Compounds for the treatment of viruses

Abstract

The present invention relates generally to novel antiviral compounds and use of the antiviral compounds for preventing and / or treating a coronavirus infection, and a method of preventing and / or treating a coronavirus infection comprising administering a Papain-like protease (PLpro) inhibitor of Formula (I), or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof.

Description

[0001] Compounds for the treatment of viruses

[0002] RELATED APPLICATION

[0003] The present application claims priority from Australian provisional patent application AU 2024900559, filed March 3rd, 2024, the entire contents of which are incorporated herein by reference.

[0004] FIELD OF THE INVENTION;

[0005] The present invention relates generally to compounds, pharmaceutical compositions, and related methods for the treatment of viral infection. The compounds, pharmaceutical compositions, and methods can modulate the protease activity associated proteases of coronaviruses, in particular the Papain-like protease (PLpro).

[0006] BACKGROUND OF THE INVENTION

[0007] Viral respiratory tract infections (VRTIs) represent a major public health concern. Respiratory viruses cause infections in all age groups and are a major contributing factor to morbidity and mortality. Disease severity can range from mild, common cold-like illness to severe, life-threatening respiratory tract infection.

[0008] Previously, a significant proportion of VRTIs could not be attributed to a specific pathogen. With the advent of molecular detection and genotyping techniques, there has been a substantial increase in the recognition of several newly identified non-influenza respiratory pathogens. These pathogens have included coronavirus (CoV), adenovirus, rhinovirus, human respiratory syncytial virus, and human bocaviruses species.

[0009] In particular, coronaviruses are ubiquitous worldwide and may present as a spectrum of symptoms severity in a subject. For example, a coronavirus infection may present in a subject as a relatively mild respiratory disease (e.g., the common cold), or may otherwise present as Severe Acute Respiratory Syndrome (SARS).

[0010] The coronaviruses are large, enveloped viruses with a positive sense, single-stranded RNA genome. Coronavirus infections are a serious threat to both humans and animals; they cause enzootic infections and are responsible for human outbreaks of SARS caused by SARS-CoV, Middle-East Respiratory Syndrome (MERS) caused by MERS-CoV, and coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2. Indeed, SARS-CoV-2 was the causative agent for the COVID-19 pandemic - a global health emergency that resulted in 163 million cases and more than 3 million deaths worldwide during the early 2020s. Symptoms of coronavirus infections greatly vary amongst subjects. On the one hand, a subject infected with a coronavirus infection may be asymptomatic. On the other hand, a subject may experience severe symptoms including acute respiratory distress syndrome (ARDS), pneumonia, and single and multi-organ failures.

[0011] In part, human coronavirus virulence is attributed to long incubation periods and the absence of, or often mild, symptoms exhibited by infected and infectious persons, meaning that many people do not identify as suffering from a coronavirus infection, therefore continuing routine behaviour and spreading the infection. Transmission of coronavirus is usually via airborne droplets to the nasal mucosa, where the virus then invades the respiratory tract. It is also possible that contaminated droplets on the hands may be transmitted to the oral and / or nasal mucosa. Currently, hygiene practices are recommended to prevent transmission.

[0012] Currently, a coronavirus infection is treated through symptom management. While the advent of coronavirus vaccines will save lives and ease suffering, it remains unclear whether and how viral variants will emerge and subvert protection by vaccination and whether and how a global protection or herd immunity is achievable in a meaningful timeframe.

[0013] Regardless, COVID-19 and SARS-CoV-2 will remain a global threat with health and socio-economic impacts in the next decade. It is therefore paramount to develop alternative, complementary, and effective coronavirus therapies.

[0014] SUMMARY OF THE INVENTION

[0015] The present inventors have surprisingly found that compounds of Formula I, demonstrate activity as Papain-like protease (PLpro) inhibitors and provide a method for preventing and / or treating a coronavirus (CoV) infection. Specifically, the present invention relates to novel oxime- like derivative compounds as disclosed herein.

[0016] In one aspect, the present disclosure provides a compound of Formula I, method of preventing and / or treating a coronavirus (CoV) infection in a subject, comprising administering to the subject a therapeutically effective amount of a Papain-like protease (PLpro) inhibitor of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer or deuterated analogue thereof:

[0017] Formula I; wherein n = 1 or 2;

[0018] R1is a 9-10 membered heterocycle or a 9-10 membered carbocycle, wherein R1may be independently substituted with one or more R4, wherein R4is independently selected from Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, halogen, 3-10 membered carbocycle, 3-10 membered heterocycle, =0, -CN, -CF3, -CF2H, -NO2, -OR8, -SR8, - S(O)R8, S(O)2R8, -C(O)R8, -C(O)OR8, -N(R8)2, -C(O)N(R8)2, S(O)N(R8)2, -S(O)2N(R8)2, - NR8C(O)R8, -NR8S(O)R8, -NR8S(O)2R8, -NR8C(O)N(R8)2, -NR8S(O)2N(R8)2, wherein R8is independently selected from hydrogen, deuterium, halogen, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, 3-10 membered carbocycle or 3-10 membered heterocycle, wherein each R8may be independently substituted with one or more R10;

[0019] R2is selected from hydrogen, Ci-s alkyl, C2-ealkenyl, C2-6 alkynyl, 3-10-membered carbocycle, 3-10-membered heterocycle, wherein R2may be independently substituted with one or more R6when valency allows, wherein R6is independently selected from deuterium, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, halogen, 3-10 membered carbocycle, 3-10 membered heterocycle, -CN, -CF3, -CF2H, - NO2, -OR9, -SR9, -S(O)R9, S(O)2R9, -C(O)R9, -C(O)OR9, -N(R9)2, -C(O)N(R9)2, S(O)N(R9)2, - S(O)2N(R9)2, -NR9C(O)R9, -NR9S(O)R9, -NR9S(O)2R9, -NR9C(O)N(R9)2, -NR9S(O)2N(R9)2, wherein each R6is optionally substituted with R9when valency allows, wherein R9is independently selected from hydrogen, halogen, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, 3-10 membered carbocycle or 3-10 membered heterocycle and wherein each R9may be independently substituted with one or more R10when valency allows; R3is selected from the group consisting of a Ci-s alkyl, C2-8 alkenyl, C2-8 alkynyl, 3-10 membered carbocycle and a 4-10 membered heterocycle, wherein R3is optionally substituted with one or more R5,wherein R5is selected from the group consisting of hydrogen, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, halogen, 3-10 carbocycle, 3-10 heterocycle, -CN, =0, =NR7, -CF3, -CF2H, -NO2, -OR7, -SR7, -S(O)R7, S(O)2R7, -C(O)R7, -C(O)OR7, -N(R7)2, -C(O)N(R7)2, S(O)N(R7)2, -S(O)2N(R7)2, -S(O)(NR7)R7, -NR7C(O)R7, -NR7S(O)R7, -NR7S(O)2R7, -NR7C(O)N(R7)2, -NR7S(O)2N(R7)2, wherein each R7is independently selected from the group consisting of hydrogen, C1.4 alkyl, C1.4 halogenated alkyl, alkoxy, a 3-10 membered carbocycle and a 3-10 membered heterocycle, wherein each R7may be independently substituted with one or more R10when valency allow, wherein when R3is a 3-10 carbocycle or 3-10 heterocycle it is optionally geminally substituted with two independent R5, and the two R5s are optionally linked to form spirocycles, and wherein when R5is 3-10 carbocycle or 3-10 heterocycle it is optionally substituted with one or more R10;

[0020] R10is selected from the group consisting of hydrogen, deuterium, halogen, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, halogen, -CH2OH, -CN, -CF3, -CF2H, -NO2, -SR11, -S(O)RU, -S(O)2Rn, - wherein R11is independently selected from hydrogen, C1.4 halogenated alkyl or Ci-6 alkyl.

[0021] In some embodiments n = 2

[0022] In some embodiments, R1is a 9-10 membered heterocycle or a 9-10 membered carbocycle selected from a group consisting of:

[0023] In some preferred embodiments, n=2 and R1is naphthyl or quinoline.

[0024] In some embodiments,

[0025] In one embodiment, n=2 and R2is selected from hydrogen, Ci-s alkyl, a 3-10 membered carbocycle or a 3-10 membered heterocycle. In one embodiment, R2is selected from hydrogen, Ci-s alkyl, preferably methyl, ethyl, isopropyl, or butyl.

[0026] In some embodiments R2is selected from methyl, ethyl, isopropyl, butyl, wherein the hydrogen of the alkyl is optionally in the form of deuterium.

[0027] In one embodiment R2is hydrogen.

[0028] In one embodiment R2is a 5-6 membered carbocycle, preferably phenyl, cyclohexane or cyclopentane or a 5-6 membered heterocycle, preferably pyrimidine, pyrazine, pyridine, or tetrahydropyran.

[0029] In some embodiments, R2is Ci-s alkyl and is independently substituted with one or more R6.

[0030] In some embodiments R3is a 5-6 membered heterocycle, preferably pyrimidine, pyrazine, pyridine, or tetrahydropyran.

[0031] In some embodiments R3is a selected from a group selected from wherein R3is optionally substituted with one or more R5wherein R5is Ci-6 alkyl or alkoxy.

[0032] In some embodiments R3is a 5-6 membered carbocycle, preferably phenyl, cyclohexane or cyclopentane.

[0033] In some embodiments, R3is Ci-s alkyl.

[0034] In some embodiments n=2 and R3is Ci-s alkyl substituted with one or more R5.

[0035] In some embodiments, n = 2, R3is a 3-8-membered heterocycle or a 3-8-membered carbocycle.

[0036] In some embodiments, n=2, R3is a 3-8-membered heterocycle selected from a group consisting of:

[0037] In some embodiments, n=2, R3is a 3-8-membered carbocycle selected from a group consisting of:

[0038] In some embodiments, n=2 and R3is:

[0039] In some embodiments, n=2 and R3is substituted with one or more R5, wherein R5is preferably is selected from -OH, -OMe, OEt, - CN, -CF3, -CHF2, methyl, ethyl, =0, =NH, -S(O)2R7, -NR7(CO)N(R7)2, -NR7(CO)R7, - NR7S(O)R7-C(O)N(R7)2, C(O)OH, or a 5-6 membered heterocycle.

[0040] In some embodiments n=2 and R3is Ci-s alkyl substituted with one or more R5,wherein R5is preferably is selected from -OH, -OMe, OEt, -CN, -CF3, -CHF2, methyl, ethyl, =0, =NH, - S(O)2R7, -NR7(CO)N(R7)2, -NR7(CO)R7, -NR7S(O)R7, -C(O)N(R7)2, C(O)OH, or a 5-6 membered heterocycle.

[0041] In some embodiments, R4is OR8,preferably wherein R8is Ci-6 alkyl, more preferably R4is -OMe, -OEt, -OCHF2, -OCD3.

[0042] In some embodiments n=2, R4is OR8,preferably wherein R8is Ci-6 alkyl, such that R4is alkoxy and wherein the hydrogen of the alkoxy is optionally in the form of deuterium.

[0043] In some embodiments, n=2, R5is selected from a group consisting of:

[0044] -OH, -OMe, OEt, -CN, -CF3, -CHF2, methyl, ethyl, =0, =NH, -S(O)2R7, -NR7(CO)N(R7)2, - NR7(CO)R7, -NR7S(O)R7-C(O)N(R7)2, C(O)OH, and a 5-6 membered heterocycle.

[0045] In some embodiments, R3is gem-substituted with two R5and optionally the two R5are linked to form a spiro[4.5]decane.

[0046] In some embodiments, n=2, R6is aryl and is optionally substituted with R9.

[0047] In some preferred embodiments, n=2, R6is aryl and is substituted with -OMe.

[0048] In some embodiments, n=2, R6is a 5-6 membered carbocycle.

[0049] In some embodiments, n=2, R6is selected from a group consisting of deuterium, -OH, -OMe, - CF3, -C(O)N(R9)2, -N(R9)2, -NH(BOC), -NR9C(O)R9, preferably R9is hydrogen.

[0050] In some embodiments R7is hydrogen.

[0051] In some embodiments R7is methyl.

[0052] In some embodiments R7is -OMe.

[0053] In some embodiments R7is -C(O)NR10.

[0054] In some embodiments R8is Ci-6 alkyl.

[0055] In some embodiments R9is hydrogen or -OMe. In some embodiments R10is selected from the group consisting of hydrogen, a 5-6 membered carbocycle, a 5-6 membered heterocycle, -NHC(O)Me and -OMe.

[0056] In some preferred embodiments n=2 and R1is a 9-10 membered heterocycle or a 9-10 carbocycle and is substituted with R4, and R4is selected from Ci- 6 alkyl, -OMe, -OCHF2, OCF3, -OCD3, =0, and R8is hydrogen, and R3is a 3-8-membered carbocycle or a 3-8-membered heterocycle, optionally substituted with one or more R5wherein R5is selected from hydrogen, -OH, -OMe, - OEt, methyl, ethyl, =0, -NR7(CO)N(R7)2, a 5-6 membered heterocycle, and R7, R9, R10and R11are hydrogen.

[0057] In some preferred embodiments R10is hydrogen. In some preferred embodiments R11is hydrogen.

[0058] In some embodiments n=2 and R1is a 9-10 membered heterocycle or a 9-10 carbocycle and is substituted with R4, and R4is selected from Ci- 6 alkyl, -OMe, -OCHF2, OCF3, -OCD3, =0, and R8is hydrogen, R3is geminally substituted with two R5and optionally the two R5are linked to form a spiro[4.5]decane and R5is selected from -OH, -OMe, OEt, methyl, ethyl, =0, - NR7(CO)N(R7)2, -NR7(CO)R7, and R9, R10and R11are each hydrogen.

[0059] In some embodiments, the compound of Formula I is selected from the group consisting of:

[0060]

[0061]

[0062] In some embodiments the compounds of Formula I are selected from the group consisting of:

[0063]

[0064] In some embodiments the Papain-like protease (PLpro) inhibitor of Formula I can be used for preparing proteolysis-targeting chimeras (PROTACs) for inducing the degradation of an RNA virus protein target, in this case a coronaviral papain-like protease (PLpro).

[0065] One embodiment the Papain-like protease (PLpro) inhibitor of Formula I can be used for preparing proteolysis-targeting chimeras (PROTACs), by combining a first moiety of Formula I disclosed herein that targets PLpro with a second moiety that binds to a protein that is a protein degrader.

[0066] In some embodiments the Papain-like protease (PLpro) inhibitor of Formula I is used for the prevention and / or treatment of a coronavirus (CoV) infection which includes preventing or reducing the likelihood of developing, or reducing the severity of, a symptom associated with a coronavirus (CoV) infection.

[0067] In some embodiments the Papain-like protease (PLpro) inhibitor of Formula I is used for the prevention and / or treatment of a COVID-19 or SARS-CoV, or MERS-CoV infection which includes preventing or reducing the likelihood of developing, or reducing the severity of, a symptom associated with a coronavirus (CoV) infection.

[0068] In some embodiments, the Papain-like protease (PLpro) inhibitor of Formula I is administered orally or intranasally. In some embodiments, the Papain-like protease (PLpro) inhibitor of Formula I is administered orally.

[0069] In some embodiments, the Papain-like protease (PLpro) inhibitor of Formula I is administered in combination with a further therapeutic agent.

[0070] Any embodiment herein shall be taken to apply mutatis mutandis to any other embodiment unless specifically stated otherwise. For instance, as the skilled person would understand, examples of proteins outlined above equally apply to the nucleic acids, vectors, and methods of the invention. The present invention is not to be limited in scope by the specific embodiments described herein, which are intended for the purpose of exemplification only. Functionally equivalent products, compositions and methods are clearly within the scope of the invention, as described herein.

[0071] Throughout this specification, unless specifically stated otherwise or the context requires otherwise, reference to a single step, composition of matter, group of steps or group of compositions of matter shall be taken to encompass one and a plurality (i.e. one or more) of those steps, compositions of matter, groups of steps or group of compositions of matter.

[0072] BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWINGS

[0073] Figure 1 shows the co-crystallography results of Cpd-22-Pkl-A.

[0074] Figure 2 shows the viral plaque assay results.

[0075] Figure 3 shows results from the in vivo mouse model.

[0076] DETAILED DESCRIPTION OF THE INVENTION

[0077] General Techniques and Definitions

[0078] Unless specifically defined otherwise, all technical and scientific terms used herein shall be taken to have the same meaning as commonly understood by one of ordinary skill in the art (e.g., chemistry, medicinal chemistry and the like). As used herein, the term “and / or”, e.g., “X and / or Y” shall be understood to mean either "X and Y" or "X or Y" and shall be taken to provide explicit support for both meanings or for either meaning.

[0079] As used herein, singular forms “a”, “an” and “the” include plural aspects, unless the context clearly indicates otherwise.

[0080] Throughout this specification, the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.

[0081] As used herein, the term “subject” refers to any organism susceptible to a viral infection. For example, the subject can be a mammal, primate, livestock (e.g., sheep, cow, horse, pig), companion animal (e.g., dog, cat), or laboratory animal (e.g., mouse, rabbit, rat, guinea pig, hamster). In one example, the subject is a mammal. In one embodiment, the subject is human. In one embodiment, the disease or condition is a coronavirus (CoV) infection.

[0082] As used herein, the term “treating” includes alleviation of the symptoms associated with a specific disorder or condition and reducing and / or eliminating said symptoms. For example, as used herein, the term “treating a coronavirus (CoV) infection” refers to alleviating the symptoms associated with a coronavirus (CoV) infection and reducing and / or eliminating the symptoms associated with a coronavirus (CoV) infection.

[0083] As used herein, the term “prevention” includes prophylaxis of the specific disorder or condition. For example, as used herein, the term “preventing a coronavirus (CoV) infection” refers to preventing the onset or duration of the symptoms associated with a coronavirus (CoV) infection. As would be understood by the person skilled in the art, a compound of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, would be administered in a therapeutically effective amount. The term “therapeutically effective amount”, as used herein, refers to a compound of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, being administered in an amount sufficient to alleviate or prevent to some extent one or more of the symptoms of the disorder or condition being treated. The result can be the reduction and / or alleviation of the signs, symptoms, or causes of a disease or condition, or any other desired alteration of a biological system. In one embodiment, the term “therapeutically effective amount” refers to a compound of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, being administered in an amount sufficient to result in a reduction of one or more symptoms associated with a coronavirus (CoV) infection. The term, “effective amount”, as used herein, refers to an amount of a compound of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, effective to achieve a desired pharmacologic effect or therapeutic improvement without undue adverse side effects or to achieve a desired pharmacologic effect or therapeutic improvement with a reduced side effect profile. By way of example only, therapeutically effective amounts may be determined by routine experimentation, including but not limited to a dose escalation clinical trial. The term “therapeutically effective amount” includes, for example, a prophylactically effective amount. In one embodiment, a prophylactically effective amount is an amount sufficient to prevent a coronavirus (CoV) infection. It is understood that “an effective amount” or “a therapeutically effective amount” can vary from subject to subject, due to variation in metabolism of the compound and any of age, weight, general condition of the subject, the condition being treated, the severity of the condition being treated, and the judgment of the prescribing physician. Thus, it is not always possible to specify an exact “effective amount”. However, an appropriate “effective amount” in any individual case may be determined by one of ordinary skill in the art using routine experimentation. Where more than one therapeutic agent is used in combination, a “therapeutically effective amount” of each therapeutic agent can refer to an amount of the therapeutic agent that would be therapeutically effective when used on its own, or may refer to an adjusted (e.g., reduced) amount that is therapeutically effective by virtue of its combination with one or more additional therapeutic agents.

[0084] The compounds of the present disclosure may contain chiral (asymmetric) centers or the molecule as a whole may be chiral. The individual stereoisomers (enantiomers and diastereoisomers) and mixtures of these are within the scope of the present disclosure. The following definitions apply to the terms as used throughout this specification, unless otherwise limited in specific instances.

[0085] As used herein, the term “halogen” means fluorine, chorine, bromine, or iodine.

[0086] As used herein, the term “alkyl” encompasses both straight chain (i.e., linear) and branched chain hydrocarbon groups. Examples of alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, t-butyl, i-butyl, sec-butyl, pentyl, and hexyl groups. In one example, the alkyl group is of one to six carbon atoms (i.e. Ci-ealkyl).

[0087] As used herein, the term “alkoxy” refers to the group -O-alkyl, where “alkyl” is as described above. Examples of alkoxy groups include methoxy, ethoxy, propoxy, and butoxy groups. In one example, the alkoxy group is of one to six carbon atoms (i.e. -O-Ci-ealkyl).

[0088] As used herein, the term “alkenyl” refers to both straight and branched chain unsaturated hydrocarbon groups with at least one carbon-carbon double bond. Examples of alkenyl groups include ethenyl, propenyl, butenyl, pentenyl, and hexenyl groups. In one example, the alkenyl group is of two to six carbon atoms (i.e. C2-ealkenyl).

[0089] As used herein, the term “alkynyl” refers to both straight and branched chain unsaturated hydrocarbon groups with at least one carbon-carbon triple bond. Examples of alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, and hexynyl groups. In one example, the alkynyl group is of two to six carbon atoms (i.e. C2-ealkynyl).

[0090] As used herein, the term “carbocycle” refers to a monovalent non-aromatic, saturated, or partially unsaturated, or aromatic ring system having 3 to 12 carbon atoms. In one example, the carbocycle is a 3-10 membered carbocycle. A carbocycle group may, for example, be monocyclic or polycyclic (i.e. bicyclic, tricyclic). A polycyclic carbocycle group may contain fused rings. Examples of monocyclic carbocycle groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, 1 -cyclopent- 1-enyl, l-cyclopent-2-enyl, 1 -cyclopent-3 -enyl, cyclohexyl, 1 -cyclohex- 1 -enyl, 1 -cyclohex -2-enyl, 1 -cyclohex-3 -enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, and the like. Examples of monocyclic, aromatic carbocycle group include, but are not limited to, phenyl. A polycyclic carbocycle group may contain fused rings. Examples of fused carbocycle groups include, but are not limited to naphthyl, tetrahydronaphthyl and dihydro- IH-indenyl.

[0091] As used herein, the term “aryl” refers to an aromatic ring having system having 3 to 12 carbon atoms (i.e., an aromatic 3-12 membered carbocycle). An aryl group may, for example, be monocyclic or polycyclic. Examples of aryl groups include, but are not limited to, phenyl and naphthyl.

[0092] As used herein, the term “heterocycle” refers to an aromatic or non-aromatic cyclic group which is analogous to a carbocycle group, but in which from one or more of the carbon atoms is / are replaced by one or more heteroatoms independently selected from nitrogen, oxygen, or sulfur. A heterocycle group may, for example, be monocyclic or polycyclic (e.g. bicyclic). A polycyclic heterocycle may for example contain fused rings. In a bicyclic heterocycle group there may be one or more heteroatoms in each ring, or heteroatoms only in one of the rings. A heteroatom may be N, O, or S. Heterocycle groups containing a suitable nitrogen atom include the corresponding N-oxides. In one example, the heterocycle group is of three to ten atoms (i.e. 3-10-membered heterocycle). Examples of monocyclic non-aromatic heterocycle groups include aziridinyl, azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidinyl, piperazinyl, tetrahydrofuranyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl and azepanyl. Examples of bicyclic heterocyclic groups in which one of the rings is non-aromatic include dihydrobenzofuranyl, indanyl, indolinyl, isoindolinyl, tetrahydroisoquinolinyl, tetrahydroquinolyl, and benzoazepanyl. Examples of monocyclic aromatic heterocyclyl groups (also referred to as monocyclic heteroaryl groups) include furanyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, oxadiazolyl, thiadiazolyl, pyridyl, triazolyl, triazinyl, pyridazyl, isothiazolyl, isoxazolyl, pyrazinyl, pyrazolyl, and pyrimidinyl. Examples of bicyclic aromatic heterocycle groups (also referred to as bicyclic heteroaryl groups) include quinoxalinyl, quinazolinyl, pyridopyrazinyl, benzoxazolyl, benzothiophenyl, benzimidazolyl, naphthyridinyl, quinolinyl, benzofuranyl, indolyl, indazolyl, benzothiazolyl, oxazolyl[4,5-b]pyridyl, pyridopyrimidinyl, isoquinolinyl, benzohydroxazole and dihydrobenzodioxine.

[0093] As used herein, the term “heteroaryl” refers to an aromatic ring having system having 3 to 12 carbon atoms in which one or more of the carbon atoms is / are replaced by one or more heteroatoms independently selected from nitrogen, oxygen, or sulfur (i.e., an aromatic 3-10 membered heterocycle). A heteroaryl group may, for example, be monocyclic or polycyclic. Examples of heteroaryl groups include, but are not limited to, imidazole and isoxazole.

[0094] As used herein, the term “saturated” refers to a group where all available valence bonds of the backbone atoms are attached to other atoms representative examples of saturated groups include, but are not limited to, butyl, cyclohexyl, piperidine, and the like.

[0095] As used herein, the term “unsaturated” refers to a group where at least one valence bond of two adjacent backbone atoms is not attached to other atoms. Representative examples include, but are not limited to, alkenes (e.g., -CH2-CH=CH-CH2-), phenyl, pyrrole, and the like.

[0096] As used herein, the term “substituted” refers to a group having one or more hydrogens or other atoms removed from a carbon or suitable heteroatom and replaced with a further group (i.e., substituent).

[0097] As used herein, the term gem-substituted refers to two groups bonded to the same carbon. The present disclosure relates to compounds of Formula I and salts thereof. Salts may be formed in the case of embodiments of the compound of Formula I, which contain a suitable acidic or basic group. Suitable salts of the compound of Formula I include those formed with organic or inorganic acids or bases.

[0098] Those skilled in the art of organic and / or medicinal chemistry will appreciate that many organic compounds can form complexes with solvents in which they are reacted or from which they are precipitated or crystallized. These complexes are known as "solvates". For example, a complex with water is known as a "hydrate". As used herein, the phrase “pharmaceutically acceptable solvate” or “solvate” refers to an association of one or more solvent molecules and a compound of the present disclosure. Examples of solvents that form pharmaceutically acceptable solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine. It will be understood that the present disclosure encompasses solvated forms, including hydrates, of the compounds of Formula I and salts thereof.

[0099] Those skilled in the art of organic and / or medicinal chemistry will appreciate that hydrogen may be substituted for its isotope deuterium. As used herein, the phrase “deuterated analogue” refers to a compound of the present disclosure where one or more hydrogens are substituted for deuterium. It will be understood that the present disclosure encompasses deuterated forms of the compounds of Formula I. Enriching for deuterium may afford certain therapeutic advantages, such as increasing in vivo half-life or reducing dosage requirements, or may provide a compound useful as a standard for characterization of biological samples.

[0100] As used herein, the term “stereoisomer” refers to compounds having the same molecular formula and sequence of bonded atoms (i.e., atom connectivity), though differ in the three- dimensional orientations of their atoms in space. Stereoisomers include geometric, optical isomers and the like which may exist due to asymmetric carbons on various substituents or may exist even in the absence of asymmetric carbons. As used herein, the term “enantiomers” refers to two compounds that are stereoisomers in that they are non-superimposable mirror images of one another. Relevant stereocenters may be denoted with (R)- or (S)- configuration as defined by the IUPAC 1974 Recommendations. Positional isomers (such as, for example, 4-pyridyl and 3-pyridyl) are also contemplated in the invention. If a compound of the invention incorporates a double bond or a fused ring, both the cis- and trans-forms, as well as mixtures, are embraced within the scope of the invention. Representation of spatial configurations using bond types such as bold, hashed, dashed and / or wedged follows lUPACs 2006 Recommendations. The single •' ' represents point of attachment of the relevant R groups. Those skilled in the art of organic and / or medicinal chemistry will appreciate that the compounds of Formula I and salts thereof may be present in amorphous form, or in a crystalline form. It will be understood that the present disclosure encompasses all forms and polymorphs of the compounds of Formula I and salts thereof.

[0101] Papain-Like Protease (PLpro)

[0102] The papain-like protease, PLpro, is encoded by coronaviruses and is an essential protein for viral replication and immune evasion. PLpro is expressed as part of a non-structural polyprotein and is cleaved by the PLpro protease activity. Mature PLpro may have de- ubiquitinase and de-ISGylase activity, efficiently cleaving posttranslational modifications on target proteins in infected cells, leading to a reduced anti-viral response to SARS-CoV-2.

[0103] It has been found that compounds of Formula I demonstrate PLpro inhibition and therefore provide a method of treating a viral infection, for example, a coronavirus infection.

[0104] Compounds of Formula I

[0105] The present disclosure provides a Papain-like protease (PLpro) inhibitor of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof:

[0106] Formula I.

[0107] In the above Formula I, n = 1 or 2.

[0108] In some embodiments n=2.

[0109] In the above Formula I, R1is a 9-10 membered heterocycle or a 9-10 membered carbocycle.

[0110] In some embodiments R1is a bicyclic 10 membered carbocycle or bicyclic 10 membered heterocycle. In some embodiments, R1is selected from a group consisting of:

[0111] In some embodiments R1is naphthalene or quinoline.

[0112] In the above Formula I, R1can be substituted with one or more R4. In some embodiments, R1is substituted with one, two, three, four, five, or more R4substituents. Each R4substituent may be the same substituent or a different substituent (e.g., the R4substituents are independently selected from one another). R4may be independently selected from Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, halogen, 3-10 membered carbocycle, 3-10 membered heterocycle, -CN, -CF3, -CF2H, - NO2, -OR8, -SR8, -S(O)R8, S(O)2R8, -C(O)R8, -C(O)OR8, -N(R8)2, -C(O)N(R8)2, S(O)N(R8)2, - S(O)2N(R8)2, -NR8C(O)R8, -NR8S(O)R8, -NR8S(O)2R8, -NR8C(O)N(R8)2, -NR8S(O)2N(R8)2.

[0113] In the above Formula I, R4may be optionally substituted with R8; wherein R8is independently selected from hydrogen, halogen, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, 3-10 membered carbocycle or 3-10 membered heterocycle; wherein each R8may be independently substituted with one or more R10.

[0114] In some embodiments R4is independently selected from the group consisting of C1.3 alkyl, cyano, alkoxy, halogen.

[0115] In some embodiments R4is alkoxy.

[0116] In some preferred embodiments at least one R4is -OCH3, -OCHF2, or -OCD3.

[0117] In the above Formula I R2is independently selected from the group consisting of hydrogen, Ci-s alkyl, C2-6 alkenyl, C2-6 alkynyl, 3-10-membered carbocycle, 3-10-membered heterocycle.

[0118] In one embodiment R2is hydrogen.

[0119] In some embodiments R2is Ci-s alkyl, preferably methyl, ethyl, isopropyl, or butyl.

[0120] In some embodiments R2is aryl or heteroaryl.

[0121] In some embodiments R2is a 5-6 membered heterocycle, preferably pyridine, pyrimidine, pyrazine or tetrahydropyran.

[0122] In some embodiments R2is a 5-6 membered carbocycle, preferably phenyl, cyclohexane or cyclopentane.

[0123] In some embodiments R2is independently substituted with one or more R6.

[0124] In the above Formula I R6is independently selected from deuterium, Ci-6 alkyl, C2-6 alkenyl, C2- 6 alkynyl, alkoxy, halogen, 3-10 membered carbocycle, 3-10 membered heterocycle, -CN, -CF3, -CF2H, -NO2, -OR9, -SR9, -S(O)R9, S(O)2R9, -C(O)R9, -C(O)OR9, -N(R9)2, -C(O)N(R9)2, S(O)N(R9)2, -S(O)2N(R9)2, -NR9C(O)R9, -NR9S(O)R9, -NR9S(O)2R9, -NR9C(O)N(R9)2, - NR9S(O)2N(R9)2; wherein each R6, is optionally substituted with R9when valency allows; wherein R9is independently selected from hydrogen, halogen, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, 3-10 membered carbocycle or 3-10 membered heterocycle; wherein each R9may be independently substituted with one or more R10when valency allows.

[0125] In some embodiments R6is selected from a group consisting of deuterium, -OR9, -OMe, -CF3, - C(O)N(R9)2, -N(R9)2, -NH(BOC), -NR9C(O)R9, 5-6 membered carbocycle, 5-6 membered heterocycle and preferably R9is hydrogen.

[0126] In some embodiments R6is a 5-6 membered carbocycle optionally independently substituted with one or more R9and preferably R9is hydrogen.

[0127] In the above Formula I R9is independently selected from hydrogen, halogen, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, 3-10 membered carbocycle or 3-10 membered heterocycle.

[0128] In some embodiments R9is hydrogen or - OMe.

[0129] In the above Formula I R3is independently selected from the group consisting of a Ci-s alkyl, C2- 6 alkenyl, C2-6 alkynyl, 3-10 -membered carbocycle and a 3-10-membered heterocycle.

[0130] In some embodiments R3is selected from the group consisting of 3-8-membered carbocycle and 3-8-membered heterocycle. In some preferred embodiments R3is selected from the group consisting of:

[0131] In some preferred embodiments R3is: 6

[0132] In the above Formula I R3is optionally substituted with one or more R5.

[0133] R5is independently selected from the group consisting of hydrogen, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, halogen, 3-10 carbocycle, 3-10 heterocycle, -CN, =0, =NR7, -CF3, -CF2H, -NO2, -OR7, -SR7, -S(O)R7, S(O)2R7, -C(O)R7, -C(O)OR7, -N(R7)2, -C(O)N(R7)2, S(O)N(R7)2, - S(O)2N(R7)2, -S(O)(NR7)R7, -NR7C(O)R7, -NR7S(O)R7, -NR7S(O)2R7, -NR7C(O)N(R7)2, - NR7S(O)2N(R7)2. 1

[0134] In some embodiments R5is selected from -OH, -OMe, OEt, -CN, -CF3, -CHF2, methyl, ethyl, =0, =NH, -S(O)2R7, -NR7(CO)N(R7)2, -NR7(CO)R7, -NR7S(O)R7-C(O)N(R7)2, C(0)0H, a 5-6 membered heterocycle.

[0135] In some preferred embodiments R5is selected from -OH, -OMe, -OEt, -CF3, -CHF2, methyl, ethyl, -NH(C0)NH2, -NH(C0)CH3

[0136] In some embodiments when R3is a 5-6 membered carbocycle R5is selected from -OH, -OMe, - OEt, methyl, ethyl, -CHF2, -CF3, -NH(C0)NH2, -NH(C0)CH3

[0137] In some embodiments R3is a 5-6 membered carbocycle or 5-6 membered heterocycle and substituted with R5or geminally substituted with two R5and the two R5s are optionally linked to form spirocycles.

[0138] In some embodiments when R3is a 5-6 membered carbocycle and is geminally substituted with two R5, the two R5may be linked to form a spiro[4.5] decane.

[0139] In the above Formula I R6is independently selected from halogen, C1.4 alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, -OH, -CN, -N(R7)2, -NR7(CO)R7, -NR7(CO)N(R7)2, -NR7S(O)2R7, - NR7S(O)N(R7)2, -(CO)N(R7)2, -(CO)N(R7)2, -(CO)OR7, -CF3, and -CF2H.

[0140] In some embodiments R6is alkoxy, -NH2, -NMe2, -NH(C0)Me, -NH(C0)0R7, -(C0)NH2.

[0141] In the above Formula I each R7is independently selected from the group consisting of hydrogen, C1.4 alkyl, C1.4 halogenated alkyl, alkoxy, 3-10 membered carbocycle or 3-10 membered heterocycle,

[0142] In some embodiments each R7is independently selected from hydrogen, C1.4 alkyl, alkoxy, aryl and heteroaryl. In some embodiments R7is hydrogen.

[0143] In some embodiments the Papain-like protease (PLpro) inhibitor of Formula I is selected from the group consisting of:

[0144]

[0145] In some embodiments the compound of Formula I consists of a stereoisomer which contains either a trans and cis cyclohexanol and / or R or 5-piperidine and / or an E and Z oximes.

[0146] Therapeutic Methods and Uses A Papain-like protease (PLpro) inhibitor of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, finds use in the treatment of diseases for which inhibition of Papain-like protease provides a therapeutic effect. In particular, the Papain-like protease (PLpro) inhibitors of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, find use in the therapy of diseases, for example, viral infections (e.g., coronavirus infections).

[0147] Accordingly, there is provided a Papain-like protease (PLpro) inhibitor of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, for use in therapy of diseases, for example a viral infection. In some embodiments, a Papain-like protease (PLpro) inhibitor of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, finds use in the prevention and / or treatment of a viral infection. In one example, a Papain-like protease (PLpro) inhibitor of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, finds use in the prevention and / or treatment of a coronavirus infection.

[0148] A Papain-like protease (PLpro) inhibitor of Formula I, or a stereoisomer, or deuterated analogue thereof, could be used for preparing proteolysis-targeting chimeras (PROTACs), by combining a first moiety of Formula I, disclosed herein that targets PLpro, with a second moiety that binds to a protein that is a protein degrader. For example, the second moiety, recruits an enzyme or a complex that catalyzes the ubiquitination of the viral protein target, PLpro, and in turn the viral protein target is degraded by the proteasome.

[0149] As used herein, the term “proteolysis targeting chimera (PROTAC)” refers to a heterobifunctional small molecule. PROTACs consist of two covalently linked protein-binding molecules: one capable of engaging an E3 ubiquitin ligase, and another as represented by a compound of Formula I, that binds to a target protein, in this case the viral protein PLpro.

[0150] Coronavirus Infection

[0151] In some embodiments, there is provided a method of preventing and / or treating a coronavirus infection in a subject, comprising administering to the subject a therapeutically effective amount of a Papain-like protease (PLpro) inhibitor of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, as defined herein, wherein the coronavirus is selected from the group consisting of Alphacoronavirus (alphaCoV), Betacoronavirus (betaCoV), Gammacoronavirus (gammaCoV), and Deltacoronavirus (deltaCoV).

[0152] In one example, there is provided a method of preventing and / or treating a Betacoronavirus (betaCoV) infection in a subject, comprising administering to the subject a therapeutically effective amount of a Papain-like protease (PLpro) inhibitor of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, as defined herein.

[0153] In one example, there is provided a method of preventing and / or treating Alphacoronavirus (alphaCoV), infection in a subject, comprising administering to the subject a therapeutically effective amount of a Papain-like protease (PLpro) inhibitor of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, as defined herein.

[0154] In some embodiments, the Alphacoronavirus (alphaCoV) is selected from coronavirus 229E (HCoV-229E), human coronavirus NL63 (HCoV-NL63), transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV), feline infectious peritonitis virus (FIPV), and canine coronavirus (CCoV).

[0155] In one example there is provided a method of preventing and / or treating Gammacoronavirus (gammaCoV) infection in a subject, comprising administering to the subject a therapeutically effective amount of a Papain-like protease (PLpro) inhibitor of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, as defined herein.

[0156] In one example there is provided a method of preventing and / or treating Deltacoronavirus (deltaCoV) infection in a subject, comprising administering to the subject a therapeutically effective amount of a Papain-like protease (PLpro) inhibitor of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, as defined herein. In one example there is provided a method of preventing or reducing fever associated with a coronavirus (CoV) infection, comprising administering to a subject in need thereof a therapeutically effective amount of a Papain-like protease (PLpro) inhibitor of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, as defined herein. In one embodiment there is provided a method of preventing or reducing cough associated with a coronavirus (CoV) infection, comprising administering to a subj ect in need thereof a therapeutically effective amount of a Papain-like protease (PLpro) inhibitor of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, as defined herein.

[0157] In one example there is provided a method of preventing or reducing viral shedding associated with a coronavirus (CoV) infection, comprising administering to a subject in need thereof a therapeutically effective amount of a Papain-like protease (PLpro) inhibitor of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, as defined herein.

[0158] In one example the prevention and / or treatment of a coronavirus (CoV) infection includes reducing the coronavirus antibody titre of the individual.

[0159] Compositions

[0160] Whilst a Papain-like protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, may, in some embodiments, be administered alone, it is more typically administered as part of a pharmaceutical composition or formulation. Thus, the present disclosure also provides a pharmaceutical composition comprising a Papain-like protease (PLpro) inhibitor of Formula I, or a salt, solvate, stereoisomer, deuterated analogue, and a pharmaceutically acceptable excipient. The pharmaceutical composition comprises one or more pharmaceutically acceptable diluents, carriers, or excipients (collectively referred to herein as “excipient” materials).

[0161] The present disclosure also provides pharmaceutical formulations or compositions, both for veterinary and for human medical use, which comprise Papain-like protease (PLpro) inhibitor of Formula I, or a salt, solvate, stereoisomers, or deuterated analogue thereof, of the present disclosure, with one or more pharmaceutically acceptable carriers, and optionally any other therapeutic ingredients, stabilisers, or the like. The carrier(s) must be pharmaceutically acceptable in the sense of being compatible with the other ingredients of the formulation and not unduly deleterious to the recipient thereof.

[0162] Examples of pharmaceutical formulations include those suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous, and intraarticular), inhalation (including fine particle dusts or mists that may be generated by means of various types of metered dose pressurised aerosols), nebulisers or insufflators, rectal, intraperitoneal and topical (including dermal, buccal, sublingual, and intraocular) administration, although the most suitable route may depend upon, for example, the condition and disorder of the recipient.

[0163] In some embodiments, that composition is formulated for oral delivery. For example, pharmaceutical formulations of the present disclosure suitable for oral administration may be presented as discrete units such as capsules, cachets, pills, or tablets each containing a predetermined amount of the active ingredient; as a powder or granules, as a solution or a suspension in an aqueous liquid or non-aqueous liquid, for example as elixirs, tinctures, suspensions or syrups; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. A Papain-like protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, may also be presented as a bolus, electuary or paste.

[0164] The Papain-like protease (PLpro) inhibitors of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, of the present disclosure may for example be formulated in compositions including those suitable for inhalation to the lung, by aerosol, or parenteral (including intraperitoneal, intravenous, subcutaneous, or intramuscular injection) administration. The compositions may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing the Papain-like protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, into association with a carrier that constitutes one or more accessory ingredients. In general, the compositions are prepared by bringing the Papain-like protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, into association with a liquid carrier to form a solution or a suspension, or alternatively, bring the Papain-like protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, into association with formulation components suitable for forming a solid, optionally a particulate product, and then, if warranted, shaping the product into a desired delivery form. Solid formulations of the present disclosure, when particulate, will typically comprise particles with sizes ranging from about 1 nanometer to about 500 microns.

[0165] It should be understood that in addition to the ingredients particularly mentioned above, the formulations may include other agents conventional in the art having regard to the type of formulation in question, for example, those suitable for oral administration may include flavouring agents or may also include polymeric excipients / additives or carriers, e.g., polyvinylpyrrolidones, derivatised celluloses such as hydroxymethylcellulose, hydroxyethylcellulose, and hydroxypropylmethylcellulose, Ficolls (a polymeric sugar), hydroxy ethyl starch (HES), dextrates (e.g., cyclodextrins, such as 2-hydroxypropyl-P- cyclodextrin and sulfobutylether-P-cyclodextrin), polyethylene glycols, and pectin. The compositions may further include diluents, buffers, citrate, trehalose, binders, disintegrants, thickeners, lubricants, preservatives (including antioxidants), inorganic salts (e.g., sodium chloride), antimicrobial agents (e.g., benzalkonium chloride), sweeteners, antistatic agents, sorbitan esters, lipids (e.g., phospholipids such as lecithin and other phosphatidylcholines, phosphatidylethanolamines, fatty acids and fatty esters, steroids (e.g., cholesterol)), and chelating agents (e.g., EDTA, zinc and other such suitable cations).

[0166] Dosage

[0167] The amount of active ingredient that is required to achieve a therapeutic effect will, of course, vary with the particular compound, the route of administration, the subject under treatment, including the type, species, age, weight, sex, and medical condition of the subject being treated, and the renal and hepatic function of the subject, and the particular condition, disorder or disease being treated, as well as its severity. An ordinary skilled physician or clinician can readily determine and prescribe the effective amount of the drug required to prevent or treat the condition, disorder or disease.

[0168] A Papain-like protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, may for example be administered as a single daily dose, or otherwise the total daily dosage may be administered in divided doses of two, three, or four times daily. In one example, the Papain-like protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, may be dosed less frequently than once per day, for example once per two days, three days, four days, five days, six days, or once per week.

[0169] If administered intravenously, an infusion of the compound over a period of time may be used, for example. Furthermore, a Papain-like protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, may be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in the art. To be administered in the form of a transdermal delivery system, the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen.

[0170] Combinations

[0171] Whilst a Papain-like protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, may be used as the sole active agent in a medicament, it is also possible for a Papain-like protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, to be used in combination with one or more further therapeutic agents. Accordingly, in one example, a Papain-like protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, is used in combination with one or more further therapeutic agents. The present disclosure therefore also provides a combination of a Papain-like protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, and a further therapeutic agent. The present disclosure also provides a pharmaceutical composition comprising a combination of a Papainlike protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, a further therapeutic agent, and a pharmaceutically acceptable excipient. Such one or more further therapeutic agents may, for example, be anti-viral agents. Drugs are often coadministered with other drugs during therapy of a viral infection. Examples of further active agent for preventing, treating, or reducing the likelihood of infection with a virus (e.g., a coronavirus) include, but are not limited to, anti-viral agents, vaccines, immunomodulators, antibacterial agents, and / or anti-inflammatory agents. In one example, a Papain-like protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, is used in combination with a further active agent selected from the group consisting of anti-viral agents, vaccines, immunomodulators, antibacterial agents, and anti-inflammatory agents.

[0172] In one example, a Papain-like protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, is used in combination with an anti-viral agent. As used herein the term “anti-viral agent” refers to a compound that is directly or indirectly effective in specifically interfering with at least one viral action selected from one or more of: virus penetration of a eukaryotic cell, virus replication in a eukaryotic cell, virus assembly, virus release from infected eukaryotic cells, or that is effective in specifically inhibiting virus titre increase or in specifically reducing a virus titre level in a eukaryotic or mammalian host system. The term also refers to an agent that prevents or reduces the likelihood of contracting a viral infection. In one example, a Papain-like protease (PLpro) inhibitor of Formula I, or salt, solvate, stereoisomer, or deuterated analogue thereof, is used in combination with a vaccine.

[0173] The further therapeutic agents, when employed in combination with a Papain-like protease (PLpro) inhibitor of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof, may be used for example in those amounts indicated in the Physicians’ Desk Reference or as otherwise determined by one of ordinary skill in the art.

[0174] EXAMPLES

[0175] Example 1. Synthesis of Compounds of Formula I

[0176] General Scheme 1 General Method 1 (GM-1)

[0177] (GM-la) To a solution of the requisite aryl OR heteroaryl halide (1 eq.) in dry THF at -78°C is added a solution of n-BuLi (2M in cyclohexane, 1.1 eq) dropwise. After stirring for 1 hour at this temperature a solution of Intermediate 1 (1.1 eq) in dry THF is added dropwise. The reaction mixture was allowed to achieve ambient temperature and stirred over 2 hours. The reaction is poured onto ice water and extracted with EtOAc. The combined organics are dried over anhydrous MgSO4 ORNa2SO4, filtered, and concentrated in vacuo. The crude material is purified by SiCb column chromatography to give exemplar ketones of formula (i)

[0178] (GM-lb) A flask was charged with Intermediate 18 (2 eq), the requisite boronic acid (1 eq), Cu(TC) (2 eq) and Pd2(dba)3 (0.05 eq) under N2 atmosphere. To this is added degassed THF followed by triethyl phosphite (0.2 eq) and the reaction stirred at ambient temperature over 5 days. The reaction is filtered through Celite® and the filter cake washed with EtOAc. The combined organics were washed with sat. NaHCCh (aq) (x2) and brine, dried over anhydrous MgSO4 OR Na2SO4, filtered, and concentrated in vacuo. The crude material is purified by reverse-phase chromatography to give exemplar ketones of formula (i)

[0179] (GM-lc) To a solution of l-(tert-butoxycarbonyl)piperidine-3 -carboxylic acid (1 eq) and 2- chloro-4,6-dimethoxy-l,3,5-triazine (1.05 eq) in dry de-gassed PhMe was added NMM (1.05 eq). The mixture was stirred for 2 hours at ambient temperature before addition of the requisite boronic acid (1 eq), K3PO4 (2 eq) and Pd(dppf)C12.DCM (1.1 eq). The reaction was heated at 100°C until complete by LCMS. The reaction was filtered through Celite® washing the filter cake with EtOAc and concentrated in vacuo. The crude material is purified by SiO2 column chromatography to give exemplar ketones of formula (i)

[0180] General Method 2 (GM-2)

[0181] A solution of a ketone of formula (i) (1 eq.) in DCM or MeOH is treated with a suitable strong acid (10 eq.) (typically TFA or HC1) at 0°C. The reaction is allowed to achieve ambient temperature and stirred until deprotection is complete by LCMS or TLC monitoring. The reaction is concentrated to dryness to give exemplar ketones of formula (ii).

[0182] General Method 3 (GM-3)

[0183] (GM-3a) A compound of formula (ii) is dissolved in DCE or DMF and treated sequentially with acetic acid (2 eq), requisite carbonyl (3 eq) and NaBH(OAc)3 (1.5eq). The reaction mixture is stirred at ambient temperature until complete by LCMS. The reaction is diluted with saturated NaHCO, (aq) and extracted with EtOAc. The combined organics were dried over anhydrous MgSCU OR Na2SO4, filtered, and concentrated in vacuo. The crude material is purified by SiO2 column chromatography to give exemplar ketones of formula (iii or vi).

[0184] (GM-3b) A compound of formula (ii) and the requisite carbonyl (3 eq.) are stirred in anhydrous THF under N2 for 30 min. After this time phenylsilane (5 eq.) is added and the reaction stirred at ambient temperature over 12 hours. The reaction mixture is poured onto water and extracted with DCM. The combined organics were washed with brine, dried over anhydrous MgSCU OR Na2SO4, filtered, and concentrated in vacuo. The crude material is purified by SiO2 column chromatography to give exemplar ketones of formula (iii or vi).

[0185] (GM-3c) To a stirred solution of a compound of formula (ii) and K2CO3 (3 eq.) in anhydrous DMF under N2 is added the requisite alkylhalide (2 eq.) and potassium iodide (0.2 eq.). The reaction is heated at 90°C until complete by LCMS monitoring. The reaction is poured onto ice water and extracted with EtOAc. The combined organics are washed with brine, dried over anhydrous MgSO4 ORNa2SO4, filtered, and concentrated in vacuo. The crude material is purified by SiO2 column chromatography to give exemplar ketones of formula (iii or vi).

[0186] General Method 4 (GM-4)

[0187] To a stirred solution of a compound of formula (iii) in EtOH OR pyridine was added the requisite hydroxylamine (3 eq.) (typically as a hydrochloride salt). The reaction is heated between 70 - 90°C until complete by LCMS or TLC monitoring. The reaction mixture is evaporated to minimal volume under reduced pressure, poured onto ice chilled water and extracted with EtOAc. The combined organics were dried over anhydrous MgSO4 OR Na2SO4, filtered, and concentrated in vacuo. The crude compound was purified by RP-HPLC to give exemplar oximes of formula (iv or v or vi).

[0188] General Method 5 (GM-5)

[0189] To an ice-cold solution of a compound of formula (v) in anhydrous THF was added sodium hydride (60% w / w, 3 eq.). The reaction is stirred for 30 min before addition of the requisite alkylhalide OR heteroarylhalide (1.5 eq.). The reaction is then heated at 60 - 80°C until complete by LCMS or TLC monitoring. The reaction mixture is evaporated to minimal volume under reduced pressure, poured onto ice chilled water and extracted with EtOAc. The combined organics were dried over anhydrous MgSC ORNa2SO4, filtered, and concentrated in vacuo. The crude compound was purified by RP-HPLC to give exemplar oximes of formula (vi).

[0190] SYNTHETIC PROCEDURES

[0191] The compositions and methods of the disclosure are illustrated further by the following examples, which are not to be construed as limiting the disclosure in the scope to the specific procedures and compounds described in them.

[0192] The following procedures using the General Methods as described above outline the synthetic pathway used to access intermediary ketones of formula iii as shown in General Scheme 2.

[0193] General Scheme 2

[0194] Int-1 - tert-butyl 3-(methoxy(methyl)carbamoyl)piperidine-l-carboxylate.

[0195] To a solution of l-(tert-butoxycarbonyl)piperidine-3 -carboxylic acid in dry THF was added CDI (1.1 eq). The mixture was stirred for 1 hour at ambient temperature before addition of DIEA (2 eq) followed by N,O-dimethylhydroxylamine hydrochloride (1.05 eq). The reaction was stirred at ambient temperature over 18 hours. The reaction was diluted with EtOAc, washed with 10% citric acidfat / / sat. NaHCCE, and brine. The organics were dried over anhydrous MgSCU, filtered, and concentrated in vacuo. The crude material was purified by SiCh column chromatography (0- 50%EtOAc / n-heptane) to afford the title compound.

[0196] Int-la - tert-butyl (3R)-3-[methoxy(methyl)carbamoyl]piperidine-l-carboxylate.

[0197] Preparation is analogous to Int-1 utilising (3R)-l-[(tert-butoxy)carbonyl]piperidine-3-carboxylic acid to afford the title compound.

[0198] Int-lb - tert-butyl (3 S)-3-[methoxy(methyl)carbamoyl]piperidine-l -carboxylate. Preparation is analogous to Int-1 utilising (3S)-l-[(tert-butoxy)carbonyl]piperidine-3-carboxylic acid to afford.

[0199] Int-2 - tert-butyl (6-m ethoxynaphthal ene-2-carbonyl)piperi dine- 1 -carboxylate.

[0200] Int-2 was prepared from Int-1 using GM- la with 2-bromo-6-methoxynaphthalene to afford the title compound. ES+ MS: (M + H) 370.3.

[0201] Int-2a - tert-butyl (3R)-3-(6-methoxynaphthalene-2-carbonyl)piperidine-l-carboxylate.

[0202] Int-2a was prepared from Int-1 a using GM- la with 2-bromo-6-methoxynaphthalene to afford the title compound. ES+ MS: (M + H) 370.3.

[0203] Int-2b - tert-butyl (3 S)-3 -(6-m ethoxynaphthal ene-2-carbonyl)piperi dine- 1 -carboxylate.

[0204] Int-2b was prepared from Int-lb using GM- la with 2-bromo-6-methoxynaphthalene to afford the title compound. ES+ MS: (M + H) 370.3.

[0205] Int-3 - 3-(6-methoxynaphthalene-2-carbonyl)piperidine.

[0206] Int-3 was prepared from Int-2 using GM-2 to afford the title compound. ES+ MS: (M + H) 270.3.

[0207] Int-3a - (3R)-3 -(6-m ethoxynaphthal ene-2-carbonyl)piperi dine.

[0208] Int-3 a was prepared from Int-2a using GM-2 to afford the title compound. ES+ MS: (M + H) 270.3.

[0209] Int-3b - (3S)-3-(6-methoxynaphthalene-2-carbonyl)piperidine.

[0210] Int-3b was prepared from Int-2b using GM-2 to afford the title compound. ES+ MS: (M + H) 270.3.

[0211] Int-4 - l-cyclopentyl-3-(6-methoxynaphthalene-2-carbonyl)piperidine.

[0212] Int-4 was prepared from Int-3 using GM-3a with cyclopentanone to afford the title compound. ES+ MS: (M + H) 338.2.

[0213] Int-4a - (3R)-l-cyclopentyl-3-(6-methoxynaphthalene-2-carbonyl)piperidine.

[0214] Int-4a was prepared from Int-3 a using GM-3a with cyclopentanone to afford the title compound. ES+ MS: (M + H) 338.2.

[0215] Int-4b - (3 S)-l-cy cl opentyl-3-(6-m ethoxynaphthal ene-2-carbonyl)piperi dine. Int-4b was prepared using GM-3a with Int-3b and cyclopentanone to afford the title compound.

[0216] ES+ MS: (M + H) 338.2.

[0217] Int-7 - 4-[(tert-butyldimethylsilyl)oxy]cyclohexan-l-one.

[0218] To a stirred solution of 4-hydroxycyclohexan-l-one (1 eq.) in DCM (25 mL) was added 1H- imidazole (1.5 eq.) and tert-butyl(chloro)dimethylsilane (1.2 eq.) under nitrogen atmosphere. The reaction mixture was stirred at ambient temperature over 5 hours. The reaction mixture was diluted with DCM and the organic layer washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude compound was purified by SiCh column chromatography (15%EtOAc / n-hexane) to afford the title compound.XH NMR (400 MHz, DMSO-d6) 64.16 - 4.11 (m, 1H), 2.45 - 2.37 (m, 2H), 2.25 - 2.18 (m, 2H), 1.93 - 1.86 (m, 2H), 1.82 - 1.74 (m, 2H), 0.88 (s, 9H), 0.84 (s, 6H). ES+ MS: (M + H) 229.2.

[0219] Int-8a - 4-[(3R)-3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexan-l-ol (Peak A). Int-8b - 4-[(3R)-3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexan-l-ol (Peak B). Int-8a & Int-8b were prepared from Intermediate 3a using GM-3a with 4-hydroxycyclohexan-l- one. Purification by RP-HPLC gave two peaks. The first eluting Peak A was assigned as Int-8a, ES+ MS: (M + H) 368.2, and the second eluting Peak B was assigned as Int-8b, ES+ MS: (M + H) 368.2.

[0220] Int-8c - 4-[(3S)-3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexan-l-ol (Peak A). Int-8d - 4-[(3S)-3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexan-l-ol (Peak B). Int-8c & Int-8d were prepared from Int-3b using GM-3a with 4-hydroxycyclohexan-l-one. Purification by RP-HPLC gave two peaks. The first eluting Peak A was assigned as Int-8c, ES+ MS: (M + H) 368.2, and the second eluting Peak B was assigned as Int-8d, ES+ MS: (M + H) 368.2.

[0221] Int-9 - l-{4-[(tert-butyldimethylsilyl)oxy]cyclohexyl}-3-(6-methoxynaphthalene-2- carbonyl)piperidine.

[0222] Int-9 was prepared from Int-3 using GM-3a with Int-7 to afford the title compound. ES+ MS: (M + H) 383.2.

[0223] Int-lOa - 3-(6-methoxynaphthalene-2-carbonyl)-l-{8-oxabicyclo[3.2. l]octan-3-yl}piperidine (Peak A). Int-lOb - 3-(6-methoxynaphthalene-2-carbonyl)-l-{8-oxabicyclo[3.2. l]octan-3-yl}piperidine (Peak B).

[0224] Int-lOa & Int-lOb were prepared from Int-3 using GM-3a with 8-oxabicyclo[3.2.1]octan-3-one. Purification by SiCb column chromatography (10%MeOH / DCM) gave two peaks. The first eluting Peak A was assigned as Int-lOa, ES+ MS: (M + H) 380.3, and the second eluting Peak B was assigned as Int-lOb, ES+ MS: (M + H) 368.2.

[0225] Int-11 - 2-(oxan-4-yloxy)-2,3-dihydro-lH-isoindole-l, 3-dione.

[0226] To an ice cold stirred solution of 2-hydroxy-2,3-dihydro-lH-isoindole-l, 3-dione (1.4 eq.), oxan- 4-ol (1 eq.) and PPI13 (1.4 eq.) in anhydrous THF under N2 was added DIAL) (1.4 eq.). The reaction was allowed to achieve ambient temperature and stirred over 16 hours. The reaction mixture was concentrated in vacuo and the crude compound was purified by SiCh column chromatography (50%EtOAc / n-hexane) to afford the title compound.XH NMR (400 MHz, DMSO-d6) 8 7.90 - 7.86 (m, 2H), 7.82 - 7.77 (m, 2H), 4.51 - 4.42 (m, 1H), 4.15 - 4.05 (m, 2H), 3.59 - 3.47 (m, 2H), 2.10 - 2.00 (m, 2H), 1.98 - 1.85 (m, 2H). ES+ MS: (M + H) 248.3.

[0227] Int-12 - O-(oxan-4-yl)hydroxylamine hydrochloride.

[0228] To a stirred solution of Int-11 (1.0 g, 4.04 mmol) in MeOH:CHCh (4: 1) was added hydrazine hydrate (3 eq.). The reaction mixture was stirred at ambient temperature over 12 hours. The reaction mixture was filtered and concentrated in vacuo. The pH of the residue was adjusted to ~5 with 4N HC1 in dioxane and the resulting solid collected via vacuum filtration washing with Et2O and DCM to afford the title compound.

[0229] Int-13 - 2-{[(l,2,3,4-tetrahydronaphthalen-l-yl)amino]oxy}-2,3-dihydro-lH-isoindole-l,3- dione.

[0230] Int-13 was prepared in an analogous method to Int-11 using 1,2,3,4-tetrahydronaphthalen-l-ol to afford the title compound.

[0231] Int-14 - O-(l,2,3,4-tetrahydronaphthalen-l-yl)hydroxylamine.

[0232] Int-14 was prepared in an analogous method to Int-12 from Intermediate 13 to afford the title compound.XH NMR (400 MHz, DMSO-d6) 6 7.38 - 7.34 (m, 1H), 7.17 - 7.12 (m, 2H), 7.07 (d, . / = 7,2Hz, 1H), 5.96 (s, 2H), 4.45 (m, 1H), 2.76 - 2.56 (m, 2H), 2.17 - 2.12 (m, 1H), 1.85 - 1.60 (m, 3H).

[0233] Int-18 - tert-butyl 3-phenylsulfanylcarbonylpiperidine-l-carboxylate. To a solution of l-tert-butoxycarbonylpiperidine-3 -carboxylic acid in dry THF (5 mL) under N2 was added CDI (1.1 eq.). The reaction was stirred at ambient temperature over 1 hour before addition of benzenethiol (1.05 eq) and DIEA (1.15 eq.). The reaction was stirred at ambient temperature overnight. The reaction was diluted with sat.NT Cl and extracted with EtOAc. The extracts were combined, dried over MgSCU, filtered, and concentrated in vacuo. Purification by SiCb column chromatography (0-10%EtOAc / n-heptane) affords the title compound.TH NMR (300 MHz, CDCh) 8 7.46 - 7.37 (m, 5H), 4.28 (s, 1H), 4.02 (d, J = 13.3 Hz, 1H), 3.01 (s, 1H), 2.86 - 2.72 (m, 2H), 2.23 - 2.11 (m, 1H), 1.85 - 1.64 (m, 3H), 1.50 (s, 9H).

[0234] Int-19 - tert-butyl 3-(2,3-dihydro-l,4-benzodioxine-6-carbonyl)piperidine-l-carboxylate.

[0235] Intermediate 19 was prepared from Int-18 using GM-lb with 2,3-dihydro-l,4-benzodioxin-6- ylboronic acid to afford the title compound. ’H NMR (300 MHz, CDCh) 5 7.57 - 7.52 (m, 2H), 6.97 - 6.92 (m, 1H), 4.41 - 4.24 (m, 4H), 4.21 - 4.07 (m, 1H), 3.42 - 3.24 (m, 1H), 3.02 - 2.86 (m, 1H), 2.75 (t, J= 12.3 Hz, 1H), 2.11 - 1.97 (m, 1H), 1.83 - 1.55 (m, 4H), 1.50 (s, 9H).

[0236] Int-20 - ( 1 -cy clopentyl-3 -piperidyl)-(2, 3 -dihydro- 1 ,4-benzodioxin-6-yl)m ethanone.

[0237] Int-20 was prepared from Inte-19 using GM-2 followed directly by GM-3a using cyclopentanone to afford the title compound. ES+ MS: (M + H) 316.2.

[0238] Int-21 - 2,3-dihydro-l,4-benzodioxin-6-yl-(l-tetrahydrothiopyran-4-yl-3-piperidyl)methanone.

[0239] Int-21 was prepared from Int-19 using GM-2 followed directly by GM-3a using tetrahydrothiopyran-4-one to afford the title compound. ES+ MS: (M + H) 348.2.

[0240] Int-22 - (3R)-l-(4-m ethoxy cy cl ohexyl)-3-(6-meth oxynaphthal ene-2-carbonyl)piperi dine.

[0241] Int-22 was prepared from Int-3a using GM-3a with 4-methoxycyclohexan-l-one to afford the title compound. ES+ MS: (M + H) 382.2.

[0242] Int-23 - (3R)-l-(4-ethoxycy cl ohexyl)-3-(6-m ethoxynaphthal ene-2-carbonyl)piperi dine.

[0243] Int-23 was prepared from Intermediate 3a using GM-3a with 4-ethoxycyclohexan-l-one to afford the title compound. ES+ MS: (M + H) 396.2.

[0244] Int-24 - 8-methyl-l,4-dioxaspiro[4.5]decan-8-ol.

[0245] To an ice cold stirred solution of l,4-dioxaspiro[4.5]decan-8-one (1 eq.) in anhydrous THF under N2 was added 3.0M methyl magnesium bromide in ether (2 eq.). The reaction was allowed to achieve ambient temperature and stirred over 3 hours. The reaction mixture was diluted with saturated NlH Cl at^ and extracted with EtOAc. The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to afford the crude title compound.

[0246] Int-25 - 4-hydroxy-4-methylcyclohexan-l-one.

[0247] To an ice cold stirred solution of Int-24 in anhydrous THF (400 mL) under N2 was added 4.0M hydrogen chloride in dioxane (2 eq.). The reaction was allowed to achieve ambient temperature and stirred over 3 hours. The reaction mixture was diluted with saturated NaHCCT / r / t / J and extracted with EtOAc. The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to afford the crude title compound.

[0248] Int-26a - 4- [3 -(6-m ethoxynaphthal ene-2-carbonyl)piperi din- 1 -y 1 ] - 1 -methylcyclohexan- 1 -ol (Peak A).

[0249] Int-26b - 4- [3 -(6-m ethoxynaphthal ene-2-carbonyl)piperi din- 1 -y 1 ] - 1 -methylcyclohexan- 1 -ol (Peak B).

[0250] Int-26a & Int-26b were prepared from Int-3 using GM-3a with Int-25. Purification by RP HPLC gave two peaks. The first eluting Peak A was assigned as Int-26a, ES+ MS: (M + H) 382.2, and the second eluting Peak B was assigned as Int-26b, ES+ MS: (M + H) 382.2.

[0251] Int-26c - 4-[(3R)-3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]-l-methylcyclohexan-l- ol.

[0252] Int-26c was prepared from Int-3 a using GM-3b with Int-25 to afford the title compound. ES+ MS: (M + H) 382.2.

[0253] Int-27 - 4-ethyl-4-hydroxycyclohexan-l-one.

[0254] Int-27 was prepared in a method analogous to Int-24 using ethyl magnesium bromide. The crude 8-ethyl-l,4-dioxaspiro[4.5]decan-8-ol was used directly in a method analogous to Int-25 to afford the title compound.

[0255] Int-28 - l-ethyl-4-[(3R)-3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexan-l-ol. Int-28 was prepared from Int-3 a using GM-3b with Int-27 to afford the title compound. ES+ MS: (M + H) 396.2.

[0256] Int-29 - l-(cyclopropylmethyl)-3-(6-methoxynaphthalene-2-carbonyl)piperidine.

[0257] In -29 was prepared from Int-3 using GM-3a with cyclopropanecarbaldehyde to afford the title compound. ES+ MS: (M + H) 324.3. Int-30 - l-(2-methoxyethyl)-3-(6-m ethoxynaphthal ene-2-carbonyl)piperi dine.

[0258] Int-30 was prepared from Int-3 using GM-3c with 1 -chi oro-3 -methoxypropane to afford the title compound. ES+ MS: (M + H) 342.3.

[0259] Intermediate 31 -4-{2-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]ethyl}pyridine.

[0260] To a stirred solution of Int-3 (free base) in isopropanol was added 4-ethenylpyridine (2 eq.) dropwise. After addition was complete the reaction was heated at 70°C over 10 hours. The reaction was concentrated in vacuo and the crude diluted with water and extracted with DCM. The combined organics were washed with water and brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude material was purified by SiCb column chromatography (5%MeOH / DCM) to afford the title compound. ES+ MS: (M + H) 375.3.

[0261] Int-32 - 3-(6-methoxynaphthalene-2-carbonyl)-l-(2-phenylethyl)piperidine.

[0262] Int-32 was prepared from Int-3 using GM-3c with (2-chloroethyl)benzene to afford the title compound. ES+ MS: (M + H) 374.3.

[0263] Int-33 - 4-{2-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]ethyl}benzamide.

[0264] Int-33 was prepared from Int-3 using GM-3c with 4-(2-chloroethyl)benzamide and acetonitrile as the solvent to afford the title compound. ES+ MS: (M + H) 417.2.

[0265] Int-34a - 3-(6-methoxynaphthalene-2-carbonyl)-l-[4-(trifluoromethyl)cyclohexyl]piperidine (Peak A).

[0266] Int-34b - 3-(6-methoxynaphthalene-2-carbonyl)-l-[4-(trifluoromethyl)cyclohexyl]piperidine (Peak B).

[0267] Int-34a & Int-34b were prepared from Int-3 using GM-3a with 4-(trifluoromethyl)cyclohexan-l- one. Purification by RP-HPLC gave two peaks. The first eluting Peak A was assigned as Int-34a, ES+ MS: (M + H) 420.4, and the second eluting Peak B was assigned as Int-34b, ES+ MS: (M + H) 420.4.

[0268] Int-36 - [4-(2-chloroethyl)phenyl]methanol.

[0269] To a stirred 0°C solution of lithium aluminium hydride (2 eq.) in anhydrous ether was added drop wise (methyl 4-(2-chloroethyl)benzoate as a solution in anhydrous ether. The reaction was allowed to achieve ambient temperature then gently warmed at 50°C over 2 hours. The reaction was cooled to ambient temperature and carefully quenched by drop wise addition of EtOAc. The inorganic precipitate was filtered and the filtrated washed with water and brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to afford the crude title compound. 'H NMR (400 MHz, DMSO-d6 with D2O) 5 7.31 - 7.10 (m, 4H), 4.45 (s, 2H), 3.90 - 3.75 (m, 2H), 3.10 - 2.95 (m, 2H).

[0270] Int-37 - (4-{2-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]ethyl}phenyl)methanol.

[0271] Int-37 was prepared from Int-3 using GM-3c with Int-36 to afford the title compound. ES+ MS: (M + H) 404.3.

[0272] Int-38 - 4-bromo-l-{ [2-(trimethylsilyl)ethoxy]methyl}-lH-pyrazole.

[0273] To a stirred solution of 4-bromopyrazole (1 eq.) and K2CO3 (2 eq.) in anhydrous DMF under N2 was added [2-(chloromethoxy)ethyl]trimethylsilane (1.1 eq.). The reaction was stirred at ambient temperature over 18 hours. The reaction mixture was poured onto ice water and extracted with EtOAc. The combined organics were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude material was purified by SiO2column chromatography (30%EtOAc / n-heptane) to afford the title compound. 'H NMR (400 MHz, DMSO-d6) 6 8.16 (s, 1H), 7.64 (s, 1H), 5.37 (s, 2H), 3.55 - 3.48 (m, 2H), 0.88 - 0.78 (m, 2H), -0.00 (s, 2H), -0.01 (s, 9H).

[0274] Int-39 - 4-{ l,4-dioxaspiro[4.5]dec-7-en-8-yl}-l-{[2-(trimethylsilyl)ethoxy]methyl}-lH- pyrazole.

[0275] To a stirred solution of Int-38 (5.0 g, 18.0 mmol), 2-{ l,4-dioxaspiro[4.5]dec-7-en-8-yl}-4, 4,5,5- tetramethyl-l,3,2-dioxaborolane (1.3 eq) and Na2CO3 (1.5 eq.) in MeCN:water (3: 1) under N2 was added Pd(PPh3)4 (0.1 eq.). The reaction mixture was de-gassed with N2 for 10 min then heated at 100°C over 12 hours. The reaction mixture was filtered through Celite ® and the filter cake washed with EtOAc. The combined organics were washed with water and brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude material was purified by SiO2column chromatography (30%EtOAc / n-heptane) to afford the title compound. ES+ MS: (M + H) 337.2.

[0276] Int-40 - 4-{ l,4-dioxaspiro[4.5]decan-8-yl}-l-{[2-(trimethylsilyl)ethoxy]methyl}-lH-pyrazole.

[0277] To a stirred solution of Inte-39 in EtOAc under N2 was added 10%Pd / C (10%w / w). The flask was evacuated and backfilled with H2. The reaction was stirred under H2over 16 hours. The reaction mixture was filtered through Celite ® and the filter cake washed with EtOAc. The combined organics were concentrated in vacuo to afford the crude title compound. ES+ MS: (M + H) 339.2.

[0278] Int-41 - 4-(l-{[2-(trimethylsilyl)ethoxy]methyl}-lH-pyrazol-4-yl)cyclohexan-l-one.

[0279] To a stirred 0°C solution of Int-40 in water was added acetic acid (40 eq.) drop wise. The reaction was allowed to achieve ambient temperature and stirred over 8 hours. The reaction was concentrated in vacuo to afford the crude title compound. ES+ MS: (M + H) 295.2.

[0280] Int-42a - 3-(6-methoxynaphthalene-2-carbonyl)-l-[4-(l-{[2-(trimethylsilyl)ethoxy]methyl}- lH-pyrazol-4-yl)cyclohexyl]piperidine (Peak A).

[0281] Int-42b - 3-(6-methoxynaphthalene-2-carbonyl)-l-[4-(l-{[2-(trimethylsilyl)ethoxy]methyl}- lH-pyrazol-4-yl)cyclohexyl]piperidine (Peak B).

[0282] Int-42a & Int-42b were prepared from Int-3 using GM-3a with Int-41. Purification by RP-HPLC gave two peaks. The first eluting Peak A was assigned as Int-42a, ES+ MS: (M + H) 548.4, and the second eluting Peak B was assigned as Int-42b, ES+ MS: (M + H) 548.4.

[0283] Int-43 - tert-butyl 3-(5-methoxy-l-benzofuran-2-carbonyl)piperidine-l-carboxylate.

[0284] To a -78°C stirred solution of 5 -methoxy- 1 -benzofuran in anhydrous THF (15 mL) under N2 was added 2.5M n-BuLi in hexane (1.5 eq.) drop wise. The reaction was stirred at this temperature for 1 hour after which Intermediate 1 (1.1 eq.) was added drop wise as a solution in anhydrous THF (5 mL). The reaction was continued at this temperature for 1 hour. The reaction was diluted with saturated NH4Cl( ^) and extracted with EtOAc. The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude material was purified by SiCh column chromatography (20%EtOAc / n-hexane) to afford the title compound. ES+ MS: (M + H) 360.2.

[0285] Int-44 - 3-(5-methoxy-l-benzofuran-2-carbonyl)piperidine hydrochloride.

[0286] Int-44 was prepared from Int-43 using GM-2 with 4M hydrogen chloride in dioxane to afford the crude title compound. ES+ MS: (M + H) 260.2.

[0287] Int-45 - l-cyclopentyl-3-(5-methoxy-l-benzofuran-2-carbonyl)piperidine.

[0288] Int-45 was prepared from Int-44 using GM-3a with cyclopentanone to afford the title compound. ES+ MS: (M + H) 328.4.

[0289] Int-46 - tert-butyl 3-(5-methoxy-l-benzothiophene-2-carbonyl)piperidine-l-carboxylate. Int-46 was prepared in an analogous method to Int-43 using 5-methoxy-l -benzothiophene to afford the title compound. ES+ MS: (M + H) 376.3.

[0290] Int-47 - 3-(5-methoxy-l-benzothiophene-2-carbonyl)piperidine hydrochloride.

[0291] Int-47 was prepared from Int-46 using GM-2 with 4M hydrogen chloride in dioxane to afford crude title compound. ES+ MS: (M + H) 276.1.

[0292] Int-48 - l-cyclopentyl-3-(5-methoxy-l-benzofuran-2-carbonyl)piperidine.

[0293] Int-48 was prepared from Int-47 using GM-3a with cyclopentanone to afford the title compound. ES+ MS: (M + H) 344.4.

[0294] Int-49 - tert-butyl 3 -(6-methoxy-l-benzothiophene-2-carbonyl)piperidine-l -carboxylate.

[0295] Int-49 was prepared in an analogous method to Intermediate 43 using 6-methoxy-l- benzothiophene to afford the title compound. ES+ MS: (M + H) 376.3.

[0296] Int-50 - 3-(6-methoxy-l-benzothiophene-2-carbonyl)piperidine hydrochloride.

[0297] Int-50 was prepared from Int-49 using GM-2 with 4M hydrogen chloride in dioxane to afford the title compound. ES+ MS: (M + H) 276.1.

[0298] Int-51 - l-cyclopentyl-3-(6-methoxy-l-benzothiophene-2-carbonyl)piperidine.

[0299] Int-51 was prepared from Int-50 using GM-3a with cyclopentanone to afford the title compound. ES+ MS: (M + H) 344.4.

[0300] Int-52 - tert-butyl 3-(6-methoxy-l,3-benzothiazole-2-carbonyl)piperidine-l-carboxylate.

[0301] Int-52 was prepared in an analogous method to Int-43 using 6-m ethoxy- 1 -benzothiophene to afford the title compound. ES+ MS: (M + H) 377.2.

[0302] Int-53 - 6-methoxy-2-(piperidine-3-carbonyl)-l,3-benzothiazole hydrochloride.

[0303] Int-53 was prepared from Int-52 using GM-2 with 4M hydrogen chloride in dioxane to afford the title compound. ES+ MS: (M + H) 277.3.

[0304] Int-54 - 2-(l-cyclopentylpiperidine-3-carbonyl)-6-methoxy-l,3-benzothiazole.

[0305] Int-54 was prepared from Int-53 using GM-3a with cyclopentanone to afford the title compound.

[0306] ES+ MS: (M + H) 345.3. Int-55a - {4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexyl}methanol (Peak

[0307] A).

[0308] Int-55b - {4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexyl}methanol (Peak

[0309] B).

[0310] Int-55a & Int-55b were prepared from Inte-3 using GM-3a with 4-(hydroxymethyl)cyclohexan- 1-one. Purification by RP-HPLC gave two peaks. The first eluting Peak A was assigned as Int- 55a, ES+ MS: (M + H) 382.4, and the second eluting Peak B was assigned as Int-55b, ES+ MS: (M + H) 382.4.

[0311] Int-56a - 4-[3-(6-m ethoxynaphthal ene-2-carbonyl)piperi din- l-yl]-l-methylcy cl ohexane-1- carbonitrile (Peak A).

[0312] Int-56b - 4-[3-(6-m ethoxynaphthal ene-2-carbonyl)piperi din- l-yl]-l-methylcy cl ohexane-1- carbonitrile (Peak B).

[0313] Int-56a & Int-56b were prepared from Int-3 using GM-3a with l-methyl-4-oxocyclohexane-l- carbonitrile. Purification by RP-HPLC gave two peaks. The first eluting Peak A was assigned as Int-55a, ES+ MS: (M + H) 391.3, and the second eluting Peak B was assigned as Int-55b, ES+ MS: (M + H) 391.3.

[0314] Int-57 - trimethyl({[8-(trifluoromethyl)-l,4-dioxaspiro[4.5]decan-8-yl]oxy})silane.

[0315] To a stirred solution of l,4-dioxaspiro[4.5]decan-8-one (1 eq.) and CS2CO3 (2 eq.) in anhydrous DME under N2 was added trimethyl(trifluoromethyl)silane (3 eq.). The reaction mixture was stirred at ambient temperature over 12 hours. After this time the reaction mixture was poured onto ice water and extracted with EtOAc. The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude material was purified by SiCh column chromatography (10%EtOAc / n-heptane) to afford the title compound. 'H NMR (400 MHz, CDCh) 5 4.00 - 3.91 (m, 4H), 1.87 - 1.79 (m, 6H), 1.71 - 1.62 (m, 2H), 0.16 (s, 9H).

[0316] Int-58 - 8-(trifluoromethyl)-l,4-dioxaspiro[4.5]decan-8-ol.

[0317] To a 0°C stirred solution of Int-57 (1 eq.) in THF under N2 was added IM tetrabutylammonium fluoride in THF (3 eq.). The reaction was allowed to achieve ambient temperature and stirred over 12 hours. The reaction mixture was poured onto ice water and extracted with EtOAc. The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to afford the title compound. 'H NMR (400 MHz, DMSO-d6) 5 5.84 (s, 1H), 3.86 (m, 4H), 1.79 - 1.51 (m, 8H). Int-59 - 4-hydroxy-4-(trifluoromethyl)cyclohexan-l-one.

[0318] To a stirred 0°C solution of Int-58 (1 eq.) in THF (4 mL) was 6N hydrochloric acid (2 eq.). The reaction was allowed to achieve ambient temperature and stirred over 12 hours. The reaction was concentrated in vacuo to afford the title compound.

[0319] Int-60a - 4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]-l-

[0320] (trifluoromethyl)cyclohexan-l-ol (Peak A).

[0321] Int-60b - 4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]-l-

[0322] (trifluoromethyl)cyclohexan-l-ol (Peak B).

[0323] Int-60a & Int-60b were prepared from Int-3 using GM-3a with Int-59. Purification by RP-HPLC gave two peaks. The first eluting Peak A was assigned as Int-60a, ES+ MS: (M + H) 436.4, and the second eluting Peak B was assigned as Int-60b, ES+ MS: (M + H) 436.4.

[0324] Int-63 - tert-butyl 3-(3-methoxyisoquinoline-7-carbonyl)piperidine-l-carboxylate.

[0325] Int-63 was prepared from Int-18 using GM-lb with (3-methoxyisoquinolin-7-yl)boronic acid to afford the title compound. ES+ MS: (M + H - tBu) 315.2.

[0326] Int-64 - 3-methoxy-7-(piperidine-3-carbonyl)isoquinoline hydrochloride.

[0327] Int-64 was prepared from Int-63 using GM-2 with 4M hydrogen chloride in dioxane to afford the title compound. ES+ MS: (M + H) 271.3.

[0328] Int-65 - 7-(l-cyclopentylpiperidine-3-carbonyl)-3-methoxyisoquinoline.

[0329] Int-65 was prepared from Int-64 using GM-3a with cyclopentanone to afford the title compound. ES+ MS: (M + H) 339.2.

[0330] Int-66a - 8-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]-l-azaspiro[4.5]decan-2-one (Peak A).

[0331] Int-66b - 8-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]-l-azaspiro[4.5]decan-2-one (Peak B).

[0332] Int-66a & Int-66b were prepared from Int-3 using GM-3a with l-azaspiro[4.5]decane-2, 8-dione. Purification by RP-HPLC gave two peaks. The first eluting Peak A was arbitrarily assigned as Int-66a, ES+ MS: (M + H) 421.3, and the second eluting Peak B was arbitrarily assigned as Int- 66b, ES+ MS: (M + H) 421.3.

[0333] Int-72 - ethyl 2-[(6-bromonaphthalen-2-yl)oxy]-2,2-difluoroacetate. To a stirred solution of ethyl 2-bromo-2,2-difluoroacetate (1.2 eq.) and 6-bromonaphthalen-2-ol (1 eq.) in DMF (9 mL) under N2 was added DBU (2.5 eq). The reaction mixture was stirred at 70°C over 12 hours. After this time the reaction mixture was poured onto ice water and extracted with EtOAc. The combined organics were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude material was purified by SiCh column chromatography (50%EtOAc / n-heptane) to afford the title compound. 'H NMR (400 MHz, DMSO-d6) 5 8.29 (s, 1H), 8.03 (d, J= 8.8 Hz, 1H), 7.98 (d, J= 8.8 Hz, 1H), 7.88 (s, 1H), 7.71 (d, J= 8.8 Hz, 1H), 7.47 (d, J= 8.8 Hz, 1H), 4.39 (q, J= 6.8 Hz, 2H), 1.25 (t, J= 7.2 Hz, 3H).

[0334] Int-73 - 2-[(6-bromonaphthalen-2-yl)oxy]-2,2-difluoroacetic acid.

[0335] To a 0°C solution of Int-72 (1 eq.) in MeOH, THF and water (3:2: 1) was added NaOH (3 eq.). The reaction was allowed to achieve ambient temperature and stirred over 12 hours. After this time the reaction mixture was poured onto ice water, the pH adjusted to 2 with IN HC1 and extracted with DCM. The combined organics were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to afford the crude title compound. 'H NMR (400 MHz, DMSO-d6) 5 8.27 (d, J= 1.6 Hz, 1H), 8.01 (d, J= 8.8 Hz, 1H), 7.96 (d, J= 8.8 Hz, 1H), 7.84 (s, 1H), 7.69 (dd, J= 2.0 Hz, 8.8 Hz, 1H), 7.46 (dd, J= 2.4 Hz, 8.8 Hz, 1H).

[0336] Int-74 - 2-bromo-6-(bromodifluoromethoxy)naphthalene.

[0337] To a 0°C solution of Int-73 (1 eq.) in DCM was added DMF (0.1 eq.) followed by drop wise addition of oxalyl chloride (4 eq.). The reaction was allowed to achieve ambient temperature and stirred over 3 hours. After this time the reaction mixture was concentrated to dryness. The crude product was dissolved in bromotrichloromethane under N2. 4-(Dimethylamino)pyridine (0.25 eq.) was added followed by sodium pyrithione (2 eq.). The resulting mixture was heated at 120°C over 2 hours. After this time the reaction was concentrated in vacuo and the crude residue purified by SiCF column chromatography (5%EtOAc / n-heptane) to afford the title compound. 'H NMR (400 MHz, CDCh) 5 8.06 (s, 1H), 7.82 (d, J= 8.8 Hz, 1H), 7.75 (d, J= 8.8 Hz, 1H), 7.69 (s, 1H), 7.64 (dd, J= 2.0 Hz, 8.8 Hz, 1H), 7.40 (dd, J= 2.0 Hz, 8.8 Hz, 1H).

[0338] Int-75 - 2-bromo-6-(trifluoromethoxy)naphthalene.

[0339] To a stirred solution of Int-74 (1 eq.) in DCM was added silver tetrafluoroborate (3 eq.). The reaction was stirred at ambient temperature over 1 hour. After this time the reaction mixture was concentrated to dryness and the crude product purified by SiCF column chromatography (l%EtOAc / pentane) to afford the title compound. 'H NMR (400 MHz, CDCh) 5 8.03 (d, J= 1.6 Hz, 1H), 7.79 (d, J= 8.8 Hz, 1H), 7.71 (d, J= 8.8 Hz, 1H), 7.69 (s, 1H), 7.66 - 7.58 (m, 2H), 7.37 (dd, J= 1.6 Hz, 8.8 Hz, 1H).

[0340] Int-76 - tert-butyl 3 -[6-(trifluoromethoxy)naphthalene-2-carbonyl]piperidine-l -carboxylate.

[0341] Int-76 was prepared from Int-65 using GM-la with Int-1 to afford the title compound. ES+ MS: (M - (*Bu) + H) 368.2.

[0342] Int-76a - tert-butyl (3R)-3-[6-(trifluoromethoxy)naphthalene-2-carbonyl]piperidine-l- carb oxy late.

[0343] Int-76 was prepared from Int-65 using GM-la with Int-la to afford the title compound. ES+ MS: (M-*Bu + H) 368.2.

[0344] Int-77 - 3-[6-(trifluoromethoxy)naphthalene-2-carbonyl]piperidine hydrochloride.

[0345] Int-77 was prepared from Int-76 using GM-2 with 4M hydrogen chloride in dioxane to afford the title compound. ES+ MS: (M + H) 324.2.

[0346] Int-77a - (3R)-3-[6-(trifluoromethoxy)naphthalene-2-carbonyl]piperidine trifluoroacetate.

[0347] Int-77a was prepared from Int-76a using GM-2 with TFA to afford the title compound. ES+ MS: (M + H) 324.2.

[0348] Int-78 - l-{4-[(tert-butyldimethylsilyl)oxy]cyclohexyl}-3-[6-(trifluoromethoxy)naphthalene-2- carbonyl]piperidine.

[0349] Int-78 was prepared from Int-77 using GM-3a with Int-7 to afford the title compound. ES+ MS: (M + H) 536.4.

[0350] Int-79 - 2-bromo-6-(difluoromethoxy)naphthalene.

[0351] To a stirred solution of 6-bromonaphthalen-2-ol (1 eq.) in acetone:water (1 : 1) was added KOH (6 eq.). The reaction mixture was stirred for 5 min before addition of (bromodifluoromethyl)tris(methyl)silane (2 eq.). The resulting mixture was heated at 50°C over 12 hours. After this time the reaction mixture was poured onto ice water and extracted with EtOAc. The combined organics were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude material was purified by SiO2 column chromatography (10%EtOAc / n-heptane) to afford the title compound. 'H NMR (400 MHz, CDCh) 5 8.00 (d, J = 1.2 Hz, 1H), 7.76 (d, J= 92 Hz, 1H), 7.67 (d, J= 8.8 Hz, 1H), 7.58 (dd, J = 2.0 Hz, 8.8 Hz, 1H), 7.48 (s, 1H), 7.30 (dd, J= 2.4 Hz, 8.8 Hz, 1H), 6.62 (t, J = 73.6 Hz, 2H). Int-80 - tert-butyl 3-[6-(difluoromethoxy)naphthalene-2-carbonyl]piperidine-l-carboxylate.

[0352] Int-80 was prepared from Int-79 using GM-la with Int-1 to afford the title compound. ES+ MS: (M + H) 406.2.

[0353] Int-80a - tert-butyl (3R)-3-[6-(difluoromethoxy)naphthalene-2-carbonyl]piperidine-l- carb oxy late.

[0354] Int-80a was prepared from Int-79 using GM-la with Int-la to afford the title compound. ES+ MS: (M + H) 406.3.

[0355] Int-81 - 3-[6-(difluoromethoxy)naphthalene-2-carbonyl]piperidine trifluoroacetate.

[0356] Int-81 was prepared from Int-80 using GM-2 with TFA to afford the title compound. ES+ MS: (M + H) 306.2.

[0357] Int-81a - (3R)-3-[6-(difluoromethoxy)naphthalene-2-carbonyl]piperidine trifluoroacetate.

[0358] Int-81 a was prepared from Int-80a using GM-2 with TFA to afford the title compound. ES+ MS: (M + H) 306.2.

[0359] Int-82 - l-{4-[(tert-butyldimethylsilyl)oxy]cyclohexyl}-3-[6-(difluoromethoxy)naphthalene-2- carbonyl]piperidine.

[0360] Int-82 was prepared from Int-81 using GM-3a with Int-7 to afford the title compound. ES+ MS: (M + H) 518.4.

[0361] Int-83#- (3R)-l-cyclopentyl-3-[6-(difluoromethoxy)naphthalene-2-carbonyl]piperidine.

[0362] Int-83 was prepared from Int-81 using GM-3a with cyclopentanone to afford the title compound. ES+ MS: (M + H) 518.4.

[0363] Int-84#- l-cyclopentyl-3-[6-(trifluoromethoxy)naphthalene-2-carbonyl]piperidine.

[0364] Int-84 was prepared from Int-77 using GM-3a with cyclopentanone to afford the title compound. ES+ MS: (M + H) 392.2.

[0365] Int-84a#- (3R)-l-cyclopentyl-3-[6-(trifluoromethoxy)naphthalene-2-carbonyl]piperidine.

[0366] Int-84a was prepared from Int-77a using GM-3a with cyclopentanone to afford the title compound. ES+ MS: (M + H) 392.2. Int-85 - 3-[(tert-butyldimethylsilyl)oxy]cyclohexan-l-one.

[0367] Int-85 was prepared in an analogous method to Intermediate 7 from 3 -hydroxy cy cl ohexan-1- one to afford the title compound. 'H NMR (400 MHz, CDC13) 6 4.17 (septet, J= 3.2 Hz, 1H), 2.57 - 2.49 (m, 1H), 2.42 - 2.22 (m, 3H), 2.13 - 2.02 (m, 1H), 1.93 - 1.85 (m, 1H), 1.78 - 1.62 (m, 2H), 0.87 (s, 9H), 0.49 (s, 6H). ES+ MS: (M + H) 229.2.

[0368] Int-86 - l-{3-[(tert-butyldimethylsilyl)oxy]cyclohexyl}-3-(6-methoxynaphthalene-2- carbonyl)piperidine.

[0369] Int-86 was prepared from Int-3 using GM-3a with Int-85 to afford the title compound. ES+ MS: (M + H) 482.2.

[0370] Int-87 - 8-(difhioromethyl)-l,4-dioxaspiro[4.5]decan-8-ol.

[0371] To a stirred solution of ethyl l,4-dioxaspiro[4.5]decane-8-carboxylate (1 eq.) and hexamethylphosphoramide (5 eq.) in anhydrous THF under N2 was added caesium fluoride (0.2 eq.) and (difluoromethyl)trimethylsilane (2 eq.). The reaction mixture was heated at 75°C over 16 hours. After cooling to ambient temperature IM tetrabutylammonium fluoride in THF (3 eq.) was added slowly and the reaction stirred over 16 hours. The reaction mixture was poured onto ice water and extracted with EtOAc. The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude material was purified by SiCh column chromatography (20%EtOAc / n-heptane) to afford the title compound.1H NMR (400 MHz, DMSO-d6) 8 5.69 (t, J= 56.4 Hz, 1H), 5.09 (s, 1H), 3.85 (s, 4H), 1.79 - 1.68 (m, 2H), 1.60 - 1.49 (m, 6H).

[0372] Int-88 - 4-(difluoromethyl)-4-hydroxycyclohexan-l-one.

[0373] Int-88 was prepared in an analogous method to Int-59 to afford the title compound.

[0374] Int-89a - l-(difluoromethyl)-4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l- yl]cyclohexan-l-ol (Peak A).

[0375] Int-89b - l-(difluoromethyl)-4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l- yl]cyclohexan-l-ol (Peak B).

[0376] Int-89a & Int-89b were prepared from Int-3 using GM-3a with Int-88. Purification by RP-HPLC gave two peaks. The first eluting Peak A was assigned as Int-89a, ES+ MS: (M + H) 418.3, and the second eluting Peak B was assigned as Int-89b, ES+ MS: (M + H) 418.3.

[0377] Int-90 - 3-bromo-l-{ [2-(trimethylsilyl)ethoxy]methyl}-lH-pyrazole. Int-90 was prepared in an analogous method to Int-38 using 3-bromopyrazole to afford the title compound.

[0378] Int-91 - 3-{ l,4-dioxaspiro[4.5]dec-7-en-8-yl}-l-{[2-(trimethylsilyl)ethoxy]methyl}-lH- pyrazole.

[0379] Int-91 was prepared in an analogous method to Int-39 to afford the title compound. ES+ MS: (M + H) 337.2.

[0380] Int-92 - 3-{ l,4-dioxaspiro[4.5]decan-8-yl}-l-{[2-(trimethylsilyl)ethoxy]methyl}-lH-pyrazole. Int-92 was prepared in an analogous method to Int-40 to afford the title compound. ES+ MS: (M + H) 339.2.

[0381] Int-93 - 3 -( 1 - { [2-(trimethyl silyl)ethoxy ]methyl } - 1 H-pyrazol-4-yl)cy clohexan- 1 -one.

[0382] Int-93 was prepared in an analogous method to Int-41 to afford the title compound. ES+ MS: (M + H) 295.2.

[0383] Int-94a - 3-(6-methoxynaphthalene-2-carbonyl)-l-[4-(l-{[2-(trimethylsilyl)ethoxy]methyl}- lH-pyrazol-3-yl)cyclohexyl]piperidine (Peak A).

[0384] Int-94b - 3-(6-methoxynaphthalene-2-carbonyl)-l-[4-(l-{[2-(trimethylsilyl)ethoxy]methyl}- lH-pyrazol-3-yl)cyclohexyl]piperidine (Peak B).

[0385] Int-94a & Int-94b were prepared from Int-3 using GM-3a with Int-93. Purification by RP-HPLC gave two peaks. The first eluting Peak A was assigned as Int-94a, ES+ MS: (M + H) 548.4, and the second eluting Peak B was assigned as Int-94b, ES+ MS: (M + H) 548.4.

[0386] Int-95 - 2-bromo-6-(2H3)m ethoxy naphthalene.

[0387] To a stirred solution of 6-bromonaphthalen-2-ol (1 eq.) in DMF was added K2CO3 (3 eq.) followed by iodomethane-d3 (1.2 eq.). The resulting mixture was stirred at ambient temperature over 12 hours. After this time the reaction mixture was poured onto ice water and extracted with EtOAc. The combined organics were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude material triturated with n-pentane to afford the title compound.XH NMR (400 MHz, DMSO-d6) 5 8.11 (d, J= 1.6 Hz, 1H), 7.82 (d, J= 8.8 Hz, 1H), 7.79 (d, J= 8.8 Hz, 1H), 7.57 (dd, J= 2.0 Hz, 8.8 Hz, 1H), 7.35 (d, J= 2.4 Hz, 1H), 7.21 (dd, J = 2.8 Hz, 9.2 Hz, 1H).

[0388] Int-96 - tert-butyl 3-[6-(2H3)methoxynaphthalene-2-carbonyl]piperidine-l-carboxylate. Int-96 was prepared from Int-95 using GM- la with Int-1 to afford the title compound. ES+ MS: (M + H) 373.4.

[0389] Int-96a - tert-butyl (3R)-3-[6-(2H3)methoxynaphthalene-2-carbonyl]piperidine-l-carboxylate. Int-96a was prepared from Int-95 using GM- la with Intermediate la to afford the title compound. ES+ MS: (M + H) 373.4.

[0390] Int-97 - 3-[6-(2H3)methoxynaphthalene-2-carbonyl]piperidine trifluoroacetate.

[0391] Int-97 was prepared from Int-96 using GM-2 with TFA to afford the title compound. ES+ MS: (M + H) 273.3.

[0392] Int-97a - (3R)-3-[6-(2H3)methoxynaphthalene-2-carbonyl]piperidine.

[0393] Int-97a was prepared from Int-96a using GM-2 with TFA to afford the title compound. ES+ MS: (M + H) 273.3.

[0394] Int-98a#- (3R)-l-cyclopentyl-3-[6-(2H3)methoxynaphthalene-2-carbonyl]piperidine.

[0395] Int-98a was prepared from Int-97a using GM-3a with cyclopentanone to afford the title compound. ES+ MS: (M + H) 392.2.

[0396] Int-99a - l-{4-[(tert-butyldimethylsilyl)oxy]cyclohexyl}-3-[6-(2H3)methoxynaphthalene-2- carbonyl]piperidine (Peak A).

[0397] Int-99b - l-{4-[(tert-butyldimethylsilyl)oxy]cyclohexyl}-3-[6-(2H3)methoxynaphthalene-2- carbonyl]piperidine (Peak B).

[0398] Int-99a & Int-99b were prepared from Int-3 using GM-3a with Int-7. Purification by RP-HPLC gave two peaks. The first eluting Peak A was assigned as Int-99a, ES+ MS: (M + H) 485.4, and the second eluting Peak B was assigned as Int-99b, ES+ MS: (M + H) 485.4.

[0399] Int-100 - 4-{ l,4-dioxaspiro[4.5]dec-7-en-8-yl}-l-methyl-lH-pyrazole.

[0400] Int-100 was prepared in an analogous method to Int-39 using 4-bromo-l-methylpyrazole to afford the title compound. ES+ MS: (M + H) 221.2.

[0401] Int-101 - 4-{ l,4-dioxaspiro[4.5]decan-8-yl}-l-{[2-(trimethylsilyl)ethoxy]methyl}-lH-pyrazole. Int-101 was prepared in an analogous method to Int-40 to afford the title compound. ES+ MS: (M + H) 223.4. Int-102 - 4-{ l,4-dioxaspiro[4.5]decan-8-yl}-l-methyl-lH-pyrazole.

[0402] Int-102 was prepared in an analogous method to Int-41 to afford the title compound. ES+ MS: (M + H) 179.0.

[0403] Int-103a - 3 -(6-m ethoxynaphthal ene-2-carbonyl)-l -[4-(l -methyl- lH-pyrazol-4- yl)cyclohexyl]piperidine (Peak A).

[0404] Int-103b - 3-(6-methoxynaphthalene-2-carbonyl)-l -[4-(l -methyl- lH-pyrazol-4- yl)cyclohexyl]piperidine (Peak B).

[0405] Int-103a & Int-103b were prepared from Int-3 using GM-3a with Int-102. Purification by RP- HPLC gave two peaks. The first eluting Peak A was assigned as Int-103a, ES+ MS: (M + H)

[0406] 432.3, and the second eluting Peak B was assigned as Int-103b, ES+ MS: (M + H) 432.3.

[0407] Int-104 - tert-butyl 3-(2-methoxyquinoline-6-carbonyl)piperidine-l-carboxylate.

[0408] Int-104 was prepared from 6-bromo-2-m ethoxy quinoline using GM- la with Int-1 to afford the title compound. ES+ MS: (M + H) 371.3.

[0409] Int-105 - 2-methoxy-6-(piperidine-3-carbonyl)quinoline trifluoroacetate.

[0410] Int-105 was prepared from Int-104 using GM-2 with TFA to afford the title compound. ES+ MS: (M + H) 271.3.

[0411] Int-106a - 6-(l-{4-[(tert-butyldimethylsilyl)oxy]cyclohexyl}piperidine-3-carbonyl)-2- methoxy quinoline (Peak A).

[0412] Int-106b - 6-(l-{4-[(tert-butyldimethylsilyl)oxy]cyclohexyl}piperidine-3-carbonyl)-2- m ethoxy quinoline (Peak B).

[0413] Int-106a & Int-106b were prepared from In-105 using GM-3a with Int-7. Purification by RP- HPLC gave two peaks. The first eluting Peak A was assigned as Int-106a, ES+ MS: (M + H)

[0414] 483.4, and the second eluting Peak B was assigned as Int-106b, ES+ MS: (M + H) 483.4.

[0415] Int-107 - l-{ l,4-dioxaspiro[4.5]decan-8-yl}-4-nitro-lH-pyrazole.

[0416] To a stirred solution of 4-nitropyrazole (1 eq.), l,4-dioxaspiro[4.5]decan-8-ol (1 eq.) and triphenylphosphine (1.5 eq.) in THF under N2 was added DIAD (1.5 eq.). The reaction was stirred at ambient temperature over 5 hours. After this time the reaction mixture was diluted with water and extracted with EtOAc. The combined organics were concentrated in vacuo. The crude material was purified by SiCb column chromatography (40%EtOAc / n-hexane) to afford the title compound. Int-108 - 4-(4-nitro-lH-pyrazol-l-yl)cyclohexan-l-one.

[0417] Intermediate 108 was prepared in an analogous method to Intermediate 41 to afford the title compound.

[0418] Int-109a - 3-(6-methoxynaphthalene-2-carbonyl)-l-[4-(4-nitro-lH-pyrazol-l- yl)cyclohexyl]piperidine (Peak A).

[0419] Int-109b - 3-(6-methoxynaphthalene-2-carbonyl)-l-[4-(4-nitro-lH-pyrazol-l- yl)cyclohexyl]piperidine (Peak B).

[0420] Int-109a & Int-109b were prepared from Int-3 using GM-3a with Int-108. Purification by RP- HPLC gave two peaks. The first eluting Peak A was assigned as Int-109a, ES+ MS: (M + H) 433.2, and the second eluting Peak B was assigned as Int-109b, ES+ MS: (M + H) 433.2.

[0421] Int-llOa - l-{4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexyl}-lH-pyrazol- 4-amine (Peak A).

[0422] To a stirred solution of Int-109a (1 eq.) and ammonium chloride (6 eq.) in EtOH:water (5: 1) was added iron powder (4 eq.). The reaction was heated at 80°C over 12 hours. After this time the reaction was filtered through a Celite® pad washing with EtOH. The filtrate was concentrated in vacuo and the crude partitioned between DCM and water. The layers were separated and the aqueous further extracted with DCM. The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to afford the crude title compound. ES+ MS: (M + H) 463.2.

[0423] Int-llOb - l-{4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexyl}-lH-pyrazol- 4-amine (Peak B).

[0424] Int-l lOb was prepared in an analogous method to Int-l lOa to afford the crude title compound. ES+ MS: (M + H) 463.2.

[0425] Int-111 - methyl 4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]butanoate.

[0426] Int-111 was prepared from Int-3 using GM-3c with methyl 4-chlorobutanoate to afford the title compound. ES+ MS: (M + H) 370.3.

[0427] Int-112 - 4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]butanoic acid.

[0428] To a solution of Intermediate 111 (1 eq.) in THF:water (5: 1) was added LiOH (5 eq.). The reaction mixture was stirred at ambient temperature over 12 hours. After this time the reaction mixture was concentrated and the pH adjusted to 4 with IN HC1 and extracted with 5%MeOH / DCM. The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to afford the title compound. ES+ MS: (M + H) 356.2.

[0429] Int-113 - 4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]-N-methylbutanamide.

[0430] To a solution of Int-112 and methylamine hydrochloride (3 eq.) in DMF under N2 was added HATU (2 eq.) followed by DIEA (4 eq.). The reaction was stirred at ambient temperature over 12 hours. After this time the reaction was poured onto ice water and extracted with EtOAc. The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude residue was purified by SiCb chromatography (5%MeOH / DCM) to give the title compound. ES+ MS: (M + H) 369.3.

[0431] Int-114 - tert-butyl N-{3-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l- yl]cyclohexyl}carbamate.

[0432] Int-114 was prepared from Int-3 using GM-3a with tert-butyl N-(3-oxocyclohexyl)carbamate to afford the title compound. ES+ MS: (M + H) 467.2.

[0433] Int-115 - 3-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexan-l -amine hydrochloride.

[0434] Int-115 was prepared from Inte-114 using GM-2 with 4M HC1 in dioxane to afford the title compound. ES+ MS: (M + H) 367.4.

[0435] Int-116a - 2-methoxy-N-{3-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l- yl]cyclohexyl}acetamide (Peak A).

[0436] Int-116b - 2-methoxy-N-{3-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l- yl]cyclohexyl}acetamide (Peak B).

[0437] Int-116a & Int-116b were prepared in an analogous method to Int-113 using 2-methoxyacetic acid. Purification by RP-HPLC gave two peaks. The first eluting Peak A was assigned as Int- 116a, ES+ MS: (M + H) 439.3, and the second eluting Peak B was assigned as Int-116b, ES+ MS: (M + H) 439.3.

[0438] Int-117 - {3-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexyl}urea.

[0439] To an ice-cold solution of Int-115 in DMF was added CDI (1 eq.) and triethylamine (2 eq.). The reaction was stirred at this temperature over 1 hour before addition of 28% ammonium hydroxide (10 eq.). The reaction was allowed to achieve ambient temperature and stirred over 3 hours. After this time the reaction was poured onto water and extracted with DCM. The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to afford the crude title compound. ES+ MS: (M + H) 410.4.

[0440] Int-118 - tert-butyl N-{4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l- yl]cyclohexyl}carbamate.

[0441] Int-118 was prepared from Int-3 using GM-3a with tert-butyl N-(4-oxocyclohexyl)carbamate to afford the title compound. ES+ MS: (M + H) 467.2.

[0442] Int-119 - 4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexan-l -amine hydrochloride.

[0443] Int-119 was prepared from Int-118 using GM-2 with 4M HC1 in dioxane to afford the title compound. ES+ MS: (M + H) 367.4.

[0444] Int-120 - N-{4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexyl}acetamide.

[0445] To an ice-cold solution of Int-119 in pyridine was slowly added acetic anhydride (2 eq.). The reaction was allowed to achieve ambient temperature and stirred over 12 hours. After this time the reaction was poured onto ice water and extracted with EtOAc. The combined organics were washed with water and brine, dried over anhydrous TsfeSCU, filtered, and concentrated in vacuo. The crude material was purified by SiCb column chromatography (5%MeOH / DCM) to afford the title compound. ES+ MS: (M + H) 409.3.

[0446] Int-121 - N-{4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l- yl]cyclohexyl}methanesulfonamide.

[0447] To an ice-cold solution of Int-119 and triethylamine (3 eq.) in DCM was slowly added methanesulfonyl chloride (1.5 eq.). The reaction was allowed to achieve ambient temperature and stirred over 12 hours. After this time the reaction was poured onto ice water and extracted with DCM. The combined organics were washed with water and brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude material was purified by SiCh column chromatography (5%MeOH / DCM) to afford the title compound. ES+ MS: (M + H) 445.3.

[0448] Int-122 - tert-butyl N-{3-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l- yl]cyclopentyl}carbamate.

[0449] Int-122 was prepared from Int-3 using GM-3a with tert-butyl N-(3-oxocyclopentyl)carbamate to afford the title compound. ES+ MS: (M + H) 453.4. Int-123a - 3 -[3 -(6-methoxynaphthalene-2-carbonyl)piperidin- 1 -yl]cy cl opentan- 1 -amine hydrochloride.

[0450] Int-123a was prepared from Int-122 using GM-2 with 4M HC1 in dioxane to afford the title compound. ES+ MS: (M + H) 353.3.

[0451] Int-123b - 3-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclopentan-l-amine (Peak

[0452] A).

[0453] Int-123c - 3-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclopentan-l-amine (Peak

[0454] B).

[0455] Additional RP-HPLC purification of Int-123a gives two peaks. The first eluting Peak A was assigned as Int-123b, ES+ MS: (M + H) 353.3, and the second eluting Peak B was assigned as Int-123c, ES+ MS: (M + H) 353.3.

[0456] Int-124a - N-{3-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclopentyl}acetamide. Int-124a was prepared from Int-123a in an analogous method to Int-120 to afford the title compound. ES+ MS: (M + H) 395.3.

[0457] Int-124b - N-{3-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclopentyl}acetamide (Peak A).

[0458] Int-124b was prepared from Int-123a in an analogous method to Int-120 to afford the title compound. ES+ MS: (M + H) 395.3.

[0459] Int-124c - N-{3-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclopentyl}acetamide (Peak B).

[0460] Int-124c was prepared from Int-123c in an analogous method to Int-120 to afford the title compound. ES+ MS: (M + H) 395.3.

[0461] Int-125 - ethyl 4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexane-l- carb oxy late.

[0462] Int-125 was prepared from Int-3 using GM-2 with ethyl 4-oxocyclohexane-l -carboxylate to afford the title compound. ES+ MS: (M + H) 424.3.

[0463] Int-126 4- [3 -(6-m ethoxynaphthal ene-2-carbonyl)piperi din- 1 -yl]cyclohexane- 1 -carboxylic acid. Int-126 was prepared in an analogous method to 112 to afford the title compound. ES+ MS: (M + H) 396.2.

[0464] Int-127 - 4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexane-l -carboxamide. Int-127 was prepared from Int-126 in an analogous method to Int-113 using ammonium chloride (2 eq.) to afford the title compound. ES+ MS: (M + H) 395.3.

[0465] Int-128a - {4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexyl}urea (Peak A). Int-128b - {4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexyl}urea (Peak B). Int-128a & Int-128b were prepared from Int-119 in an analogous method to Int-117. Purification by RP-HPLC gave two peaks. The first eluting Peak A was assigned as Int-128a, ES+ MS: (M + H) 410.3, and the second eluting Peak B was assigned as Int-128b, ES+ MS: (M + H) 410.3.

[0466] Int-129 - 3-(6-methoxynaphthalene-2-carbonyl)-l -(thian-4-yl)piperidine.

[0467] Int-129 was prepared from Int-3 using GM-3a with tetrahydrothiopyran -4-one to afford the title compound. ES+ MS: (M + H) 370.3.

[0468] Int-130 - 1 -imino-4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]-lx6-thian-l-one.

[0469] To a stirred solution of Int-129 in MeOH was added ammonium carbonate (3.5 eq.) followed by (di acetoxy iodo)benzene (2 eq.). The reaction was stirred at ambient temperature over 2 hours. After this time the reaction was concentrated and the crude partitioned between water and DCM. The layers were separated and the aqueous further extracted with DCM. The combined organics were dried over anhydrous TsfeSCU, filtered, and concentrated in vacuo to afford the crude title compound. ES+ MS: (M + H) 401.2.

[0470] Int-131 - 4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexane-l -carbonitrile.

[0471] Int-131 was prepared from Int-3 using GM-3a with 4-oxocyclohexane-l -carbonitrile to afford the title compound. ES+ MS: (M + H) 377.4.

[0472] Int-132a - 3-(6-methoxynaphthalene-2-carbonyl)-l-[4-(2H-l,2,3,4-tetrazol-5- yl)cyclohexyl]piperidine (Peak A).

[0473] Int-132b - 3-(6-methoxynaphthal ene-2-carbonyl)-l-[4-(2H- 1,2,3, 4-tetrazol-5- yl)cyclohexyl]piperidine (Peak B).

[0474] To a stirred solution of Int-131 and sodium azide (1.2 eq.) in isopropanol: water (1 :2) was added ZnBn (0.5 eq.). The reaction was heated at 110°C over 24 hours. After this time the reaction was poured onto water and extracted with EtOAc. The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. Purification by RP-HPLC gave two peaks. The first eluting Peak A was assigned as Int-132a, ES+ MS: (M + H) 410.3, and the second eluting Peak B was assigned as Int-132b, ES+ MS: (M + H) 410.3.

[0475] Int-133 - l-{ l,4-dioxaspiro[4.5]decan-8-yl}-lH-pyrazole.

[0476] Int-133 was prepared from l,4-dioxaspiro[4.5]decan-8-yl 4-m ethylbenzene- 1 -sulfonate using GM-5 with pyrazole to afford the title compound. ES+ MS: (M + H) 209.0.

[0477] Int-134 - 4-(lH-pyrazol-l-yl)cyclohexan-l-one.

[0478] Int-134 was prepared from Int-133 using GM-2 with 4M HC1 in dioxane to afford the title compound. ES+ MS: (M + H) 165.0.

[0479] Int-135a - 3-(6-methoxynaphthalene-2-carbonyl)-l-[4-(lH-pyrazol-l-yl)cyclohexyl]piperidine (Peak A).

[0480] Int-135b - 3-(6-methoxynaphthalene-2-carbonyl)-l-[4-(lH-pyrazol-l-yl)cyclohexyl]piperidine (Peak B).

[0481] Int-135a & Int-135b were prepared from Int-3 using GM-3a with Int-134. Purification by RP- HPLC gave two peaks. The first eluting Peak A was assigned as Int-135a, ES+ MS: (M + H) 418.2, and the second eluting Peak B was assigned as Int-135b, ES+ MS: (M + H) 418.2.

[0482] Int-136 - 3-{ l,4-dioxaspiro[4.5]dec-7-en-8-yl}pyridine.

[0483] Int-136 was prepared in an analogous method to Int-39 using 3 -bromopyridine to afford the title compound. ES+ MS: (M + H) 218.2.

[0484] Int-137 - l-{ l,4-dioxaspiro[4.5]decan-8-yl}-lH-pyrazole.

[0485] Int-137 was prepared in an analogous method to Int-40 from Int-136 to afford the title compound. ES+ MS: (M + H) 220.2.

[0486] Int-138 - 4-(pyridin-3-yl)cyclohexan-l-one.

[0487] Int-138 was prepared from Int-137 using GM-2 with 4M HC1 in dioxane to afford the title compound.

[0488] Int-139a 3-{4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexyl}pyridine

[0489] (Peak A). Int-139b - 3-{4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexyl}pyridine (Peak B).

[0490] Intermediate 139a & 139b were prepared from Intermediate 3 using GM-3a with Intermediate 138. Purification by RP-HPLC gives two peaks. The first eluting peak was isolated to afford 3- [trans-4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexyl]pyridine. ES+ MS: (M + H) 429.2. The second eluting peak was isolated to afford 3-[cis-4-[3-(6- methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexyl]pyridine. ES+ MS: (M + H) 429.2.

[0491] ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++

[0492] General Scheme 3

[0493] The following examples in Table 1 were prepared from the indicated intermediary ketones described above of formula iii (General Scheme 3) using GM-4 and the appropriate hydroxylamine. The table provides exemplar compound numbers and nomenclature, with reference to its molecular structure, and is adhered to herein. Compound numbers designated “Pkl”, “Pk2”, etc. were separated via standard RP-chromatography techniques with the number indicating elution order. Epimerisation was observed alpha to the ketoxime during the synthetic sequence. As such some examples were further purified by chiral -HPLC. These compound numbers are designated with an additional “A” or “B” with the letter sequence indicating elution order.

[0494] Table 1: Route 1

[0495] The following procedures use the General Methods and Intermediates as described above to outline the synthetic pathway used to access intermediary ketoximes of formula iv as shown in General Scheme 4.

[0496] General Scheme 4

[0497] Int-5a - methoxy [(6-methoxynaphthalen-2-yl)(piperidin-3-yl)methylidene]amine

[0498] Int-5a was prepared from Int-3 using GM-4 with O-methylhydroxylamine hydrochloride to afford the title compound. ES+ MS: (M + H) 299.2.

[0499] Int-5b - methoxy[(6-methoxynaphthalen-2-yl)[(3R)-piperidin-3-yl]methylidene]amine (Peak A) Int-5c - methoxy[(6-methoxynaphthalen-2-yl)[(3R)-piperidin-3-yl]methylidene]amine (Peak B) Int-5b & Int-5c were prepared from Int-3a using GM-4 with O-methylhydroxylamine hydrochloride. Purification by RP-HPLC gave two peaks. The first eluting Peak A was arbitrarily assigned as Int-5b, ES+ MS: (M + H) 299.2, and the second eluting Peak B was arbitrarily assigned as Int-5c, ES+ MS: (M + H) 299.2.

[0500] Int-5d - methoxy[(6-methoxynaphthalen-2-yl)(piperidin-3-yl)methylidene]amine (Peak A). Int-5e - methoxy[(6-methoxynaphthalen-2-yl)(piperidin-3-yl)methylidene]amine (Peak B). Int-5d & Int-5e were prepared from Int-3 using GM-4 with O-methylhydroxylamine hydrochloride. Purification by SFC chromatography gave two peaks. The first eluting Peak A was arbitrarily assigned as Int-5d, ES+ MS: (M + H) 299.2, and the second eluting Peak B was arbitrarily assigned as Int-5e, ES+ MS: (M + H) 299.2.

[0501] Int-6a - ethoxy[(6-methoxynaphthalen-2-yl)(piperidin-3-yl)methylidene]amine (Peak A).

[0502] Int-6b - ethoxy[(6-methoxynaphthalen-2-yl)(piperidin-3-yl)methylidene]amine (Peak B).

[0503] Int-6a & Int-6b were prepared from Int-3 using GM-4 with O-ethylhydroxylamine hydrochloride. Purification by SFC chromatography gave two peaks. The first eluting Peak A was arbitrarily assigned as Int-6a, ES+ MS: (M + H) 313.2, and the second eluting Peak B was arbitrarily assigned as Int-6b, ES+ MS: (M + H) 313.2.

[0504] Int-6c - ethoxy[(6-methoxynaphthalen-2-yl)[(3R)-piperidin-3-yl]methylidene]amine (Peak A). Int-6d - ethoxy[(6-methoxynaphthalen-2-yl)[(3R)-piperidin-3-yl]methylidene]amine (Peak B). Int-6c & Int-6d were prepared from Int-3 a using GM-4 with O-ethylhydroxylamine hydrochloride. Purification by SFC chromatography gave two peaks. The first eluting Peak A was arbitrarily assigned as Int-6c, ES+ MS: (M + H) 313.2, and the second eluting Peak B was arbitrarily assigned as Int-6d, ES+ MS: (M + H) 313.2.

[0505] Int-6e - ethoxy[(6-methoxynaphthalen-2-yl)[(3R)-piperidin-3-yl]methylidene]amine (Peak A). Int-6f - ethoxy[(6-methoxynaphthalen-2-yl)[(3R)-piperidin-3-yl]methylidene]amine (Peak B). Int-6e & Int-6f were prepared from Int-3b using GM-4 with O-ethylhydroxylamine hydrochloride. Purification by SFC chromatography gave two peaks. The first eluting Peak A was arbitrarily assigned as Int-6e, ES+ MS: (M + H) 313.2, and the second eluting Peak B was arbitrarily assigned as Int-6f, ES+ MS: (M + H) 313.2.

[0506] General Scheme 5

[0507] The following examples in Table 2 were prepared from the indicated intermediary ketoximes described above of formula iv (General Scheme 4) using GM-3 (a, b or c) and the appropriate carbonyl compound. The table provides exemplar compound numbers and nomenclature, with reference to its molecular structure, and is adhered to herein. Compound numbers designated “Pkl”, “Pk2”, etc. were separated via standard RP-chromatography techniques with the number indicating elution order. Epimerisation was observed alpha to the ketoxime during the synthetic sequence. As such some examples were further purified by chiral -HPLC. These compound numbers are designated with an additional “A” or “B” with the letter sequence indicating elution order. Table 2: Route 2

[0508] General Scheme 6

[0509] The following examples in Table 3 were prepared from Cpd-107 (Table 1) using GM-5 and the appropriate alkyl or heteroaryl halide. The table provides exemplar compound numbers and nomenclature, with reference to its molecular structure, and is adhered to herein. Compound numbers designated “Pkl”, “Pk2”, etc. were separated via standard RP-chromatography techniques with the number indicating elution order. Epimerisation was observed alpha to the ketoxime during the synthetic sequence. As such some examples were further purified by chiral- HPLC. These compound numbers are designated with an additional “A” or “B” with the letter sequence indicating elution order.

[0510] Table 3: Route 3.

[0511] General Scheme 7 The following procedures describe examples of formula vii-a obtained through further chemical manipulation of groups present on R2on compounds of formula vi as shown in General Scheme 7. Cpd-138

[0512] To an ice-cold solution of Cpd-11 in DCM was added TFA (3 eq.). The reaction was allowed to achieve ambient temperature and stirred over 12 hours. After this time the reaction was concentrated in vacuo and the crude residue purified by RP-HPLC to afford the title compound. ES+ MS: (M + H) 396.3.XH NMR (400 MHz, DMSO d6): 8 7.91 - 7.78 (m, 3H), 7.51 - 7.39 (m, 1H), 7.32 (d, J= 2.4 Hz, 1H), 7.18 (dd, J= 2.4 Hz, 8.8 Hz, 1H), 4.11 - 3.92 (m, 2H), 3.88 (s, 3H), 3.00 - 2.87 (m, 2H), 2.81 - 2.76 (m, 3H), 2.46 - 2.39 (m, 2H), 1.94 - 1.67 (m, 6H), 1.63 - 1.41 (m, 6H), 1.32 - 1.14 (m, 3H).

[0513] Cpd-139-Pkl & 139-Pk2

[0514] To an ice-cold solution of Cpd-137 and tri ethylamine (3 eq.) in DCM was added acetyl chloride (2 eq.). The reaction was allowed to achieve ambient temperature and stirred over 12 hours. After this time the reaction was poured onto ice water and extracted with DCM. The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. Purification by RP-HPLC gives two peaks. The first eluting peak was arbitrarily assigned as Cpd-139-Pkl. ES+ MS: (M + H) 438.3. 'H NMR (400 MHz, DMSO d6): 6 7.85 - 7.80 (m, 4H), 7.42 - 7.40 (m, 1H), 7.33 (d, J = 2.0 Hz, 1H), 7.19 (dd, J= 2.4 Hz, 9.2 Hz, 1H), 3.95-3.93 (t, J= 5.6 Hz, 2H), 3.88 (s, 3H), 3.28 - 3.24 (m, 2H), 3.07 - 2.85 (m, 3H), 1.95 - 177 (m, 6H), 1.75 - 1.68 (m, 2H), 1.55 - 1.45 (m, 7H), 1.31 - 1.21 (m, 3H).

[0515] And the second eluting peak arbitrarily assigned as Cpd-139-Pk2. ES+ MS: (M + H) 438.3. 'H

[0516] NMR (400 MHz, DMSO d6): 6 7.97-7.94 (m, 1H), 7.93 (d, J= 12.0 Hz, 1H), 7.86 (s, 1H), 7.81 (d, J= 8.0, 1H), 7.52 (dd, J= 1.8 Hz, 8.8 Hz, 1H), 7.33 (d, J= 2.4 Hz, 1H), 7.19 (dd, J= 2.4 Hz, 8.8 Hz, 1H), 4.10 - 4.07 (t, J= 5.6 Hz, 2H), 3.88 (s, 3H), 3.87 (s, 3H), 3.46 - 3.40 (m, 2H), 2.96 - 2.89 (m, 2H), 2.49 - 2.44 (m, 1H), 2.19 - 2.14 (m, 1H), 1.85 - 1.80 (m, 1H), 1.79 - 1.71 (m, 4H), 1.66 - 1.43 (m, 7H), 1.31 - 1.23 (m, 2H).

[0517] General Scheme 8

[0518] The following procedures describe the examples of formula vii-b obtained through further chemical manipulation of groups present on R3on compounds of formula vi as shown in General Scheme 8.

[0519] Int-67 - ethyl 8-hydroxy-l,4-dioxaspiro[4.5]decane-8-carboxylate.

[0520] To a 0°C stirred solution of ethyl l,4-dioxaspiro[4.5]decane-8-carboxylate (1 eq.) in anhydrous THF under N2 was added lithium bis(trimethylsilyl)amide (1 eq.). The reaction mixture was stirred at this temperature over 1 hour before bubbling ozone through the solution over 2 hours. The reaction mixture was diluted with saturated TsfeSCh and extracted with EtOAc. The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude material was purified by SiCh column chromatography (40%EtOAc / n-heptane) to afford the title compound. 'H NMR (400 MHz, DMSO-d6) 5 5.24 (s, 1H), 4.08 (q, J= 7.2 Hz, 2H), 3.89 - 3.81 (m, 5H), 1.93 - 1.80 (m, 2H), 1.78 - 1.59 (m, 3H), 1.59 - 1.44 (m, 3H), 1.19 (t, J= 7.2 Hz, 3H).

[0521] Int-68 - ethyl l-hydroxy-4-oxocyclohexane-l -carboxylate

[0522] To a 0°C stirred solution of Int-67 (1 eq.) in THF (20 mL) was added 2M HC1 (2 eq.). The reaction mixture was allowed to achieve ambient temperature and stirred over 2 hours. After this time the reaction mixture was diluted with water and extracted with EtOAc. The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to afford the crude title compound. Int-69a - ethyl l-hydroxy-4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexane- 1 -carboxylate (Peak A).

[0523] Int-69b - ethyl l-hydroxy-4-[3-(6-methoxynaphthalene-2-carbonyl)piperidin-l-yl]cyclohexane- 1 -carboxylate (Peak B). -

[0524] Int-69a & Int-69b were prepared from Int-3 using GM-3a with Int-68. Purification by RP-HPLC gave two peaks. The first eluting Peak A was assigned as Int-69a, ES+ MS: (M + H) 440.3, and the second eluting Peak B was assigned as Int-69b, ES+ MS: (M + H) 440.3.

[0525] Int-70a - ethyl l-hydroxy-4-{3-[(methoxyimino)(6-methoxynaphthalen-2-yl)methyl]piperidin- 1 -yl } cyclohexane- 1 -carboxylate (Peak A).

[0526] Int-70a was prepared from Int-69a using GM-4 with O-methylhydroxylamine to afford the title compound. ES+ MS: (M + H) 469.4.

[0527] Int-70b - ethyl l-hydroxy-4-{3-[(methoxyimino)(6-methoxynaphthalen-2-yl)methyl]piperidin- 1 -yl } cyclohexane- 1 -carboxylate (Peak B).

[0528] Int-70b was prepared from Int-69b using GM-4 with O-methylhydroxylamine to afford the title compound. ES+ MS: (M + H) 469.4.

[0529] Int-71a - l-hydroxy-4-{3-[(methoxyimino)(6-methoxynaphthalen-2-yl)methyl]piperidin-l- yl} cyclohexane- 1 -carboxylic acid (Peak A).

[0530] Int-71a was prepared in an analogous method to Int-112 to afford the title compound. ES+ MS: (M + H) 441.3.

[0531] Int-71b - l-hydroxy-4-{3-[(methoxyimino)(6-methoxynaphthalen-2-yl)methyl]piperidin-l- yl} cyclohexane- 1 -carboxylic acid (Peak B).

[0532] Int-71b was prepared in an analogous method to Int-112 to afford the title compound. ES+ MS: (M + H) 441.3. Cpd-140-Pkl & Cpd-140-Pk2

[0533] To a stirred solution of Int-71a (1 eq.) in anhydrous DMF under N2 was added HATU (1.5 eq.), DIPEA (3 eq.) and ammonium chloride (5 eq.). The reaction was stirred at ambient temperature over 12 hours. After this time the reaction was poured onto ice water and extracted with EtOAc. The combined organics were washed with H2O and brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. Purification by RP-HPLC affords two peaks. The first eluting peak arbitrarily assigned as 140-Pkl ES+ MS: (M + H) 440.3.XH NMR (400 MHz, DMSO-d6) 5 7.91 (d, = 9.2 Hz, 1H), 7.85 (s, 1H), 7.80 (d, J = 8.4 Hz, 1H), 7.50 (dd, J= 1.2 Hz, 8.4 Hz, 1H), 7.33 (d, J= 2.0 Hz, 1H), 7.18 (dd, J= 2.8 Hz, 9.2 Hz, 1H), 7.02 (s, 1H), 6.85 (s, 1H), 5.09 (br s, 1H), 3.93 - 3.87 (m, 6H), 2.91 (t, J= 11.6 Hz, 2H), 2.25 - 2.15 (m, 2H), 2.02 - 1.85 (m, 3H), 1.79 - 1.45 (m, 9H), 1.29 - 1.17 (m, 2H).

[0534] And the second eluting peak arbitrarily assigned as 140-Pk2. ES+ MS: (M + H) 440.4. 'H NMR (400 MHz, DMSO-d6) 5 7.85 (d, J= 8.8 Hz, 1H), 7.80 (d, J = 8.8 Hz, 1H), 7.77 (s, 1H), 7.37 - 7.30 (m, 2H), 7.18 (dd, J = 2.4 Hz, 8.8 Hz, 1H), 7.00 (s, 1H), 6.84 (s, 1H), 5.06 (s, 1H), 3.88 (s, 3H), 3.70 (s, 3H), 2.97 (d, J= 11.2 Hz, 1H), 2.84 (d, J= 10.0 Hz, 1H), 2.79 - 2.71 (m, 1H), 2.20 - 2.12 (m, 1H), 2.02 - 1.85 (m, 4H), 1.81 - 1.73 (m, 1H), 1.70 - 1.37 (m, 6H), 1.33 - 1.95 (m, 3H).

[0535] Cpd-141-Pkl & Cpd-141-Pk2

[0536] The title compounds were prepared in an analogous method to Cpd-140-Pkl & Cpd-140-Pk2 using Int-71b. Purification by RP-HPLC affords two peaks. The first eluting peak arbitrarily assigned as 141-Pkl. ES+ MS: (M + H) 440.3. 'H NMR (400 MHz, DMSO-d6) 5 7.91 (d, J = 9.2 Hz, 1H), 7.85 (s, 1H), 7.81 (d, J= 8.8 Hz, 1H), 7.50 (d, J= 9.2 Hz, 1H), 7.34 (d, J= 2.8 Hz, 1H), 7.19 (dd, J= 2.4 Hz, 8.8 Hz, 1H), 7.10 (s, 1H), 6.97 (s, 1H), 4.96 (s, 1H), 3.93 - 3.87 (m, 6H), 2.85 - 2.75 (m, 2H), 2.49 - 2.39 (m, 1H), 2.32 - 2.21 (m, 1H), 2.20 - 2.11 (m, 1H), 1.78 - 1.59 (m, 4H), 1.59 - 1.42 (m, 9H).

[0537] And the second eluting peak arbitrarily assigned as 141-Pk2. ES+ MS: (M + H) 440.4. 'H NMR (400 MHz, DMSO-d6) 5 7.85 (d, J= 8.8 Hz, 1H), 7.80 (d, J = 8.8 Hz, 1H), 7.77 (s, 1H), 7.37 - 7.31 (m, 2H), 7.18 (dd, J= 2.4 Hz, 9.2 Hz, 1H), 7.08 (s, 1H), 6.96 (s, 1H), 4.93 (s, 1H), 3.88 (s, 3H), 3.71 (s, 3H), 2.85 (d, J= 10.0 Hz, 1H), 2.79 - 2.69 (m, 2H), 2.29 - 2.11 (m, 3H), 1.78 (d, J = 11.2 Hz, 1H), 1.72 - 1.58 (m, 3H), 1.57 - 1.40 (m, 7H), 1.37 - 1.24 (m, 1H).

[0538] Int-61a - (4-{3-[(methoxyimino)(6-methoxynaphthalen-2-yl)methyl]piperidin-l- yl}cyclohexyl)methanol (Peak A).

[0539] Int-61a was prepared from Int-55a using GM-4 with O-methylhydroxylamine to afford the title compound. ES+ MS: (M + H) 411.4.

[0540] Int-61b - (4-{3-[(methoxyimino)(6-methoxynaphthalen-2-yl)methyl]piperidin-l- yl}cyclohexyl)methanol (Peak B).

[0541] Int-61b was prepared from Int-55b using GM-4 with O-methylhydroxylamine to afford the title compound. ES+ MS: (M + H) 411.4.

[0542] Int-62a - 4- {3 -[(methoxyimino)(6-m ethoxynaphthal en-2-yl)methyl]piperi din- 1- yl} cyclohexane- 1-carbaldehy de (Peak A).

[0543] To a -78°C solution of Intermediate 61a in DCM (5 mL) was added Dess-Martin periodinane (2 eq.). The reaction was stirred at this temperature over 2 hours before allowing the reaction to achieve ambient temperature and stirring for a further 6 hours. The reaction mixture poured onto ice water and extracted with DCM. The combined organics were washed with saturated NaHCOs (aq) and brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude compound was purified by SiCb column chromatography (5%MeOH / DCM) to afford the title compound. ES+ MS: (M + H) 409.3.

[0544] Int-62b - 4-{3-[(methoxyimino)(6-methoxynaphthalen-2-yl)methyl]piperidin-l- yl} cyclohexane- 1-carbaldehy de (Peak B).

[0545] Intermediate 62b was prepared in an analogous method to Intermediate 62a using Intermediate 61b to afford the title compound. ES+ MS: (M + H) 409.3. Cpd-142-Pkl & Cpd-142-Pk2

[0546] To a -78°C solution of Intermediate 62a (1 eq.) in DCM was added (diethylamino)sulfur trifluoride (2 eq.). The reaction was stirred at this temperature over 2 hours and then at ambient temperature over 16 hours. After this time the reaction was poured onto ice water and extracted with DCM. The combined organics were washed with saturated NaHCCT / r / t / J and brine, dried over anhydrous MgSC , filtered, and concentrated in vacuo. Purification by RP-HPLC affords two peaks. The first eluting peak arbitrarily assigned as 142-Pkl. ES+ MS: (M + H) 431.3.TH NMR (400 MHz, DMSO d6): 8 7.97 - 7.83 (m, 3H), 7.60 - 7.50 (m, 1H), 7.34 (s, 1H), 7.20 (d, J = 8.0 Hz, 1H), 5.97 - 5.67 (m, 1H), 3.88 (s, 6H), 3.72 - 3.71 (m, 1H), 3.06 - 3.04 (m, 1H), 2.83 -

[0547] 2.82 (m, 2H), 2.12 - 2.10 (m, 2H), 1.89 - 1.76 (m, 8H), 1.64 - 1.63 (m, 1H), 1.52 - 1.42 (m, 2H), 1.17 - 1.10 (m, 2H).

[0548] And the second eluting peak arbitrarily assigned as 142-Pk2. ES+ MS: (M + H) 431.3. 'H NMR (400 MHz, DMSO d6): 6 7.85 (d, J= 8.8 Hz, 1H), 7.80 (d, J= 8.0 Hz, 1H), 7.76 (s, 1H), 7.35 - 7.32 (m, 2H), 7.18 (dd, J= 2.4 Hz, 8.8 Hz, 1H), 5.94 - 5.65 (m, 1H), 3.88 (s, 3H), 3.70 (s, 3H),

[0549] 2.83 - 2.74 (m, 2H), 2.21 - 2.11 (m, 3H), 1.75 - 1.65 (m, 8H), 1.42 - 1.33 (m, 2H), 1.29 - 1.09 (m, 4H).

[0550] Cpd-143-Pkl

[0551] The title compounds were prepared in an analogous method to Cpd-142-Pkl & Cpd-142-Pk2 using Int-62b. Purification by RP-HPLC affords two peaks. The first eluting peak arbitrarily assigned as 143-Pkl. ES+ MS: (M + H) 431.3.XH NMR (400 MHz, DMSO d6): 6 7.90 (d, J = 8.8 Hz, 1H), 7.89 - 7.80 (m, 2H), 7.61 - 7.52 (m, 1H), 7.36 - 7.34 (m, 1H), 7.19 (dd, J= 2.4 Hz, 8.8 Hz, 1H), 5.96 - 5.67 (m, 1H), 3.88 (s, 6H), 2.85 - 2.84 (m, 2H), 1.81 - 1.68 (m, 7H), 1.56 - 1.50 (m, 2H), 1.23 - 1.09 (m, 7H), 0.86 - 0.85 (m, 1H).

[0552] And the second eluting peak arbitrarily assigned as 143-Pk2. ES+ MS: (M + H) 431.3. 'H NMR (400 MHz, DMSO d6): 6 7.85 (d, J= 9.2 Hz, 1H), 7.80 (d, J= 8.4 Hz, 1H), 7.76 (s, 1H), 7.35 - 7.32 (m, 2H), 7.19 (dd, J= 2.4 Hz, 8.8 Hz, 1H), 5.94 - 5.65 (m, 1H), 3.88 (s, 3H), 3.70 (s, 3H), 2.83 - 2.73 (m, 2H), 2.23 - 2.20 (m, 4H), 1.76 - 1.65 (m, 8H), 1.42 - 1.41 (m, 1H), 1.23 - 1.22 (m, 1H), 1.12 - 0.98 (m, 3H).

[0553] Int-15 - tert-butyl 3 -[3 -(cyclopentyloxy)benzoyl]piperidine-l -carboxylate.

[0554] Int-15 was prepared using GM- 1c with [3-(cyclopentyloxy)phenyl]boronic acid to afford the title compound. ES+ MS: (M - ‘Bu + H) 318.2.

[0555] Cpd-144

[0556] Preparation was from Int-15 using GM-2 directly followed by GM-3a with cyclopentanone directly followed by GM-4 with O-ethylhydroxylamine hydrochloride to afford the title compound. ES+ MS: (M + H) 385.2. 'H NMR (400 MHz, DMSO-d6) 5 7.89 (d, J= 9.2 Hz, 1H), 7.85 (s, 1H), 7.80 (d, = 8.4 Hz, 1H), 7.50 (d, J= 8.4 Hz, 1H), 7.34 (d, J= 2.4 Hz, 1H), 7.19 (dd, J= 2.4 Hz, 8.8 Hz, 1H), 3.93 - 3.85 (m, 6H), 3.46 - 3.34 (m, 1H), 2.98 (t, J= 11.2 Hz, 2H), 2.39 - 2.26 (m, 1H), 2.22 (br s, 1H), 2.12 (t, J = 10.8 Hz, 1H), 1.91 - 1.77 (m, 3H), 1.77 - 1.58 (m, 4H), 1.58 - 1.46 (m, 4H), 1.46 - 1.33 (m, 2H).

[0557] Cpd-145

[0558] Preparation was from Int-2 using GM-2 directly followed by GM-3a with 1,4- dioxaspiro[4.5]decan-8-one directly followed by GM-4 with O-ethylhydroxylamine hydrochloride to afford the title compound. ES+ MS: (M + H) 395.2. 'H NMR (300 MHz, CDCh)

[0559] 5 7.81 - 7.67 (m, 3H), 7.53 - 7.34 (m, 1H), 7.19 - 7.11 (m, 2H), 3.98 - 3.79 (m, 6H), 3.20 - 2.65 (m, 4H), 2.57 - 1.66 (m, 13H), 1.47 - 1.28 (m, 1H).

[0560] Table 4: Summary of chiral purification and assessment methods.

[0561] Example 2: Biological Evaluation of Compounds of Formula

[0562] PLpro activity was evaluated with a homogenous fluorescence intensity assay using the substrate Ub-Rhodaminel lOGly (UbiQ bio, hereafter referred to as Ub-Rhodamine). Experiments were performed in either 384-well or 1,536-well black non-binding plates (Aurora ABA000000A and ABI000000A, respectively) with a final reaction volume of 6 pL. The assay buffer contained 20 mM Tris (pH 8), 1 mM GSH, 0.03% (w / v) BSA, and 0.01% (v / v) Triton-X. Assay-ready plates were prepared by dry-spotting compounds in DMSO using an Echo® Acoustic Dispenser (LabCyte). PLpro at a final concentration of 5 nM was added to the plates and incubated at room temperature for 10 min. Ub-Rhodamine (final concentration 100 nM) was added to start the reaction and incubated for 120 min at room temperature. For end-point assays, the reaction was stopped by the addition of aqueous citric acid (1 pL) at a final concentration of 10 mM. All reagents were dispensed using the CERTUS FLEX (v2.0.1, Gyger), and microplates were centrifuged using a microplate centrifuge (Agilent). The reaction was monitored by an increase in fluorescence (excitation 485 nm and emission 520 nm) on a PHERAstar FSX microplate reader (v5.41, BMG Labtech) using the FI 485 520 optic module.

[0563] Table 5: Results of biological activity (IC50s from Example 2) of some exemplar compounds of this invention.

[0564] Comparative data

[0565] A series of ketone like compounds were made to further demonstrate the importance and robustness of some of the substitutions of the current invention.

[0566] Table 6 summarises some of these structures and their IC50 activity ranges. Example 3: Assessing the interaction of an exemplar compounds with PLpro from crystallography.

[0567] Prior efforts to develop compounds that inhibit PLpro have focused on improving GRL-scaffold inhibitors (such as GRL0617), which bind PLpro in a binding groove near the active site, forming extensive interactions with blocking loop 2 (BL2) (Osipiuk, J. et al. Nat. Commun. 12, 743 (2021)). The work performed in this experiment was to assesses the interaction of an exemplar compound and how it differed from the GRL scaffold inhibitors.

[0568] Crystallography studies, including the provision of purified PLpro, were performed according to the methods described in Komander et al (2024) https: / / www.researchsquare.com / article / rs- 4899442 / vl.

[0569] A co-crystal structure of Cpd-22-Pkl-A (2.8A) in a P4s 3 2 space group showed the compound binding with a closed BL2-loop, Figure 1. In this structure, Cpd-22-Pkl-A displayed interactions including the side chain of BL2-loop, Tyr268 stacked against the oxime group, the backbone carbonyl of Tyr268 forming a hydrogen bond with the hydroxy group of the cyclohexanol moiety and the methoxynaphthalene interacting with Pro247. The Met208 side chain was protracted into the protease main chain which accommodated compound binding by exposing a novel binding pocket. Figure 1 collectively reveals how the compounds of this disclosure achieve their high potency through exploiting an unexpected PLpro binding site, ensuring a greater interaction between inhibitor and protease and thus heightened compound binding, compared to the GRL scaffold inhibitors. This increased interaction results in a compound that is a superior PLpro inhibitor with respect to efficacy and activity (<0.1 pM level in this and similar exemplar compounds).

[0570] The co-crystal structure results also help elucidate why particular isomers (as represented by different peaks in Tables 1-4 and for example the trans cyclohexanol and 7?-piperidine and E- oxime of Cpd-22-Pkl-A) result in greater inhibition than the alternative isomers, owing to the orientation of these groups into the PLpro pockets. The activity of Cpd-22-Pkl-B is in the range of 0.1-1.0 pM, of Cpd-23 Pkl-B is in the range of 1-4.9 pM in comparison. Other stereoisomers may give rise to differing interactions depending on the various substitutions of any given compound and its target protease (nsp-3 for example) which may have slightly different structures and binding pockets.

[0571] Example 4 Broad-spectrum PLpro inhibitor data.

[0572] Genetic diversity among CoVs is significant, but activity in PLpro enzymes is a conserved feature across CoVs. aCoVs harbour two PLpro domains in nsp-3, of which the PL2pro domain shows highest similarity to SARS-CoV-2 PLpro.

[0573] The next set of experiments were designed to demonstrate pan-PLpro activity.

[0574] PLpro expression and purification for these studies were followed according to the method outlined in https: / / www.researchsquare.com / article / rs-4899442 / vl. In summary, bacterial pOPIN-B expression vectors for SARS-CoV-2 PLpro (QHD43415), SARS-CoV PLproWT (RefSeq: NP_828849.7), MERS-CoV PLproWT (RefSeq: YP_009047202), HKUl-CoV PLproWT (RefSeq: YP 009944268), OC43-CoV PLproWT (RefSeq: AY391777), 229E-CoV PL2proWT (RefSeq: NP_073549), NL63-CoV PL2proWT (RefSeq: YP_003766) were codon optimised and cloned into pOPIN-B 765 and then digested.

[0575] The protein expression vectors were transformed into E. coli Rosetta competent cells and grown in medium. Expression was induced with 0.3 mM IPTG. Cells were harvested 16 h post induction. For purification, cells were lysed, and lysates removed by centrifugation. The clarified lysate was filtered, and His-tagged proteins were captured using a HisTrap HP column. The captured protein was washed with buffer, and then the His tag removed. The extracted PLpro was further purified by size exclusion chromatography.

[0576] Cpd-22-Pkl-A inhibited PLpro of SARS-CoV-2 and SARS-CoV and showed weak activity (IC50 52 pM) against PL2pro of the aCoV NL63, which only has 20% sequence identity to SARS-CoV-2. Replacing the cyclohexanol with a 3-substituted pyrazole in Cpd-63-Pkl-B strengthened pan activity, inhibiting four of the seven PLpro enzymes with nanomolar activity against SARS-CoV-2 and SARS-CoV (IC5098 and 94 nM, respectively), submicromolar activity against NL63 (IC50 648 nM) and weak activity against 229E (IC50 26 pM).

[0577] PLpro activity was monitored in a fluorescence intensity assay using the substrate Ub-Rhodamine 110 (UbRh), that upon cleavage becomes fluorescent. The assay buffer contained 20 mM Tris (pH 8), 1 mM TCEP, 0.03% BSA (w / v) and 0.01% (v / v) Triton-X. Experiments were performed in 1536-well black non-binding plates (Greiner 782900) with a final reaction volume of 6 pL. PLpro enzyme was added to the plates (50 nM or 5 nM) and incubated at ambient temperature for 10 min. UbRh (final concentration 100 nM) was added to start the reaction and incubated for 12 min (50 nM PLpro), or 2 h (5 nM PLpro), at room temperature. For endpoint assays the reaction was stopped by addition of citric acid (1 pL) at a final concentration of 10 mM. All additions were performed using the CERTUS FLEX (v2.0.1, Gyger). The reaction was monitored by an increase in fluorescence (excitation 485 nm and emission 520 nm) on a PHERAstar® (v5.41, BMG Labtech) using the FI 485 520 optic module. Data was normalised to 1% (v / v) DMSO (negative control, 0% inhibition) and 100 pM compound 5c (positive control, 100% inhibition). Table 7: Results of biological activity (IC50s from Example 4) of some exemplar compounds

[0578] ++++: < 0.5 pM; +++: > 0.5-1.0 pM; ++: > 1.0-10 pM; +: > 10 pM;

[0579] Example 5 Antiviral activity, in vitro and in vivo

[0580] In vitro antiviral activity was assessed using a viral plaque assay, adapted and performed based on protocols previously described (Mendoza et al. Curr. Protoc. Microbiol. 57, cpmcl05 (2020). Briefly, cells were seeded in flat bottom well plates and left to adhere overnight. Cells were washed twice with PBS and transferred to serum-free DMEM containing TPCK trypsin. Cells were infected with 150 pL of a SARS-CoV- clinical isolate VIC001 (TCID50 2.6 x 1031268 / mL) and incubated for 30 minutes. 150 pL of 1 :2 serial dilutions of the exemplar compounds ranging from final concentrations of 5 pM to 0.0098 pM with or without 2 pM of the P- glycoprotein inhibitor CP100356 were transferred to the infected cells and incubated at 37°C / 5% CO2 for 30 minutes. Cells were then overlayed with 1.5% (w / v) methylcellulose and 4% FCS (v / v) in DMEM and incubated for 4 days. At 4 dpi the overlay was removed, and cells were washed once with PBS before fixation. Wells were then stained, washed and air dried before plaque counting and calculation of antiviral EC50 for each compound using four-parameter logistic regression.

[0581] For in vivo evaluation, mouse models were developed based on a mouse-adapted SARS-CoV-2 strain that was obtained by serially passaging a naturally occurring variant identified during a 2020 COVID-19 outbreak in Victoria, Australia (Bader et al, PNAS 2023). C57BL / 6 (WT) mice that were intranasally infected with this virus present with high lung viral titres, significant weight loss and an influx of immune cells and inflammatory cytokines in the lung, thereby reflecting key disease outcomes seen in human patients with severe COVID-19. To test exemplary compounds, C57BL / 6 mice were infected with the mouse adapted virus and treated with either vehicle (10% DMSO in com oil), 100 mg / kg of Cpd-118-Pk2-A at 6, 24 and 48h post-infection and body weights measured daily. At the end of experiment (day 3), lungs were harvested. Lung TCID50 was performed as previously described (Hierholzer. & Killington. Virology Methods Manual. Sect. : Class. 604 Tech. 25-46 (1996). Briefly, vero cells were seeded in flat bottom well plates and left to adhere overnight. Cells were washed twice with PBS and transferred to serum-free DMEM containing TPCK trypsin. Infected organs homogenised, clarified by centrifugation and supernatant was added. Cells were incubated for 4 days until virus-induced cytopathic effect (CPE) was scored.

[0582] Figure 2 shows that Cpd-118-Pk2-A displayed an antiviral EC50 of 360 nM with slight improvement upon PgP inhibition (290 nM). Figure 3 shows Cpd- 118 -Pk2- A effectively reduced viral burden and prevented the significant weight loss (surrogate marker for severe disease) that is observed in vehicle treated mice.

Claims

CLAIMS1. A compound of Formula I or a pharmaceutically acceptable salt, solvate, stereoisomer, or deuterated analogue thereof:Formula I; wherein n = 1 or 2;R1is a 9-10 membered heterocycle or a 9-10 membered carbocycle, wherein R1may be independently substituted with one or more R4, wherein R4is independently selected from Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, halogen, 3-10 membered carbocycle, 3-10 membered heterocycle, =0, -CN, -CF3, -CF2H, -NO2, -OR8, -SR8, -S(O)R8, S(O)2R8, -C(O)R8, -C(O)OR8, -N(R8)2, -C(O)N(R8)2, S(O)N(R8)2, -S(O)2N(R8)2 , -NR8C(O)R8, -NR8S(O)R8, -NR8S(O)2R8, -NR8C(O)N(R8)2, -NR8S(O)2N(R8)2, wherein R8is independently selected from hydrogen, deuterium, halogen, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, 3-10 membered carbocycle or 3-10 membered heterocycle, wherein each R8may be independently substituted with one or more R10;R2is selected from hydrogen, Ci-s alkyl, C2-ealkenyl, C2-6 alkynyl, 3-10-membered carbocycle, 3-10-membered heterocycle, wherein R2may be independently substituted with one or more R6when valency allows, wherein R6is independently selected from deuterium, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, halogen, 3-10 membered carbocycle, 3-10 membered heterocycle, -CN, -CF3, -CF2H, -NO2, - OR9, -SR9, -S(O)R9, S(O)2R9, -C(O)R9, -C(O)OR9, -N(R9)2, -C(O)N(R9)2, S(O)N(R9)2, - S(O)2N(R9)2, -NR9C(O)R9, -NR9S(O)R9, -NR9S(O)2R9, -NR9C(O)N(R9)2, -NR9S(O)2N(R9)2, wherein each R6is optionally substituted with R9when valency allows,148 wherein R9is independently selected from hydrogen, halogen, Cue alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, 3-10 membered carbocycle or 3-10 membered heterocycle, wherein each R9may be independently substituted with one or more R10when valency allows;R3is selected from the group consisting of a Ci-s alkyl, C2-8 alkenyl, C2-8 alkynyl, 3-10 membered carbocycle or a 4-10 membered heterocycle, wherein R3is optionally substituted with one or more R5, wherein R5is selected from the group consisting of hydrogen, Cue alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, halogen, 3-10 carbocycle, 3-10 heterocycle, -CN, =0, =NR7, -CF3, -CF2H, -NO2, -OR7, -SR7, -S(O)R7, S(O)2R7, -C(O)R7, -C(O)OR7, -N(R7)2, -C(O)N(R7)2, S(O)N(R7)2, -S(O)2N(R7)2, -S(O)(NR7)R7, -NR7C(O)R7, -NR7S(O)R7, -NR7S(O)2R7, -NR7C(O)N(R7)2, -NR7S(O)2N(R7)2, wherein each R7is independently selected from the group consisting of hydrogen, C1.4 alkyl, C1.4 halogenated alkyl, alkoxy, 3-10 membered carbocycle or 3-10 membered heterocycle, wherein each R7may be independently substituted with one or more R10when valency allows, wherein when R3is a 3-10 carbocycle or 3-10 heterocycle it is optionally geminally substituted with two independent R5, and the two R5s are optionally linked to form spirocycles, and wherein when R5is 3-10 carbocycle or 3-10 heterocycle it is optionally substituted with one or more R10;R10is selected from the group consisting of hydrogen, deuterium, halogen, C1-6alkyl, C2-6 alkenyl, C2-6 alkynyl, alkoxy, halogen, -CH2OH, -CN, -CF3, -CF2H, -NO2, -SR , -S(O)R11-S(O)2R11- C(O)R11-C(O)OR11-N(R112, -C(O)N(R112, S(O)N(R112, -S(O)2N(R112, -NRnC(O)Rn, - NR11(O)R11-NR11(O)2R11-NR11(O)N(R112, -NR11(O)2N(R112, wherein R11is independently selected from hydrogen, C1-4halogenated alkyl2. A compound of claim 1 wherein n=2.

3. A compound of claim 1 or 2 wherein R1is a 10 membered carbocycle or a 10 membered heterocycle.

4. A compound of claim 1 or 2 wherein R1is selected from5. A compound of any one of claims 1 -4 wherein R1is6. A compound of claim 1 or 2 wherein R1isand wherein R1is substituted with one or more R4.

7. A compound of claim 1 or 2 wherein R1isand wherein R1is substituted with R4wherein R4is -OR8.

8. A compound of any one of claims 1 to 7 wherein R2is hydrogen, Ci-s alkyl, a 3-10 membered carbocycle or 3-10 membered heterocycle.9 A compound of any one of claims 1 to 8 wherein R2is hydrogen, methyl, ethyl, isopropyl, or butyl.

10. A compound of any one of claims 1 to 9 wherein R3is a 3-8-membered heterocycle or a 3- 8-membered carbocycle.

11. A compound of any one of claims 1 to 10 wherein R3is selected from the group consisting12. A compound of claim 11 wherein R3is13. A compound of any one of claims 1 to 12 wherein R3is substituted with one or more R5, wherein each R5is independently selected from hydrogen, -OH, -OMe, OEt, -CN, -CF3, -CHF2, methyl, ethyl, =0, =NH, -S(O)2R7, -NR7(CO)N(R7)2, -NR7(CO)R7, -NR7S(O)R7, -C(O)N(R7)2, C(0)0H, or a 5-6 membered heterocycle.

14. A compound of any one of claims 1 to 13 wherein R3is substituted with one or more R5, wherein each R5is independently selected from hydrogen, -OH, -OMe, OEt, -CN, -CF3, -CHF2, -NR7(CO)N(R7)2, -NR7(CO)R7, or a 5-6 membered heterocycle.

15. A compound of any one of claims 1 to 14 wherein R6is alkoxy.

16. A compound of any one of claims 1 to 15 wherein R7is hydrogen.

17. A compound of claim 1 wherein the compound is selected from the group consisting of:

19. A compound of claim 1 wherein the compound is selected from any of the compounds listed in Tables 1 to 3.

20. A composition for the prevention and / or treatment of a coronavirus (CoV) infection comprising a compound of claim 1.

21. A composition of claim 20 wherein the composition further comprises a therapeutic agent, preferably a further antiviral agent.

22. A method of preventing and / or treating a coronavirus (CoV) infection in a subject, comprising administering to the subject a therapeutically effective amount of the composition of claims 20 or 21.

Images