RU2837702C2 - Amino acid derivatives for treating inflammatory diseases - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, which have inhibitory activity on nuclear factor-kappa B (NF-kB). In formula (I) n is equal to 0, 1 or 2; dotted line is an optional link; Het is pyridin-3-yl; R¹ is hydrogen; R² is hydrogen; R³ is hydrogen, alkyl, cycloalkylalkyl or aralkyl; R⁴ is hydrogen and R⁵ is -C(O)R⁶, R⁶ is aryl or heteroaryl, where aryl or heteroaryl is optionally substituted with R^g, R^h and/or Rⁱ independently selected from alkyl, C₁-C₆ alkoxy, halogen, halogen-C₁-C₆ alkyl, nitro, amino, C₁-C₆ alkylamino and acylamino; acylamino is -NR'C(O)R, where R' is hydrogen or alkyl and where R is C₁-C₆ alkyl or C₃-C₁₀ cycloalkyl, where C₁-C₆ alkyl can be substituted with amino or 2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl; or R⁴ and R⁵ together with the nitrogen atom to which they are attached form 1,3-dioxoisoindolin-2-yl. "Alkyl" denotes a linear saturated monovalent hydrocarbon radical of one to six carbon atoms or a branched saturated monovalent hydrocarbon radical of three to six carbon atoms; "alkylene" denotes a linear saturated divalent hydrocarbon radical of one to six carbon atoms or a branched saturated divalent hydrocarbon radical of three to six carbon atoms; "heteroaryl" denotes a monovalent monocyclic or bicyclic aromatic radical of 5-10 ring atoms, where one or two ring atoms are heteroatoms selected from N and O, and the remaining ring atoms are carbon; "aryl" denotes a monocyclic or bicyclic aromatic hydrocarbon radical of 6-10 ring atoms; "cycloalkyl" denotes a cyclic saturated monovalent hydrocarbon radical of three to ten carbon atoms; "cycloalkylalkyl" denotes a radical -(alkylene)-R, where R is cycloalkyl as defined above. Invention also relates to a pharmaceutical composition containing said compound and a method of treating a disease which can be treated by inhibiting NF-kB activation.

EFFECT: compound can be useful for treating inflammatory diseases.

21 cl, 3 tbl, 68 ex

Description

Field of technology

The present invention provides certain amino acid derivatives that inhibit the activation of nuclear factor-kappa B (NF-kB) induced by inflammatory stimuli. Accordingly, the compounds according to the present invention are used to treat inflammatory diseases such as rheumatoid arthritis, multiple sclerosis, asthma and inflammatory bowel disease. Also provided are pharmaceutical compositions containing such compounds and methods for their preparation.

State of the art

Inflammation is an important host response to infection or injury. However, dysregulation of this response, resulting in persistent or inappropriate inflammation, contributes to a wide range of pathological processes. Inflammatory disorders include autoimmune diseases, allergies, asthma, chronic obstructive pulmonary disease, and sepsis, which are major causes of illness and death. It is also becoming apparent that chronic low-grade inflammation underlies many diseases, including diabetes, cancer, cardiovascular disease, and neurodegenerative disorders. Therefore, the identification of new drugs to suppress inflammation is an area of great interest.

NF-kB is a ubiquitously expressed transcription factor that regulates the expression of genes involved in inflammation and pain. NF-kB activation plays a central role in inflammation through its ability to induce the transcription of proinflammatory genes (see Tak et al. NF-kB: a key role in inflammatory diseases. J Clin Inv 2001; 107:7–11; and Liu et al. NF-κB signaling in inflammation. Signal Transduct Target Ther. 2017;2. pii: 17023). The synthesis of proinflammatory cytokines such as TNF-α, IL-1β, IL-6, and IL-8 is mediated by NF-kB, as is the expression of cyclooxygenase 2 (COX-2), inducible nitric oxide synthase (iNOS), adhesion molecules (ICAM-1, E-selectin, and VCAM-1), chemokines (MCP-1, KC, MIP-1), and metalloproteinases. Similarly, cytokines such as TNF-α and IL-1β or prostaglandins produced by COX-2 trigger NF-kB activation, creating an inflammatory cascade.

NF-kB is activated at sites of inflammation in various diseases, including rheumatoid arthritis, osteoarthritis, atherosclerosis, multiple sclerosis, asthma, inflammatory bowel disease, diabetes, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, osteoporosis, systemic lupus erythematosus, chronic obstructive pulmonary disease, stroke, acute kidney injury, glomerulonephritis, psoriasis, atopic dermatitis, etc. (Pai et al. Immune deficiency or hyperactivity-Nf-kappab illuminates autoimmunity. J Autoimmun 2008; 31:245-251; Zhang et al. NFkB in inflammation and renal diseases. Cell Biosci 2015; 5:63; Roman-Blas et al. NF-kB as a potential therapeutic target in osteoarthritis and rheumatoid arthritis. OsteoArthritis and Cartilage 2006; 14:839-848).

Corticosteroids were developed to treat inflammatory conditions and have many side effects that can be mild or severe, especially when corticosteroids are used for a long period of time. For example, corticosteroids can cause weight gain, swelling, high blood pressure, glaucoma, hypokalemia, cataracts, mental disorders, osteoporosis, and a weakened immune system. Nonsteroidal anti-inflammatory drugs (NSAIDs), which act through inhibition of cyclooxygenase (COX) isoenzymes, are widely prescribed, but chronic use of NSAIDs is associated with common adverse effects, including cardiovascular events (hypertension, myocardial infarction, stroke, and heart failure), gastrointestinal adverse effects (diffuse gastritis and isolated gastric and duodenal ulcers) that can be fatal, renal adverse effects (interstitial nephritis), and hepatic adverse reactions (transaminitis, hepatitis). Biologics have been developed to target inflammatory cytokines. Although biologics have been shown to alleviate the pathology and progression of inflammatory diseases, their use has been limited due to serious adverse reactions and the fact that they abolish host defenses against infection (Rider et al. Biologics for Targeting Inflammatory Cytokines, Clinical Uses, and Limitations. Int J Cell Biol. 2016; 2016:9259646).

Therefore, there is a need to develop other anti-inflammatory drugs with fewer side effects for chronic use. The present invention satisfies this and related needs.

The essence of the invention

In one aspect, a compound of formula (I) is provided:

Where:

n is 0, 1, 2, or 3;

the dotted line is an optional connection;

Het is heteroaryl optionally substituted with R ^a , R ^b and/or R ^c independently selected from alkyl, alkoxy, halogen, haloalkyl, haloalkoxy, cyano, hydroxy, amino, alkylamino, dialkylamino, carboxy and alkoxycarbonyl;

R ¹ is hydrogen or alkyl;

R ² is hydrogen or alkyl;

R ³ is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, hydroxyalkyl, alkoxyalkyl, thioalkyl, alkylthioalkyl, aminoalkyl, acylaminoalkyl, carboxyalkyl, alkoxycarbonylalkyl, aminocarbonylalkyl, aryl, aralkyl, heteroaryl or heteroaralkyl, wherein aryl or heteroaryl, by itself or as part of aralkyl or heteroaralkyl, is optionally substituted with R ^d , R ^e and/or R ^f, independently selected from alkyl, alkoxy, hydroxy, halogen, haloalkyl, haloalkoxy, cyano, nitro, carboxy, alkoxycarbonyl, amino, alkylamino, and dialkylamino;

R ⁴ is hydrogen or alkyl; and

R ⁵ is -C(O)R ⁶ , where R ⁶ is alkyl, aryl, aralkyl, heteroaryl, heteroaralkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl or heterocycloalkylalkyl, wherein aryl or heteroaryl by itself or as part of aralkyl or heteroaralkyl is optionally substituted with R ^g , ^Rh and/or R ⁱ , independently selected from alkyl, alkoxy, aminoalkoxy, hydroxyalkoxy, alkoxyalkoxy, cycloalkyloxy, cycloalkylalkyloxy, optionally substituted aryloxy, optionally substituted aralkyloxy, optionally substituted heteroaryloxy, optionally substituted heteroaralkyloxy, optionally substituted heterocycloalkyloxy, optionally substituted heterocycloalkylalkyloxy, halogen, haloalkyl, haloalkoxy, cyano, nitro, hydroxy, carboxy, alkoxycarbonyl, amino, alkylamino, dialkylamino, acylamino and sulfonylamino; or

R ⁴ and R ⁵ together with the nitrogen atom to which they are attached form a 5-7 membered heterocycloamino ring; or

its pharmaceutically acceptable salt.

In a second aspect, the present disclosure relates to a pharmaceutical composition comprising a compound of formula (I) (or any of its embodiments described herein) or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable excipient.

In a third aspect, the present disclosure relates to a method of treating a disease treatable through inhibition of NF-kB activation, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof in a pharmaceutical composition comprising a compound of formula (I) (or any of its embodiments described herein) and a pharmaceutically acceptable excipient. In one embodiment, the diseases are inflammatory diseases, including autoimmune diseases, pain, allergy, asthma, chronic obstructive pulmonary disease and sepsis. In another embodiment, the diseases are rheumatoid arthritis, osteoarthritis, atherosclerosis, multiple sclerosis, asthma, inflammatory bowel disease, diabetes, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, osteoporosis, systemic lupus erythematosus, chronic obstructive pulmonary disease, cystic fibrosis, stroke, acute kidney injury, glomerulonephritis, psoriasis, atopic dermatitis, Behcet's disease, tuberculosis, Crohn's disease, colitis, Paget's disease, pancreatitis, periodonitis, inflammatory lung disease, and lupus nephritis.

In a fourth aspect, the disclosure relates to a compound of formula (I) (or any of its embodiments described herein) or a pharmaceutically acceptable salt thereof for use as a medicine. In one embodiment, the compound of formula (I) (or any of its embodiments described herein) or a pharmaceutically acceptable salt thereof is used to treat inflammatory diseases, including autoimmune diseases, pain, allergy, asthma, chronic obstructive pulmonary disease, and sepsis. In another embodiment, the diseases are rheumatoid arthritis, osteoarthritis, atherosclerosis, multiple sclerosis, asthma, inflammatory bowel disease, diabetes, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, osteoporosis, systemic lupus erythematosus, chronic obstructive pulmonary disease, cystic fibrosis, stroke, acute kidney injury, glomerulonephritis, psoriasis, atopic dermatitis, Behcet's disease, tuberculosis, Crohn's disease, colitis, Paget's disease, pancreatitis, periodonitis, inflammatory lung disease, and lupus nephritis.

In a fifth aspect, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof (and any embodiments thereof described herein) for use in the treatment of a disease in a patient in which NF-kB activation plays a role in the pathology and/or symptoms of the disease. In one embodiment, the diseases are inflammatory diseases, including autoimmune diseases, pain, allergy, asthma, chronic obstructive pulmonary disease, and sepsis. In another embodiment, the diseases are rheumatoid arthritis, osteoarthritis, atherosclerosis, multiple sclerosis, asthma, inflammatory bowel disease, diabetes, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, osteoporosis, systemic lupus erythematosus, chronic obstructive pulmonary disease, cystic fibrosis, stroke, acute kidney injury, glomerulonephritis, psoriasis, atopic dermatitis, Behcet's disease, tuberculosis, Crohn's disease, colitis, Paget's disease, pancreatitis, periodonitis, inflammatory lung disease, and lupus nephritis.

In the sixth aspect, an intermediate compound of formula (II) is provided:

wherein the dotted line, n, Het, R ¹ , R ² and R ³ are as defined for formula (I) above (including embodiments described herein). In one aspect, the intermediate compound has the structure (IIB):

wherein n, Het, R ¹ , R ² and R ³ are as defined for formula (I) (including embodiments described herein). In another aspect, the intermediate compound has the structure (IIB'):

wherein R ¹ , R ² and R ³ are as defined for formula (I) (including the embodiments described herein) and pyridyl is optionally substituted with R ^a , R ^b and/or R ^c .

In the seventh aspect, a method for producing a compound of formula (IB) is provided:

where n, Het, R ¹ , R ² and R ³ are as defined above, R ⁴ is hydrogen or alkyl and R ⁵ is -COR ⁶ , where R ⁶ is as defined above, comprising:

interaction of the compound of formula (IIB):

where the dotted line, n, Het, R ¹ , R ² and R ³ are as defined above;

(i) with a compound of formula R ⁶ COLG, where R ⁶ is as defined in formula (I) above and LG is a leaving group, under acylation reaction conditions; or

(ii) with a compound of formula R ⁶ COOH under amino acid coupling conditions;

(iii) optionally converting the compound of formula (IB) obtained in step (i) or (ii) into an acid addition salt; or

(iv) optionally converting the compound of formula (IB) obtained in step (i) or (ii) into the free base.

Detailed description

Definitions:

Unless otherwise specified, the following terms used in the description and claims are defined for the purposes of this application and have the following meaning:

"Alkyl" means a linear saturated monovalent hydrocarbon radical containing from one to six carbon atoms or a branched saturated monovalent hydrocarbon radical containing from three to six carbon atoms, such as methyl, ethyl, propyl, 2-propyl, butyl, pentyl and the like.

"Alkylene" means a linear saturated divalent hydrocarbon radical containing from one to six carbon atoms or a branched saturated divalent hydrocarbon radical containing from three to six carbon atoms, unless otherwise specified, such as methylene, ethylene, propylene, 1-methylpropylene, 2-methylpropylene, butylene, pentylene and the like.

"Alkylthio" means an -SR radical where R is alkyl as defined above, such as methylthio, ethylthio and the like.

"Alkylsulfonyl" means a _-SO2R radical where R is alkyl as defined above, such as methylsulfonyl, ethylsulfonyl and the like.

"Amino" means -NH ₂ .

"Alkylamino" means a -NHR radical where R is alkyl as defined above, such as methylamino, ethylamino, propylamino or 2-propylamino and the like.

"Aminoalkyl" means a linear monovalent hydrocarbon radical of one to six carbon atoms or a branched monovalent hydrocarbon radical of three to six carbon atoms substituted with -NR'R", where R' and R" are independently hydrogen or alkyl as defined above, such as aminomethyl, aminoethyl, methylaminomethyl and the like.

"Alkoxy" means an -OR radical where R is alkyl as defined above, such as methoxy, ethoxy, propoxy or 2-propoxy, n- , iso- or tert- butoxy and the like.

"Alkoxyalkyl" means a linear monovalent hydrocarbon radical containing from one to six carbon atoms, or a branched monovalent hydrocarbon radical containing from three to six carbon atoms, substituted by at least one alkoxy group, such as one or two alkoxy groups as defined above, for example, 2-methoxyethyl, 1-, 2- or 3-methoxypropyl, 2-ethoxyethyl and the like.

"Alkoxyalkyloxy" means a -(O)R radical where R is alkoxyalkyl as defined above, such as methoxyethoxy, ethoxyethoxy and the like.

“Aminoalkyloxy” means an -O-alkylene-R radical where R is -NR’R”, where R’ and R” are independently hydrogen or alkyl as defined above, such as aminoethyloxy, methylaminoethyloxy, dimethylaminoethyloxy, diethylaminoethyloxy and the like.

"Thioalkyl" means a linear monovalent hydrocarbon radical of one to six carbon atoms or a branched monovalent hydrocarbon radical of three to six carbon atoms substituted with a -SH group, such as thioethyl, 1-, 2-, or 3-thiopropyl, and the like.

"Alkylthioalkyl" means a linear monovalent hydrocarbon radical of one to six carbon atoms or a branched monovalent hydrocarbon radical of three to six carbon atoms substituted with an alkylthio group as defined above, such as 2-methylthioethyl, 1-, 2- or 3-ethylthiopropyl and the like.

"Alkoxycarbonyl" means a -C(O)OR radical where R is alkyl as defined above, such as methoxycarbonyl, ethoxycarbonyl and the like.

"Alkoxycarbonylalkyl" means a linear monovalent hydrocarbon radical of one to six carbon atoms or a branched monovalent hydrocarbon radical of three to six carbon atoms substituted with an alkoxycarbonyl group as defined above, such as 2-methylcarboxyethyl, methylcarboxymethyl, 1-, 2- or 3-ethylcarboxypropyl and the like.

"Acyl" means a -C(O)R radical where R is alkyl as defined above, such as methylcarbonyl, ethylcarbonyl and the like.

"Acylamino" means a -NHC(O)R radical wherein R is alkyl as defined above, such as methylcarbonylamino, ethylcarbonylamino and the like, or R may be optionally substituted with one, two or three substituents independently selected from substituents as described herein, such as alkyl, hydroxyl, cycloalkyl, heterocycloalkyl, carboxy, alkoxycarbonyl, hydroxy, alkoxy, alkylthio, alkylsulfonyl, amino, alkylamino, dialkylamino, halogen, haloalkyl, haloalkoxy and cyano.

"Acylaminoalkyl" means a linear monovalent hydrocarbon radical of one to six carbon atoms or a branched monovalent hydrocarbon radical of three to six carbon atoms substituted with an acylamino group as defined above, such as 2-acetylaminoethyl, 1-, 2- or 3-ethanoylaminopropyl and the like.

"Aminocarbonylalkyl" means a linear monovalent hydrocarbon radical of one to six carbon atoms or a branched monovalent hydrocarbon radical of three to six carbon atoms substituted by a -CONRR' group, wherein R is hydrogen or alkyl and R' is hydrogen, alkyl, optionally substituted aryl, optionally substituted heteroaryl, or R and R' together with the nitrogen atom to which they are attached form a heterocycloamino, for example, aminocarbonylethyl, methylaminocarbonylethyl, methylaminocarbonylmethyl, 1-, 2- or 3-ethylaminocarbonylpropyl, pyrrolidinylmethyl, piperidinylethyl and the like.

"Aryl" means a monocyclic or bicyclic aromatic hydrocarbon radical of 6 to 10 ring atoms, such as phenyl or naphthyl.

"Aralkyl" means a -(alkylene)-R radical where R is aryl as defined above, such as benzyl, phenethyl, and the like.

"Cycloalkyl" means a cyclic saturated monovalent hydrocarbon radical of three to ten carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl and the like.

"Cycloalkyloxy" means -O-R where R is cycloalkyl as defined above, such as cyclopropyloxy, cyclobutyloxy, cyclopentyloxy and the like.

"Cycloalkylalkyl" means a -(alkylene)-R radical where R is cycloalkyl as defined above, such as cyclopropylmethyl, cyclohexylmethyl and the like.

"Cycloalkylalkyloxy" means an -O-cycloalkylalkyl radical as defined above, such as cyclopropylmethyloxy, 2-cyclopropylethyloxy, 1-, 2-, 3-cyclobutylpropoxy and the like.

"Carboxy" means -COOH.

"Carboxyalkyl" means a linear monovalent hydrocarbon radical of one to six carbon atoms or a branched monovalent hydrocarbon radical of three to six carbon atoms substituted with a carboxy group as defined above, such as 2-carboxyethyl, 1-, 2- or 3-carboxypropyl and the like.

“Dialkylamino” means a -NRR’ radical where R and R’ are alkyl as defined above, such as dimethylamino, methylethylamino and the like.

"Halogen" means fluorine, chlorine, bromine or iodine, preferably fluorine or chlorine.

"Haloalkyl" means an alkyl radical as defined above that is substituted by one or more halogen atoms, for example, one to five halogen atoms such as fluorine or chlorine, including those substituted by different halogens, for example, _{-CH2Cl , -CF3, -CHF2, -CH2CF3, -CF2CF3 , -CF ( CH3 ) 2} and the _like . When alkyl is substituted only by fluorine, it may be referred to in this application as fluoroalkyl.

"Haloalkoxy" means an -OR radical where R is a haloalkyl as defined above, such as _-OCF3 , _-OCHF2 , and the like. If R is a haloalkyl where the alkyl is substituted only by fluorine, it may be referred to in this application as fluoroalkoxy.

"Hydroxyalkyl" means a linear hydrocarbon radical of one to six carbon atoms or a branched monovalent hydrocarbon radical of three to six carbon atoms substituted by one or two hydroxy groups, with the proviso that if two hydroxy groups are present, they cannot both be on the same carbon atom. Representative examples include, but are not limited to, hydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, 1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl, 3-hydroxybutyl, 4-hydroxybutyl, 2,3-dihydroxypropyl, 1-(hydroxymethyl)-2-hydroxyethyl, 2,3-dihydroxybutyl, 3,4-dihydroxybutyl and 2-(hydroxymethyl)-3-hydroxypropyl, preferably 2-hydroxyethyl, 2,3-dihydroxypropyl and 1-(hydroxymethyl)-2-hydroxyethyl.

"Hydroxyalkyloxy" means an -O-R radical where R is hydroxyalkyl as defined above, such as hydroxymethyloxy, 2-hydroxyethyloxy, 1-, 2-, 3-hydroxypropoxy and the like.

"Heterocycloalkyl" or "heterocyclyl" means a saturated or unsaturated monovalent monocyclic group having 4-8 ring atoms, in which one, two or three of the atoms are heteroatoms selected from N, O or S(O) _n , wherein n is an integer from 0 to 2, the remaining ring atoms being C. Additionally, one or two ring carbon atoms in the heterocyclyl ring may be optionally substituted with a -CO- group, and the heterocycloalkyl is optionally fused with phenyl or a 5- or 6-membered heteroaryl as defined above. More specifically, the term heterocyclyl includes, but is not limited to, pyrrolidino, piperidino, homopiperidino, 2-oxopyrrolidinyl, 2-oxopiperidinyl, morpholino, piperazino, tetrahydropyranyl, thiomorpholino and the like. If the heterocyclyl ring is unsaturated, it may contain one or two double bonds, provided that the ring is not aromatic. If the heterocyclyl group contains at least one nitrogen atom, it is also referred to herein as heterocycloamino and is a subset of the heterocyclyl group. Unless otherwise specified, the heterocycloalkyl ring is optionally substituted with one, two, or three substituents independently selected from alkyl, hydroxyl, alkoxy, amino, alkylamino, and dialkylamino.

^" Heterocycloalkylalkyl" means a -(alkylene)-R radical where R is a heterocycloalkyl ring as defined above, such as tetrahydrofuranylmethyl, piperazinylmethyl, morpholinylethyl and the like.

"Heterocycloamino" means a saturated or unsaturated monovalent monocyclic group of 4 to 8 carbon atoms in which one or two ring atoms are heteroatoms selected from N, O or S(O) _n , where n is an integer from 0 to 2, the remaining ring atoms being C, provided that at least one of the ring atoms is N, unless otherwise specified. Additionally, one or two carbon atoms in the heterocycloamino ring may optionally be substituted with a -CO- group. Heterocycloamino is optionally fused with phenyl or a 5- or 6-membered heteroaryl as defined above. More specifically, the term heterocyclyl includes, but is not limited to, pyrrolidino, piperidino, homopiperidino, 2-oxopyrrolidinyl, 2-oxopiperidinyl, morpholino, piperazino, thiomorpholino and the like. Unless otherwise specified, the heterocycloamino ring may be optionally substituted with one, two, or three substituents independently selected from alkyl, halogen, hydroxyl, alkoxy, amino, alkylamino, dialkylamino, hydroxyalkyl, alkoxyalkyl, or aminoalkyl, each as defined herein.

"Heteroaryl" means a monovalent monocyclic or bicyclic radical of 5 to 10 ring atoms, unless otherwise specified, wherein one or more (in one embodiment, one, two or three) of the ring atoms are heteroatoms selected from N , O or S, wherein the remaining atoms are carbon. Representative examples include, but are not limited to, pyrrolyl, thienyl, thiazolyl, imidazolyl, furanyl, indolyl, isoindolyl, oxazolyl, isoxazolyl, benzothiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazolyl, triazolyl, tetrazolyl and the like. As defined herein, the terms "heteroaryl" and "aryl" are mutually exclusive. If a heteroaryl ring contains 5 or 6 ring atoms, it is also referred to herein as a 5- or 6-membered heteroaryl.

"Heteroaralkyl" means a -(alkylene)-R radical where R is a heteroaryl as defined above, such as pyridinylmethyl and the like. If the heteroaryl ring in a heteroaralkyl contains 5 or 6 ring atoms, it is also referred to herein as a 5- or 6-membered heteroaralkyl.

"Patient" means a mammal, preferably a human.

The present disclosure also includes protected derivatives of the compounds of the present disclosure (I). For example, when the compounds of the present invention contain groups such as hydroxy, carboxy, thiol, or any group containing nitrogen atom(s), these groups can be protected with suitable protecting groups. A complete list of suitable protecting groups can be found in T. W. Greene, Protective Groups in Organic Synthesis , John Wiley & Sons, Inc. (1999), the disclosure of which is incorporated herein by reference in its entirety. Protected derivatives of the compounds of the present invention can be prepared by methods well known in the art.

The present disclosure also includes polymorphic forms and deuterated forms of a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

A "pharmaceutically acceptable salt" of a compound means a salt that is pharmaceutically acceptable and that possesses the desired pharmacological activity of the parent compound. Such salts include:

acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like; or formed with organic acids such as formic acid, acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, glucoheptonic acid, 4,4'-methylenebis(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, mucic acid and the like; or

salts formed when an acidic proton present in the parent compound is either replaced by a metal ion, such as an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinated to an organic base, such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N -methylglucamine, and the like. It is understood that pharmaceutically acceptable salts are non-toxic. Further information on suitable pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences , 17th ed., Mack Publishing Company, Easton, PA, 1985, which is incorporated herein by reference in its entirety.

The compounds of the present disclosure may have asymmetric centers. The compounds of the present disclosure containing an asymmetrically substituted atom may be isolated in optically active or racemic forms. It is well known in the art to prepare optically active forms, for example, by resolution of materials. All chiral, diastereomeric, all mixtures of chiral or diastereomeric forms and racemic forms are included within the scope of this disclosure unless the specific stereochemistry or isomeric form is specifically indicated. It will also be understood by one skilled in the art that when a compound is designated as an (R) or (S) stereoisomer, it may contain the corresponding (S) or (R) stereoisomer as an impurity, preferably the undesired enantiomer is present in an amount of less than about 10%, preferably 5% w/w. About means + or - 10% of the original value.

Some compounds of the present invention may exist as tautomers and/or geometric isomers. All possible tautomers and cis and trans isomers in individual forms and mixtures thereof are included within the scope of the present disclosure. Additionally, the term alkyl as used herein includes all possible isomeric forms of the said alkyl group, although only a few examples are given. Similarly, when cyclic groups such as aryl, heteroaryl, heterocyclyl are substituted, they include all positional isomers, although only a few examples are given. Additionally, all hydrates of a compound of the present disclosure are included within the scope of this disclosure.

"Oxo" or "carbonyl" means =(O) group.

"Optionally substituted alkyl" means alkyl, as defined above, that is optionally substituted with one, two or three substituents independently selected from, for example, alkyl, hydroxyl, cycloalkyl, heterocycloalkyl, carboxy, alkoxycarbonyl, hydroxy, alkoxy, alkylthio, alkylsulfonyl, amino, alkylamino, dialkylamino, halogen, haloalkyl, haloalkoxy and cyano.

“Optionally substituted aryl” means aryl, as defined above, that is optionally substituted with one, two, or three substituents independently selected from alkyl, hydroxyl, cycloalkyl, carboxy, alkoxycarbonyl, hydroxy, alkoxy, alkylthio, alkylsulfonyl, amino, alkylamino, dialkylamino, halogen, haloalkyl, haloalkoxy, and cyano.

"Optionally substituted aryloxy" means an -O-alkylene-R radical wherein R is an optionally substituted aryl as defined above, such as benzyloxy, methoxybenzyloxy, halobenzyloxy, 2-phenethyloxy and the like.

"Optionally substituted aralkyl" means -alkylene-R, wherein R is optionally substituted aryl, each as defined above. "Optionally substituted aralkyloxy" means -O-alkylene-R radical, wherein R is optionally substituted aryl as defined above, for example, benzyloxy, methoxybenzyloxy, halobenzyloxy, 2-phenethyloxy and the like.

"Optionally substituted heteroaryl" means heteroaryl, as defined above, that is optionally substituted with one, two, or three substituents independently selected from alkyl, alkylthio, alkylsulfonyl, hydroxyl, cycloalkyl, carboxy, alkoxycarbonyl, hydroxy, alkoxy, halogen, haloalkyl, haloalkoxy, amino, alkylamino, dialkylamino, and cyano.

"Optionally substituted heteroaryloxy" means an -O-R radical where R is an optionally substituted heteroaryl as defined above, such as pyridinyloxy, furanyloxy, thienyloxy and the like.

"Optionally substituted heteroaralkyloxy" means an -O-alkylene-R radical, wherein R is an optionally substituted heteroaryl as defined above.

"Optionally substituted heterocycloalkyl" means heterocycloalkyl, as defined above, that is optionally substituted with one, two, or three substituents independently selected from alkyl, alkylthio, alkylsulfonyl, hydroxyl, cycloalkyl, carboxy, alkoxycarbonyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, aminoalkyl, halogen, haloalkyl, haloalkoxy, and cyano.

"Optionally substituted heterocycloalkylalkyl" means an -alkylene-R radical wherein R is optionally substituted heterocycloalkyl as defined above, such as piperidinylmethyl, pyrrolidinylethyl, piperazin-1-ylethyl and the like.

"Optionally substituted heterocycloalkyloxy" means an -O-R radical where R is an optionally substituted heterocycloalkyl as defined above, such as piperidinyloxy, pyrrolidinyloxy, tetrahydrofuranyloxy and the like.

"Optionally substituted heterocycloalkylalkyloxy" means an -O-alkylene-R radical wherein R is optionally substituted heterocycloalkyl as defined above, such as piperidinylmethyloxy, pyrrolidinylethyloxy, piperazin-1-ylethyloxy and the like.

"Optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "a heterocyclyl group optionally substituted with an alkyl group" means that alkyl may, but need not, be present and the description includes instances where the heterocyclyl group is substituted with an alkyl group and instances where the heterocyclyl group is not substituted with alkyl.

"Pharmaceutically acceptable carrier or excipient" means a carrier or excipient that is used in the preparation of a pharmaceutical composition that is generally safe, non-toxic, and not biologically or otherwise undesirable, and includes a carrier or excipient that is acceptable for veterinary use as well as pharmaceutical use. "Pharmaceutically acceptable carrier/excipient" as used in the specification and claims includes both one and more than one such excipient.

"Sulfonylamino" means _a -NRSO2R' radical where R is hydrogen or alkyl, and R' is alkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, or optionally substituted heteroaralkyl, each as defined above.

"Treatment" or "treatment" of a disease includes:

(1) prevention of disease, i.e., prevention of the development of clinical symptoms of a disease in a mammal that may be susceptible or predisposed to the disease but does not yet experience or exhibit symptoms of the disease;

(2) inhibition of the disease, i.e. stopping or reducing the progression of the disease or its clinical symptoms; or

(3) relief of the disease, i.e. regression of the disease or its clinical symptoms.

"Therapeutically effective amount" means the amount of a compound of the present invention and/or a pharmaceutically acceptable salt thereof that, when administered to a patient for the treatment of a disease, is sufficient to effect such treatment of the disease. "Therapeutically effective amount" will vary depending on the compound, the disease and its severity, and the age, weight, etc. of the mammal being treated.

Implementation options:

Implementation option 1:

In embodiment 1, the compounds of formula (I) are as defined in the Summary of the Invention section above.

Implementation option 2:

In embodiment 2, the compounds of formula (I) have structure (IA):

where Het, R ¹ , R ² , R ³ , R ⁴ , R ⁵ and n are as defined in the section “Essence of the invention”.

Implementation option 3:

In embodiment 3, the compounds of formula (I) have the structure (IB):

where Het, R ¹ , R ² , R ³ , R ⁴ , R ⁵ and n are as defined in the section “Essence of the invention”.

Implementation option 4:

In embodiment 4, the compounds of any one of embodiments 1, 2 and 3 are such that n is 1, 2 or 3.

Implementation option 5:

In embodiment 5, the compounds of any one of embodiments 1, 2, 3 and 4 are such that n is 1.

Implementation option 6:

In embodiment 6, the compounds of any one of embodiments 1, 2 and 3 are such that n is 0 or 2. In one sub-embodiment of embodiment 6, the compounds of any one of embodiments 1, 2 and 3 are such that n is 0. In a second sub-embodiment of embodiment 6, the compounds of any one of embodiments 1, 2 and 3 are such that n is 2.

Implementation option 7:

In embodiment 7, the compounds of any one of embodiments 1 through 6 and subembodiments contained herein, those wherein R ⁴ is hydrogen or alkyl and R ⁵ is -C(O)R ⁶ , wherein R ⁶ is alkyl, aryl, aralkyl, heteroaryl, heteroaralkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl or heterocycloalkylalkyl, wherein the aryl or heteroaryl by itself or as part of an aralkyl or heteroaralkyl is optionally substituted with R ^g , ^Rh and/or R ⁱ , independently selected from alkyl, alkoxy, aminoalkoxy, hydroxyalkoxy, alkoxyalkoxy, cycloalkyloxy, cycloalkylalkyloxy, optionally substituted aryloxy, optionally substituted aralkyloxy, optionally substituted heteroaryloxy, optionally substituted heteroaralkyloxy, optionally substituted heterocycloalkyloxy, optionally substituted heterocycloalkylalkyloxy, halogen, haloalkyl, haloalkoxy, cyano, nitro, hydroxy, carboxy, alkoxycarbonyl, amino, alkylamino, dialkylamino, acylamino and sulfonylamino. In one sub-embodiment of Embodiment 7, the compounds of any one of Embodiments 1-6 are those wherein ^R6 is aryl or heteroaryl (e.g., thienyl, furanyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, indolyl or indazolyl), optionally substituted with ^Rg , ^Rh and/or ^Ri as defined above (hereinafter, above, means in Embodiment 7). In a second sub-embodiment of embodiment 7, the compounds of any one of embodiments 1-6 are those wherein R ⁶ is aralkyl or heteroaralkyl optionally substituted with R ^g , ^Rh and/or R ⁱ as defined above. In a third sub-embodiment of embodiment 7, the compounds of any one of embodiments 1-6 are those wherein R ⁶ is phenyl optionally substituted with R ^g , ^Rh and/or R ⁱ as defined above. In a fourth sub-embodiment of embodiment 7, the compounds of any one of embodiments 1-6 are those wherein R ⁶ is heteroaryl (e.g., thienyl, furanyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, indolyl or indazolyl) optionally substituted with R ^g , ^Rh and/or R ⁱ as defined above. In a fifth sub-embodiment of embodiment 7, the compounds of any one of embodiments 1 to 6 and the compounds of sub-embodiment one to four of embodiment 7 are those wherein R ^g is alkyl, alkoxy, halogen, haloalkyl, haloalkoxy, hydroxy or cyano, and R ^h and R ⁱ are independently selected from alkyl, alkoxy, aminoalkoxy, hydroxyalkoxy, alkoxyalkoxy, cycloalkyloxy, cycloalkylalkyloxy, optionally substituted aralkyloxy, optionally substituted heteroaryloxy, optionally substituted heterocycloalkyloxy, optionally substituted heterocycloalkylalkyloxy, halogen, haloalkyl, haloalkoxy, cyano, hydroxy, carboxy, alkoxycarbonyl, amino, alkylamino, dialkylamino, acylamino and sulfonylamino. In a sixth sub-embodiment of embodiment 7, the compounds of any one of embodiments 1 to 6 and the compounds of the first to fifth sub-embodiments of embodiment 7 are those wherein R ^g , R ^h and/or R ⁱ are independently selected from alkyl, alkoxy, halogen, haloalkyl, haloalkoxy, cyano, hydroxy, amino, acylamino, preferably methyl, ethyl, methoxy, ethoxy, chloro, fluoro, trifluoromethyl, trifluoromethoxy, hydroxy, acetylamino, butanoylamino and pentanoylamino.

Implementation option 8:

In embodiment 8, the compounds of any one of embodiments 1-6 and subembodiments contained herein are those wherein ^R4 and ^R5 , together with the nitrogen atom to which they are attached, form a 5-7-membered heterocycloamino ring. In one subembodiment of embodiment 8, the heterocycloamino ring is pyrrolidino, piperidino, homopiperidino, 2-oxopyrrolidinyl, 2-oxopiperidinyl, morpholino, piperazino, thiomorpholino, isoindolinyl, or 1,3-dioxoisoindlin-2-yl, optionally substituted with one, two, or three substituents independently selected from alkyl, hydroxyl, alkoxy, hydroxyalkyl, alkoxyalkyl, and aminoalkyl.

Implementation option 9:

In embodiment 9, the compounds of any one of embodiments 1-8 and sub-embodiments contained herein are those wherein R ¹ and R ² are independently hydrogen or methyl, preferably hydrogen.

Implementation option 10:

In embodiment 10, the compounds of any one of embodiments 1-9 and subembodiments contained herein are those wherein when both R ¹ and R ² are alkyl, they are not bonded to the same ring carbon.

Implementation option 11:

In embodiment 11, the compounds of any one of embodiments 1-10 and subembodiments contained herein are those wherein Het is pyridinyl, pyrimidinyl, pyrazinyl, furanyl, thienyl, quinolinyl, isoquinolinyl, pyrazolyl or indolyl, wherein each ring is optionally substituted with ^Ra , ^Rb and/or ^Rc independently selected from alkyl, alkoxy, halogen, haloalkyl, haloalkoxy, cyano, hydroxy, amino, alkylamino, dialkylamino, carboxy and alkoxycarbonyl; preferably ^Ra , ^Rb and/or ^Rc independently selected from methyl, ethyl, methoxy, ethoxy, chloro, fluoro, trifluoromethyl, trifluoromethoxy, cyano, amino, methylamino and dimethylamino. In one sub-embodiment, Het is pyridin-2-yl.

Implementation option 12:

In embodiment 12, the compounds of any one of embodiments 1-11 and subembodiments contained herein are those wherein R ³ is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, hydroxyalkyl, alkoxyalkyl, thioalkyl, alkylthioalkyl, aminoalkyl, acylaminoalkyl, carboxyalkyl, alkoxycarbonylalkyl, aminocarbonylalkyl, aryl, aralkyl, heteroaryl, or heteroaralkyl, wherein aryl or heteroaryl, by itself or in aralkyl and heteroaralkyl, is optionally substituted with R ^d , R ^e , and/or R ^f , independently selected from alkyl, alkoxy, hydroxy, halogen, haloalkyl, haloalkoxy, cyano, nitro, carboxy, alkoxycarbonyl, amino, alkylamino, or dialkylamino. In one sub-embodiment of embodiment 12, the compounds of any one of embodiments 1-11 are those wherein R ³ is hydrogen or alkyl, preferably methyl, ethyl, propyl, isopropyl, sec-propyl, n-, sec-, iso- or tert-butyl.

In a second sub-embodiment of embodiment 12, the compounds of any of embodiments 1-11 are those wherein R ³ is aralkyl optionally substituted with R ^d , R ^e and/or R ^f as defined above, preferably R ³ is benzyl or phenethyl, more preferably benzyl optionally substituted with R ^d , R ^e and R ^f as defined above, even more preferably benzyl.

In a third sub-embodiment of embodiment 12, the compounds of any one of embodiments 1-11 are those wherein R ³ is cycloalkylalkyl optionally substituted with R ^d , R ^e and/or R ^f as defined above, preferably cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl or cyclohexylmethyl optionally substituted with R ^d , R ^e and R ^f as defined above.

In a fourth sub-embodiment of embodiment 12, the compounds of any one of embodiments 1-11 are those wherein R ³ is heteroaralkyl (e.g., thienylmethyl, furanylmethyl, pyridinylmethyl, quinolinylmethyl, isoquinolinylmethyl, indolylmethyl, or indazolylmethyl) optionally substituted with R ^d , R ^e and/or R ^f , as defined above.

In a fifth sub-embodiment of embodiment 12, the compounds of any one of embodiments 1-11 are those wherein R ³ is hydroxyalkyl, alkoxyalkyl or aminoalkyl, preferably hydroxymethyl, hydroxyethyl, methoxymethyl, methoxyethyl, aminomethyl or aminobutyl.

In one embodiment, the stereochemistry at the carbon atom to which R ³ is attached is (S).

In another embodiment, the stereochemistry at the carbon atom to which R ³ is attached is (R).

Typical compounds of formula (I) where R ¹ and R ² are H, n is 1 and other groups are as shown in Table 1 below, include:

Table 1
Conn. No. R R ³ R ⁴ R ⁵ = -COR ⁶ where R ⁶ is: NR ⁴ R ⁵ 1 pyridin-3-yl S-methyl 1,3-dioxoisoindolin-2-yl 2 S-pyridin-3-yl S-methyl 1,3-dioxoisoindolin-2-yl 3 S-pyridin-3-yl S-isopropyl 1,3-dioxoisoindolin-2-yl 4 R-pyridin-3-yl S-isopropyl 1,3-dioxoisoindolin-2-yl 5 mixture of S-pyridin-3-yl and R-pyridin-3-yl S-ethyl and R-ethyl 1,3-dioxoisoindolin-2-yl 6 mixture of S-pyridin-3-yl and R-pyridin-3-yl R-ethyl and S-ethyl 1,3-dioxoisoindolin-2-yl 7 pyridin-3-yl benzyl 1,3-dioxoisoindolin-2-yl 8 pyridin-3-yl hydrogen 1,3-dioxoisoindolin-2-yl 9 pyridin-3-yl S-methyl H 3-nitrophenyl 10 pyridin-3-yl S-methyl H 3-methoxyphenyl 11 pyridin-3-yl S-methyl H 3,4-diethoxyphenyl 12 pyridin-3-yl S-methyl H 3,4-dimethoxyphenyl 13 pyridin-3-yl S-methyl H 4-nitrophenyl 14 pyridin-3-yl S-methyl H 4-chlorophenyl 15 pyridin-3-yl benzyl H 3-nitrophenyl 16 pyridin-3-yl benzyl H 3-methoxyphenyl 17 pyridin-3-yl R-methyl 1,3-dioxoisoindolin-2-yl 18 pyridin-3-yl benzyl H 3-chlorophenyl 19 pyridin-3-yl benzyl H 3-fluorophenyl 20 pyridin-3-yl benzyl H 3-ethoxyphenyl 21 pyridin-3-yl benzyl H 3,4-dimethoxyphenyl 23 pyridin-3-yl benzyl H 3-trifluoromethylphenyl 24 pyridin-3-yl benzyl H 3-fluoro-5-methoxyphenyl 25 pyridin-3-yl R-benzyl H 3-methoxyphenyl 26 pyridin-3-yl R-benzyl H 3-ethoxy-4-methoxyphenyl 27 pyridin-3-yl R-benzyl H 2-chloro-5-methoxyphenyl 28 pyridin-3-yl R-benzyl H 3,5-dimethoxyphenyl 29 pyridin-3-yl R-benzyl H 5-methoxy-2-methylphenyl 30 pyridin-3-yl R-benzyl H 4-chloro-2-methoxyphenyl 31 pyridin-3-yl R-benzyl H 4-fluoro-3-methoxyphenyl 32 pyridin-3-yl R-benzyl H 2-methoxyphenyl 33 pyridin-3-yl S-benzyl H 3-ethoxyphenyl 34 pyridin-3-yl S-benzyl H 4-fluoro-3-methoxyphenyl 35 pyridin-3-yl S-benzyl H 3-methoxyphenyl 36 pyridin-3-yl benzyl H 3-pentanoylaminophenyl 37 pyridin-3-yl benzyl H 3-butanoylaminophenyl 38 pyridin-3-yl benzyl H 3-aminophenyl 39 pyridin-3-yl benzyl H 3-acetylaminophenyl 40 quinolin-3-yl benzyl H 3-Butyramidophenyl 41 pyridin-3-yl benzyl H 3-biotinylaminophenyl 42 pyridin-3-yl S-benzyl H 3-(methylbutanoyl amino)phenyl 43 pyridin-3-yl S-benzyl H 3-(2,2-dimethylpropanoyl amino)phenyl 44 pyridin-3-yl S-benzyl H 3-(cyclopropanyl methaneylamino)phenyl 45 pyridin-3-yl S-benzyl H 3-(cyclobutanyl methaneylamino)phenyl 46 pyridin-3-yl S-benzyl H 3-(cyclopentanyl methaneylamino)phenyl 47 pyridin-3-yl S-benzyl H 2-(butanoylamino)-phenyl 48 pyridin-3-yl S-benzyl H 4-(butanoylamino)-phenyl 49 pyridin-3-yl S-benzyl H quinolin-2-yl 50 pyridin-3-yl S-benzyl H pyrazin-2-yl 51 pyridin-3-yl S-benzyl H pyridin-3-yl 52 pyridin-3-yl S-benzyl H furan-2-yl 53 pyridin-3-yl S-benzyl H 3-(2-amino acetamido)phenyl 54 pyridin-3-yl S-benzyl H 3-(3-aminopropanoyl amino)phenyl 55 pyridin-3-yl S-benzyl H 3-(4-aminobutanoyl amino)phenyl 56 pyridin-3-yl S-benzyl H pyrrol-2-yl 57 pyridin-3-yl S-benzyl H (S)-3-(2-methylbutanoylamino)phenyl 58 pyridin-3-yl S-benzyl H (R)-3-(2-methylbutanoylamino)phenyl 59 pyridin-3-yl S-benzyl H 3-(N-methylbutanoylamino)phenyl 60 pyridin-3-yl S-benzyl H 3-(butylamino)phenyl 61 pyridin-3-yl S-benzyl H N-butyl-1H-pyrrol-2-yl 63 pyridin-3-yl S-benzyl H 3-(butanoylamino)phenyl 64 pyridin-3-yl S-naphth-1-yl H 3-(butanoylamino)phenyl 65 pyridin-3-yl S-naphth-2-yl H 3-(butanoylamino)phenyl 66 pyridin-3-yl S-cyclohexyl H 3-(butanoylamino)phenyl 67 2-chloroquinolin-3-yl benzyl H 3-(butanoylamino)phenyl 68 quinolin-3-yl methyl H 3-(butanoylamino)-phenyl

Additional compounds of formula (I) under consideration are presented in Table 2 below:

Table 2
Conn. No. R R ³ R ⁶ (R ⁵ =COR ⁶ ) n 101 pyridin-3-yl benzyl 3-butanoylaminophenyl 0 102 pyridin-3-yl benzyl 3-butanoylaminophenyl 2 103 pyridin-3-yl 4-methoxybenzyl 3-butanoylaminophenyl 1 104 pyridin-3-yl 3,4-dimethoxybenzyl 3-butanoylaminophenyl 1 105 pyridin-3-yl benzyl tetrahydroquinolin-7-yl 1 106 pyridin-3-yl benzyl indol-6-yl 1 107 pyridin-3-yl benzyl 3-propylsulfonylaminophenyl 1 108 pyridin-3-yl benzyl 3-isopropoxyphenyl 1 109 pyridin-3-yl benzyl 3-cyclopentyloxyphenyl 1 110 pyridin-3-yl benzyl 3-(2-acetylamino-ethyloxy)phenyl 1 111 pyridin-3-yl benzyl 3-(2-dimethylamino-ethyloxy)phenyl 1 112 pyridin-3-yl benzyl 3-(2-pyrrolidin-1-ylethyloxy)phenyl 1 113 pyridin-3-yl benzyl 3-(1-methylpiperidin-4-yloxy)phenyl 1 114 pyridin-3-yl benzyl 3-[2-(1-methylpiperazin-4-yl)ethyloxy]phenyl 1 115 pyridin-3-yl benzyl 3-(2-morpholin-4-ylethyloxy)phenyl 1 116 pyridin-3-yl benzyl pyridin-3-yl 1 117 pyridin-3-yl benzyl 2-aminothiadiazol-5-yl 1 118 pyridin-3-yl benzyl thien-2-yl 1 119 pyridin-3-yl benzyl 5-chlorothien-5-yl 1 120 pyridin-3-yl benzyl 3-(2-methylaminoethyloxy)phenyl 1

General scheme of synthesis

The compounds of the present description can be prepared by the methods depicted in the reaction schemes shown below.

The starting materials and reagents used in the preparation of these compounds are either available from commercial suppliers such as Aldrich Chemical Co., (Milwaukee, Wis.), Bachem (Torrance, Calif.), or Sigma (St. Louis, Mo.) or are prepared by methods known to those skilled in the art according to procedures provided in references such as Fieser and Fieser's Reagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5 and Supplementals (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991), March's Advanced Organic Chemistry, (John Wiley and Sons, 4th Edition) and Larock's Comprehensive Organic Transformations (VCH Publishers Inc., 1989). These schemes are merely illustrative of some of the ways in which the compounds of this description may be synthesized, and various modifications to these schemes may be made and will be suggested to one skilled in the art reading this description. The starting materials, intermediates, and final reaction products may be isolated and purified, if desired, using conventional techniques, including, but not limited to, filtration, distillation, crystallization, chromatography, and the like. Such materials may be characterized using conventional means, including physical constants and spectral data. Unless otherwise indicated, the reactions described herein are carried out at atmospheric pressure in the temperature range from about -78°C to about 150°C, such as from about 0°C to about 125°C, and also, for example, at about room (or ambient) temperature, such as about 20°C.

Compounds of formulae (IA) and (IB), wherein R ¹ , R ² , R ³ , Het are as defined in the Summary of the Invention section, R ⁴ is hydrogen or alkyl and R ⁵ is -COR ⁶ , wherein R ⁶ is as defined in the Summary of the Invention section, can be prepared as described in Scheme 1 below.

Scheme 1

Treatment of compound (I′), where R ⁴ and R ⁵ together with the nitrogen atom form a phthalimido ring, prepared as described in Scheme 2 below, with an aqueous hydrazine solution gives the amino compound of formula 1. The reaction can be carried out in alcoholic solvents such as methanol, ethanol or isopropyl alcohol and the like.

Coupling a compound of formula 1 with an acid of formula 2, wherein R ⁶ is as defined in the Summary of the Invention section, in the presence of a coupling agent such as HOBt, EDC and the like, and a non-nucleophilic base such as triethylamine, diisopropylamine, i-Pr ₂ NEt and the like, in an organic solvent such as dichloromethane, chloroform and the like, at room temperature, yields compound (I”).

Alternatively, compounds of formula (I) can be prepared by reacting a compound of formula 1 with an acetyl of formula R ⁶ CO(LG), where LG is a halogen, preferably chlorine, under acylation reaction conditions.

Compounds of formula 2 are commercially available or they can be prepared by methods well known in the art from readily available starting materials. For example, benzoic acid, o-, m- and p-nitrobenzoic acid, phenylacetic acid, trifluorobenzoic acid, picolinic acid, nicotinic acid, methoxybenzoic acids, fluorobenzoic acids, chlorobenzoic acids, hydroxybenzoic acids, 3-fluoro-5-methoxybenzoic acid, 3-aminobenzoic acid, acetic acid, propionic acids, cyclopropanecarboxylic acid, cyclopentanecarboxylic acid are commercially available. Compounds of formula 2 can also be prepared from commercially available compounds. For example, compounds of formula 2 wherein R ⁶ is aryl, aralkyl, heteroaryl, or heteroaralkyl substituted with hydroxy can be reacted with an optionally substituted aralkyl halide, an optionally substituted heterocycloalkyl halide, and an optionally substituted heterocycloalkylalkyl halide under alkylation reaction conditions to yield the corresponding compound of formula 2 wherein R ⁶ is substituted with an optionally substituted aralkyloxy, an optionally substituted heterocycloalkyloxy, or an optionally substituted heterocycloalkylalkyloxy, respectively. Similarly, compounds of formula 2 wherein R ⁶ is substituted with an amino group can be reacted with acid halides or sulfonyl halides under acylation and sulfonylation reaction conditions to yield the corresponding compounds of formula 2 wherein R ⁶ is substituted with acylamino or sulfonylamino groups, respectively.

Alternatively, the above transformations can be carried out on compounds of formula (I′′) or (I′′′), wherein R ⁶ is aryl, aralkyl, heteroaryl or heteroaralkyl substituted with hydroxy or amino.

Compounds of formulae (IA) and (IB), where the dotted line R ¹ , R ² , R ³ , Het are as defined in the Summary of the Invention section and R ⁴ and R ⁵ together with the nitrogen atom to which they are attached form a phthalimido ring, can be prepared as described in Scheme 2 below.

Scheme 2

Compounds of formula 4, wherein R ³ is as defined in the Summary of the Invention, can be prepared by reacting a compound of formula 3 with phthalic anhydride under basic conditions (see J. C. Sheehan, D. W. Chapman, and R. W. Roth, J. Am. Chem. Soc. , 74 , 3822 (1952)) or acidic conditions (see R. Mahboub, int. J. Chem. Sci, 7(1), 2009, 28-36) at elevated temperatures. If basic reaction conditions are used, the reaction is carried out in the presence of organic bases such as Et ₃ N, iPr ₂ Net, and the like, in an aromatic organic solvent such as benzene, toluene, and the like. Compounds of formula 3 are commercially available or readily prepared by methods known in the art.

Compound (I) can be prepared by adding a solution of a compound of formula 6 with an acid chloride of formula 5 under acylation reaction conditions. Suitable solvents include aromatic or halogenated organic solvents such as benzene, toluene or DCM in their anhydrous forms.

Compounds of formula 5 can be prepared by reacting a compound of formula 4 with a chlorinating agent such as _SOCl2 , (COCl) ₂ or _PCl5 under conditions well known in the art. Compounds of formula 6, such as ( R )-2-(piperidin-2-yl)pyridine and S-2-(piperidin-2-yl)pyridine, are commercially available or they can be prepared by methods well known in the art.

Compounds of formulae (IA) and (IB), where the dotted line, R ¹ , R ² , R ³ , Het are as defined in the Summary of the Invention section and R ⁴ and R ⁵ together with the nitrogen atom to which they are attached form pyrrolidinyl, piperidinyl or homopiperidinyl, can be prepared as described in Scheme 3 below.

Scheme 3

Protection of the amino group in a compound of formula 1 with a suitable amino protecting group such as Boc, benzyl or Ns, followed by treatment of the resulting compound of formula 7 with a compound of formula 8, wherein each X is a leaving group such as a halogen (such as chlorine or bromine), tosylate or mesylate, and m is 0-2, yields a compound of formula 9. The reaction is carried out in the presence of NaH in THF. Removal of the amino protecting group followed by cyclization of the resulting amine in the presence of a base such as potassium hydroxide, sodium hydroxide or the like, in a suitable organic solvent such as dichloromethane, yields a compound of formula (I).

It will be appreciated by one skilled in the art that by treatment as described above, but replacing compound 8 with other starting materials such as bis(2-chloroethyl)amine, oxybis(ethane-2,1-diyl)bis(4-methylbenzenesulfonate), (Z)-1,4-dichlorobut-2-ene, 2-(bromomethyl)benzoic acid, 2-(2-bromoethyl)benzoic acid, 2-(bromomethyl)-5-methoxybenzoic acid and 2-(bromomethyl)-4-fluorobenzoic acid, compounds of formula (I) can be synthesized wherein NR ⁴ R ⁵ form other heterocycloamino rings.

Introduction and pharmaceutical composition

In general, the compounds of the present description are administered in a therapeutically effective amount by any of the acceptable routes of administration for agents that serve similar purposes. Therapeutically effective amounts of the compounds of the description can range from about 0.01 to about 500 mg per kg of patient body weight per day, which can be administered in single or multiple doses. A suitable dosage level can be from about 0.1 to about 250 mg/kg per day; or from about 0.5 to about 100 mg/kg per day. A suitable dosage level can be from about 0.01 to about 250 mg/kg per day, from about 0.05 to about 100 mg/kg per day, or from about 0.1 to about 50 mg/kg per day. Within this range, the dosage can be from about 0.05 to about 0.5, from about 0.5 to about 5, or from about 5 to about 50 mg/kg per day. For oral administration, the compositions may be presented in the form of tablets containing from about 1.0 to about 1000 milligrams of the active ingredient, in particular about 1, 5, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900 and 1000 milligrams of the active ingredient. The actual amount of the compound of the present description, i.e. the active ingredient, will depend on a variety of factors such as the severity of the disease being treated, the age and relative health of the patient, the potency of the compound used, the route and form of administration and other factors.

In general, the compounds of the present disclosure will be administered as pharmaceutical compositions by any of the following routes: oral, systemic (e.g., transdermal, intranasal, or suppository), or parenteral (e.g., intramuscular, intravenous, or subcutaneous). The preferred route of administration is oral administration using a convenient daily dosage regimen that can be adjusted depending on the degree of damage. The compositions may be in the form of tablets, pills, capsules, semi-solids, powders, sustained-release formulations, solutions, suspensions, elixirs, aerosols, or any other suitable compositions.

The choice of formulation depends on various factors such as the route of administration of the drug (e.g., for oral administration, formulations in the form of tablets, pills or capsules are preferred, including enteric-coated or sustained-release tablets, pills or capsules) and the bioavailability of the drug. Recently, pharmaceutical formulations have been developed specifically for drugs that exhibit low bioavailability, based on the principle that bioavailability can be increased by increasing the surface area, i.e., decreasing the particle size. For example, U.S. Patent No. 4,107,288 describes a pharmaceutical formulation having particles ranging in size from 10 to 1000 nm, in which the active material is supported by a cross-linked matrix of macromolecules. U.S. Patent No. 5,145,684 describes the production of a pharmaceutical formulation in which a drug substance is atomized into nanoparticles (average particle size 400 nm) in the presence of a surface modifier and then dispersed in a liquid medium to produce a pharmaceutical formulation that exhibits significantly increased bioavailability.

The compositions typically comprise a compound of the present disclosure in combination with at least one pharmaceutically acceptable excipient. Acceptable excipients are non-toxic, facilitate administration, and do not adversely affect the therapeutic effect of the compound of the present disclosure. Such excipients may be any solid, liquid, semi-solid, or, in the case of an aerosol composition, gaseous excipients that are generally available to those skilled in the art. Solid pharmaceutical excipients include starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, dry skim milk, and the like. Liquid and semi-solid excipients can be selected from glycerol, propylene glycol, water, ethanol and various oils, including oils of petroleum, animal, vegetable or synthetic origin, for example, peanut oil, soybean oil, mineral oil, sesame oil, etc. Carriers, especially for injection solutions, include water, saline, aqueous dextrose and glycols.

Compressed gases can be used to disperse the compound of the present description in the form of an aerosol. Inert gases suitable for this purpose are nitrogen, carbon dioxide, etc.

Other suitable pharmaceutical excipients and their formulations are described in Remington's Pharmaceutical Sciences, edited by E. W. Martin (Mack Publishing Company, 20th ed., 2000).

The level of the compound in the formulation may vary within the full range usable by one skilled in the art. Typically, the formulation contains, in weight percent, from about 0.01-99.99% by weight of the compound of the present disclosure, based on the total weight of the formulation, with the balance being one or more suitable pharmaceutical excipients. For example, the compound is present at a level of about 1-80% by weight.

The compounds of the present invention may be used in combination with one or more other drugs in the treatment of diseases or conditions for which the compounds of the present invention or other drugs may be used. Such other drugs may be administered in a manner and in an amount normally used therefor, simultaneously or sequentially with the compound of the present invention. When a compound of the present disclosure is used simultaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such other drugs and a compound of the present disclosure is preferred. However, combination therapy may also include therapy in which the compound of the present invention and one or more other drugs are administered on different overlapping schedules. It is also contemplated that when used in combination with one or more other active ingredients, the compounds of the present invention and the other active ingredients may be used in lower doses than when each is used alone.

Examples

The following preparations of compounds of formula (I) are provided to enable those skilled in the art to more clearly understand and practice the present disclosure. They are not to be construed as limiting the scope of the description, but merely as illustrative and representative.

Examples of synthesis

Example 1

Synthesis of 2-((2S)-1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione (Compound 1)

Stage 1

A 1-neck round-bottom flask (1 L) equipped with a condenser and a Dean-Stark apparatus was charged with L-alanine (17.9 g, 0.2 mol), phthalic anhydride (29.6 g, 0.2 mol), toluene (300 mL), and Et ₃ N (2.6 mL), in that order. The resulting mixture was refluxed for 16 h until H ₂ O was formed. H ₂ O (300 mL) was added to the cooled solution, and the layers were separated. The organic layer was further washed with brine, dried, and concentrated. The crude product was crystallized from EtOAc/hexane (80 mL/200 mL) to give (S)-2-(1,3-dioxoisoindolin-2-yl)propanoic acid (34.5 g, 79%) as a white solid.

Stage 2

To a solution of (S)-2-(1,3-dioxoisoindolin-2-yl)propanoic acid (2.3 g, 10 mmol) in hexane/benzene (20 mL/20 mL) was added oxalyl chloride (1.7 mL, 20 mmol) dropwise, followed by DMF (0.01 mL). After 4 h, all volatiles were completely removed by rotary evaporation. The resulting (S)-2-(1,3-dioxoisoindolin-2-yl)propanoyl chloride was used in the next step as crude product without further purification.

Stage 3

To a solution of (S)-2-(1,3-dioxoisoindolin-2-yl)propanoyl chloride (~10 mmol) in anhydrous DCM (60 mL) was added (+/-)-anatabine (1.4 g, 9 mmol) and Et ₃ N (1.7 mL, 12 mmol). The resulting solution was stirred at rt for 20 h and quenched with H ₂ O (60 mL). The layers were separated and the aqueous layer was extracted with DCM (60 mL x 3). The combined organic layer was washed with 2N HCl, sat. NaHCO ₃ and brine, dried and concentrated. Flash chromatography on a normal phase column (hexane/EtOAc=50/50) gave the title compound (1.0 g, 31%) as a white foam. MS (ESI, pos. ion) m/z: 362.1 (M+1).

Example 2

Synthesis of 2-((S)-1-oxo-1-((S)-6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione (Compound 2)

To a solution of (S)-2-(1,3-dioxoisoindolin-2-yl)propanoyl chloride (4.5 g, 20 mmol) in anhydrous benzene (120 mL) was slowly added (+/-)-anatabine (3.0 g, 19 mmol). The resulting solution was stirred at rt for 20 h and quenched with sat. NaHCO ₃ (100 mL). The layers were separated and the benzene layer was washed with brine, dried and concentrated. Crude 2-((2S)-1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione (4.7 g, 68%) was obtained as a slightly yellow foam in a mixture of two diastereomers: (A) 2-((S)-1-oxo-1-((S)-6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione; (B) 2-((S)-1-oxo-1-((R)-6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione, which was separated by CombiFlash® using an HP C18 Aq GOLD column with MeCN/buffered _H2O as the eluting mobile phase. The assignment was based on hydrolysis experiments: A diastereomer gave S-anatabine and B diastereomer gave R-anatabine.

Example 3

Synthesis of 2-((S)-3-methyl-1-oxo-1-((S)-6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)butan-2-yl)isoindolin-1,3-dione (Compound 3) and 2-((S)-3-methyl-1-oxo-1-((R)-6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)butan-2-yl)isoindolin-1,3-dione (Compound 4)

Stage 1

A 1-necked round-bottomed flask (1 L) equipped with a condenser and a Dean-Stark apparatus was charged with L-valine (23.4 g, 0.2 mol), phthalic anhydride (29.6 g, 0.2 mol), toluene (300 mL), and Et ₃ N (2.6 mL), in that order. The resulting mixture was refluxed for 16 h until H ₂ O was formed. H ₂ O (300 mL) was added to the cooled solution and the layers were separated. The organic layer was then washed with brine, dried, and concentrated. The crude product was crystallized from EtOAc/hexane (80 mL/200 mL) to give (S)-2-(1,3-dioxoisoindolin-2-yl)-3-methylbutanoic acid (39.0 g, 80%) as a white solid.

Stage 2

To a solution of (S)-2-(1,3-dioxoisoindolin-2-yl)-3-methylbutanoic acid (5.0 g, 20 mmol) in hexane/benzene (40 mL/40 mL) was added oxalyl chloride (3.5 mL, 40 mmol) dropwise, followed by DMF (0.02 mL). After 4 h, all volatiles were completely removed by rotary evaporation. The resulting (S)-2-(1,3-dioxoisoindolin-2-yl)-3-methylbutanoyl chloride was used in the next step as crude product without further purification.

Stage 3

To a solution of (S)-2-(1,3-dioxoisoindolin-2-yl)-3-methylbutanoyl chloride (20 mmol) in anhydrous benzene (120 mL) was slowly added (+/-)-anatabine (3.0 g, 19 mmol). The resulting solution was stirred at rt for 20 h and quenched with sat. NaHCO ₃ (100 mL). The layers were separated and the benzene layer was washed with brine, dried and concentrated. The crude product was purified by CombiFlash® using an HP C18 Aq GOLD column with MeCN/buffered H ₂ O as the eluting mobile phase. (A): The first peak, 2-((S)-3-methyl-1-oxo-1-((S)-6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)butan-2-yl)isoindolin-1,3-dione (1.2 g, 15%) as a white foam. MS (ESI, poll. ion) m/z: 390.1 (M+1); (B): The second peak, 2-((S)-3-methyl-1-oxo-1-((R)-6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)butan-2-yl)isoindolin-1,3-dione (1.0 g, 12%) as a white foam. MS (ESI, poll. ion) m/z: 390.1 (M+1).

Example 4

Synthesis of a mixture of 2-((S)-1-oxo-1-((S)-6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)butan-2-yl)isoindolin-1,3-dione and 2-((R)-1-oxo-1-((R)-6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)butan-2-yl)isoindolin-1,3-dione (Compound 5) and a mixture of 2-((S)-1-oxo-1-((R)-6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)butan-2-yl)isoindolin-1,3-dione and 2-((R)-1-oxo-1-((S)-6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)butan-2-yl)isoindolin-1,3-dione (Compound 6)

Stage 1

A 1-neck round-bottomed flask (200 mL) equipped with a condenser was charged with DL-2-aminobutanoic acid (3.5 g, 34 mmol), phthalic anhydride (5.0 g, 33.8 mmol), and glacial acetic acid (75 mL) in that order. The resulting mixture was heated under reflux for 6 h. The solvent was completely removed in vacuo to give 2-(1,3-dioxoisoindolin-2-yl)butanoic acid (7.3 g, 92%) as a clear oil.

Stage 2

To a solution of 2-(1,3-dioxoisoindolin-2-yl)butanoic acid (7.0 g, 29.3 mmol) in hexane/benzene (100 mL/100 mL) was added oxalyl chloride (5.1 mL, 58.6 mmol) dropwise, followed by DMF (0.02 mL). After 4 h, all volatiles were completely removed by rotary evaporation. The resulting 2-(1,3-dioxoisoindolin-2-yl)butanoyl chloride was used in the next step as crude product without further purification.

Stage 3

To a solution of 2-(1,3-dioxoisoindolin-2-yl)butanoyl chloride (29 mmol) in anhydrous benzene (120 mL) was slowly added (+/-)-anatabine (4.2 g, 27 mmol). The resulting solution was stirred at rt for 20 h and quenched with sat. NaHCO ₃ (100 mL). The layers were separated and the benzene layer was washed with brine, dried and concentrated to give 2-(1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)butan-2-yl)isoindolin-1,3-dione as a crude product (5.5 g, 54%). Racemic diastereomers were separated on a CombiFlash® using an HP C18 Aq GOLD column with MeCN/buffered _H2O as the eluting mobile phase. (A): The first peak, 2-((S)-1-oxo-1-((S)-6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)butan-2-yl)isoindolin-1,3-dione and 2-((R)-1-oxo-1-((R)-6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)butan-2-yl)isoindolin-1,3-dione as a white foam. MS (ESI, poles ion) m/z: 376.1 (M+1); (B): Second peak, 2-((S)-1-oxo-1-((R)-6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)butan-2-yl)isoindolin-1,3-dione and 2-((R)-1-oxo-1-((S)-6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)butan-2-yl)isoindolin-1,3-dione as a white foam. MS (ESI, pos. ion) m/z: 376.1 (M+1).

Example 5

Synthesis of 2-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione (Compound 7)

Stage 1

A 1-necked round-bottomed flask (200 mL) equipped with a condenser was charged with DL-phenylalanine (5.6 g, 34 mmol), phthalic anhydride (5.0 g, 33.8 mmol), and glacial acetic acid (100 mL) in that order. The resulting mixture was heated under reflux for 6 h. The solvent was completely removed in vacuo, and the residue was precipitated from a mixture of ethyl acetate and hexane to give 2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoic acid (7.0 g, 70%) as a white solid.

Stage 2

To a solution of 2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoic acid (5.0 g, 16.9 mmol) in hexane/benzene (100 mL/100 mL) was added oxalyl chloride (3.5 mL, 40.2 mmol) dropwise, followed by DMF (0.02 mL). After 4 h, all volatiles were completely removed by rotary evaporation. The resulting 2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoyl chloride was used in the next step as crude product without further purification.

Stage 3

To a solution of 2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoyl chloride (16.9 mmol) in anhydrous benzene (100 mL) was slowly added (+/-)-anatabine (2.4 g, 15.7 mmol). The resulting solution was stirred at rt for 20 h and quenched with sat. NaHCO ₃ (100 mL). The layers were separated and the benzene layer was washed with brine, dried and concentrated to give 2-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione (4.5 g, 65%) as a white foam as a mixture of 2 diastereomers after CombiFlash® purification using an HP C18 Aq GOLD column with MeCN/buffered _H2O as the eluting mobile phase. MS (ESI, poles ion) m/z: 438.1 (M+1).

Example 6

Synthesis of 2-(2-oxo-2-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)ethyl)isoindolin-1,3-dione (Compound 8)

Stage 1

A 1-necked round-bottomed flask (200 mL) equipped with a condenser was charged with glycine (5.1 g, 68 mmol), phthalic anhydride (10.0 g, 67.5 mmol), and glacial acetic acid (120 mL) in that order. The resulting mixture was heated under reflux for 6 h. The solvent was completely removed in vacuo, and the residue was precipitated from a mixture of ethyl acetate and hexane to give 2-(1,3-dioxoisoindolin-2-yl)acetic acid (10.8 g, 80%) as a white solid.

Stage 2

To a solution of 2-(1,3-dioxoisoindolin-2-yl)acetic acid (4.1 g, 20 mmol) in hexane/benzene (40 mL/40 mL) was added oxalyl chloride (3.5 mL, 40 mmol) dropwise, followed by DMF (0.02 mL). After 4 h, all volatiles were completely removed by rotary evaporation. The resulting 2-(1,3-dioxoisoindolin-2-yl)acetyl chloride was used in the next step as crude product without further purification.

Stage 3

To a solution of 2-(1,3-dioxoisoindolin-2-yl)acetyl chloride (20 mmol) in anhydrous benzene (120 mL) was slowly added (+/-)-anatabine (2.9 g, 19.0 mmol). The resulting solution was stirred at rt for 20 h and quenched with sat. NaHCO ₃ (100 mL). The layers were separated and the benzene layer was washed with brine, dried and concentrated to give 2-(2-oxo-2-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)ethyl)isoindolin-1,3-dione (3.8 g, 58%) as a white foam after CombiFlash® purification using an HP C18 Aq GOLD column with MeCN/buffered H ₂ O as the eluting mobile phase. MS (ESI, pos. ion) m/z: 348.1 (M+1).

Example 7

Synthesis of 2-((2R)-1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione (Compound 17)

Stage 1

A 1-necked round-bottomed flask (200 mL) equipped with a condenser was charged with D-alanine (5.0 g, 56.2 mmol), phthalic anhydride (18.2 g, 56.2 mmol), and glacial acetic acid (100 mL) in that order. The resulting mixture was heated under reflux for 3 h. The solvent was completely removed in vacuo, and the residue was precipitated from a mixture of ethyl acetate and hexane to give (R)-2-(1,3-dioxoisoindolin-2-yl)propanoic acid (8.0 g, 64%) as a white solid.

Stage 2

To a solution of (R)-2-(1,3-dioxoisoindolin-2-yl)propanoic acid (7.5 g, 33.3 mmol) in anhydrous DCM (120 mL) was added PCl ₅ (7.6 g, 36.5 mmol) in portions. The resulting reaction mixture was stirred for 3 h at rt. All volatiles were completely removed by rotary evaporation. The residue (R)-2-(1,3-dioxoisoindolin-2-yl)propanoyl chloride as a clear oil was used in the next step without further purification.

Stage 3

To a solution of (R)-2-(1,3-dioxoisoindolin-2-yl)propanoyl chloride (33 mmol) in anhydrous benzene (120 mL) was slowly added (+/-)-anatabine (5.3 g, 34.7 mmol). The resulting solution was stirred at rt for 20 h and quenched with sat. NaHCO ₃ (100 mL). The layers were separated and the benzene layer was washed with brine, dried and concentrated to give 2-((2R)-1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione (9.7 g, 81%) as a foam in a mixture of two diastereomers. MS (ESI, pos. ion) m/z: 362.1 (M+1).

Example 8

Synthesis of 3-nitro-N-((2S)-(1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (Compound 9)

Stage 1

To a solution of (S)-2-(1,3-dioxoisoindolin-2-yl)propanoic acid (11.2 g, 50.0 mmol) in anhydrous DCM (150 mL) was added PCl ₅ (11.4 g, 55 mmol) in portions. The resulting reaction mixture was stirred for 5 h at rt. All volatiles were completely removed by rotary evaporation. The residue (S)-2-(1,3-dioxoisoindolin-2-yl)propanoyl chloride as a clear oil was used in the next step without further purification.

Stage 2

To a solution of (S)-2-(1,3-dioxoisoindolin-2-yl)propanoyl chloride (~50 mmol) in anhydrous benzene (120 mL) was slowly added (+/-)-anatabine (7.6 g, 50 mmol). The resulting solution was stirred at rt for 20 h and quenched with sat. NaHCO ₃ (100 mL). The layers were separated and the benzene layer was washed with brine, dried and concentrated to give (S)-2-(1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione (13.5 g, 74%) as a crude product in a mixture of two diastereomers. MS (ESI, pos. ion) m/z: 362.4 (M+1).

Stage 3

To a solution of (S)-2-(1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione (13.5 g, 37 mmol) in EtOH (200 mL) was added hydrazine monohydrate (3.6 mL). The reaction solution was refluxed for 2 h and the resulting suspension was filtered at rt. The filtrate was concentrated to give (S)-2-amino-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (9.0 g, 77% from step 2) as a crude product in a mixture of two diastereomers.

Stage 4

To a solution of 3-nitrobenzoic acid (0.50 g, 3.0 mmol) and (S)-2-amino-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (0.7 g, 3.0 mmol) were added EDC hydrochloride (0.72 g, 3.8 mmol) and i- Pr ₂ NEt (0.70 mL, 3.9 mmol) in DCM (10 mL). The reaction mixture was stirred at rt overnight and quenched with H ₂ O (10 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give 3-nitro-N-(1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.2 g, 17%) as a film in the flask existing as a mixture of two diastereomers. MS (ESI, poles ion) m/z: 381.3 (M+1).

The compounds in Examples 9-13 were prepared according to the procedure described in Example 8, starting from the corresponding benzoic acids.

Example 9: 3-methoxy-N-((2S)-1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.5 g, 46%). (Compound 10)

MS (ESI, pos. ion) m/z: 366.1 (M+1).

Example 10: 3,4-diethoxy-N-((2S)-1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.6 g, 47%). (Compound 11)

MS (ESI, pos. ion) m/z: 424.2 (M+1).

Example 11: 3,4-dimethoxy-N-((2S)-1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.7 g, 59%). (Compound 12)

MS (ESI, pos. ion) m/z: 396.1 (M+1).

Example 12: 4-nitro-N-((2S)-1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.1 g, 8%), which was crystallized from MeCN. (Compound 13)

MS (ESI, pos. ion) m/z: 381.1 (M+1).

Example 13. 4-chloro-N-((2S)-1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (150 mg, 13%), which was crystallized from MeCN. (Compound 14)

MS (ESI, pos. ion) m/z: 370.1 (M+1).

Example 14

Synthesis of 3-nitro-N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (Compound 15)

Stage 1

A 1-necked round-bottomed flask (200 mL) equipped with a condenser was charged with DL-phenylalanine (11.2 g, 68 mmol), phthalic anhydride (10.0 g, 67.6 mmol), and glacial acetic acid (200 mL) in that order. The resulting mixture was heated under reflux for 2 h. The solvent was completely removed in vacuo, and the residue was precipitated from a mixture of ethyl acetate and hexane to give 2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoic acid (18.1 g, 90%) as a white solid.

Stage 2

To a solution of 2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoic acid (19.8 g, 67.1 mmol) in anhydrous DCM (300 mL) was added PCl ₅ (15.3 g, 73.8 mmol) portionwise. The reaction mixture was stirred for 3 h at rt. All volatiles were completely removed by rotary evaporation to give 2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoyl chloride as a crude product used in the next step without further purification.

Stage 3

To a solution of 2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoyl chloride (67 mmol) in anhydrous benzene (300 mL) was slowly added (+/-)-anatabine (12.0 g, 75.0 mmol). The resulting solution was stirred at rt overnight and quenched with sat. NaHCO ₃ (300 mL). The layers were separated and the benzene layer was washed with brine, dried and concentrated to give 2-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione (25.0 g, 86%) as a foam as a mixture of 2 diastereomers without further purification.

Stage 4

To a solution of 2-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione (25.0 g) in EtOH (300 mL) was added hydrazine monohydrate (5.0 mL). The reaction solution was refluxed for 3 h and the resulting suspension was filtered at rt. The filtrate was dissolved in a mixture of EtOAc and hexane and kept overnight. The resulting solid was filtered off and the filtrate was concentrated to give 2-amino-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (16.7 g, 54%) as a crude product in a mixture of two diastereomers.

Stage 5

To a solution of 3-nitrobenzoic acid (0.5 g, 3.0 mmol) and 2-amino-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (0.9 g, 3.0 mmol) was added EDC hydrochloride (0.7 g, 3.8 mmol) and i- Pr ₂ NEt (0.7 mL, 3.9 mmol) in DCM (10 mL). The reaction mixture was stirred at rt overnight and quenched with H ₂ O (10 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give 3-nitro-N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.6 g, 43%) as a film in the flask existing as a mixture of two diastereomers after CombiFlash® purification using an HP C18 Aq GOLD column with MeCN/buffered _H2O as the eluting mobile phase. MS (ESI, poles ion) m/z: 457.1 (M+1).

Alternative stage 5

To a solution of 3-nitrobenzoic acid (0.8 g, 5 mmol) in DCM (20 mL) was added PCl ₅ (1.2 g). The reaction mixture was stirred at rt for 2 h, concentrated and diluted with anhydrous DCM, which was taken up in a solution of 2-amino-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (1.5 g, 5 mmol) and Et ₃ N (7.5 mmol). Workup and purification gave the title product as described in Step 5 above.

Example 15

Synthesis of 3-methoxy-N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (Compound 16)

To a solution of 3-methoxybenzoic acid (0.46 g, 3.0 mmol) and 2-amino-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (0.92 g, 3.0 mmol) was added EDC hydrochloride (0.72 g, 3.8 mmol) and i- Pr ₂ NEt (0.70 mL, 3.9 mmol) in DCM (10 mL). The reaction mixture was stirred at rt overnight and quenched with H ₂ O (10 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give 3-methoxy-N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.60 g, 45%) as a film in the flask, existing as a mixture of two diastereomers after CombiFlash® purification using a normal phase silica gel column with EtOAc and hexane as the eluting mobile phase. MS (ESI, poles ion) m/z: 442.2 (M+1).

Example 16

Synthesis of 3-chloro-N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (Compound 18)

Stage 1

To a purple solution of _KMnO4 (4.0 g, 25.0 mmol) and _Na2HPO4 (6.7 g, 25 mmol) in _H2O (100 mL) was added a solution _of 3-chlorobenzaldehyde (3.5 g, 25 mmol) in MeOH (100 mL) dropwise at rt. The reaction mixture was stirred for about 30 min until a brown suspension formed. The resulting suspension was filtered through a pad of Celite® and the filtrate was concentrated by rotary evaporation and diluted with _H2O (100 mL). The diluted filtrate was acidified with 1N HCl until pH 3-4 was reached. The resulting precipitate was filtered and dried in vacuo at 76 °C overnight to give 3-chlorobenzoic acid (3.5 g, 90%) as a white solid.

Stage 2

To a solution of 3-chlorobenzoic acid (0.47 g, 3.0 mmol) and 2-amino-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (0.92 g, 3.0 mmol) was added EDC hydrate (0.75 g, 3.8 mmol) and i- Pr ₂ NEt (0.72 mL, 3.9 mmol) in DCM (10 mL). The reaction mixture was stirred at rt overnight and quenched with H ₂ O (10 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give 3-chloro-N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.30 g, 20%) as a film in the flask existing as a mixture of two diastereomers after CombiFlash® purification using an HP C18 Aq GOLD column with MeCN/buffered _H2O as the eluting mobile phase. MS (ESI, poles ion) m/z: 446.1 (M+1).

Example 17

Synthesis of 3-fluoro-N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (Compound 19)

Stage 1

To a purple solution of _KMnO4 (4.0 g, 25.0 mmol) and _Na2HPO4 (6.7 g, 25 mmol) in _H2O (100 mL) was added a solution _of 3-fluorobenzaldehyde (3.1 g, 25 mmol) in MeOH (100 mL) dropwise at rt. The reaction mixture was stirred for about 30 min until a brown suspension formed. The resulting suspension was filtered through a pad of Celite® and the filtrate was concentrated by rotary evaporation and diluted with _H2O (100 mL). The diluted filtrate was acidified with 1N HCl until pH 3-4 was reached. The resulting precipitate was filtered and dried in vacuo at 76 °C overnight to give 3-fluorobenzoic acid (2.0 g, 57%) as a white solid.

Stage 2

To a solution of 3-fluorobenzoic acid (0.46 g, 3.0 mmol) and 2-amino-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (0.92 g, 3.0 mmol) was added EDC hydrochloride (0.72 g, 3.8 mmol) and i- Pr ₂ NEt (0.70 mL, 3.9 mmol) in DCM (10 mL). The reaction mixture was stirred at rt overnight and quenched with H ₂ O (10 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give 3-fluoro-N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.50 g, 39%) as a film in the flask existing as a mixture of two diastereomers after CombiFlash® purification using an HP C18 Aq GOLD column with MeCN/buffered _H2O as the eluting mobile phase. MS (ESI, poles ion) m/z: 430.1 (M+1).

Example 18

Synthesis of 3-ethoxy-N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (Compound 20)

Stage 1

To a solution of 3-hydroxybenzaldehyde (6.1 g, 50 mmol) in DMF (200 mL) were added ethyl iodide (9.4 g, 60 mmol) and K ₂ CO ₃ (10.3 g, 75 mmol), in that order. The resulting suspension was heated to 120 °C for 4 h and cooled to rt. The reaction was quenched with H ₂ O (200 mL) and extracted with ether (200 mL). The ether layers were washed with H ₂ O (200 mL x 3), dried, and concentrated to give 3-ethoxybenzaldehyde (6.2 g, 83%) as a clear oil.

Stage 2

To a purple solution of _KMnO4 (6.2 g, 39.0 mmol) and _Na2HPO4 (10.5 g, 39 mmol) in _H2O (150 mL) was added a solution _of 3-ethoxybenzaldehyde (5.8 g) in MeOH (150 mL) dropwise at rt. The reaction mixture was stirred for about 30 min until a brown suspension formed. The resulting suspension was filtered through a pad of Celite® and the filtrate was concentrated by rotary evaporation and diluted with _H2O (100 mL). The diluted filtrate was acidified with 1N HCl until pH 3-4 was reached. The resulting precipitate was filtered and dried in vacuo at 76 °C overnight to give 3-ethoxybenzoic acid (2.9 g, 50%) as a white solid.

Stage 3

To a solution of 3-ethoxybenzoic acid (0.55 g, 3.0 mmol) and 2-amino-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (0.92 g, 3.0 mmol) was added EDC hydrochloride (0.72 g, 3.8 mmol) and i- Pr ₂ NEt (0.70 mL, 3.9 mmol) in DCM (10 mL). The reaction mixture was stirred at rt overnight and quenched with H ₂ O (10 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give 3-ethoxy-N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.66 g, 48%) as a film in the flask, existing as a mixture of two diastereomers after CombiFlash® purification using an HP C18 Aq GOLD column with MeCN/buffered _H2O as the eluting mobile phase. MS (ESI, poles ion) m/z: 456.2 (M+1).

By processing according to the procedure similar to Example 18 above, the compounds of Examples 19-22 are synthesized using the corresponding obtained or commercial benzoic acids.

Example 19: 3,4-dimethoxy-N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.35 g, 25%). (Compound 21)

MS (ESI, pos. ion) m/z: 472.2 (M+1).

Example 21: N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)-3-(trifluoromethyl)benzamide (0.55 g, 38%). (Compound 23)

MS (ESI, pos. ion) m/z: 480.1 (M+1).

Example 22: 3-fluoro-5-methoxy-N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.57 g, 41%). (Compound 24)

MS (ESI, pos. ion) m/z: 460.2 (M+1).

Example 23

Synthesis of 3-methoxy-N-((2R)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (Compound 25)

Stage 1

A 1-necked round-bottomed flask (500 mL) equipped with a condenser was charged with D-phenylalanine (25.0 g, 151.3 mmol), phthalic anhydride (22.2 g, 151.3 mmol), and glacial acetic acid (300 mL) in that order. The resulting mixture was heated under reflux for 4 h. The solvent was completely removed in vacuo, and the residue was precipitated from a mixture of ethyl acetate and hexane to give (R)-2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoic acid (38.4 g, 86%) as a white solid.

Stage 2

To a solution of (R)-2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoic acid (37.9 g, 128.0 mmol) in anhydrous DCM (500 mL) was added PCl ₅ (29.4 g, 141.0 mmol) in portions. The reaction mixture was stirred for 3 h at rt. All volatiles were completely removed by rotary evaporation to give (R)-2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoyl chloride as a crude product used in the next step without further purification.

Stage 3

To a solution of (R)-2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoyl chloride (128 mmol) in anhydrous benzene (500 mL) was slowly added (+/-)-anatabine (20.6 g, 128.7 mmol). The resulting solution was stirred at rt overnight and quenched with sat. NaHCO ₃ (500 mL). The layers were separated and the benzene layer was washed with brine, dried and concentrated to give 2-((2R)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione (43.3 g, 79%) as a foam as a mixture of 2 diastereomers without further purification.

Stage 4

To a solution of 2-((2R)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione (43.3 g) in EtOH (500 mL) was added hydrazine monohydrate (8.7 mL). The reaction solution was refluxed for 3 h and the resulting suspension was filtered at rt. The filtrate was dissolved in a mixture of EtOAc and hexane and kept overnight. The resulting solid was filtered off and the filtrate was concentrated to give (2R)-2-amino-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (32.3 g, 82%) as a crude product in a mixture of two diastereomers.

Stage 5

To a solution of 3-methoxybenzoic acid (0.46 g, 3.0 mmol) and (2R)-2-amino-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (0.92 g, 3.0 mmol) were added EDC hydrochloride (0.72 g, 3.8 mmol) and i- Pr ₂ NEt (0.70 mL, 3.9 mmol) in DCM (10 mL). The reaction mixture was stirred at rt overnight and quenched with H ₂ O (10 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give 3-methoxy-N-((2R)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.73 g, 55%) as a film in the flask existing as a mixture of two diastereomers after CombiFlash® purification using an HP C18 Aq GOLD column with MeCN/buffered _H2O as the eluting mobile phase. MS (ESI, poles ion) m/z: 442.2 (M+1).

Example 24

Synthesis of 3-ethoxy-4-methoxy-N-((2R)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (Compound 26)

Stage 1

To a solution of 3-ethoxy-4-hydroxybenzaldehyde (8.6 g, 50 mmol) in DMF (200 mL) was added NaH (3.0 g, 75 mmol, 60 wt%) in portions at 0 °C, followed by MeI (3.35 mL, 60 mmol). The reaction mixture was stirred at rt overnight and quenched with water. Extracted with ether (100 mL x 3). The ether layer was washed, dried and concentrated to give 3-ethoxy-4-methoxybenzaldehyde (5.9 g, 66%) as crude product.

Stage 2

To a purple solution of _KMnO4 (5.1 g, 32.0 mmol) and _Na2HPO4 (8.4 g, 32 mmol) in _H2O (120 mL) was added a solution _of 3-ethoxy-4-methoxybenzaldehyde (5.6 g) in MeOH (120 mL) dropwise at rt. The reaction mixture was stirred for about 30 min until a brown suspension formed. The resulting suspension was filtered through a pad of Celite® and the filtrate was concentrated by rotary evaporation and diluted with _H2O (100 mL). The diluted filtrate was acidified with 1N HCl until pH 3-4 was reached. The resulting precipitate was filtered and dried in vacuo at 76 °C overnight to give 3-ethoxy-4-methoxybenzoic acid (4.0 g, 66%) as a white solid.

Stage 3

To a solution of 3-ethoxy-4-methoxybenzoic acid (0.59 g, 3.0 mmol) and (2R)-2-amino-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (0.92 g, 3.0 mmol) was added EDC hydrochloride (0.72 g, 3.8 mmol) and i- Pr ₂ NEt (0.70 mL, 3.9 mmol) in DCM (10 mL). The reaction mixture was stirred at rt overnight and quenched with H ₂ O (10 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give 3-ethoxy-4-methoxy-N-((2R)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as a film in the flask existing as a mixture of two diastereomers after CombiFlash® purification using an HP C18 Aq GOLD column with MeCN/buffered _H2O as the eluting mobile phase. MS (ESI, poles ion) m/z: 486.2 (M+1).

By a procedure similar to that described in Example 24 above, the compounds of Examples 25-30 are prepared by ether substitution with synthesized or commercially available benzoic acids.

Example 25: 2-chloro-5-methoxy-N-((2R)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.76 g, 53%). (Compound 27)

MS (ESI, pos. ion) m/z: 476.1 (M+1).

Example 26: 3,5-dimethoxy-N-((2R)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.86 g, 61%). (Compound 28)

MS (ESI, pos. ion) m/z: 472.2 (M+1).

Example 27: 5-methoxy-2-methyl-N-((2R)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.76 g, 56%). (Compound 29)

MS (ESI, pos. ion) m/z: 456.2 (M+1).

Example 28: 4-chloro-2-methoxy-N-((2R)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.56 g, 41%). (Compound 30)

MS (ESI, pos. ion) m/z: 476.1 (M+1).

Example 29: 4-fluoro-3-methoxy-N-((2R)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.65 g, 47%). (Compound 31)

MS (ESI, pos. ion) m/z: 460.2 (M+1).

Example 30: 2-methoxy-N-((2R)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide. (Compound 32)

MS (ESI, pos. ion) m/z: 442.3 (M+1).

Example 31

Synthesis of 3-ethoxy-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (Compound 33)

Stage 1

A 1-necked round-bottomed flask (500 mL) equipped with a condenser was charged with L-phenylalanine (25.0 g, 151.3 mmol), phthalic anhydride (22.2 g, 151.3 mmol), and glacial acetic acid (300 mL) in that order. The resulting mixture was heated under reflux for 4 h. The solvent was completely removed in vacuo, and the residue was precipitated from a mixture of ethyl acetate and hexane to give (S)-2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoic acid (42.8 g, 96%) as a white solid.

Stage 2

To a solution of (S)-2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoic acid (42.3 g, 142.0 mmol) in anhydrous DCM (550 mL) was added PCl ₅ (32.8 g, 157.0 mmol) portionwise. The reaction mixture was stirred for 3 h at rt. All volatiles were completely removed by rotary evaporation to give (S)-2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoyl chloride as a crude product used in the next step without further purification.

Stage 3

To a solution of (S)-2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoyl chloride (142 mmol) in anhydrous benzene (550 mL) was slowly added (+/-)-anatabine (23.0 g, 142.9 mmol). The resulting solution was stirred at rt overnight and quenched with sat. NaHCO ₃ (550 mL). The layers were separated and the benzene layer was washed with brine, dried and concentrated to give 2-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione (57.7 g, 100%) as a foam as a mixture of 2 diastereomers without further purification.

Stage 4

To a solution of 2-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)isoindolin-1,3-dione (57.7 g) in EtOH (500 mL) was added hydrazine monohydrate (11.6 mL). The reaction solution was refluxed overnight and the resulting suspension was filtered at rt. The filtrate was dissolved in a mixture of EtOAc and hexane and kept overnight. The resulting solid was filtered off and the filtrate was concentrated to give (2S)-2-amino-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (39.0 g, 99%) as a crude product in a mixture of two diastereomers.

Stage 5

To a solution of 3-ethoxybenzoic acid (0.83 g, 5.0 mmol) and (2S)-2-amino-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (1.5 g, 5.0 mmol) was added EDC hydrochloride (1.2 g, 6.3 mmol) and i- Pr ₂ NEt (1.1 mL, 6.5 mmol) in DCM (30 mL). The reaction mixture was stirred at rt overnight and quenched with H ₂ O (30 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give 3-ethoxy-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (1.3 g, 63%) as a film in the flask existing as a mixture of two diastereomers after CombiFlash® purification using an HP C18 Aq GOLD column with MeCN/buffered _H2O as the eluting mobile phase. MS (ESI, poles ion) m/z: 456.2 (M+1).

By a procedure similar to that described in Example 31 above, the compounds of Examples 32-33 are prepared by substitution of the ether with either synthesized or commercially available benzoic acids.

Example 32: 4-fluoro-3-methoxy-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.4 g, 29%). (Compound 34)

MS (ESI, pos. ion) m/z: 460.2 (M+1).

Example 33: 3-methoxy-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (0.8 g, 36%). (Compound 35)

MS (ESI, pos. ion) m/z: 442.2 (M+1).

By a procedure similar to that described in Example 31 above, the compounds of Examples 40-56 are prepared by substitution of the ether with commercially available carboxylic acids or with carboxylic acids that can be readily prepared from aminobenzoic acids and the corresponding acid chlorides or acids using coupling reactions.

Example 40: 3-(3-methylbutanamido)-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as a tan solid (0.90 g, 60%). (Compound 42)

MS (ESI, pos. ion) m/z: 511.2 (M+1).

Example 41: N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)-3-pivalamidobenzamide as a gray solid (0.93 g, 62%). (Compound 43)

MS (ESI, pos. ion) m/z: 511.2 (M+1).

Example 42: 3-(cyclopropanecarboxamido)-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as a white solid (1.01 g, 69%). (Compound 44)

MS (ESI, pos. ion) m/z: 495.2 (M+1).

Example 43: 3-(cyclobutanecarboxamido)-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as an orange solid (1.05 g, 69%). (Compound 45)

MS (ESI, pos. ion) m/z: 509.2 (M+1).

Example 44: 3-(cyclopentanecarboxamido)-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as a white solid (0.69 g, 64%). (Compound 46)

MS (ESI, pos. ion) m/z: 523.2 (M+1).

Example 45: 2-butyramido-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as a white solid (0.12 g, 12%). (Compound 47)

MS (ESI, pos. ion) m/z: 497.2 (M+1).

Example 46: 4-butyramido-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as a white solid (0.45 g, 45%). (Compound 48)

MS (ESI, pos. ion) m/z: 497.2 (M+1).

Example 47: N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)quinoline-2-carboxamide as a white solid (0.50 g, 53%). (Compound 49)

MS (ESI, pos. ion) m/z: 463.2 (M+1).

Example 48: N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)pyrazine-2-carboxamide as a yellow solid (0.50 g, 61%). (Compound 50)

MS (ESI, pos. ion) m/z: 414.1 (M+1).

Example 49: N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)nicotinamide as a white solid (0.41 g, 50%). (Compound 51)

MS (ESI, pos. ion) m/z: 413.1 (M+1).

Example 50: N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)furan-2-carboxamide as a white solid (0.50 g, 63%). (Compound 52)

MS (ESI, pos. ion) m/z: 402.1 (M+1).

Example 51: N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)-1H-pyrrole-2-carboxamide as a light yellow solid (0.20 g, 25%). (Compound 56)

MS (ESI, pos. ion) m/z: 401.1 (M+1).

Example 52: 3-((S)-2-methylbutanamido)-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as a light yellow solid (0.60 g, 56%). (Compound 57)

MS (ESI, pos. ion) m/z: 511.2 (M+1).

Example 53: 3-((R)-2-methylbutanamido)-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as a light yellow solid (0.67 g, 65%). (Compound 58)

MS (ESI, pos. ion) m/z: 511.2 (M+1).

Example 54: 3-(N-methylbutyramido)-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as a light yellow solid (0.80 g, 78%). (Compound 59)

MS (ESI, pos. ion) m/z: 511.2 (M+1).

Example 55: 3-(butylamino)-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as a light yellow solid (0.82 g, 85%). (Compound 60)

MS (ESI, pos. ion) m/z: 483.2 (M+1).

Example 56: 1-butyl-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)-1H-pyrrole-2-carboxamide as a yellow oil (0.21 g, 23%). (Compound 61)

MS (ESI, pos. ion) m/z: 457.2 (M+1).

Example 34

Synthesis of N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)-3-pentanamidobenzamide (Compound 36)

Stage 1

To a suspension of 3-aminobenzoic acid (5.0 g, 36.5 mmol) in DCM (120 mL) were added pyridine (6.5 mL) and valeroyl chloride (5.5 mL, 45.6 mmol) in that order at 0 °C. The resulting solution was stirred at rt overnight and quenched with water. The layers were separated and the organic layer was washed, dried and concentrated to give 3-pentanamidobenzoic acid (7.2 g, 90%) as a white solid.

Stage 2

To a solution of 3-pentanamidobenzoic acid (0.66 g, 3.0 mmol) and 2-amino-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (0.92 g, 3.0 mmol) was added EDC hydrochloride (0.72 g, 3.8 mmol) and i- Pr ₂ NEt (0.70 mL, 3.9 mmol) in DCM (10 mL). The reaction mixture was stirred at rt overnight and quenched with H ₂ O (10 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)-3-pentanamidobenzamide (0.81 g, 59%) as a film in the flask existing as a mixture of two diastereomers after CombiFlash® purification using an HP C18 Aq GOLD column with MeCN/buffered _H2O as the eluting mobile phase. MS (ESI, poles ion) m/z: 511.2 (M+1).

By a procedure similar to that described in Example 34 above, the compounds of Examples 35-37 are prepared by substituting the ether with synthesized or commercially available benzoic acids.

Example 35: 3-butyramido-N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide. (Compound 37)

MS (ESI, pos. ion) m/z: 497.2 (M+1).

Example 36: 3-amino-N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide. (Compound 38)

MS (ESI, pos. ion) m/z: 427.2 (M+1).

Example 37: 3-acetamido-N-(1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (169 mg, 12%). (Compound 39)

MS (ESI, pos. ion) m/z: 469.2 (M+1).

Example 38

Synthesis of 3-butyramido- N -(1-oxo-3-phenyl-1-(2-(quinolin-3-yl)-3,6-dihydropyridin-1(2 H )-yl)propan-2-yl)benzamide (Compound 40)

Scheme of synthesis of 3-butyramido-N-(1-oxo-3-phenyl-1-(2-(quinolin-3-yl)-3,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (1,2-combo compound):

Stage 1

Synthesis of N -phenylacetamide .

To a suspension of alanine (5 g, 53.7 mmol) in 2N NaOH (40.3 mL) was added acetyl chloride (4.63 g, 59.07 mmol, 1.1 equiv.) in 40.3 mL dichloromethane dropwise over 1 h in an ice bath. The resulting mixture was stirred for an additional hour at room temperature. The organic layer was washed with diluted ice, NaHCO ₃ and brine. The organic layer was concentrated under reduced pressure to afford the desired product as crude acetanilide product. The crude acetanilide was recrystallized by dissolution with distilled water to afford pure and dry acetanilide as a white crystalline solid (6.3 g, 87% isolated yield).

Stage 2

Synthesis of 2-chloroquinoline-3-carbaldehyde

To DMF (10.82 g, 148 mmol, 2.5 equiv) was added phosphorus oxychloride (63.5 g, 414.13 mmol, 7 equiv) dropwise and cooled in an ice bath (30 min). The mixture was stirred for an additional 30 min at 0 ^o C, then acetanilide (8 g, 59.18 mmol, 1 equiv) was added in one portion. The white suspension turned into a greenish solution, which was heated overnight. The mixture was poured onto ice. The yellow precipitate was filtered and dried to give a yellow-orange solid (6.8 g, 60% isolated yield).

Stage 3

Synthesis of quinoline-3-carbaldehyde

To 2-chloroquinoline-3-carbaldehyde (3 g, 15.69 mmol, 1 equiv) in DMF (18 mL) at 23 °C under nitrogen atmosphere were added triethylamine (19.05 g, 188.28 mmol, 12 equiv), tetrakis(triphenylphosphine)palladium(0) (0.9 g, 0.78 mmol, 5% mol) and formic acid (3.9 g, 84.72 mmol, 5.4 equiv). After stirring for 3 h at 110 °C, the reaction mixture was cooled to 23 °C and treated with water (90 mL) and ethyl acetate (3×60 mL). The combined organic phase was washed with brine (50 mL), dried over anhydrous Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo to give the crude product, which was purified by flash chromatography using 50% EtOAc/hexane to give the desired product as a reddish-brown solid (1.5 g, 60.8% isolated yield).

Stage 4

Synthesis of ( E )- N -allyl-1-(quinolin-3-yl)methanamine

In a 250 mL round-bottomed flask, allylamine (1.3 g, 22.92 mmol, 1.2 equiv) was added to a stirred solution of quinoline-3-carbaldehyde (3 g, 19.1 mmol, 1 equiv) in anhydrous dichloromethane (96 mL) containing activated 4°A MS (5 g). The resulting mixture was then stirred at room temperature for 24 h. Subsequently, the molecular sieves were filtered, washed with DCM, and the solvent was evaporated to give the imine product as a brown solid (4.3 g, 95% isolated yield).

Stage 5

Synthesis of ( N -allyl-1-(quinolin-3-yl)but-3-ene-1-amine

In a 250 mL 2-neck flask, allyl bromide (3.95 g, 32.63 mmol, 2 equiv) was added dropwise to a cold (0 °C) anhydrous ethanol (83 mL) solution of ( E ) -N- allyl-1-(quinolin-3-yl)methanamine (3.2 g, 16.31 mmol, 1 equiv) and indium powder (2.79 g, 24.46 mmol, 1.5 equiv). The resulting mixture was stirred at the same temperature for 30 min. Then the temperature was maintained until it reached room temperature, and the system was stirred for additional 3.5 h. Then the solvent was evaporated in vacuo, the residue was dissolved in EtOAc (60 mL) and treated with sat. aq. NaHCO ₃ (40 mL). The two layers were separated and the aqueous layer was extracted with EtOAc (3×20 mL). The combined organic layers were dried using _{anhydrous Na2SO4} , filtered and the solvent was evaporated under reduced pressure. The resulting crude product was purified by column chromatography using 50% EtOAc/hexane as eluent to afford the pure product as a tan oil (3.4 g, 88% isolated yield).

Stage 6

Synthesis of 3-(1,2,3,6-tetrahydropyridin-2-yl)quinoline .

In a 100 mL 2-necked pear-shaped flask equipped with a magnetic stirrer, N -allyl-1-(quinolin-3-yl)but-3-ene-1-amine (1.56 g, 6.55 mmol, 1 equiv) was dissolved in anhydrous dichloromethane (64 mL). p-Toluenesulfonic acid monohydrate (2.74 g, 14.41 mmol, 2.2 equiv) was added and the mixture was stirred for 10 min. Grubbs was added until the pH of the solution was 9-10. The organic layer was separated and dried using _{anhydrous Na2SO4} , filtered, and the solvent was evaporated under reduced pressure. The obtained crude product was purified, second generation catalyst (0.555 g, 0.65 mmol, 10% mol) was added and the resulting mixture was stirred at room temperature for 18 h. The reaction mixture was neutralized with 50 mL 2N NaOH and stirred for 20 min. The organic layer was collected and washed with water (3 x 50 mL). The organic layer was dried with anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The residue was dissolved in 100 mL EtOAc and then 30 mL 6N HCl, 70 mL deionized water, 20 mL brine were added and the mixture was stirred for 20 min. The aqueous layer was collected and the aqueous layer was then extracted (3 x 50 mL) with EtOAc. The aqueous layer was transferred to a 500 mL round bottom flask and cooled at 0 °C. 120 ml of methyl tert-butyl ether was added to the aqueous layer and anhydrous potassium carbonate was subjected to column chromatography packed with a mixture of hexane and ammonium hydroxide in a ratio of 20:1 and using 4% methanol/chloroform as eluent to give the pure product as a reddish-brown oil (0.325 g, 28% isolated yield).

Stage 7

Synthesis of 2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoyl chloride

Stage 7(i)

An oven-dried round-bottomed flask (500 mL) equipped with a condenser and a Dean-Stark apparatus was charged with DL-phenylalanine (20 g, 121 mmol, 1 equiv), phthalic anhydride (17.9 g, 121 mmol, 1 equiv), toluene (180 mL), and Et ₃ N (1.6 mL), in that order. The resulting mixture was refluxed for 16 h until water formation ceased. H ₂ O (180 mL) was added to the cooled solution and the product precipitated. The product was filtered and dried under high vacuum overnight. The dried product 2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoic acid was collected as a white solid (29.5 g, 82.6% isolated yield). ¹ H NMR (CDCl ₃ ): 7.80-7.78 (m, 2H), 7.70-7.69 (m, 2H), 7.27-7.13 (m, 5H), 5.23 (t, J=8.2 Hz, 1H), 3.61 (d, J=7.5 Hz, 2H); ¹³ C NMR (CDCl ₃ ): 174.1, 167.3, 136.4, 134.1, 131.4, 128.8, 128.5, 126.9, 123.5, 53.0, 34.4.

Stage 7(ii)

To a solution of 2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoic acid (2.5 g, 8.47 mmol, 1 equiv) in anhydrous DCM (32 mL) was added PCl ₅ (1.93 g, 9.28 mmol, 1.09 equiv) portionwise. The resulting reaction mixture was stirred for 3 h at rt. All volatiles were completely removed under reduced pressure. The residual 2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoyl chloride (2.12 g) as a white solid was used in the next step without further purification.

Stage 8

Synthesis of 2-(1-oxo-3-phenyl-1-(2-(quinolin-3-yl)-3,6-dihydropyridin-1( 2H )-yl)propan-2-yl)isoindolin-1,3-dione

To a solution of 2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoyl chloride (6.78 mmol) in anhydrous benzene (25 mL) was slowly added 3-(1,2,3,6-tetrahydropyridin-2-yl)quinoline (1.5 g, 7.13 mmol, 1.05 equiv). The resulting solution was stirred at room temperature for 20 h and quenched with sat. NaHCO ₃ (20 mL). The layers were separated and the benzene layer was washed with brine, dried and concentrated to give 3-(1,2,3,6-tetrahydropyridin-2-yl)quinoline as a crude product. The crude product was purified by column chromatography using 50% EtOAc/hexane as eluent to give the desired product as a white solid (1.4 g, 42.3%) in a mixture of diastereomers. MS (ESI, poles ion) m/z: 488.2 (M+1).

Stage 9

Synthesis of 2-amino-3-phenyl-1-(2-(quinolin-3-yl)-3,6-dihydropyridin-1(2 H )-yl)propan-1-one

To a solution of 2-(1-oxo-3-phenyl-1-(2-(quinolin-3-yl)-3,6-dihydropyridin-1( 2H )-yl)propan-2-yl)isoindolin-1,3-dione (1.4 g, 2.9 mmol) in EtOH (20 mL) was added hydrazine monohydrate (0.45 mL). The reaction solution was refluxed overnight at 85 ^o C, and the resulting suspension was filtered at room temperature. The filtrate was concentrated and then dissolved with 20 mL of 1:1 EtOAc/hexane and stored at 0 ^o C overnight. The mixture was filtered and the filtrate was concentrated to give 2-amino-3-phenyl-1-(2-(quinolin-3-yl)-3,6-dihydropyridin-1(2 H )-yl)propan-1-one (0.8 g, 77%) as a brown oil in a mixture of diastereomers. MS (ESI, poles ion) m/z: 358.19 (M+1).

Stage 10

Synthesis of 2-amino-3-phenyl-1-(2-(quinolin-3-yl)-3,6-dihydropyridin-1(2 H )-yl)propan-1-one

To a solution of 3-butyramidobenzoic acid (0.47 g, 2.15 mmol, 1 equiv) and 2-amino-3-phenyl-1-(2-(quinolin-3-yl)-3,6-dihydropyridin-1(2 H )-yl)propan-1-one (0.47 g, 2.15 mmol, 1 equiv) were added EDC hydrochloride (0.56 g, 2.8 mmol, 1.3 equiv) and i-Pr ₂ NEt (0.5 mL, 2.80 mmol, 1.3 equiv) in DCM (10 mL). The reaction mixture was stirred at room temperature overnight and quenched with H ₂ O (10 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give the crude product and the crude product was crystallized using acetonitrile as solvent. The crystals were filtered and collected as a white solid (0.45 g, 38.3%) existing as a mixture of diastereomers. MS (ESI, poles ion) m/z: 547.5 (M+1).

By processing according to a procedure similar to that described in Example 38 above, the compounds of Examples 65-66 are obtained starting from the corresponding amino acids.

Example 65: 3-butyramido-N-(1-(2-(2-chloroquinolin-3-yl)-3,6-dihydropyridin-1(2H)-yl)-1-oxo-3-phenylpropan-2-yl)benzamide as a pale yellow solid (0.05 g, 17.2%). (Compound 67)

MS (ESI, pos. ion) m/z: 581.3(M+1).

Example 66: 3-butyramido-N-(1-oxo-1-(2-(quinolin-3-yl)-3,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as a creamy white solid (0.13 g, 38.6%). (Compound 68)

MS (ESI, pos. ion) m/z: 471.4 (M+1).

Example 39

Synthesis of 3-{5-[(3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentanamido}-N-[(2S)-1-oxo-3-phenyl-1-{1,2,3,6-tetrahydro-[2,3'-bipyridin]-1-yl}propan-2-yl]benzamide (Compound 41)

Stage 1

To a solution of 3-nitrobenzoic acid (8.4 g, 50 mmol) in MeOH (150 mL) was slowly added SOCl ² (7.1 mL, 100 mmol) at rt. The reaction mixture was refluxed for 2 h. Removal of all solvents gave methyl 3-nitrobenzoate (9.1 g, 100%) as a crude product.

Stage 2

To a solution of methyl 3-nitrobenzoate (9.1 g, 50 mmol) in EtOH (150 mL) was added _{SnCl2 2H2O} (22.5 g, 100 mmol) in one portion. The reaction mixture was heated under reflux for 2 h and then the solvent was removed. The residue was extracted with EtOAc. The organic layer was washed, dried and concentrated to give methyl 3-aminobenzoate (8.4 g, 100%) as a brown oil.

Stage 3

To a solution of methyl 3-aminobenzoate (0.87 g, 5.7 mmol) and biotin (1.4 g, 5.7 mmol) in DCM (15 mL) were added EDC (1.3 g, 7.0 mmol) and i-Pr2NEt (1.2 mL, 7.5 mmol). The reaction mixture was stirred at rt overnight and quenched with _H2O (10 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give methyl 3-(5-(2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)benzoate (1.13 g, 52%).

Stage 4

To a solution of methyl 3-(5-(2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)benzoate (1.13 g, 2.9 mmol) in a mixture of THF (15 mL) and H ₂ O (15 mL) was added LiOH (0.36 g, 14 mmol) in one portion and stirred at rt overnight. The reaction mixture was further diluted with 1N NaOH and the aqueous layer was washed with EtOAc, neutralized with 1N HCl. The resulting solid was collected to give 3-(5-(2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)benzoic acid (0.7 g, 83%) as a crude product.

Stage 5

To a solution of 3-(5-(2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)benzoic acid (0.7 g, 1.9 mmol) and (2S)-2-amino-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (0.6 g, 1.9 mmol) was added EDC hydrochloride (0.5 g, 2.4 mmol) and i-Pr2NEt (0.4 mL, 2.5 mmol) in DCM (20 mL). The reaction mixture was stirred at rt overnight and quenched with _H2O (30 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give 3-{5-[(3aS,4S,6aR)-2-oxo-hexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentanamido}-N-[(2S)-1-oxo-3-phenyl-1-{1,2,3,6-tetrahydro-[2,3'-bipyridin]-1-yl}propan-2-yl]benzamide (80 mg, 7%) as a powder after CombiFlash® purification using an HP C18 Aq GOLD column with MeCN/buffered _H2O as the eluting mobile phase. MS (ESI, poles ion) m/z: 653.5 (M+1).

Example 57

Synthesis of 3-(3-aminopropanamido)-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide hydrochloride (Compound 54)

Scheme of synthesis of 3-(3-aminopropanamido)-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide hydrochloride:

To a solution of 3-(3-(( tert -butoxycarbonyl)amino)propanamido)benzoic acid (0.62 g, 2.0 mmol) and (2S)-2-amino-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-1-one (0.62 g, 2.0 mmol) was added EDC hydrochloride (0.57 g, 2.9 mmol) and i-Pr ₂ NEt (0.47 mL, 3.9 mmol) in DCM (10 mL). The reaction mixture was stirred at rt overnight and quenched with H ₂ O (10 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give 3-(3-aminopropanamido)-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as a film in the flask, existing as a mixture of two diastereomers after CombiFlash® purification using an HP C18 Aq GOLD column with MeCN/buffered _H2O as the eluting mobile phase. The benzamide was dissolved in 5 mL of 1,4-dioxane and converted to benzamide hydrochloride by stirring with 2 eq. 4N HCl in 1,4-dioxane for 1 h, then concentrated the reaction mixture to give 3-(3-aminopropanamido)-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide hydrochloride as a brown solid (0.89 g, 84%) MS (ESI, pol. ion) m/z: 498.2 (M+1).

By a procedure similar to that described in Example 57 above, the compounds of Examples 58-59 are obtained starting from the corresponding benzoic acids.

Example 58: 3-(2-aminoacetamido)-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide hydrochloride as a beige solid (0.83 g, 83%). (Compound 53)

MS (ESI, pos. ion) m/z: 484.2 (M+1).

Example 59: 3-(4-aminobutanamido)-N-((2S)-1-oxo-3-phenyl-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide hydrochloride as a beige solid (0.40 g, 36%). (Compound 55)

MS (ESI, pos. ion) m/z: 512.2 (M+1).

Example 61

Synthesis of 3-butyramido-N-((2S)-1-oxo-3-phenyl-1-(2-(pyridin-3-yl)piperidin-1-yl)propan-2-yl)benzamide (Compound 63)

Synthesis scheme for 3-butyramido-N-((2S)-1-oxo-3-phenyl-1-(2-(pyridin-3-yl)piperidin-1-yl)propan-2-yl)benzamide:

Stage 1

To a solution of (S)-2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoic acid (1.3 g, 4.4 mmol) in anhydrous DCM (20 mL) was added PCl ₅ (1.0 g, 4.8 mmol) portionwise. The reaction mixture was stirred for 3 h at rt. All volatiles were completely removed by rotary evaporation to give (S)-2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoyl chloride, which was used in the next step without further purification.

Stage 2

To a solution of (S)-2-(1,3-dioxoisoindolin-2-yl)-3-phenylpropanoyl chloride (4.8 mmol) in anhydrous benzene (20 mL) was slowly added (+/-)-anabasine (0.8 g, 4.9 mmol). The resulting solution was stirred at rt overnight and quenched with saturated NaHCO ₃ (20 mL). The layers were separated and the benzene layer was washed with brine, dried and concentrated to give 2-((2S)-1-oxo-3-phenyl-1-(2-(pyridin-3-yl)-piperidin-1-yl)propan-2-yl)isoindolin-1,3-dione as a foam (1.5 g), existing as a mixture of two diastereomers, which was used in the next step without further purification.

Stage 3

To a solution of 2-((2S)-1-oxo-3-phenyl-1-(2-(pyridin-3-yl)-piperidin-1-yl)propan-2-yl)isoindolin-1,3-dione (1.5 g) in EtOH (30 mL) was added hydrazine monohydrate (0.3 mL). The reaction solution was refluxed overnight and the resulting suspension was filtered at rt. The filtrate was dissolved in a mixture of EtOAc and hexane and kept overnight. The resulting solid was filtered off and the filtrate was concentrated to give (2S)-2-amino-3-phenyl-1-(2-(pyridin-3-yl)piperidin-1-yl)propan-1-one (0.82 g) as a mixture of two diastereomers, which was used in the next step without further purification.

Stage 4

To a solution of 3-butyramidobenzoic acid (0.56 g, 2.7 mmol) and (2S)-2-amino-3-phenyl-1-(2-(pyridin-3-yl)piperidin-1-yl)propan-1-one (0.82 g, 2.7 mmol) was added EDC hydrochloride (0.78 g, 4.0 mmol) and i-Pr2NEt (0.65 mL, 4.1 mmol) in DCM (20 mL). The reaction mixture was stirred at rt overnight and quenched with _H2O (20 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give 3-butyramido-N-((2S)-1-oxo-3-phenyl-1-(2-(pyridin-3-yl)piperidin-1-yl)propan-2-yl)benzamide as a light yellow solid (0.64 g, 32%) MS (ESI, poll. ion) m/z: 499.2 (M+1), existing as a mixture of two diastereomers after CombiFlash® purification using an HP C18 Aq GOLD column with MeCN/buffered _H2O as the eluting mobile phase.

Example 62

Synthesis of 3-butyramido-N-((2S)-3-(naphthalen-1-yl)-1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide (Compound 64)

Stage 1

A 1-necked round-bottomed flask (100 mL) equipped with a condenser was charged with 3-(1-naphthyl)-L-alanine (1.0 g, 4.6 mmol), phthalic anhydride (0.7 g, 4.6 mmol), and glacial acetic acid (10 mL) in that order. The resulting mixture was heated under reflux for 4 h. The solvent was completely removed in vacuo, and the residue was precipitated from a mixture of ethyl acetate and hexane to give (S)-2-(1,3-dioxoisoindolin-2-yl)-3-(naphthalene-1-yl)propanoic acid (0.52 g) as a beige solid.

Stage 2

To a solution of (S)-2-(1,3-dioxoisoindolin-2-yl)-3-(naphthalen-1-yl)propanoic acid (0.52 g, 1.4 mmol) in anhydrous DCM (10 mL) was added PCl ₅ (0.32 g, 1.6 mmol) portionwise. The reaction mixture was stirred for 3 h at rt. All volatiles were completely removed by rotary evaporation to give (S)-2-(1,3-dioxoisoindolin-2-yl)-3-(naphthalen-1-yl)propanoyl chloride, which was used in the next step without further purification.

Stage 3

To a solution of (S)-2-(1,3-dioxoisoindolin-2-yl)-3-(naphthalen-1-yl)propanoyl chloride (0.5 g, 1.3 mmol) in anhydrous benzene (10 mL) was slowly added (+/-)-anatabine (0.2 g, 1.3 mmol). The resulting solution was stirred at rt overnight and quenched with saturated NaHCO ₃ (10 mL). The layers were separated and the benzene layer was washed with brine, dried and concentrated to give 2-((2S)-1-oxo-3-phenyl-1-(2-(pyridin-3-yl)piperidin-1-yl)propan-2-yl)isoindolin-1,3-dione as a brown solid (0.33 g) existing as a mixture of two diastereomers, which was used in the next step without further purification.

Stage 4

To a solution of 2-((2S)-1-(3,6-dihydro-[2,3'-bipyridine]-1(2H)-yl)-3-(naphthalen-1-yl)-1-oxopropan-2-yl)isoindolin-1,3-dione (0.33 g) in EtOH (10 mL) was added hydrazine monohydrate (0.1 mL). The reaction solution was refluxed for 3 h and the resulting suspension was filtered at rt. The filtrate was dissolved in a mixture of EtOAc and hexane and kept overnight. The resulting solid was filtered off and the filtrate was concentrated to give (2S)-2-amino-1-(3,6-dihydro-[2,3'-bipyridin]-1(2H)-yl)-3-(naphthalen-1-yl)propan-1-one as a brown oil (0.38 g), existing as a mixture of two diastereomers, which was used in the next step without further purification.

Stage 5

To a solution of 3-butyramidobenzoic acid (0.21 g, 1.0 mmol) and (2S)-2-amino-1-(3,6-dihydro-[2,3'-bipyridin]-1(2H)-yl)-3-(naphthalen-1-yl)propan-1-one (0.36 g, 1.0 mmol) were added EDC hydrochloride (0.28 g, 1.4 mmol) and i-Pr2NEt (0.23 mL, 1.4 mmol) in DCM (10 mL). The reaction mixture was stirred at rt overnight and quenched with _H2O (10 mL). The layers were separated and the organic layer was washed with brine, dried and concentrated to give 3-butyramido-N-((2S)-3-(naphthalen-1-yl)-1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as a yellow solid (0.19 g) MS (ESI, pol. ion) m/z: 547.2 (M+1), existing as a mixture of two diastereomers after CombiFlash® purification using an HP C18 Aq GOLD column with MeCN/buffered _H2O as the eluting mobile phase.

By a procedure similar to that described in Example 62 above, the compounds of Examples 63-64 are obtained starting from the corresponding benzoic acids.

Example 63: 3-butyramido-N-((2S)-3-(naphthalen-2-yl)-1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as a yellow solid (0.35 g). (Compound 65)

MS (ESI, pos. ion) m/z: 547.2 (M+1).

Example 64: 3-butyramido-N-((2S)-3-cyclohexyl-1-oxo-1-(6-(pyridin-3-yl)-5,6-dihydropyridin-1(2H)-yl)propan-2-yl)benzamide as a yellow solid (0.06 g). (Compound 66)

MS (ESI, pos. ion) m/z: 503.3 (M+1).

Biological examples

Example 1

Determination of inhibition of NF-kB activation induced by TNFα… in vitro assay

The ability of the compound described herein to inhibit TNFα-induced NFkB activation was determined using the TNFα Luciferase assay. The HEK293/NF-kB luciferase cell line was generated by cotransfection of pNFkB-luc vector with pHyg (Panomics, CA, USA), followed by hygromycin selection. HEK293/NFkB luciferase cells were grown in DMEM supplemented with 10% fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin/Fungizone, and 100 μg/ml hygromycin B as a selection agent. HEK293 NFkB Luciferase cells were grown in T75 flasks (Nunc™ Cell Culture Treated EasYFlasks™) in a cell culture incubator at 37°C and 5% CO ₂ .

Culture flasks with confluent T75 cells (approximately 8-10 million cells per flask) are washed with 10 ml sterile PBS (calcium and magnesium free) in a microbiological safety cabinet. After aspirating the PBS with a sterile glass pipette, the cell layer (for each T75 flask) is overlaid with 1 ml TrypLE™, phenol red free expression (Gibco) at room temperature. After one minute of incubation, HEK293 are mechanically resuspended in TrypLE by tapping the side of the flask. The cells are then resuspended with 10 ml pre-warmed complete culture medium and transferred to a conical tube. The cells are centrifuged for 5 minutes at 4000 rpm (1000g) at room temperature. The cell pellet was resuspended with 24 ml of pre-warmed cell culture medium and 200 ml of the cell suspension (approximately 80,000 cells per well) was seeded per well in 96-well cell culture plates (Costar #3599, Corning, NY, USA) using a multichannel pipette and reverse pipetting to prevent formation of air bubbles. After seeding, the cells were incubated overnight in a cell culture incubator and then processed. For each 96-well culture plate, 8 wells were used as “control wells” (only culture medium was added) and 8 wells were added with TNFα only. The NF-κB luciferase reporter cell line was infected with 25 ng/ml TNF-α for four hours in the presence and absence (control conditions) of a range of doses of the different test compounds. For each compound, a range of concentrations is tested in quadruplicate and used to determine the IC ₅₀ .

Luciferase activity was monitored using the Luc-Screen® Extended-Glow Luciferase Reporter Gene Assay System via chemiluminescence according to the manufacturer's instructions (Invitrogen) and a BioTek Synergy HT plate reader (BioTek Instruments, VT, USA). IC ₅₀ , the concentration at which NF-kB activation is reduced by 50%, for typical amounts of compounds are presented in Table 3 below.

Table 3 Conn. No. (see conn. from table 1 above) IC ₅₀
(µM) Conn. No. (see conn. from table 1 above) IC ₅₀
(µM) Conn. No. (see conn. from table 1 above) IC ₅₀
(µM) 1 375 24 4.0 46 0.05855 2 425 25 2.5 47 0,1587 3 750 26 2.8 48 0.1139 4 550 27 4.0 49 0,1741 5 925 28 6.5 50 0,1308 6 750 29 8.0 51 0.1138 7 225 30 4.3 52 0.1339 8 550 31 1.7 53 0.2124 9 200 32 8.6 54 0,1862 10 125 33 1.8 55 0.1699 11 250 34 2.5 56 0.9675 12 470 35 1.6 57 0,1208 13 276 36 0.15 58 0.1851 14 170 37 0.125 59 0.2541 15 4.3 38 0.25 60 0.2165 16 2.0 39 0.21 61 0.7082 17 140 40 0.04 63 0.0584 18 7.0 41 0.25 64 0.285 19 5.4 42 0.0869 65 0.7036 20 1.5 43 0.1288 66 0.3457 21 4.5 44 0,0802 67 1.7 23 6.5 45 0.0754 68 0.85

Example 2

Determination of anti-inflammatory activity using an LPS acute inflammation model in mice… in vivo model

The ability of the compounds described herein to reduce inflammation is determined in vivo using the above mouse model. The effect of the compounds on LPS-induced proinflammatory cytokine production can be assessed in various tissues (e.g., plasma, brain, intestine, spleen, lung, etc.). Adult wild-type C57Bl6/J mice are used to assess the effect of a compound described herein (the "test" compound) on cytokine production induced by intraperitoneal injection of LPS (lipopolysaccharide) [LPS from Escherichia coli O111:B4, Sigma-Aldrich # L4391]. Prior to LPS treatment (1 mg/kg (ip) dissolved in sterile PBS), mice were randomly assigned to a placebo/control group receiving an intraperitoneal (IP) injection of vehicle used to solubilize the test compound (50% PEG4000/50% DMSO) or a treatment group (receiving 20 mg/kg test compound IP). Mice were injected with test compound or vehicle 15 min prior to LPS injection.

Mice were then humanely sacrificed four hours after intraperitoneal injection of LPS. Following euthanasia, blood was collected by intracardiac puncture using EDTA as an anticoagulant. Blood was immediately centrifuged at 1500 g for 4 minutes, and plasma was collected and snap frozen in liquid nitrogen. All other tissues were rapidly dissected and snap frozen in liquid nitrogen. Samples were stored at -80°C.

Tissue homogenates were prepared by sonication in ice-cold M-PER Reagent (Pierce Biotechnology, Rockford, IL, USA) containing 1 mM phenylmethanesulfonyl fluoride, 1X protease inhibitor cocktail (Roche, Inc., USA), and 1 mM sodium orthovanadate (Sigma-Aldrich, MO, USA). Cytoxins were quantified by electrochemiluminescence using MULTI-SPOT plates from the V-Plex assay kit from the Pro-Inflammatory Panel 1 (mouse) kit (Mesoscale discovery, USA). All LPS-treated samples were diluted 10X with kit diluent 41, and control samples were assayed without dilution. The amount of proinflammatory cytokines interferon-gamma, IL-1β, and TNFα produced was lower in the treatment group compared to the control group.

Examples of composition

The following are typical pharmaceutical formulations containing the compound of the present description.

Tablet composition

The following ingredients are thoroughly mixed and pressed into single-scored tablets.

Ingredient Quantity per tablet mg The connection according to the present description 400 Cornstarch 50 Croscarmellose sodium 25 Lactose 120 Magnesium stearate 5

Capsule composition

The following ingredients are thoroughly mixed and loaded into hard gelatin capsules.

Ingredient Quantity per capsule mg The connection according to the present description 200 Dried lactose spray 148 Magnesium stearate 2

Injection composition

Compound as described (e.g., Compound 1) in 2% HPMC, 1% Tween 80 in DI water, pH 2.2 with MSA, as needed to at least 20 mg/mL.

Composition for inhalation

To prepare a pharmaceutical composition for inhalation delivery, 20 mg of the compound described herein is mixed with 50 mg of anhydrous citric acid and 100 ml of 0.9% sodium chloride solution. The mixture is introduced into an inhalation delivery device, such as a nebulizer, which is suitable for inhalation administration.

Gel composition for local use

To prepare a pharmaceutical gel composition for topical use, 100 mg of the compound described herein is mixed with 1.75 g of hydroxypropyl cellulose, 10 ml of propylene glycol, 10 ml of isopropyl myristate and 100 ml of purified alcohol USP. The resulting gel solution is then placed in containers, such as tubes, that are suitable for topical use.

Composition of ophthalmic solution

To prepare a pharmaceutical ophthalmic solution composition, 100 mg of the compound described herein is mixed with 0.9 g of NaCl in 100 ml of purified water and filtered using a 0.2 micron filter. The resulting isotonic solution is then introduced into ophthalmic delivery vehicles, such as eye drop containers, that are suitable for ophthalmic administration.

Solution for nasal spray

To prepare a pharmaceutical solution for nasal spray, 10 g of the compound described herein is mixed with 30 ml of a 0.05 M phosphate buffer solution (pH 4.4). The solution is placed in a nasal dispenser designed to deliver 100 µl of spray per application.

Claims (41)

1. Compound of formula (I) , (I) where n is 0, 1 or 2; the dotted line is an optional connection; Het is pyridin-3-yl; R ¹ is hydrogen; R ² is hydrogen; R ³ is hydrogen, alkyl, cycloalkylalkyl or aralkyl; R ⁴ is hydrogen and R ⁵ is -C(O)R ⁶ , wherein R ⁶ is aryl or heteroaryl, wherein aryl or heteroaryl is optionally substituted with R ^g , ^Rh and/or R ⁱ , independently selected from alkyl, C ₁ -C ₆ alkoxy, halogen, halo-C ₁ -C ₆ alkyl, nitro, amino, C ₁ -C ₆ alkylamino and acylamino; wherein acylamino is -NR'C(O)R, wherein R' is hydrogen or alkyl and wherein R is _C1 - _C6 alkyl or _C3 - _C10 cycloalkyl, wherein _C1 - _C6 alkyl may be substituted with amino or 2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl; or R ⁴ and R ⁵ together with the nitrogen atom to which they are attached form 1,3-dioxoisoindolin-2-yl; or its pharmaceutically acceptable salt; where "alkyl" means a linear saturated monovalent hydrocarbon radical of one to six carbon atoms or a branched saturated monovalent hydrocarbon radical of three to six carbon atoms; where "alkylene" means a linear saturated divalent hydrocarbon radical of one to six carbon atoms or a branched saturated divalent hydrocarbon radical of three to six carbon atoms; where "heteroaryl" means a monovalent monocyclic or bicyclic aromatic radical of 5 to 10 ring atoms, where one or two ring atoms are heteroatoms selected from N and O, and the remaining ring atoms are carbon; where "aryl" means a monocyclic or bicyclic aromatic hydrocarbon radical of 6-10 ring atoms; where "cycloalkyl" means a cyclic saturated monovalent hydrocarbon radical of three to ten carbon atoms; where "cycloalkylalkyl" means the radical -(alkylene)-R, where R is cycloalkyl as defined above. 2. The compound according to claim 1, wherein the compound has the structure (IA) . 3. The compound according to claim 1, wherein the compound has the structure (IB) . 4. A compound according to any one of paragraphs 1-3, where n is 1 or 2. 5. A compound according to any one of paragraphs 1-4, where n is equal to 1. 6. A compound according to any one of paragraphs 1-3, where n is 0 or 2. 7. A connection according to any one of paragraphs 1-3, where n is equal to 0. 8. A compound according to any one of paragraphs 1-3, where n is equal to 2. 9. A compound according to any one of claims 1 to 8, wherein R ⁵ is phenyl optionally substituted with R ^g , ^Rh and/or R ⁱ . 10. A compound according to any one of claims 1 to 9, wherein R ^g is alkyl, C ₁ -C ₆ alkoxy, halogen, halo-C ₁ -C ₆ alkyl and R ^h and R ⁱ are independently selected from alkyl, C ₁ -C ₆ alkoxy, halogen, halo-C ₁ -C ₆ alkyl, amino, C ₁ -C ₆ alkylamino and acylamino. 11. A compound according to any one of claims 1 to 9, wherein R ^g , ^Rh and R ⁱ are independently selected from alkyl, C ₁ -C ₆ alkoxy, halogen, halo-C ₁ -C ₆ alkyl, amino and acylamino, preferably methyl, ethyl, methoxy, ethoxy, chlorine, fluorine, trifluoromethyl, acetylamino, butanoylamino and pentanoylamino. 12. A compound according to any one of claims 1 to 8, wherein R ⁴ and R ⁵ together with the nitrogen atom to which they are attached form 1,3-dioxoisoindolin-2-yl. 13. A compound according to any one of claims 1 to 12, wherein R ³ is hydrogen or alkyl, preferably methyl, ethyl, propyl, isopropyl, sec-propyl, n-, sec-, iso-, tert-butyl. 14. A compound according to any one of claims 1 to 12, wherein R ³ is aralkyl, preferably R ³ is benzyl or phenethyl, more preferably R ³ is benzyl. 15. A compound according to any one of claims 1 to 12, wherein R ³ is cycloalkylalkyl, preferably R ³ is cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl or cyclohexylmethyl. 16. A compound according to any one of claims 1 to 15, wherein the stereochemistry at the carbon to which R ³ is attached is (S). 17. A compound according to any one of claims 1 to 15, wherein the stereochemistry at the carbon to which R ³ is attached is (R). 18. A pharmaceutical composition having inhibitory activity against nuclear factor-kappa B (NF-kB), containing an effective amount of a compound according to any one of claims 1-17 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. 19. A method for treating a disease that can be treated by inhibiting NF-kB activation, comprising administering to a patient in need thereof a pharmaceutical composition according to claim 18, wherein the disease is an inflammatory disease. 20. The method according to claim 19, wherein the disease is selected from the group consisting of an autoimmune disease, pain, allergies, asthma, chronic obstructive pulmonary disease and sepsis. 21. The method of claim 19, wherein the disease is selected from the group consisting of rheumatoid arthritis, osteoarthritis, atherosclerosis, multiple sclerosis, asthma, inflammatory bowel disease, diabetes, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, osteoporosis, systemic lupus erythematosus, chronic obstructive pulmonary disease, cystic fibrosis, stroke, acute kidney injury, glomerulonephritis, psoriasis, atopic dermatitis, Behcet's disease, tuberculosis, Crohn's disease, colitis, Paget's disease, pancreatitis, periodontitis, inflammatory lung disease, and lupus nephritis.