The disclosure relates to compounds of formula (I), which are formyl peptide 2 (FPR2) receptor agonists and/or formyl peptide 1 (FPR1) receptor agonists. The disclosure also provides compositions and methods of using the compounds, for example, for the treatment of atherosclerosis, heart failure, and related diseases.

BACKGROUND OF THE INVENTION

The present invention relates to novel piperidinone compounds, which are formyl peptide 2 (FPR2) receptor agonists and/or formyl peptide 1 (FPR1) receptor agonists, compositions containing them, and methods of using them, for example, for the treatment of atherosclerosis, heart failure, chronic obstructive pulmonary disease (COPD), and related diseases.

Formyl peptide receptor 2 (FPR2) belongs to small group of seven-transmembrane domain, G protein-coupled receptors that are expressed mainly by mammalian phagocytic leukocytes and are known to be important in host defense and inflammation. FPR2 shares significant sequence homology with FPR1 and FPR3. Collectively, these receptors bind large number of structurally diverse group of agonists, including N-formyl and nonformyl peptides which act as chemo attractants and activate phagocytes. The endogenous anti-inflammatory peptide Annexin A1 and its N-terminal fragments also bind human FPR1 and FPR2. Importantly, anti-inflammatory eicosanoid lipoxin A4, which belongs to newly discovered class of small pro-resolution mediators (SPMs), has been identified as an agonist for FPR2 (Ye R D., et al., Pharmacol. Rev., 2009, 61, 119-61).

Endogenous FPR2 pro-resolution ligands, such as lipoxin A4and Annexin A1 trigger a wide array of cytoplasmatic cascades such as Gi coupling, Ca2+mobilization and □-arrestin recruitment via FPR2. Activation of FPR2 by lipoxin A4modifies the effects of peptidic agonists, such as serum amyloid A (SAA), and has alternative effects on phosphorylation pathways depending on the cell type. Lipoxins regulate components of both innate and adaptive immune systems including neutrophils, macrophages, T-, and B-cells. In neutrophils, lipoxins modulate their movement, cytotoxicity and life span. In macrophages, lipoxins prevent their apoptosis and enhance efferocytosis. In most inflammatory cells, lipoxins also down-regulate expression of several pro-inflammatory cytokines, such as IL-6, IL-1□ and IL-8 as well as up-regulate expression of anti-inflammatory cytokine IL-10 (Chandrasekharan J A, Sharma-Walia N., J. Inflamm. Res., 2015, 8, 181-92). The primary effects of lipoxin on neutrophils and macrophages are termination of inflammation and initiation of resolution of inflammation. The latter is primarily responsible for enhancing anti-fibrotic wound healing and returning of the injured tissue to homeostasis (Romano M., et al., Eur. J. Pharmacol., 2015, 5, 49-63).

Chronic inflammation is part of the pathway of pathogenesis of many human diseases and stimulation of resolution pathways with FPR2 agonists may have both protective and reparative effects. Ischaemia-reperfusion (I/R) injury is a common feature of several diseases associated with high morbidity and mortality, such as myocardial infarction and stroke. Non-productive wound healing associated with cardiomyocyte death and pathological remodeling resulting from ischemia-reperfusion injury leads to scar formation, fibrosis, and progressive lost of heart function. FPR2 modulation is proposed to enhance myocardial wound healing post injury and diminish adverse myocardial remodeling (Kain V., et al., J. Mol. Cell. Cardiol., 2015, 84, 24-35). In addition, FPR2 pro-resolution agonists, in the central nervous system, may be useful therapeutics for the treatment of a variety of clinical I/R conditions, including stroke in brain (Gavins F N., Trends Pharmacol. Sci., 2010, 31, 266-76) and I/R induced spinal cord injury (Liu Z Q., et al., Int. J. Clin. Exp. Med., 2015, 8, 12826-33).

In addition to beneficial effects of targeting the FPR2 with novel pro-resolution agonists for treatment of I/R induced injury therapeutic, utility of these ligands can also be applied to other diseases. In the cardiovascular system both the FPR2 receptor and its pro-resolution agonists were found to be responsible for atherogenic-plaque stabilization and healing (Petri M H., et al., Cardiovasc. Res., 2015, 105, 65-74; and Fredman G., et al., Sci. Trans. Med., 2015, 7(275); 275ra20). FPR2 agonists also have been shown to be beneficial in preclinical models of chronic inflammatory human diseases, including: infectious diseases, psoriasis, dermatitis, occular inflammation, sepsis, pain, metabolic/diabetes diseases, cancer, COPD, asthma and allergic diseases, cystic fibrosis, acute lung injury and fibrosis, rheumatoid arthritis and other joint diseases, Alzheimer's disease, kidney fibrosis, and organ transplantation (Romano M., et al., Eur. J. Pharmacol., 2015, 5, 49-63, Perrett, M., et al., Trends in Pharm. Sci., 2015, 36, 737-755).

DESCRIPTION OF THE INVENTION

The invention encompasses compounds of formula I, which are formyl peptide 2 (FPR2) receptor agonists and/or formyl peptide 1 (FPR1) receptor agonists, compositions containing them, and methods of using them, for example, in the treatment of atherosclerosis, heart failure, chronic obstructive pulmonary disease (COPD), and related diseases.

One aspect of the invention is a compound of formula I

Another aspect of the invention is a compound of formula I where:

Another aspect of the invention is a compound of formula I where Ar1is phenyl or pyridinyl and is substituted with 1-3 substituents selected from cyano, halo, alkyl, fluoroalkyl, alkoxy, fluoroalkoxy, alkylthio, and alkylsulfonyl.

Another aspect of the invention is a compound of formula I where Ar2is phenyl or pyridinyl and is substituted with 0-3 substituents selected from cyano, halo, alkyl, fluoroalkyl, alkoxy, and fluoroalkoxy.

For a compound of Formula I, the scope of any instance of a variable substituent, including Ar1, Ar2, R1, R2, R3, and X can be used independently with the scope of any other instance of a variable substituent. As such, the invention includes combinations of the different aspects.

Unless specified otherwise, these terms have the following meanings. “Alkyl” means a straight or branched alkyl group composed of 1 to 6 carbons. “Alkenyl” means a straight or branched alkyl group composed of 2 to 6 carbons with at least one double bond. “Alkynyl” means a straight or branched alkyl group composed of 2 to 6 carbons with at least one triple bond. “Cycloalkyl” means a monocyclic ring system composed of 3 to 7 carbons. Terms with a hydrocarbon moiety (e.g. alkoxy) include straight and branched isomers for the hydrocarbon portion. “Halo” includes fluoro, chloro, bromo, and iodo. “Haloalkyl” and “haloalkoxy” include all halogenated isomers from monohalo to perhalo. “Aryl” means a monocyclic or bicyclic aromatic hydrocarbon group having 6 to 12 carbon atoms. Bicyclic ring systems can consist of a phenyl group fused to a aromatic or non-aromatic carbocyclic ring. Representative examples of aryl groups include but are not limited to phenyl, indanyl, indenyl, naphthyl, and tetrahydronaphthyl. “Heteroaryl” means a 5 to 7 membered monocyclic or 8 to 11 membered bicyclic aromatic ring system with 1-5 heteroatoms independently selected from nitrogen, oxygen, and sulfur. Where a bonding attachment location is not specified, the bonding may be attached at any appropriate location as understood by practitioners in the art. Combinations of substituents and bonding patterns are only those that result in stable compounds as understood by practitioners in the art. Parenthetic and multiparenthetic terms are intended to clarify bonding relationships to those skilled in the art. For example, a term such as ((R)alkyl) means an alkyl substituent further substituted with the substituent R.

The invention includes all pharmaceutically acceptable salt forms of the compounds. Pharmaceutically acceptable salts are those in which the counter ions do not contribute significantly to the physiological activity or toxicity of the compounds and as such function as pharmacological equivalents. These salts can be made according to common organic techniques employing commercially available reagents. Some anionic salt forms include acetate, acistrate, besylate, bromide, chloride, citrate, fumarate, glucouronate, hydrobromide, hydrochloride, hydroiodide, iodide, lactate, maleate, mesylate, nitrate, pamoate, phosphate, succinate, sulfate, tartrate, tosylate, and xinofoate. Some cationic salt forms include ammonium, aluminum, benzathine, bismuth, calcium, choline, diethylamine, diethanolamine, lithium, magnesium, meglumine, 4-phenylcyclohexylamine, piperazine, potassium, sodium, tromethamine, and zinc.

Some of the compounds of the invention exist in stereoisomeric forms. The invention includes all stereoisomeric forms of the compounds including enantiomers and diastereomers. Methods of making and separating stereoisomers are known in the art. The invention includes all tautomeric forms of the compounds. The invention includes atropisomers and rotational isomers.

The invention is intended to include all isotopes of atoms occurring in the compounds. Isotopes include those atoms having the same atomic number but different mass numbers. By way of general example and without limitation, isotopes of hydrogen include deuterium and tritium. Isotopes of carbon include13C and14C. Isotopically-labeled compounds of the invention can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described herein, using an appropriate isotopically-labeled reagent in place of the non-labeled reagent otherwise employed. Such compounds may have a variety of potential uses, for example as standards and reagents in determining biological activity. In the case of stable isotopes, such compounds may have the potential to favorably modify biological, pharmacological, or pharmacokinetic properties.

Biological Methods

N-formyl peptide receptors (FPRs) are a family of chemo attractant receptors that facilitate leukocyte response during inflammation. FPRs belong to the seven-transmembrane G protein-coupled receptor superfamily and are linked to inhibitory G-proteins (Gi). Three family members (FPR1, FPR2 and FPR3) have been identified in humans and are predominantly found in myeloid cells with varied distribution and have also been reported in multiple organs and tissues. After agonist binding, the FPRs activate a multitude of physiological pathways, such as intra cellular signaling transduction, Ca2+ mobilization and transcription. The family interacts with a diverse set of ligands that includes proteins, polypeptides and fatty acid metabolites which activate both pro-inflammatory and pro-resolution downstream responses.

The FPR2 receptor binds multiple ligands to invoke both inflammatory and anti-inflammatory responses. Inflammation mediator release by FPR2 is promoted by endogenous protein ligands such as Serum amyloid A (SAA) and Amyloid □ (1-42), whereas resolution of inflammation is induced by ligands that include arachidonic acid metabolites, lipoxin A4 (LXA4) and Epi-lipoxin (ATL), and a docosahexenoic acid metabolite, resolvin D1 (RvD1). The pro-resolving fatty acid metabolites mediate inhibition and resolution of inflammation through the FPR2 receptor by stimulating phagocytosis of apototic neutrophils by macrophages. Removal of the apototic neutrophils induces the release of cytokines that activate pro-resolution pathways.

The FPR1 receptor was originally isolated as a high affinity receptor for N-Formylmethionine containing peptides, such as N-Formylmethionine-leucyl-phenylalanine (FMLP). The protein directs mammalian phagocytic and blood leukocyte cells to sites of invading pathogens or inflamed tissues and activates these cells to kill pathogens or to remove cellular debris.

FPR2 and FPR1 Cyclic Adenosine Monophosphate (cAMP) Assays. A mixture of forskolin (5 μM final for FPR2 or 10 μM final for FPR1) and IBMX (200 μM final) were added to 384-well Proxiplates (Perkin-Elmer) pre-dotted with test compounds in DMSO (1% final) at final concentrations in the range of 1.7 nM to 100 μM. Chinese Hamster Ovary cells (CHO) overexpressing human FPR1 or human FPR2 receptors were cultured in F-12 (Ham's) medium supplemented with 10% qualified FBS, 250 μg/ml zeocin and 300 μg/ml hygromycin (Life Technologies). Reactions were initiated by adding 2,000 human FPR2 cells per well or 4,000 human FPR1 cells per well in Dulbecco's PBS (with calcium and magnesium) (Life Technologies) supplemented with 0.1% BSA (Perkin-Elmer). The reaction mixtures were incubated for 30 min at room temperature. The level of intracellular cAMP was determined using the HTRF HiRange cAMP assay reagent kit (Cisbio) according to manufacturer's instruction. Solutions of cryptate conjugated anti-cAMP and d2 flurorophore-labelled cAMP were made in a supplied lysis buffer separately. Upon completion of the reaction, the cells were lysed with equal volume of the d2-cAMP solution and anti-cAMP solution. After a 1-h room temperature incubation, time-resolved fluorescence intensity was measured using the Envision (Perkin-Elmer) at 400 nm excitation and dual emission at 590 nm and 665 nm. A calibration curve was constructed with an external cAMP standard at concentrations ranging from 1 μM to 0.1 pM by plotting the fluorescent intensity ratio from 665 nm emission to the intensity from the 590 nm emission against cAMP concentrations. The potency and activity of a compound to inhibit cAMP production was then determined by fitting to a 4-parametric logistic equation from a plot of cAMP level versus compound concentrations.

The exemplified Examples disclosed below were tested in the FPR2 and FPR1 cAMP assay described above and found having FPR2 and/or FPR1 agonist activity. A range of IC50values of ≤1 μM (1000 nM) in one of the assays was observed. Table 1 below lists EC50values in the FPR2 and FPR1 cAMP assays measured for the following examples.

Pharmaceutical Compositions and Methods of Use

The compounds of the present invention may be administered to patients for the treatment of a variety of conditions and disorders, including atherosclerosis, heart failure, lung diseases including asthma, COPD, cystic fibrosis, neuroinflammatory diseases including multiple sclerosis, Alzheimer's disease, stroke, and chronic inflammatory diseases such as inflammatory bowel disease, rheumatoid arthritis, psoriasis, sepsis, lupus and kidney fibrosis.

Another aspect of the invention is a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I in combination with a pharmaceutical carrier.

Another aspect of the invention is a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I in combination with at least one other therapeutic agent and a pharmaceutical carrier.

Unless otherwise specified, the following terms have the stated meanings. The term “patient” means a subject suitable for therapy as determined by practitioners in the field and encompasses all suitable mammalian species including humans that could potentially benefit from treatment with a FPR2 and/or FPR1 agonist as understood by practioners in this field. Common risk factors include, but are not limited to, age, sex, weight, family history, sleep apnea, alcohol or tobacco use, physical inactivity arrthymia or signs of insulin resistance such as acanthosis nigricans, hypertension, dyslipidemia, or polycystic ovary syndrome (PCOS). “Treating” or “treatment” encompass the treatment of a patient as understood by practitioners in the art and include inhibiting the disease-state, i.e., arresting it development; relieving the disease-state, i.e., causing regression of the disease state; and/or preventing the disease-state from occurring in a patient. “Therapeutically effective amount” is intended to include an amount of a compound that is effective or beneficial as understood by practitioners in this field.

“Pharmaceutical composition” means a composition comprising a compound of the invention in combination with at least one additional pharmaceutically acceptable carrier. A “pharmaceutically acceptable carrier” refers to media for the delivery of biologically active agents as understood by practitioners in the art, such as diluents, preserving agents, fillers, flow regulating agents, disintegrating agents, wetting agents, emulsifying agents, suspending agents, sweetening agents, flavoring agents, perfuming agents, anti-bacterial agents, anti-fungal agents, lubricating agents, and dispensing agents. Pharmaceutically acceptable carriers are formulated according to a number of factors known to those of ordinary skill in the art. These include, without limitation, the type and nature of the active agent being formulated; the subject to which the agent-containing composition is to be administered; the intended route of administration of the composition; and the therapeutic indication being targeted. Descriptions of suitable pharmaceutically acceptable carriers and factors involved in their selection are known in the art in such references as Allen, L. V., Jr. et al.,Remington: The Science and Practice of Pharmacy(2 Volumes), 22nd Edition, Pharmaceutical Press (2012).

Solid compositions are normally formulated in dosage units and compositions providing form about 1 to 1000 mg of the active ingredient per dose are preferred. Some examples of dosages are 1 mg, 10 mg, 100 mg, 250 mg, 500 mg, and 1000 mg.

Liquid compositions are usually in dosage unit ranges. Generally, the liquid composition will be in a unit dosage range of 1-100 mg/mL. Some examples of dosages are 1 mg/mL, 10 mg/mL, 25 mg/mL, 50 mg/mL, and 100 mg/mL.

Another aspect of the invention is a method for treating heart disease comprising administering a therapeutically effective amount of a compound of formula I to a patient.

Another aspect of the invention is a method for treating heart disease wherein the heart disease is selected from the group consisting of angina pectoris, unstable angina, myocardial infarction, heart failure, acute coronary disease, acute heart failure, chronic heart failure, and cardiac iatrogenic damage.

Another aspect of the invention is a method for treating heart disease wherein the treatment is post myocardial infarction.

Another aspect of the invention is the method wherein the heart disease is associated with chronic heart failure.

Another aspect of the invention is the method wherein the treatment is to improve myocardial wound healing.

Another aspect of the invention is the method wherein the treatment is to diminish myocardial fibrosis.

The invention encompasses all conventional modes of administration; oral and parenteral methods are preferred. Generally, the dosing regimen will be similar to other cardiovascular agents used clinically. The dosage regimen and mode for administration for the compounds of the present invention will depend on known factors known by practitioners in the art and include age, sex, health, medical condition, and weight of the recipient; the nature and extent of the symptoms; the kind of concurrent treatment; the frequency of treatment; the route of administration, and the effect desired. Typically, the daily dose will be 0.1-100 mg/kg body weight daily. Generally, more compound is required orally and less parenterally. The specific dosing regimen, however, will be determined by a physician using sound medical judgment.

Another aspect of the invention is a method for treating heart disease comprising administering a therapeutically effective amount of a compound of formula I to a patient in conjunction with at least one other therapeutic agent.

The compounds of the present invention are also useful as standard or reference compounds, for example as a quality standard or control, in tests or assays involving the FPR2. Such compounds may be provided in a commercial kit, for example, for use in pharmaceutical research involving FPR2 activity. For example, a compound of the present invention could be used as a reference in an assay to compare its known activity to a compound with an unknown activity. This would ensure the experimenter that the assay was being performed properly and provide a basis for comparison, especially if the test compound was a derivative of the reference compound. When developing new assays or protocols, compounds according to the present invention could be used to test their effectiveness. The compounds of the present invention may also be used in diagnostic assays involving FPR2.

Chemical Methods

Abbreviations as used herein, are defined as follows: “1×” for once, “2×” for twice, “3×” for thrice, “° C.” for degrees Celsius, “aq” for aqueous, “Col” for column, “eq” for equivalent or equivalents, “g” for gram or grams, “mg” for milligram or milligrams, “L” for liter or liters, “mL” for milliliter or milliliters, “μL” for microliter or microliters, “N” for normal, “M” for molar, “nM” for nanomolar, “mol” for mole or moles, “mmol” for millimole or millimoles, “min” for minute or minutes, “h” for hour or hours, “rt” for room temperature, “RT” for retention time, “ON” for overnight, “atm” for atmosphere, “psi” for pounds per square inch, “conc.” for concentrate, “aq” for “aqueous”, “sat” or “sat'd” for saturated, “MW” for molecular weight, “mw” or “μwave” for microwave, “mp” for melting point, “Wt” for weight, “MS” or “Mass Spec” for mass spectrometry, “ESI” for electrospray ionization mass spectroscopy, “HR” for high resolution, “HRMS” for high resolution mass spectrometry, “LCMS” for liquid chromatography mass spectrometry, “HPLC” for high pressure liquid chromatography, “RP HPLC” for reverse phase HPLC, “TLC” or “tlc” for thin layer chromatography, “NMR” for nuclear magnetic resonance spectroscopy, “nOe” for nuclear Overhauser effect spectroscopy, “1H” for proton, “δ” for delta, “s” for singlet, “d” for doublet, “t” for triplet, “q” for quartet, “m” for multiplet, “br” for broad, “Hz” for hertz, and “α”, “β”, “R”, “S”, “E”, and “Z” are stereochemical designations familiar to one skilled in the art.

The compounds of this invention can be made by various methods known in the art including those of the following schemes and in the specific embodiments section. The structure numbering and variable numbering shown in the synthetic schemes are distinct from, and should not be confused with, the structure or variable numbering in the claims or the rest of the specification. The variables in the schemes are meant only to illustrate how to make some of the compounds of this invention.

The disclosure is not limited to the foregoing illustrative examples and the examples should be considered in all respects as illustrative and not restrictive, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced.

It will also be recognized that another major consideration in the planning of any synthetic route in this field is the judicious choice of the protecting group used for protection of the reactive functional groups present in the compounds described in this invention. An authoritative account describing the many alternatives to the trained practitioner is Greene, T. W. et al.,Protecting Groups in Organic Synthesis,4th Edition,Wiley(2007)).

Compounds having the general Formula (IA): wherein rings A, B and C are defined above as Ar1, Ar2and R1, respectively, and can be prepared by the following one or more of the synthetic Schemes.

1-Arylpiperidinone compounds of this invention wherein rings A and B are substituted phenyl or heteroaryl rings and ring C is a substituted saturated heterocycle can be prepared by the general route shown in Scheme 1, starting from a suitably protected 3-aminopiperidin-2-one 1a, where PG is a protecting group such as Boc or Cbz. Copper-catalyzed coupling of 1a to a substituted iodobenzene 1b or other suitable halo aryl or heteroaryl compound in a suitable solvent such as butanol or dioxane, in the presence of a base such as potassium carbonate and a suitable ligand such as N,N′-dimethylethylenediamine, can afford 1-arylpiperidinones 1c. Additional methods for this transformation include other variations of Ullmann, Goldberg, and Buchwald copper-catalyzed amidation or Buchwald Pd-catalyzed amidation depending on the nature of ring B, using methods known to one skilled in the art for these types of couplings (see for example Yin & BuchwaldOrganic Lett.2000, 2, 1101; Klapers et al.JACS,2001, 123, 7727; Klapars et al.JACS,2002, 124, 7421; Yin & BuchwaldJACS.2002, 124, 6043; Kiyomor, Madoux & Buchwald,Tet. Lett.,1999, 40, 2657, Surry and BuchwaldAngew. Chem. Int. Ed.,2008, 47, 6338). Subsequent palladium-catalyzed amination of 1c to a suitably substituted amine 1d can provide compound 1e. Other methods for forming this bond can be found in the literature and can used by those skilled in the art. (Surry & Buchwald Chem Sci. 2011; 2(1): 27-50; Shaughnessy, Ciganek & DeVasher,Organic Reactions.2014, 85:1:1-668). Removal of the protecting group from 1e, followed by condensation of the resulting free amine with a suitably substituted phenyl isocyanate, 1g or phenylcarbamate 1h can provide ureas 1f. Suitable isocyanates or 4-nitrophenylcarbamates are either commercially available or can be readily obtained from the corresponding aniline by methods known to one skilled in the art. Alternately, the ureas 1f can be obtained by treatment of the deprotected 3-aminopiperidinone intermediate with 4-nitrophenylchloroformate to form the carbamate, followed by condensation with an appropriately substituted aniline 1j. It will also be recognized by one skilled in the art that additional compounds of this invention wherein rings A and B are heteroaryl rings, such as pyridine, pyrimidine, thiazole, etc., can also be prepared using the methods outlined in Scheme 1 by substituting the appropriate heteroaryl iodide or bromine for 1b and heteroaryl amine, isocyanate or p-nitrophenylcarbamate for 1e.

Alternatively as described in Scheme 2, compounds of this invention can be prepared from intermediate 1c by first deprotecting the amine and forming the urea linkage to ring A using the conditions described above for the conversion of 1e to 1f to provide compounds 2a. Compound 2a can then be coupled with amine under Pd-catalysis or Cu-catalysis conditions as shown in Scheme 1 for the transformation of 1c to 1e.

Additionally, compounds of this invention can be prepared from intermediate 2a by conversion to boronate 3b using palladium-catalyzed borylation according to the method of Suzuki and Miyaura followed by coupling of the resulting pinacolatoboron species with an amine copper catalyzed Chan-Lam coupling to provide compounds 1f (J. Org. Chem.,2016, 81 (9), pp 3942-3950).

Alternatively, compounds of this invention can be prepared from intermediate 4b by nucleophilic displacement of the arylfluoride with cyclic amines (1d) to form intermediate 4c. Deprotection and installation of the urea, as shown in the above Schemes, results in the synthesis of some compounds described by this invention.

Even though rings A and B are shown as phenyl and C is shown as piperidine in Schemes 1-4, those skilled in the art can use analogous chemistry to make other compounds claimed in this patent. For example, the chemistry for introducing Ring C can also used with other cyclic amines.

The following methods were used in the exemplified Examples, except where noted otherwise. Purification of intermediates and final products was carried out via either normal or reverse phase chromatography. Normal phase chromatography was carried out using pre-packed SiO2cartridges eluting with either gradients of hexanes and EtOAc or DCM and MeOH unless otherwise indicated. Reverse phase preparative HPLC was carried out using C18 columns eluting with gradients of Solvent A (10 mM ammonium acetate in water) and Solvent B (ACN, UV 220 nm) or with gradients of Solvent A (10 mM ammonium acetate in water) and Solvent B (MeOH, UV 220 nm) or with gradients of Solvent A (0.1% TFA in water) and Solvent B (ACN, UV 220 nm) (or) SunFire Prep C18 OBD 5μ. 19×150 mm, 25 min gradient from 0-100% B. A=10 mM ammonium acetate in water. B=ACN/MeOH (or) Waters XBridge C18, 19×1500 mm, 5-μm particles; A=10 mM ammonium acetate in water. B=ACN/MeOH; Gradient: 0-100% B over 25 minutes, then a 5-minute hold at 100% B; Flow: 15 mL/min.

Unless otherwise stated, analysis of final products was carried out by reverse phase analytical HPLC.

SFC and Chiral Purity Methods

NMR Employed in Characterization of Examples. 1H NMR spectra were obtained with Bruker or JEOL® Fourier transform spectrometers operating at frequencies as follows: 1H NMR: 400 MHz (Bruker or JEOL®) or 500 MHz (Bruker or JEOL®).13C NMR: 100 MHz (Bruker or JEOL®). Spectra data are reported in the format: chemical shift (multiplicity, coupling constants, number of hydrogens). Chemical shifts are specified in ppm downfield of a tetramethylsilane internal standard (δ units, tetramethylsilane=0 ppm) and/or referenced to solvent peaks, which in 1H NMR spectra appear at 2.49 ppm for CD2HSOCD3, 3.30 ppm for CD2HOD, 1.94 for CD3CN, and 7.24 ppm for CHCl3, and which in13C NMR spectra appear at 39.7 ppm for CD3SOCD3, 49.0 ppm for CD3OD, and 77.0 ppm for CDCl3. All13C NMR spectra were proton decoupled.

(R)-pyrrolidin-3-ol (0.24 g, 2.7 mmol), and Cs2CO3(1.8 g, 5.4 mmol) were added to a stirred solution of tert-butyl (R)-(1-(4-bromophenyl)-2-oxopiperidin-3-yl)carbamate (1.0 g, 2.7 mmol)) in toluene (10 mL). The reaction mixture was purged with nitrogen for 5 min and charged with Pd(OAc)2(0.061 g, 0.27 mmol) and 2-dicyclohexylphosphino-2′,6′-dipropoxy-1,1′-biphenyl (0.25 g, 0.54 mmol). The reaction mixture was again purged with nitrogen for 3 min and heated to 100° C. for 16 h. The reaction mixture was cooled, filtered through celite, and the filtrate was concentrated under reduced pressure. The crude mixture was purified using column chromatography to afford tert-butyl (1-(4-((R)-3-hydroxypyrrolidin-1-yl)phenyl)-2-oxopiperidin-3-yl)carbamate (250 mg, 0.66 mmol, 25% yield) as a pale yellow solid. MS(ESI) m/z: 376.2 (M+H)+.

4N HCl in 1,4-dioxane (1.7 mL, 6.7 mmol) was added to an ice cooled solution of tert-butyl (1-(4-((R)-3-hydroxypyrrolidin-1-yl)phenyl)-2-oxopiperidin-3-yl)carbamate (0.25 g, 0.67 mmol) in 1,4-dioxane (1 mL), and the mixture was stirred at rt for two hours. The solvent was evaporated under reduced pressure to obtain a gummy solid. The solid was triturated with diethyl ether (2×20 mL) and dried to afford 3-amino-1-(4-((R)-3-hydroxypyrrolidin-1-yl)phenyl)piperidin-2-one hydrochloride (0.20 g, 0.64 mmol, 96% yield) as a off white solid. MS(ESI) m/z: 275.9 (M+H)+.

Additional examples of compounds of this invention shown in Table 1 were prepared using combinations of the procedures described in Example 1 or modifications thereof known to one skilled in the art of organic synthesis.

To a stirred solution of Example 54A (1.0 g, 3.2 mmol) in DMF (1 mL) under nitrogen atmosphere at room temperature, morpholine (0.34 g, 3.9 mmol), and DIPEA (1.7 mL, 9.7 mmol) were added. The reaction mixture was gradually warmed to 130° C. and stirred for 16 hours. The reaction mixture was then cooled to rt, filtered through celite, and washed with EtOAc. The combined filtrates were concentrated under reduced pressure, and the crude compound was purified using by column chromatography to afford Example 54B (0.50 g, 1.3 mmol, 41% yield) as a pale yellow solid. MS(ESI) m/z: 377.5 [M+H]+.

To an ice cooled solution of Example 54B (0.80 g, 2.1 mmol) in 1,4-dioxane (5 mL), was added 4N HCl in 1,4-dioxane (5.3 mL, 21 mmol), and the mixture was stirred at rt for two hours. The solvent was evaporated, and the sample was dried under reduced pressure to obtain a gummy solid. The solid was triturated with diethyl ether (2×20 mL) and dried to afford Example 54C (0.66 g, 2.1 mmol, 99% yield) as a off white solid. The product was used in the subsequent step without purification.

Additional examples of compounds of this invention shown in Table 2 below were prepared using combinations of the procedures described in Example 54 or modifications thereof known to one skilled in the art of organic synthesis.

tert-Butyl trans-5-hydroxy-2-oxopiperidin-3-yl)carbamate was synthesized using the procedures found in Gordon, Sandra et al, Farmaco, 52(10), 603-608: 1997. Acetic anhydride (20 mL, 210 mmol) was added to a solution of tert-butyl trans-(5-hydroxy-2-oxopiperidin-3-yl)carbamate (8.0 g, 35 mmol) in pyridine (20 mL) at rt and the reaction mixture was stirred for 12 h under nitrogen atmosphere. The mixture was concentrated under reduced pressure to give Example 65A (8.0 g, 29 mmol, 85% yield) as a white solid.

A mixture of Example 65A (6.0 g, 22 mmol), 4-bromophenylboronic acid (5.3 g, 26 mmol), copper (II) acetate (4.8 g, 26 mmol) and TEA (9.2 mL, 66 mmol) in DCM (30 mL) was stirred for 2 h under an oxygen atmosphere at rt. The reaction mixture was filtered through celite and washed with ethyl acetate. The filtrate was concentrated under reduced pressure to give the crude product, which was purified via column chromatography (1:4 ethyl acetate/hexanes) to yield Example 65B (3.0 g, 7.0 mmol, 32% yield). MS(ESI) m/z: 427.0/429.0 (M+H)+.

LiOH (0.19 g, 8.0 mmol) was added to a solution of Example 65B (3.4 g, 8.0 mmol) in THF/water (1:1, 30 mL). The reaction mixture was stirred for 2 h at rt. The mixture was concentrated under reduced pressure to remove the volatiles and acidified with aqueous citric acid solution. The solution was filtered and the precipitate was washed with water and dried in vacuo. The crude compound purified by prep HPLC to yield Example 65C (2.0 g, 5.2 mmol, 65% yield). MS(ESI) m/z: 385.0/3870 (M+H)+.

To a stirred solution of Example 65C (0.30 g, 0.78 mmol) in toluene (2 mL), were added morpholine (0.14 g, 0.78 mmol), and Cs2CO3(0.51 g, 1.56 mmol). The reaction mixture was purged with nitrogen for 5 min and charged with PdOAc2(0.017 g, 0.078 mmol) and 2-dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl (0.073 g, 0.16 mmol). The reaction mixture was again purged with nitrogen for 3 min and heated to 100° C. for 16 hours. The reaction mixture was cooled, filtered through celite and the filtrate was concentrated under reduced pressure. The crude mixture was purified by column chromatography to afford a mixture of trans enantiomers of Example 65D (0.12 g, 0.22 mmol, 27.6%) as pale yellow solid. MS(ESI) m/z: 392.3 [M+H]+.

To an ice cooled solution Example 65D (0.15 g, 0.38 mmol) in 1,4-dioxane (0.5 mL), was added 4N HCl in 1,4-dioxane (0.96 mL, 3.8 mmol), and the mixture was stirred at rt for two hours. The solvent was evaporated under reduced pressure to obtain a gummy solid.

Additional examples of compounds of this invention shown in Table 3 below were prepared using combinations of the procedures described previous examples or modifications thereof known to one skilled in the art of organic synthesis.