31%) as an orange solid.

1H MR (500 MHz, CDC13) δ 8.33 (d, J = 2.4 Hz, 1H), 8.01 (s, 1H), 7.54 (dd, J = 9.1, 2.3

Hz, 1H), 6.79 (d, J = 9.1 Hz, 1H), 3.54 - 3.47 (m, 2H), 2.53 (t, J = 6.9 Hz, 2H), 2.13 - 2.04 (m, 2H). HRMS calc. for CioHioBrNsOi, 284.00292 found, 284.00322.

4-Bromo-N-(3-methylsulfonylpropyl)-2-ni roaniline (NP-RSV-012)

NP-RSV-012 was synthesized according to general procedure A as described above. The following quantities were employed: 4-bromo-2-nitroaniline (400 mg, 1.84 mmol), 60% sodium hydride (111 mg, 2.77 mmol) and l-bromo-3 -methyl sulfonylpropane (556 mg, 2.77 mmol) in DCM (10 mL). This procedure yielded NP-RSV-012 (316 mg, 0.937 mmol, 51%) as an orange solid.

¾ NMR (500 MHz, CDC13) δ 8.31 (d, J = 2.4 Hz, 1H), 8.07 (s, 1H), 7.53 (dd, J = 9.2, 2.2 Hz, 1H), 6.81 (d, J = 9.1 Hz, 1H), 3.59 - 3.53 (m, 2H), 3.15 (t, J = 7.3 Hz, 2H), 2.97 (s, 3H), 2.31 - 2.23 (m, 2H).

General procedure for nitro reduction of the N-alkylated 4-bromo-2-nitroaniline (Procedure B)

Fe(s), AcOH

Iron powder (5 eq) was suspended in a solution of the N-alkylated 4-bromo-2-nitroaniline (1 eq) in ethanol, acetic acid and water (2:2: 1) in a single-necked 25 mL round bottomed flask. The flask was subsequently placed in a sonicator and irradiated at room temperature for 1 hour. Once TLC confirmed that all starting material had been consumed the iron powder was filtered off over celite and the resulting filtrate was basified with a 1M solution of NaOH. The crude product was then extracted from the basic solution into DCM three times. The organic phases were combined, dried over MgS0₄ and concentrated in vacuo. The resulting crude material was then purified by column chromatography or taken crude to the subsequent step.

Lmino-4-bromo-anilino)butanenitrile NP-RSV-005)

NP-RSV-005 was synthesized according to general procedure B as described above. The following quantities were employed: NP-RSV-003 (399 mg, 1.40 mmol) and iron powder (392 mg, 7.02 mmol) in EtOH (3 mL), AcOH (3 mL) and H₂0 (1.5 mL). This procedure yielded NP- RSV-005 (240 mg, 0.944 mmol, 67%) as a pale orange solid.

¾ NMR (500 MHz, CDCL3) δ 6.90 (dd, J = 8.3, 2.2 Hz, 1H), 6.84 (d, J = 2.2 Hz, 1H), 6.50 (d, J = 8.4 Hz, 1H), 3.39 (s, 3H), 3.26 (t, J = 6.6 Hz, 2H), 2.49 (t, J = 7.0 Hz, 2H), 2.03 - 1.95 (m, 2H). ¹³C NMR (126 MHz, CDC13) δ 136.4, 136.0, 123.1, 119.7, 119.2, 113.3, 111.4, 42.8, 25.2, 15.2. HRMS calc. for CioHisBrNs, 254.02874 found, 254.02899.

4-Bromo-Nl-(3-methylsulfonylpropyl)b nzene-l,2-diamine (NP-RSV-013)

NP-RSV-013 was synthesized according to general procedure B as described above. The following quantities were employed: NP-RSV-012 (250 mg, 0.741 mmol) and iron powder (207 mg, 3.71 mmol) in EtOH (2 mL), AcOH (2 mL) and H2O (1 mL). This procedure yielded crude NP-RSV-013 (205 mg, 0.667 mmol) as a tan solid. ¾ NMR (500 MHz, CDC13) δ 6.89 (dd, J = 8.3, 1.8 Hz, 1H), 6.84 (d, J = 1.7 Hz, 1H), 6.48 (d, J = 8.4 Hz, 1H), 3.45 (s, 2H), 3.27 (t, J = 6.5 Hz, 2H), 3.18 (t, J = 7.4 Hz, 2H), 2.93 (s, 3H), 2.24 - 2.16 (m, 2H).

General procedure for 2-chloromethyl benzimidazole formation (Procedure C)

A solution of the N-alkylated 4-bromobenzene-l,2-diamine (1.0 eq.), 2-chl oro- 1, 1,1- trimethoxy-ethane (3.0 eq.) and p-toluenesulfonyl chloride monohydrate (0.1 eq.) in DCM was stirred vigorously at room temperature, under an atmosphere of argon for approximately 4 hours, until TLC confirmed full consumption of the starting material. The reaction was then concentrated in vacuo and the resulting crude product was dry-loaded onto silica gel. Purification was undertaken with column chromatography.

4-[5-Bromo-2-(chloromethyl)benzimidazole-l- l]butanenitrile (NP-RSV-007)

NP-RSV-007 was synthesized according to general procedure C as described above. The following quantities were employed: NP-RSV-5 (200 mg, 0.787 mmol), 2-chl oro- 1, 1,1- trimethoxyethane (0.32 mL, 2.36 mmol) and p-TsOH H₂0 (15 mg, 0.079 mmol) in DCM (4 mL). This procedure yielded NP-RSV-007 (215 mg, 0.688 mmol, 87%) as a pale-yellow solid.

¾ MR (400 MHz, CDCL3) δ 7.89 (d, J = 1.7 Hz, 1H), 7.44 (dd, J = 8.6, 1.8 Hz, 1H), 7.30 - 7.23 (m, 1H), 4.83 (s, 2H), 4.38 (t, J = 7.4 Hz, 2H), 2.45 (t, J = 6.9 Hz, 2H), 2.31 - 2.23 (m, 2H). ¹³C NMR (101 MHz, CDC13) δ 149.8, 143.6, 134.2, 127.4, 123.6, 118.4, 116.2, 110.9, 42.8, 36.6, 25.5, 15.0. HRMS calc. for Ci₂Hi₂BrClN₃, 311.98976 found, 311.99105. 4-Bromo-2-(chloromethyl)-l-(3-methylsulfonylpropyl)benzimidazole (NP-RSV-014)

NP-RSV-014 was synthesized according to general procedure C as described above. The following quantities were employed: NP-RSV-013 (150 mg, 0.488 mmol), 2-chl oro- 1, 1,1 - trimethoxyethane (0.20 mL, 1.47 mmol) and p-TsOH H₂0 (9.0 mg, 0.049 mmol) in DCM (3 mL). This procedure yielded NP-RSV-014 (138 mg, 0.377 mmol, 77%).

¾ NMR (500 MHz, CDCh) δ 7.90 (d, J = 1.6 Hz, 1H), 7.44 (dd, J = 8.6, 1.8 Hz, 1H), 7.32 (d, J = 8.6 Hz, 1H), 4.85 (s, 2H), 4.53 - 4.40 (m, 2H), 3.09 (t, J = 6.9 Hz, 2H), 2.96 (s, 3H), 2.55 - 2.35 (m, 2H). ¹³C NMR (126 MHz, CDCh) δ 149.9, 143.7, 134.2, 127.3, 123.6, 116.2, 111.2, 51.2, 42.8, 41.4, 36.6, 22.5.

General cou ling procedure (Procedure D)

Potassium carbonate (1.2 eq.) and the respective 2-chloromethyl benzimidazole (1.2 eq.) were added to a solution of RJW-2-010 (1 eq.) in DMF and the reaction was heated to 50 °C. After approximately 18 hours the reaction was cooled to room temperature, quenched with a saturated solution of NH4CI and extracted into DCM three times. The organic phases were combined, dried over MgS0₄ and concentrated in vacuo. The resulting crude material was then purified by column chromatography and then recrystallized from hot MeOH. 4- [5-Bromo-2- [(2-oxo-l-tetrahydropyran-4-yl-imidazo [4,5-c]pyridin-3- yl)methyl]benzimidazol-l-yl]butanenitrile (NP-RSV-010)

NP-RSV-010 was synthesized according to general procedure D as described above. The following quantities were employed: RJW-2-010 (100 mg, 0.456 mmol), NP-RSV-007 (171 mg, 0.547 mmol) and K2CO3 (76 mg, 0.55 mmol) in DMF (3 mL). This procedure yielded NP-RSV- 010 (139 mg, 0.281 mmol, 62%) as a yellow solid.

¹H MR (500 MHz, DMSO) δ 8.46 (s, 1H), 8.24 (d, J = 5.4 Hz, 1H), 7.80 (d, J = 1.7 Hz, 1H), 7.62 (d, J = 8.7 Hz, 1H), 7.45 (d, J = 5.4 Hz, 1H), 7.41 (dd, J = 8.6, 1.8 Hz, 1H), 5.46 (s, 2H), 4.54 - 4.46 (m, 1H), 4.45 - 4.38 (m, 2H), 3.99 (dd, J = 11.2, 4.0 Hz, 2H), 3.48 (t, J = 11.3 Hz, 2H), 2.62 (t, J = 7.4 Hz, 2H), 2.42 - 2.30 (m, 2H), 2.10 - 2.00 (m, 2H), 1.73 - 1.66 (m, 2H). ¹³C MR (101 MHz, DMSO) δ 152.3, 150.5, 143.3, 142.6, 134.4, 134.3, 129.9, 126.5, 125.3, 121.7, 120.0, 114.3, 112.3, 104.5, 66.5, 50.2, 42.4, 37.6, 29.5, 25.3, 13.9. HRMS calc. for

495.11386 found, 495.11472.

3-[[5-Bromo-l-(3-methylsulfonylpropyl)benzimidazol-2-yl]methyl]-l- tetrahydr opyr an-4-yl-imidazo [4,5-c] ridin-2-one (NP-RS V- 16)

NP-RSV-016 was synthesized according to general procedure D as described above. The following quantities were employed: RJW-2-010 (48 mg, 0.22 mmol), NP-RSV-014 (96 mg, 0.26 mmol) and K2CO3 (36 mg, 0.26 mmol) in DMF (1 mL). This procedure yielded NP-RSV-016 (19 mg, 0.035 mmol, 16%) as a white solid.

1H MR (500 MHz, CDCh) δ 8.72 (s, 1H), 8.28 (d, J = 5.3 Hz, 1H), 7.86 (s, 1H), 7.34 (d,

J = 10.0 Hz, 1H), 7.23 (d, J = 8.6 Hz, 1H), 7.12 (d, J = 5.3 Hz, 1H), 5.34 (s, 2H), 5.27 (s, 1H), 4.55 - 4.46 (m, 2H), 4.11 (dd, J = 11.5, 3.9 Hz, 2H), 3.52 (t, J = 11.6 Hz, 2H), 3.07 (t, J = 7.2 Hz, 2H), 2.90 (s, 3H), 2.47 - 2.34 (m, 2H), 2.29 - 2.18 (m, 2H), 1.81 - 1.73 (m, 2H). ¹³C MR (126 MHz, CDCh) δ 152.7, 148.9, 143.6, 134.4, 134.0, 131.0, 126.9, 126.4, 123.3, 115.9, 111.1, 110.1, 104.6, 67.4, 51.2, 50.8, 42.4, 41.2, 38.4, 30.1, 22.7. HRMS calc. for CisHivBrNsC S, 548.09616 found, 548.09723.

Example 2: Biological Activity for exemplary compounds

To quantify viral resistance, dose-response curves were generated for exemplary compounds of this disclosure and, for comparison a developmentally advanced RSV inhibitor, BMS-433711 (also labeled SP-RSV-21), against wild type and RSV-F489 mutants. The compounds of Formula A were all potent against wild type RSV. Of special note is that compounds of Formula A were unexpectedly potent against wildtype RSV as compared to BMS-433771. BMS-433771 is reported in the literature as having a 21 nM IC50. (Meanwell, N. A. et al. Bioorg. Med. Chem. Lett., 2007, 17, 895-901). However, upon assaying BMS-433771 in an assay technique developed by Plemper et al., it was discovered that exemplary compounds EJ2140, EJ2143, RJW2011 and TK3-118 were 110-, 93-, 242- and 55x- more potent than BMS-433771. This unexpected potency for these exemplary compounds of Formula A can be seen in that ZD2- 160 was 1.6-fold less potent than BMS-433771, but ZD2-160 was expected to be as potent as the rest of the compounds of Formula A. Mutations in the microdomain resulted in a surprisingly high fold increased extrapolated IC50 concentration for the exemplary compounds of the disclosure confirming robust resistance. (See Figures 4 and 5 and Table 1)

The assay used for the generation of data was developed by the group of Richard Plemper at Georgia State University. Recombinant RSV strain expressing renilla luciferase were generated as reporter strains for robust automated drug discovery assays, which proposes major advantages over conventional RSV-based assays and a higher propensity to identify early and intermediate stage inhibitors of the viral life cycle (i.e. inhibitors of viral attachment, fusion, and viral polymerase activity) than blockers of viral assembly and egress, because the latter would act downstream of the luciferase reporter expression. (Yan et al. 2014, Cross-resistance mechanism of respiratory syncytial virus against structurally diverse entry inhibitors, PNAS, E3441-E3449).

Table 1: IC₅o data against Wild Type and RSV-F489 mutant RSV Virus

Claims

A compound accordin to Formula A or salt thereof,

Formula A

wherein R¹ is an optionally substituted alkyl and when X is N then R² is a heterocyclyl, and R³ is absent; and when X is C, then R² and R³ are linked to form a carbocyclyl and R⁴ is selected from H, alkoxy, halogen, amino or nitro group.

2. The compound as claimed in claim 1 wherein R¹ is an optionally substituted C₃ to C₆ alkyl and R² is a C₃ to C₆ heterocyclyl when X is N.

3. The compound as claimed in either claim 1 or claim 2 wherein R¹ is an optionally substituted C₄ alkyl and R² is a C₃ or C₄ heterocyclyl when X is N.

4. The compound as claimed in any one of claims 1 to 3 wherein R¹ is substituted with≡N, - S0₂(alkyl) or -OH.

5. The compound as claimed in any one of claims 1 to 4 wherein the heteroatoms of the R² heterocyclyl is selected from O, N, S or S0₂.

6. The compound as claimed in either claim 1 or claim 4 wherein R² and R³ are linked to form a C₃-C₆ carbocycle when X is C.

7. The compound as claimed in any one of claims 1, 4 or 6 wherein R² and R³ are linked to form a C₃ carbocyclyl when X is C.

8. The compound as claimed in any one of claims 1 to 7 wherein R⁴ is H or halogen.

9. The compound as claimed in any one of claims 1 to 8 which is selected from Formula Al or Formula A2, or salts thereof

Formula Al Formula A2 wherein n is 1, 2, 3, or 4; Y is selected from N, O, S, or SO2; R is H or halogen and R is

OH, S0₂(alkyl) or≡N.

11. A compound of Formula A or salts thereof for use in the treatment of RSV infections.

12. A method of treating viral RSV infections in a subject in need thereof, the method comprising administering an effective amount of the compound of Formula A or salts thereof to the subject.

13. The method of treating as claimed in claim 12, wherein the compound is administered in combination with one or more further active antiviral agent.

14. A pharmaceutical composition comprising a compound of Formula A or salts thereof and a pharmaceutically acceptable excipient.

15. The pharmaceutical composition as claimed in claim 14 in the form of a tablet, capsule, pill, gel, granules, aerosol, or aqueous buffer or a nanoparticle formulation, emulsion, or liposome, and which optionally includes one or more further active antiviral agents.