Apoptosis signal-regulating kinase 1 inhibitors and methods of use thereof

The present invention discloses compounds of Formula (I), and pharmaceutically acceptable salts and esters thereof: which inhibit the Apoptosis signal-regulating kinase 1 (ASK-1), which associated with autoimmune disorders, neurodegenerative disorders, inflammatory diseases, chronic kidney disease, cardiovascular disease. The present invention further relates to pharmaceutical compositions comprising the aforementioned compounds for administration to a subject suffering from ASK-1 related disease. The invention also relates to methods of treating an ASK-1 related disease in a subject by administering a pharmaceutical composition comprising the compounds of the present invention. The present invention specifically relates to methods of treating ASK-1 associated with hepatic steatosis, including non-alcoholic fatty liver disease (NAFLD) and non-alcohol steatohepatitis disease (NASH).

TECHNICAL FIELD

The present invention relates generally to compounds and pharmaceutical compositions useful as ASK-1 inhibitors. Specifically, the present invention relates to compounds useful as inhibitors of ASK-1 and methods for their preparation and use.

BACKGROUND OF THE INVENTION

More specifically, ASK-1 has been associated with hepatic steatosis, including non-alcoholic fatty liver disease (NAFLD) and non-alcohol steatohepatitis (NASH). In a mouse model, high fat diets have caused induction of hepatic steatosis, ultimately causing fat accumulation and fatty acid oxidation. This led to the generation of ROS which caused hepatocyte dysfunction and death (S. K. Mantena, et al.,Free Radic. Biol. Med.2008, 44, 1259-1272; S. K. Mantena, et al.,Biochem. J.2009, 417, 183-193). Moreover, TNF was shown to be critical for apoptosis of hepatocytes through the ASK-1-JNK pathway, and TNF deficient mice showed reduced hepatic steatosis and fibrosis (W. Zhang, et al.,Biochem. Biophys. Res. Commun.2010, 391, 1731-1736).

There is a need for the development of ASK-1 inhibitors for the treatment and prevention of disease. The present invention has identified compounds which inhibit ASK-1 as well as methods of using these compounds to treat disease.

SUMMARY OF THE INVENTION

In one aspect, the invention provides compounds represented by Formula I, or a pharmaceutically acceptable salt or ester thereof:

wherein:
X1and X2are each independently C(R8) or N;
X3is C(R9) or N, wherein R9is selected from the group consisting of hydrogen, optionally substituted —C1-C8alkyl, optionally substituted —C1-C8alkoxy and halo;
R1is selected from

R4is selected from the group consisting of:

9) Substituted or unsubstituted heteroaryl; and

R2, R5and R8are each independently selected from the group consisting of:

R3is selected from the group consisting of:

R10and R11are each independently selected from the group consisting of hydrogen, hydroxyl, and optionally substituted —C1-C8alkyl; alternatively, R10and R11are taken together with the carbon atom to which they are attached to form C(O), spiro-C3-C8cycloalkyl, or spiro-3- to 8-membered heterocycloalkyl;

R12and R13are each independently selected from the group consisting of hydrogen, halo, and optionally substituted —C1-C8alkyl; and

n is 0, 1 or 2; preferably n is 0 or 1.

In another embodiment, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound or combination of compounds of the present invention, or a pharmaceutically acceptable salt form, stereoisomer, solvate, hydrate or combination thereof, in combination with a pharmaceutically acceptable carrier or excipient.

In another embodiment, the present invention provides a method for the prevention or treatment of an ASK-1 mediated disease or condition. The method comprises administering a therapeutically effective amount of a compound of Formula (I) to a subject in need thereof. The present invention also provides the use of a compound of Formula (I) for the preparation of a medicament for the prevention or treatment of an ASK-1 mediated disease or condition. Such diseases include autoimmune disorders, neurodegenerative disorders, inflammatory diseases, chronic kidney disease, cardiovascular disease, metabolic disorders, and acute and chronic liver diseases.

DETAILED DESCRIPTION OF THE INVENTION

A first embodiment of the invention is a compound represented by Formula I as described above, or a pharmaceutically acceptable salt or esterthereof.

In a certain embodiment, the present invention relates to compounds of Formula I, or a pharmaceutically acceptable salt or ester thereof, wherein R4is selected from the groups below:

wherein each of the above shown groups is optionally substituted. Preferably, R4is selected from

In certain embodiments, the present invention relates to compounds of Formula I, or a pharmaceutically acceptable salt or ester thereof, wherein R2is hydrogen or halogen.

In certain embodiments, the present invention relates to compounds of Formula I, or a pharmaceutically acceptable salt or ester thereof, wherein R5is hydrogen.

In certain embodiments, the present invention relates to compounds of Formula I, or a pharmaceutically acceptable salt or ester thereof, wherein R2is hydrogen, R5is hydrogen, and n is 0 or 1.

In certain embodiments, the present invention relates to compounds of Formula I, or a pharmaceutically acceptable salt or ester thereof, wherein R3is

wherein R6is selected from the groups below:

wherein each of the above shown groups is optionally substituted.

In certain embodiments, the present invention relates to compounds of Formula I, or a pharmaceutically acceptable salt or ester thereof, wherein R3is

R6and R7, together with the nitrogen atom to which they are they attached, form an optionally substituted heterocycloalkyl is selected from the groups below:

In another embodiment, the present invention relates to compounds of Formula I, or a pharmaceutically acceptable salt or ester thereof, wherein R3is

wherein R6is selected from the groups below:

In certain embodiments, the present invention relates to compounds of Formula I, and pharmaceutically acceptable salts and esters thereof, wherein R3is selected from the groups below:

wherein each of these groups is optionally substituted.

In certain embodiments, the present invention relates to compounds of Formula I, or a pharmaceutically acceptable salt or ester thereof, wherein X3is selected from C—H, C—F, C—OMe, and N.

In certain embodiments, the compound of Formula I is represented by Formula Ia-1, Ia-2, Ib-1, Ib-2, Ic-1, Ic-2, or Ie, or a pharmaceutically acceptable salt or ester thereof:

In certain embodiments, the compound of Formula I is represented by Formula II or a pharmaceutically acceptable salt or ester thereof:

In certain embodiments, the compound of Formula I is represented by Formula III or a pharmaceutically acceptable salt or ester thereof:

In certain embodiments, the compound of Formula I is represented by Formula IV or a pharmaceutically acceptable salt or ester thereof:

In certain embodiments, the compound of Formula I is represented by Formula V or a pharmaceutically acceptable salt or ester thereof:

In certain embodiments, the compound of Formula I is represented by Formula VI or a pharmaceutically acceptable salt or ester:

In certain embodiments, the compound of Formula I is represented by Formula VII or a pharmaceutically acceptable salt or ester thereof:

In certain embodiments, the compound of Formula I is represented by Formula VIII or a pharmaceutically acceptable salt or ester thereof:

wherein R3, X3, and n are as previously defined.

Representative compounds of the invention include, but are not limited to, the following compounds (compound 1 to compound 300 in Table 1) according to Formula VIII, and pharmaceutically acceptable salts thereof, wherein R3, X3, and n are delineated for each compound in Table 1.

In certain embodiments, the compound of Formula I is represented by Formula IX or a pharmaceutically acceptable salt or ester thereof:

In certain embodiments, the compound of Formula I is represented by Formula X or a pharmaceutically acceptable salt or ester thereof:

In certain embodiments, the compound of Formula I is represented by Formula XI or a pharmaceutically acceptable salt or ester thereof:

wherein R3, X3, and n are as previously defined.

Representative compounds of the invention include, but are not limited to, the following compounds (compound 301 to compound 600 in Table 2) according to Formula XI, and pharmaceutically acceptable salts thereof, wherein R3, X3, and n are delineated for each compound in Table 2.

In another embodiment of the invention, the compound of Formula I is represented by Formula XII-a, Formula XII-b, or a pharmaceutically acceptable salt or ester thereof:

In another embodiment of the invention, the compound of Formula I is represented by Formula XIII-a, Formula XIII-b, or a pharmaceutically acceptable salt or ester thereof:

In another embodiment of the invention, the compound of Formula I is represented by Formula XIV-a, Formula XIV-b, or a pharmaceutically acceptable salt or ester thereof:

In another embodiment of the invention, the compound of Formula I is represented by Formula XV-a, Formula XV-b, or a pharmaceutically acceptable salt or ester thereof:

In certain embodiments, the present invention provides a method for the treatment of an ASK-1 mediated disease or condition. The method comprises administering a therapeutically effective amount of a compound of Formula (I). The present invention also provides the use of a compound of Formula (I) for the preparation of a medicament for the treatment of an ASK-1 mediated disease or condition.

In certain embodiments, the ASK-1 mediated disease or condition is an autoimmune disorder, a neurodegenerative disorder, an inflammatory disease, chronic kidney disease, renal disease, cardiovascular disease, a metabolic disease, or an acute or chronic liver disease.

In certain embodiments, the renal disease is diabetic nephropathy, focal segmental glomerulosclerosis (FSGS), hypertensive nephrosclerosis, chronic glomerulonephritis, chronic transplant glomerulopathy, chronic interstitial nephritis, or polycystic kidney disease.

In certain embodiments, the cardiovascular disease is atherosclerosis, arteriosclerosis, reperfusion/ischemia in stroke, cardiac hypertrophy, respiratory diseases, heart attacks, myocardial ischemia.

In certain embodiments, the metabolic disease is insulin resistance, Type I and Type II diabetes, or obesity.

In certain embodiments, the chronic kidney disease is polycystic kidney disease, pyelonephritis, kidney fibrosis and glomerulonephritis.

Yet a further aspect of the present invention is a process of making any of the compounds delineated herein employing any of the synthetic means delineated herein.

Definitions

The term “alkyl” as used herein, refers to saturated, straight- or branched-chain hydrocarbon radicals. “C1-C3alkyl,” “C1-C6alkyl,” “C1-C10alkyl” C2-C4alkyl,” or “C3-C6alkyl,” refer to alkyl groups containing from one to three, one to six, one to ten carbon atoms, 2 to 4 and 3 to 6 carbon atoms respectively. Examples of C1-C8alkyl radicals include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl, heptyl and octyl radicals.

The term “alkenyl” as used herein, refers to straight- or branched-chain hydrocarbon radicals having at least one carbon-carbon double bond by the removal of a single hydrogen atom. “C2-C10alkenyl,” “C2-C8alkenyl,” “C2-C4alkenyl,” or “C3-C6alkenyl,” refer to alkenyl groups containing from two to ten, two to eight, two to four or three to six carbon atoms respectively. Alkenyl groups include, but are not limited to, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, heptenyl, octenyl, and the like.

The term “alkynyl” as used herein, refers to straight- or branched-chain hydrocarbon radicals having at least one carbon-carbon triple bond by the removal of a single hydrogen atom. “C2-C10alkynyl,” “C2-C8alkynyl,” “C2-C4alkynyl,” or “C3-C6alkynyl,” refer to alkynyl groups containing from two to ten, two to eight, two to four or three to six carbon atoms respectively. Representative alkynyl groups include, but are not limited to, for example, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl, and the like.

The term “cycloalkyl”, as used herein, refers to a monocyclic or polycyclic saturated carbocyclic ring or a bi- or tri-cyclic group fused, bridged or spiro system, and the carbon atoms may be optionally oxo-substituted or optionally substituted with exocyclic olefinic, iminic or oximic double bond. Preferred cycloalkyl groups include C3-C12cycloalkyl, C3-C6cycloalkyl, C3-C8cycloalkyl and C4-C7cycloalkyl. Examples of C3-C12cycloalkyl include, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl, cyclooctyl, 4-methylene-cyclohexyl, bicyclo[2.2.1]heptyl, bicyclo[3.1.0]hexyl, spiro[2.5]octyl, 3-methylenebicyclo[3.2.1]octyl, spiro[4.4]nonanyl, and the like.

The term “cycloalkenyl”, as used herein, refers to monocyclic or polycyclic carbocyclic ring or a bi- or tri-cyclic group fused, bridged or spiro system having at least one carbon-carbon double bond and the carbon atoms may be optionally oxo-substituted or optionally substituted with exocyclic olefinic, iminic or oximic double bond. Preferred cycloalkenyl groups include C3-C12cycloalkenyl, C3-C8cycloalkenyl or C5-C7cycloalkenyl groups. Examples of C3-C12cycloalkenyl include, but not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, bicyclo[2.2.1]hept-2-enyl, bicyclo[3.1.0]hex-2-enyl, spiro[2.5]oct-4-enyl, spiro[4.4]non-1-enyl, bicyclo[4.2.1]non-3-en-9-yl, and the like.

The term “aryl,” as used herein, refers to a mono- or polycyclic carbocyclic ring system comprising at least one aromatic ring, including, but not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, and indenyl. A polycyclic aryl is a polycyclic ring system that comprises at least one aromatic ring. Polycyclic aryls can comprise fused rings, covalently attached rings or a combination thereof.

The term “heteroaryl,” as used herein, refers to a mono- or polycyclic aromatic radical having one or more ring atom selected from S, O and N; and the remaining ring atoms are carbon, wherein any N or S contained within the ring may be optionally oxidized. Heteroaryl includes, but is not limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isoxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl, quinoxalinyl. A polycyclic heteroaryl can comprise fused rings, covalently attached rings or a combination thereof.

In accordance with the invention, aromatic groups can be substituted or unsubstituted. The term “bicyclic aryl” or “bicyclic heteroaryl” refers to a ring system consisting of two rings wherein at least one ring is aromatic; and the two rings can be fused or covalently attached.

As used herein, the term “arylalkyl” means a functional group wherein an alkylene chain is attached to an aryl group, e.g., —CH2CH2-phenyl. The term “substituted arylalkyl” means an arylalkyl functional group in which the aryl group is substituted. Similarly, the term “heteroarylalkyl” means a functional group wherein an alkylene chain is attached to a heteroaryl group. The term “substituted heteroarylalkyl” means a heteroarylalkyl functional group in which the heteroaryl group is substituted.

The term “alkylene” as used herein, refers to a diradical of a branched or unbranched saturated hydrocarbon chain, typically having from 1 to 20 carbon atoms (e.g. 1-10 carbon atoms, or 1, 2, 3, 4, 5, or 6 carbon atoms). This term is exemplified by groups such as methylene (—CH2—), ethylene (—CH2CH2—), the propylene isomers (e.g., —CH2CH2CH2— and —CH(CH3)CH2—), and the like.

In accordance with the invention, any of the aryls, substituted aryls, heteroaryls and substituted heteroaryls described herein, can be any aromatic group. Aromatic groups can be substituted or unsubstituted.

It is understood that any alkyl, alkenyl, alkynyl, cycloalkyl and cycloalkenyl moiety described herein can also be an aliphatic group, an alicyclic group or a heterocyclic group. An “aliphatic group” is non-aromatic moiety that may contain any combination of carbon atoms, hydrogen atoms, halogen atoms, oxygen, nitrogen or other atoms, and optionally contain one or more units of unsaturation, e.g., double and/or triple bonds. An aliphatic group may be straight chained, branched or cyclic and preferably contains between about 1 and about 24 carbon atoms, more typically between about 1 and about 12 carbon atoms. In addition to aliphatic hydrocarbon groups, aliphatic groups include, for example, polyalkoxyalkyls, such as polyalkylene glycols, polyamines, and polyimines, for example. Such aliphatic groups may be further substituted. It is understood that aliphatic groups may be used in place of the alkyl, alkenyl, alkynyl, alkylene, alkenylene, and alkynylene groups described herein.

The term “alicyclic” as used herein, denotes a monovalent group derived from a monocyclic or polycyclic saturated carbocyclic ring compound by the removal of a single hydrogen atom. Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo[2.2.1]heptyl, and bicyclo[2.2.2]octyl. Such alicyclic groups may be further substituted.

As used herein, the term “alkoxy” employed alone or in combination with other terms means, unless otherwise stated, an alkyl group having the designated number of carbon atoms connected to the rest of the molecule via an oxygen atom, such as, for example, methoxy, ethoxy, 1-propoxy, 2-propoxy (isopropoxy) and the higher homologs and isomers. Preferred alkoxy are (C1-C3) alkoxy.

The term “aryloxy” refers to the group aryl-O— wherein the aryl group is as defined above, and includes optionally substituted aryl groups as also defined above. The term “arylthio” refers to the group R—S—, where R is as defined for aryl.

The terms “heterocyclic” or “heterocycloalkyl” can be used interchangeably and refer to a non-aromatic ring or a bi- or tri-cyclic group fused, bridged or spiro system, where (i) the ring system contains at least one heteroatom independently selected from oxygen, sulfur and nitrogen, (ii) the ring system can be saturated or unsaturated (iii) the nitrogen and sulfur heteroatoms may optionally be oxidized, (iv) the nitrogen heteroatom may optionally be quaternized, (v) any of the above rings may be fused to an aromatic ring, and (vi) the remaining ring atoms are carbon atoms which may be optionally oxo-substituted or optionally substituted with exocyclic olefinic, iminic or oximic double bond. Representative heterocycloalkyl groups include, but are not limited to, 1,3-dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, 2-azabicyclo[2.2.1]-heptyl, 8-azabicyclo[3.2.1]octyl, 5-azaspiro[2.5]octyl, 1-oxa-7-azaspiro[4.4]nonanyl, 7-oxooxepan-4-yl, and tetrahydrofuryl. Such heterocyclic groups may be further substituted. Heteroaryl or heterocyclic groups can be C-attached or N-attached (where possible).

It is understood that any alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic and cycloalkenyl moiety described herein can also be an aliphatic group or an alicyclic group.

It will be apparent that in various embodiments of the invention, the substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, arylalkyl, heteroarylalkyl, and heterocycloalkyl are intended to be monovalent or divalent. Thus, alkylene, alkenylene, and alkynylene, cycloaklylene, cycloalkenylene, cycloalkynylene, arylalkylene, heteroarylalkylene and heterocycloalkylene groups are to be included in the above definitions, and are applicable to provide the Formulas herein with proper valency.

The term “optionally substituted”, as used herein, means that the referenced group may be substituted or unsubstituted. In one embodiment, the referenced group is optionally substituted with zero substituents, i.e., the referenced group is unsubstituted. In another embodiment, the referenced group is optionally substituted with one or more additional group(s) individually and independently selected from groups described herein.

The term “hydrogen” includes hydrogen and deuterium. In addition, the recitation of an atom includes other isotopes of that atom so long as the resulting compound is pharmaceutically acceptable.

In certain embodiments, the compounds of each formula herein are defined to include isotopically labelled compounds. An “isotopically labelled compound” is a compound in which at least one atomic position is enriched in a specific isotope of the designated element to a level which is significantly greater than the natural abundance of that isotope. For example, one or more hydrogen atom positions in a compound can be enriched with deuterium to a level which is significantly greater than the natural abundance of deuterium, for example, enrichment to a level of at least 1%, preferably at least 20% or at least 50%. Such a deuterated compound may, for example, be metabolized more slowly than its non-deuterated analog, and therefore exhibit a longer half-life when administered to a subject. Such compounds can synthesize using methods known in the art, for example by employing deuterated starting materials. Unless stated to the contrary, isotopically labelled compounds are pharmaceutically acceptable.

The term “subject” as used herein refers to a mammal. A subject therefore refers to, for example, dogs, cats, horses, cows, pigs, guinea pigs, and the like. Preferably the subject is a human. When the subject is a human, the subject may be referred to herein as a patient.

As used herein, the term “pharmaceutically acceptable salt” refers to those salts of the compounds formed by the process of the present invention which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art.

The term “hydroxy activating group,” as used herein, refers to a labile chemical moiety which is known in the art to activate a hydroxyl group so that it will depart during synthetic procedures such as in a substitution or an elimination reaction. Examples of hydroxyl activating group include, but not limited to, mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate and the like.

The term “activated hydroxyl,” as used herein, refers to a hydroxy group activated with a hydroxyl activating group, as defined above, including mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups, for example.

The term “hydroxy protecting group,” as used herein, refers to a labile chemical moiety which is known in the art to protect a hydroxyl group against undesired reactions during synthetic procedures. After said synthetic procedure(s) the hydroxy protecting group as described herein may be selectively removed. Hydroxy protecting groups as known in the art are described generally in T. H. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis,3rd edition, John Wiley & Sons, New York (1999). Examples of hydroxyl protecting groups include benzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, tert-butoxy-carbonyl, isopropoxycarbonyl, diphenylmethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, t-butyl, 2,2,2-trichloroethyl, 2-trimethylsilyl ethyl, allyl, benzyl, triphenyl-methyl (trityl), methoxymethyl, methylthiomethyl, benzyloxymethyl, 2-(trimethylsilyl)-ethoxymethyl, methanesulfonyl, trimethylsilyl, triisopropylsilyl, and the like.

The term “protected hydroxy,” as used herein, refers to a hydroxy group protected with a hydroxy protecting group, as defined above, including benzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl groups, for example.

The term “hydroxy prodrug group,” as used herein, refers to a promoiety group which is known in the art to change the physicochemical, and hence the biological properties of a parent drug in a transient manner by covering or masking the hydroxy group. After said synthetic procedure(s), the hydroxy prodrug group as described herein must be capable of reverting back to hydroxy group in vivo. Hydroxy prodrug groups as known in the art are described generally in Kenneth B. Sloan,Prodrugs, Topical and Ocular Drug Delivery, (Drugs and the Pharmaceutical Sciences; Volume 53), Marcel Dekker, Inc., New York (1992) and in “Prodrugs of Alcohols and Phenols” by S. S. Dhareshwar and V. J. Stella, inProdrugs Challenges and RewardsPart-2, (Biotechnology: Pharmaceutical Aspects), edited by V. J. Stella, et al, Springer and AAPSPress, 2007, pp 31-99.

The term “amino” as used herein, refers to the group —NH2.

The term “substituted amino” as used herein, refers to the group —NRR where each R is independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, heteroaryl and heterocycloalkyl provided that both R groups are not hydrogen, or a group —Y—Z, in which Y is optionally substituted alkylene and Z is alkenyl, cycloalkenyl, or alkynyl.

The term “amino protecting group” as used herein, refers to a labile chemical moiety which is known in the art to protect an amino group against undesired reactions during synthetic procedures. After said synthetic procedure(s) the amino protecting group as described herein may be selectively removed. Amino protecting groups as known in the are described generally in T. H. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis,3rd edition, John Wiley & Sons, New York (1999). Examples of amino protecting groups include, but are not limited to, t-butoxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyloxycarbonyl, and the like.

The term “leaving group” means a functional group or atom which can be displaced by another functional group or atom in a substitution reaction, such as a nucleophilic substitution reaction. By way of example, representative leaving groups include chloro, bromo and iodo groups; sulfonic ester groups, such as mesylate, tosylate, brosylate, nosylate and the like; and acyloxy groups, such as acetoxy, trifluoroacetoxy and the like.

The term “pharmaceutically acceptable prodrugs” as used herein refers to those prodrugs of the compounds formed by the process of the present invention which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals with undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the present invention. “Prodrug”, as used herein means a compound, which is convertible in vivo by metabolic means (e.g. by hydrolysis) to afford any compound delineated by the Formulae of the instant invention. Various forms of prodrugs are known in the art, for example, as discussed in Bundgaard, (ed.),Design of Prodrugs, Elsevier (1985); Widder, et al. (ed.),Methods in Enzymology, Vol. 4, Academic Press (1985); Krogsgaard-Larsen, et al., (ed). “Design and Application of Prodrugs,Textbook of Drug Design and Development, Chapter 5, 113-191 (1991); Bundgaard, et al.,Journal of Drug Deliver Reviews,8:1-38(1992); Bundgaard,J. of Pharmaceutical Sciences,77:285 et seq. (1988); Higuchi and Stella (eds.) Prodrugs as Novel Drug Delivery Systems,American Chemical Society(1975); and Bernard Testa & Joachim Mayer, “Hydrolysis In Drug And Prodrug Metabolism: Chemistry, Biochemistry And Enzymology,” John Wiley and Sons, Ltd. (2002).

The term “treating”, as used herein, means relieving, lessening, reducing, eliminating, modulating, or ameliorating, i.e. causing regression of the disease state or condition. Treating can also include inhibiting, i.e. arresting the development, of an existing disease state or condition, and relieving or ameliorating, i.e. causing regression of an existing disease state or condition, for example when the disease state or condition may already be present.

The term “preventing”, as used herein means, to completely or almost completely stop a disease state or condition, from occurring in a patient or subject, especially when the patient or subject is predisposed to such or at risk of contracting a disease state or condition.

The term “aprotic solvent,” as used herein, refers to a solvent that is relatively inert to proton activity, i.e., not acting as a proton-donor. Examples include, but are not limited to, hydrocarbons, such as hexane and toluene, for example, halogenated hydrocarbons, such as, for example, methylene chloride, ethylene chloride, chloroform, and the like, heterocyclic compounds, such as, for example, tetrahydrofuran and N-methylpyrrolidinone, and ethers such as diethyl ether, bis-methoxymethyl ether. Such solvents are well known to those skilled in the art, and individual solvents or mixtures thereof may be preferred for specific compounds and reaction conditions, depending upon such factors as the solubility of reagents, reactivity of reagents and preferred temperature ranges, for example. Further discussions of aprotic solvents may be found in organic chemistry textbooks or in specialized monographs, for example:Organic Solvents Physical Properties and Methods of Purification,4th ed., edited by John A. Riddick et al., Vol. II, in theTechniques of Chemistry Series, John Wiley & Sons, N Y, 1986.

The terms “protogenic organic solvent” or “protic solvent” as used herein, refer to a solvent that tends to provide protons, such as an alcohol, for example, methanol, ethanol, propanol, isopropanol, butanol, t-butanol, and the like. Such solvents are well known to those skilled in the art, and individual solvents or mixtures thereof may be preferred for specific compounds and reaction conditions, depending upon such factors as the solubility of reagents, reactivity of reagents and preferred temperature ranges, for example. Further discussions of protogenic solvents may be found in organic chemistry textbooks or in specialized monographs, for example:Organic Solvents Physical Properties and Methods of Purification,4th ed., edited by John A. Riddick et al., Vol. II, in theTechniques of Chemistry Series, John Wiley & Sons, N Y, 1986.

The synthesized compounds can be separated from a reaction mixture and further purified by a method such as column chromatography, high pressure liquid chromatography, or recrystallization. Additionally, the various synthetic steps may be performed in an alternate sequence or order to give the desired compounds. In addition, the solvents, temperatures, reaction durations, etc. delineated herein are for purposes of illustration only and variation of the reaction conditions can produce the desired isoxazole products of the present invention. Synthetic chemistry transformations and protecting group methodologies (protection and deprotection) useful in synthesizing the compounds described herein include, for example, those described in R. Larock,Comprehensive Organic Transformations, VCH Publishers (1989); T. W. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis,2d. Ed., John Wiley and Sons (1991); L. Fieser and M. Fieser,Fieser and Fieser's Reagents for Or anic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995).

The compounds of this invention may be modified by appending various functionalities via synthetic means delineated herein to enhance selective biological properties. Such modifications include those which increase biological penetration into a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral availability, increase solubility to allow administration by injection, alter metabolism and alter rate of excretion.

Pharmaceutical Compositions

Unless otherwise defined, all technical and scientific terms used herein are accorded the meaning commonly known to one with ordinary skill in the art. All publications, patents, published patent applications, and other references mentioned herein are hereby incorporated by reference in their entirety.

Abbreviations

Abbreviations which have been used in the descriptions of the schemes and the examples that follow are:Alloc for allyloxycarbonyl;Alloc-Cl for allyl chloroformate;ASK1 for apoptosis signal-regulating kinase 1;ATP for adenosine triphosphate;Boc for tert-butyloxycarbonyl;BOP—Cl for bis(2-oxo-3-oxazolidinyl)phosphinic chloride;Cbz for benzyloxycarbonyl;Cbz-Cl for benzyl chloroformate;CDI for carbonyldiimidazole;(COCl)2for oxalyl chloride;DBU for 1,8-diazabicycloundec-7-ene;DCC for N,N-dicyclohexylcarbodiimide;1,2-DCE for 1,2-dichloroethane;DCM for dichloromethane;DIPEA or Hunig's base or i-Pr2NEt for N,N-diisopropylethylamine;DMAc for N,N-dimethylacetamide;DMAP for N,N-dimethylaminopyridine;DMF for N,N-dimethyl formamide;EDC for 1-(3-diethylaminopropyl)-3-ethylcarbodiimide hydrochloride;EGTA for ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid;ESI for electrospray ionization;Et3N or TEA for triethylamine;Et2O for diethylether;EtOAc for ethyl acetate;Ghosez's Reagent for 1-chloro-N,N,2-trimethyl-1-propenylamine;HATU for 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate;HEPES for 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid, N-(2-Hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid);IC50for half maximal inhibitory concentration;KOt-Bu for potassium tert-butoxide;LCMS for liquid chromatography-mass spectrometry;MeCN for acetonitrile;MTBE for methyl tert-butyl ether;m/z for mass-to-charge ratio;NaOt-Bu for sodium tert-butoxide;NMP for 1-methyl-2-pyrrolidinone;NMR for nuclear magnetic resonance spectroscopy;OMs or mesylate for methanesulfonate;OTf or triflate for trifluoromethanesulfonate;OTs or tosylate for para-toluenesulfonate;Pd2(dba)3for tris(dibenzylideneacetone)dipalladium(0);P(o-tolyl)3for tri(o-tolyl)phosphine;PyAOP for 7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate;PyBOP for benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate;STK3 for serine/threonine-protein kinase 3TEA for triethylamine;THF for tetrahydrofuran.
Synthetic Methods

As shown in Scheme 1, compounds of Formula (Ie) are prepared from the compound of Formula (1-1) wherein X3is as previously defined. Thus, the compound of Formula (1-1) is reacted with Cbz-Cl to afford a compound of Formula (1-2) using a suitable base such as, but not limited to, Et3N, DIPEA, DMAP, or pyridine. The reaction solvent can be, but is not limited to, THF or DCM. The reaction temperature is from −20° C. to 40° C. The compound of Formula (1-2) is hydrolyzed to afford a compound of Formula (1-3) using a suitable hydroxide source such as, but not limited to, NaOH or LiOH. The compound of Formula (1-3) is reacted with a suitable chlorinating reagent such as, but not limited to, oxalyl chloride in combination with a catalytic quantity of DMF, thionyl chloride, or Ghosez's reagent to afford a compound of Formula (1-4). The reaction solvent can be, but is not limited to, THF or DCM. The reaction temperature is from −20° C. to 40° C. For the preparation of compounds of Formula (1-5), see US 2014/0018370. The compound of Formula (1-4) is reacted with a compound of Formula (1-5), wherein X1, X2, R1and R2are as previously defined, to afford a compound of Formula (1-6) using a suitable base such as, but not limited to, Et3N, DMAP, pyridine, or DIPEA. The reaction solvent can be, but is not limited to, THF, DCM, pyridine and toluene. The reaction temperature is from −20° C. to 40° C. Alternatively, the compound of Formula (1-3) is reacted with a compound of Formula (1-5) to afford a compound of Formula (1-6) using a suitable coupling reagent such as, but not limited to, BOP—Cl, CDI, DCC, EDC, HATU, PyAOP or PyBOP in the presence of a suitable base such as, but not limited to, Et3N or DIPEA. The reaction solvent can be, but is not limited to, THF, DCM and DMF. The reaction temperature is from −20° C. to 40° C. The compound of Formula (1-6) is reacted with palladium on carbon in the presence of hydrogen gas to afford a compound of Formula (1-7). The reaction solvent can be, but is not limited to, MeOH, EtOH, EtOAc, and THF. Compounds of Formula (1-7) are reacted with a suitable combination of reagents to afford compounds of Formula (Ie). The reagent combinations may be, but are not limited to:1) An aldehyde in combination with a suitable reducing agent, such as, but not limited to, NaBH4, NaBH(OAc)3, or NaBH3CN. The reaction solvent can be, but is not limited to, DCM, 1,2-DCE, or THF.2) A ketone in combination with a suitable reducing agent, such as, but not limited to, NaBH4, NaBH(OAc)3, or NaBH3CN. The reaction solvent can be, but is not limited to, DCM, 1,2-DCE, or THF.3) An alkyl halide, alkyl mesylate, or alkyl tosylate in combination with a suitable base such as, but not limited to, NaH, NaOt-Bu, KOt-Bu, Et3N, or DIPEA. The reaction solvent can be, but is not limited to, DCM or THF.4) An aryl-, heteroaryl-, or alkenyl-halide, or an aryl- or heteroaryl-, or alkenyl-triflate in combination with a suitable base, palladium(0) catalyst, ligand, and solvent. The base can be, but is not limited to, NaOt-Bu or KOt-Bu. The palladium(0) catalyst can be, but is not limited to, Pd(PPh3)4or Pd2(dba)3. The ligand can be, but is not limited to, P(o-tolyl)3or (2-biphenyl)di-tert-butylphosphine. The solvent can be, but is not limited to, toluene or THF.5) An acyl chloride in the presence of a suitable base such as, but not limited to, Et3N, DIPEA, or DMAP. The reaction solvent can be, but is not limited to, DCM or THF.6) A chloroformate in the presence of a suitable base such as, but not limited to, Et3N, DIPEA, or DMAP. The reaction solvent can be, but is not limited to, DCM or THF.7) A sulfonyl chloride in the presence of a suitable base such as, but not limited to, Et3N, DIPEA, or DMAP. The reaction solvent can be, but is not limited to, DCM or THF.8) An isocyanate in the presence of a suitable base such as, but not limited to, Et3N, DIPEA, or DMAP. The reaction solvent can be, but is not limited to, DCM or THF.9) A primary or secondary amine in the presence of a suitable activating reagent such as, but not limited to, phosgene, triphosgene, or CDI. The reaction solvent can be, but is not limited to, DCM or THF.

As shown in Scheme 2, novel analogs of the compound of Formula (Ie) are prepared from the compound of Formula (1-1) wherein X3is as previously defined. Thus, the compound of Formula (1-1) is reacted with Alloc-Cl to afford a compound of Formula (2-1) using a suitable base such as, but not limited to, Et3N, DIPEA, DMAP, or pyridine. The reaction solvent can be, but is not limited to, THF or DCM. The reaction temperature is from −20° C. to 40° C. The compound of Formula (2-1) is hydrolyzed to afford a compound of Formula (2-2) using a suitable hydroxide source such as, but not limited to, NaOH or LiOH. The compound of Formula (2-2) is reacted with a suitable chlorinating reagent such as, but not limited to, oxalyl chloride in combination with a catalytic quantity of DMF, thionyl chloride, or Ghosez's reagent to afford a compound of Formula (2-3). The reaction solvent can be, but is not limited to, THF or DCM. The reaction temperature is from −20° C. to 40° C. For the preparation of compounds of Formula (1-5), see US 2014/0018370. The compound of Formula (2-3) is reacted with a compound of Formula (1-5), wherein X1, X2, R1and R2are as previously defined, to afford a compound of Formula (2-4) using a suitable base such as, but not limited to, Et3N, DMAP, pyridine, or DIPEA. The reaction solvent can be, but is not limited to, THF, DCM, pyridine and toluene. The reaction temperature is from −20° C. to 40° C. Alternatively, the compound of Formula (2-2) is reacted with a compound of Formula (1-5) to afford a compound of Formula (2-4) using a suitable coupling reagent such as, but not limited to, BOP—Cl, CDI, DCC, EDC, HATU, PyAOP or PyBOP in the presence of a suitable base such as, but not limited to, Et3N or DIPEA. The reaction solvent can be, but is not limited to, THF, DCM and DMF. The reaction temperature is from −20° C. to 40° C. The compound of Formula (2-4) is reacted with a suitable palladium(0) catalyst in the presence of a suitable nucleophile to afford a compound of Formula (1-7). The palladium(0) catalyst can be, but is not limited to, Pd(PPh3)4or Pd2(dba)3in combination with a catalytic quantity of 1,4-bis(diphenylphosphino)butane. The nucleophile can be, but is not limited to, Et3SiH or 1,3-dimethylbarbituric acid. The solvent can be, but is not limited to, DCM or THF. Compounds of Formula (1-7) are reacted with a suitable combination of reagents to afford compounds of Formula (Ie), as described previously in Scheme 1.

As shown in Scheme 3, novel analogs of the compound of Formula (3-1) are prepared from the compound of Formula (2-4) wherein X1, X2, X3, R1, R2, and n are as previously defined. Thus, the compound of Formula (2-4) is reacted with a suitable palladium(0) catalyst such as, but not limited to, Pd(PPh3)4or Pd2(dba)3in combination with a catalytic quantity of 1,4-bis(diphenylphosphino)butane to afford a compound of Formula (3-1). The solvent can be, but is not limited to, THF or DCM.

As shown in Scheme 4, novel analogs of the compound of Formula (Ie) are prepared from the compound of Formula (4-1) wherein X3is as previously defined. Thus, the compound of Formula (4-1) is hydrolyzed to afford a compound of Formula (4-2) using a suitable hydroxide source such as, but not limited to, NaOH or LiOH. The compound of Formula (4-2) is reacted with a suitable chlorinating reagent such as, but not limited to, oxalyl chloride in combination with a catalytic quantity of DMF, thionyl chloride, or Ghosez's reagent to afford a compound of Formula (4-3). The reaction solvent can be, but is not limited to, THF or DCM. The reaction temperature is from −20° C. to 40° C. For the preparation of compounds of Formula (1-5), see US 2014/0018370. The compound of Formula (4-3) is reacted with a compound of Formula (1-5), wherein X1, X2, R1and R2are as previously defined, to afford a compound of Formula (4-4) using a suitable base such as, but not limited to, Et3N, DMAP, pyridine, or DIPEA. The reaction solvent can be, but is not limited to, THF, DCM, pyridine and toluene. The reaction temperature is from −20° C. to 40° C. Alternatively, the compound of Formula (4-2) is reacted with a compound of Formula (1-5) to afford a compound of Formula (4-4) using a suitable coupling reagent such as, but not limited to, BOP—Cl, CDI, DCC, EDC, HATU, PyAOP or PyBOP in the presence of a suitable base such as, but not limited to, Et3N or DIPEA. The reaction solvent can be, but is not limited to, THF, DCM and DMF. The reaction temperature is from −20° C. to 40° C. The compound of Formula (4-4) is reacted with a suitable acid to afford a compound of Formula (1-7). The acid can be, but is not limited to HCl or TFA. The solvent can be, but is not limited to, DCM or MeOH. Compounds of Formula (1-7) are reacted with a suitable combination of reagents to afford compounds of Formula (Ie), as described previously in Scheme 1.

EXAMPLES

A solution of LiOH (0.22 g, 6.4 mmol, 5.0 eq) in H2O (2.7 mL) was added to a solution of compound (4-5) (300 mg, 1.0 mmol, 1.0 eq) in MeOH (4.0 mL) and the reaction was stirred for 2 hrs. The reaction was partitioned between Et2O and H2O. The layers were separated and the aqueous layer was adjusted to an acidic pH (˜5) with 0.1M aqueous HCl. The resulting mixture was extracted with EtOAc (2×). The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure to afford pure compound (4-6) (286 mg, 1.0 mmol, 100%) as a colorless gum.

HATU (471 mg, 1.2 mmol, 1.2 eq) was added to a suspension of crude compound (4-6) (286 mg, 1.0 mmol, 1.0 eq) in DMF (2.7 mL). Compound (1-12) (210 mg, 1.0 mmol, 1.0 eq), prepared according to US 2014/0018370 and Hunig's base (0.45 mL, 2.6 mmol, 2.5 eq) were added and the reaction was stirred overnight. The reaction was diluted with EtOAc and the organic layer was washed with 10% citric acid (2×), H2O (1×), sat. NaHCO3(2×), and brine (1×). The organic layer was dried (MgSO4), filtered, and concentrated under reduced pressure. The resultant yellow gum was purified by column chromatography eluting with hexanes/EtOAc (0% EtOAc→50% EtOAc) to afford pure compound 183a (275 mg, 0.60 mmol, 58%) as a colorless amorphous solid.

Example 1: Synthesis of 6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-amine

Step 1. Synthesis of N-isopropyl-6-nitropicolinamide

Step 2. Synthesis of 2-(1-isopropyl-1H-tetrazol-5-yl)-6-nitropyridine

Step 3. Synthesis of 6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-amine

Step 4. Synthesis of methyl 2-isopropyl-3-oxoisoindoline-5-carboxylate

Step 5. Synthesis of 2-isopropyl-3-oxoisoindoline-5-carboxylic acid

To a solution of methyl 2-isopropyl-3-oxoisoindoline-5-carboxylate (80 mg, 0.343 mmol) in MeOH (0.86 mL) was added lithium hydroxide, 1 M solution (0.86 mL, 0.857 mmol). After stirring at rt for several hours, the reaction mixture was concentrated to remove most of the MeOH, acidified with 1N HCl, and extracted with EtOAc (3×). The combined organic layers were dried over Na2SO4and concentrated to afford the desired 2-isopropyl-3-oxoisoindoline-5-carboxylic acid (70 mg, 93% yield), which was directly used in the next step without further purification.

Step 6. Synthesis of 2-isopropyl-3-oxoisoindoline-5-carbonyl chloride

To a solution of 2-isopropyl-3-oxoisoindoline-5-carboxylic acid (70 mg, 0.319 mmol) in dry DCM (1.5 mL) at rt was added one drop of DMF and oxalyl chloride (0.117 mL, 0.234 mmol, 2 M in DCM). The suspension was stirred at rt for 45 min and turned into a clear solution. The mixture was then concentrated under reduced pressure to provide the crude 2-isopropyl-3-oxoisoindoline-5-carbonyl chloride, which was directly used in the next step.

Step 7. Synthesis of 2-isopropyl-N-(6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-yl)-3-oxoisoindoline-5-carboxamide

To a solution of 2-isopropyl-3-oxoisoindoline-5-carbonyl chloride (73.2 mg, 0.308 mmol) from the previous step in dry DCM (2 mL) at 0° C. was added 6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-amine (0.061 g, 0.30 mmol), followed by the addition of pyridine (0.24 ml, 3.00 mmol). The reaction mixture was allowed to warm to rt and stirred overnight. The reaction mixture was concentrated under reduced pressure, and then partitioned between EtOAc/H2O. The organic layer was separated, washed with water, brine, dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by SiO2column chromatography (100% hexanes to 70% EtOAc/hexanes) to provide the desired 2-isopropyl-N-(6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-yl)-3-oxoisoindoline-5-carboxamide (82 mg, 72% yield).

Example 3: Synthesis of benzyl 5-((6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)carbamoyl)isoindoline-2-carboxylate

Step 2. Synthesis of 2-((benzyloxy)carbonyl)isoindoline-5-carboxylic acid

Step 3. Synthesis of benzyl 5-(chlorocarbonyl)isoindoline-2-carboxylate

Representative Procedure for Acid Chloride Formation with Oxalyl Chloride

To a mixture of 2-((benzyloxy)carbonyl)isoindoline-5-carboxylic acid (0.75 g, 2.5 mmol) in DCM (7.2 ml) and DMF (0.02 ml, 0.25 mmol) was charged oxalyl chloride (0.38 mL, 4.3 mmol). The mixture was stirred for 3 h. The reaction was concentrated under reduced pressure and the resultant orange oil was used without purification.

Step 4. Synthesis of benzyl 5-((6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)carbamoyl)isoindoline-2-carboxylate

6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-amine was prepared according to the method disclosed in WO 2016106384, the entire contents of which are incorporated herein by reference.

Representative Procedure 1 for Amide Formation

A solution of crude benzyl 5-(chlorocarbonyl)isoindoline-2-carboxylate (398 mg, 1.26 mmol) in DCM (1.8 mL) was added to a suspension of 6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-amine (256 mg, 1.26 mmol) in pyridine (1.8 mL) and the reaction was stirred overnight. The reaction was concentrated under reduced pressure. The resultant brown residue was purified by column chromatography eluting with CH2Cl2/MeOH (0% MeOH→4% MeOH) to give benzyl 5-((6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)carbamoyl)isoindoline-2-carboxylate (454 mg, 0.941 mmol, 75% yield) as a pale yellow amorphous solid.

Example 2 was prepared according to the procedure for the synthesis of Example 3, utilizing 6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-amine in Step 4 as the amine coupling partner.

Example 9: Synthesis of N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)isoindoline-5-carboxamide

Representative Procedure for Hydrogenolysis

Pd—C (90 mg, 10% loading) was added to a solution of 5-((6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)carbamoyl)isoindoline-2-carboxylate (450 mg, 0.933 mmol) in MeOH (18.7 mL). The reaction was evacuated and backfilled with H2(3×) and the reaction was stirred overnight under a balloon of H2. The reaction was filtered through Celite, rinsing with MeOH, EtOAc, and DCM and concentrated under reduced pressure. The resultant clear residue was dissolved in DCM/EtOAc and filtered through Celite, rinsing with DCM and EtOAc. The filtrate was concentrated under reduced pressure to give N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)isoindoline-5-carboxamide (123 mg, 0.35 mmol, 38% yield) as a colorless amorphous solid.

Example 8 was prepared according to the representative procedure for hydrogenolysis.

Representative Procedure for Sulfonamide Formation

Examples 6, 14, and 15 were prepared according to the representative procedure for sulfonamide formation.

Example 5: Synthesis of N2-ethyl-N5-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)-N2-methylisoindoline-2,5-dicarboxamide

Representative Procedure for Secondary Urea Formation

Examples 11, 12, 13, and 17 were prepared according to the representative procedure for secondary urea formation.

Example 7: Synthesis of ethyl 5-((6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)carbamoyl)isoindoline-2-carboxylate

Representative Procedure for Carbamate Formation

Example 16 was prepared according to the representative procedure for carbamate formation.

Example 10: Synthesis of 2-(N,N-dimethylsulfamoyl)-N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)isoindoline-5-carboxamide

Representative Procedure for Secondary Sulfonyl Urea Formation

Step 1. Synthesis of methyl 2-chloro-5-methylisonicotinate

Step 2. Synthesis of methyl 5-(bromomethyl)-2-chloroisonicotinate

Step 3. Synthesis of 2-chloro-5-(hydroxymethyl)-N-isopropylisonicotinamide

Step 4. Synthesis of 6-chloro-2-isopropyl-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one

Representative Procedure for Palladium Catalyzed Carbonylation

Step 5. Synthesis of ethyl 2-isopropyl-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine-6-carboxylate

Representative Procedure for Amide Formation with Trimethylaluminum

Trimethylaluminum (0.15 mL, 0.29 mmol of a 2.0M solution in toluene) was added dropwise to DCM (0.46 mL) at 0° C. A solution of 6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-amine (30.0 mg, 0.15 mmol) in DCM (0.46 mL) was added at 0° C., and the mixture was stirred for 20 min at 0° C. then for 1 h at rt. A solution of ethyl 2-isopropyl-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine-6-carboxylate (36.5 mg, 0.15 mmol) in DCM (0.37 mL) was added and the reaction was heated at 35° C. overnight. The reaction was quenched with sat. potassium sodium tartrate and diluted with CH2Cl2. The layers were separated and the aqueous layer was extracted with CH2Cl2(2×). The combined organic layers were washed with brine, dried (MgSO4), filtered, and concentrated under reduced pressure. The resultant yellow gum was purified by column chromatography eluting with CH2Cl2/MeOH (0% MeOH→8% MeOH) to give 2-isopropyl-N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine-6-carboxamide (38.6 mg, 0.095 mmol, 65% yield) as a tan solid.

Example 18: Synthesis of (R)—N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-2-isopropyl-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine-6-carboxamide

Step 2. Synthesis of (R)-2-(6-nitropicolinamido)propyl acetate

Step 3. Synthesis of (R)-2-(5-(6-nitropyridin-2-yl)-1H-tetrazol-1-yl)propyl acetate

Step 4. Synthesis of (R)-2-(5-(6-aminopyridin-2-yl)-1H-tetrazol-1-yl)propyl acetate

Step 5. Synthesis of (R)-2-(5-(6-aminopyridin-2-yl)-1H-tetrazol-1-yl)propan-1-ol

Step 6. Synthesis of (R)—N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-2-isopropyl-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine-6-carboxamide

Trimethylaluminum (0.221 mL, 0.441 mmol of a 2.0M solution in toluene) was added dropwise to DCM (0.457 mL) at 0° C. A solution of (R)-2-(5-(6-aminopyridin-2-yl)-1H-tetrazol-1-yl)propan-1-ol (32.4 mg, 0.147 mmol) in DCM (0.457 mL) was added at 0° C., and the mixture was stirred for 20 mins at 0° C. then for 1 h at rt. A solution of ethyl 2-isopropyl-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine-6-carboxylate (36.5 mg, 0.147 mmol) in DCM (0.365 mL) was added and the reaction was heated at 35° C. overnight. The reaction was quenched with sat. potassium sodium tartrate and diluted with CH2Cl2. The layers were separated and the aqueous layer was extracted with CH2Cl (2×). The combined organic layers were washed with brine, dried (MgSO4), filtered, and concentrated under reduced pressure. The resultant yellow gum was purified by column chromatography eluting with CH2Cl2/MeOH (0% MeOH→5% MeOH) to give (R)—N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-2-isopropyl-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine-6-carboxamide (45.6 mg, 0.108 mmol, 73% yield) as a tan solid.

Example 53: Synthesis of benzyl 6-fluoro-7-((6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)carbamoyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate

Step 1. Synthesis of 7-bromo-6-fluoro-1,2,3,4-tetrahydroisoquinoline

Lithium triethylborohydride (9.73 ml, 9.73 mmol of a 1.0M solution in THF) was added dropwise to a solution of 7-bromo-6-fluoroisoquinoline (1.0 g, 4.42 mmol) in THF (27.6 ml) and the reaction was stirred for 5 hr. The reaction was quenched with MeOH until gas evolution ceased. The reaction was diluted with 1 M HCl and MTBE, and the layers were separated. The aqueous layer was extracted with MTBE (2×). The aqueous layer was made basic (pH 14) with 1 M NaOH, then extracted with DCM (5×). The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure to give 823 mg of crude 7-bromo-6-fluoro-1,2,3,4-tetrahydroisoquinoline as a yellow oil that was used without further purification:1H NMR (400 MHz, Chloroform-d) δ 7.18 (d, J=6.9 Hz, 1H), 6.85 (d, J=9.2 Hz, 1H), 3.95 (s, 2H), 3.10 (t, J=6.0 Hz, 2H), 2.73 (t, J=6.0 Hz, 2H).

Step 2. Synthesis of benzyl 7-bromo-6-fluoro-3,4-dihydroisoqunine-2(1H)-carboxylate

Step 3. Synthesis of 2-((benzyloxy)carbonyl)-6-fluoro-1,2,3,4-tetrahydroisoquinoline-7-carboxylic acid

Isopropylmagnesium chloride (1.4 mL, 2.7 mmol of a 2.0M solution in THF) was added dropwise (maintaining internal temperature below 5° C.) to a solution of 7-bromo-6-fluoro-3,4-dihydroisoquinoline-2(1H)-carboxylate (704 mg, 1.9 mmol) in THF (8.4 mL) at −10° C. The reaction was stirred for 1 h at −10° C. CO2was passed through a tube containing drierite and bubbled slowly through the reaction mixture (exothermic) for 15 min. The cold bath was removed and the reaction was stirred for 1 h at rt. The reaction was quenched with sat. NH4Cl and diluted with EtOAc. The layers were separated and the aqueous layer was extracted with EtOAc (2×). The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure. The resultant pale yellow oil was purified by column chromatography eluting with CH2C1/MeOH (0% MeOH→10% MeOH) to give 2-((benzyloxy)carbonyl)-6-fluoro-1,2,3,4-tetrahydroisoquinoline-7-carboxylic acid (265 mg, 0.805 mmol, 42% yield) as a pale yellow amorphous solid:1H NMR (500 MHz, Chloroform-d) δ 7.77 (br s, 1H), 7.43-7.29 (comp, 6H), 6.96 (d, J=11.0 Hz, 1H), 5.19 (s, 2H), 4.66 (s, 2H), 3.74 (br s, 2H), 2.90 (br s, 2H).

The synthesis of example 53 was completed using the representative procedure for acid chloride formation with oxalyl chloride, followed by the representative procedure 1 for amide formation.

Example 31 was prepared according to the procedure for the synthesis of example 53, utilizing 6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-amine as the amine coupling partner during the representative procedure 1 for amide formation.

Example 109 was prepared according to the procedure for the synthesis of example 53, utilizing (R)-2-(5-(6-aminopyridin-2-yl)-1H-tetrazol-1-yl)propyl acetate as the amine coupling partner during the representative procedure 1 for amide formation.

Example 83: Synthesis of benzyl 6-fluoro-7-((6-(5-isopropyl-1H-tetrazol-1-yl)pyridin-2-yl)carbamoyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate

Step 1. Synthesis of N-(6-chloropyridin-2-yl)isobutyramide

Step 2. Synthesis of (Z)—N-(6-chloropyridin-2-yl)isobutyrimidoyl chloride

To a solution of N-(6-chloropyridin-2-yl)isobutyramide (1.5 g, 7.67 mmol) in DCM (38 mL) was added PCl5(1.76 g, 8.05 mmol). The resulting clear solution was stirred at rt for 1 hr. The reaction was concentrated under reduced pressure and directly used in the next step without any purification.

Step 3. Synthesis of 2-chloro-6-(5-isopropyl-1H-tetrazol-1-yl)pyridine

Step 4. Synthesis of N-(6-(5-isopropyl-1H-tetrazol-1-yl)pyridin-2-yl)-1,1-diphenylmethanimine

Step 5. Synthesis of 6-(5-isopropyl-1H-tetrazol-1-yl)pyridin-2-amine

Example 83 was prepared according to the procedure for the synthesis of example 53, utilizing 6-(5-isopropyl-1H-tetrazol-1-yl)pyridin-2-amine as the amine coupling partner during the representative procedure 1 for amide formation.

Examples 27, 58, 71, and 81 were prepared according to the representative procedure for hydrogenolysis.

Example 20: Synthesis of 2-(N-cyclopropylsulfamoyl)-6-fluoro-N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)-1,2,3,4-tetrahydroisoquinoline-7-carboxamide

Representative Procedure for Primary Sulfonyl Urea Formation

Example 22 was prepared according to the representative procedure for primary sulfonyl urea formation.

Example 24: Synthesis of 6-fluoro-N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)-2-(3-methoxypropanoyl)-1,2,3,4-tetrahydroisoquinoline-7-carboxamide

Representative Procedure 2 for Amide Formation

Examples 25, 26, 30, and 57 were prepared according to the representative procedure for carbamate formation.

Example 29: Synthesis of N2-ethyl-6-fluoro-N7-(6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-yl)-3,4-dihydroisoquinoline-2,7(1H)-dicarboxamide

Representative Procedure for Primary Urea Formation

Examples 51, 52, and 56 were prepared according to the representative procedure for primary urea formation.

Examples 49 and 89 were prepared according to the representative procedure for secondary sulfonyl urea formation.

Example 54: Synthesis of 6-fluoro-N-(6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-yl)-2-phenyl-1,2,3,4-tetrahydroisoquinoline-7-carboxamide

Representative Procedure for C—N Coupling

A solution of Pd2(dba)3(2.9 mg, 3.15 μmol) and (±)-BINAP (3.9 mg, 6.29 μmol) in toluene (0.25 mL) was sparged with N2for 10 min. The mixture was heated at 110° C. for 10 min. The reaction was cooled to rt whereupon sodium tert-butoxide (13.3 mg, 0.138 mmol), bromobenzene (13.0 μL, 0.126 mmol), and 6-fluoro-N-(6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-yl)-1,2,3,4-tetrahydroisoquinoline-7-carboxamide (24 mg, 0.063 mmol) were added and the reaction was heated at 110° C. overnight. The reaction was filtered through Celite, rinsing with EtOAc. The filtrate was concentrated under reduced pressure and purified by column chromatography eluting with hexanes/EtOAc (0% EtOAc→50% EtOAc) to give 6-fluoro-N-(6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-yl)-2-phenyl-1,2,3,4-tetrahydroisoquinoline-7-carboxamide (6.3 mg, 0.014 mmol, 22% yield) as a pale yellow solid.

Example 75: Synthesis of (R)-6-fluoro-N7-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-N2-methyl-N2-(2,2,2-trifluoroethyl)-3,4-dihydroisoquinoline-2,7(1H)-dicarboxamide

Representative Procedure for Acetate Deprotection

Lithium hydroxide (0.056 mL, 0.056 mmol of a 1.0M solution in H2O) was added to a solution of (R)-2-(5-(6-(6-fluoro-2-(methyl(2,2,2-trifluoroethyl)carbamoyl)-1,2,3,4-tetrahydroisoquinoline-7-carboxamido)pyridin-2-yl)-1H-tetrazol-1-yl)propyl acetate (27 mg, 0.047 mmol) in MeOH (0.78 mL) and the reaction was stirred for 1 h. The reaction was concentrated under reduced pressure and the residue was diluted with DCM/EtOAc. The mixture was dried (MgSO4), filtered, and concentrated under reduced pressure. The resultant colorless residue was purified by column chromatography eluting with CH2Cl2/MeOH (0% MeOH→6% MeOH) to give (R)-6-fluoro-N7-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-N2-methyl-N2-(2,2,2-trifluoroethyl)-3,4-dihydroisoquinoline-2,7(1H)-dicarboxamide (19.7 mg, 0.037 mmol, 79% yield) as a colorless amorphous solid.

Examples 76 and 82 were prepared according to the representative procedure for acetate deprotection.

Example 67: Synthesis of benzyl 7-((6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)carbamoyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate

Step 2. Synthesis of 2-((benzyloxy)carbonyl)-1,2,3,4-tetrahydroisoquinoline-7-carboxylic acid

Example 37 was prepared according to the representative procedure for hydrogenolysis.

Examples 32 and 60 were prepared according to the representative procedure 2 for amide formation.

Examples 33, 61, 62, and 63 were prepared according to the representative procedure for primary urea formation.

Examples 34, 35, 36, 64, 65, and 66 were prepared according to the representative procedure for carbamate formation.

Representative Procedure for Reductive Alkylation

Sodium triacetoxyborohydride (85 mg, 0.40 mmol, 5.0 eq) was added to a solution of N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)-1,2,3,4-tetrahydroisoquinoline-7-carboxamide (29 mg, 0.08 mmol, 1.0 eq) and 3-methoxypropanal (35 μL, 0.40 mmol, 5.0 eq) in 1,2-DCE (1 mL) and the reaction was stirred overnight. The reaction was quenched with sat. NaHCO3and diluted with DCM. The layers were separated and the aqueous layer was extracted with DCM (2×). The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure. The resultant yellow residue was purified by column chromatography eluting with DCM/MeOH (0% MeOH→20% MeOH) to afford N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)-2-(3-methoxypropyl)-1,2,3,4-tetrahydroisoquinoline-7-carboxamide (4.1 mg, 9.4 μmol, 12%) as a yellow gum.

Example 59: Synthesis of allyl 7-((6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)carbamoyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate

Step 2. Synthesis of 2-((allyloxy)carbonyl)-1,2,3,4-tetrahydroisoquinoline-7-carboxylic acid

Step 3. Synthesis of allyl 7-(chlorocarbonyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate

Oxalyl chloride (0.17 mL, 1.9 mmol, 1.7 eq) was added dropwise to a mixture of 2-((allyloxy)carbonyl)-1,2,3,4-tetrahydroisoquinoline-7-carboxylic acid (291 mg, 1.1 mmol, 1.0 eq) and DMF (8.6 μL, 0.11 mmol, 0.1 eq) in DCM (3.1 mL). The mixture was stirred for 1 hr. The reaction was concentrated under reduced pressure to afford crude allyl 7-(chlorocarbonyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate which was used without purification.

Step 4. Synthesis of allyl 7-((6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)carbamoyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate

A solution of 6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-amine (205 mg, 1.0 mmol, 1.0 eq), prepared according to US 2014/0018370, in pyridine (2.7 mL) was added to crude allyl 7-(chlorocarbonyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (312 mg, 1.1 mmol, 1.1 eq) and the reaction was stirred overnight. The reaction was concentrated under reduced pressure. The residue was dissolved in 7N NH3in MeOH (7.9 mL) and stirred for 1 hr. The reaction was concentrated under reduced pressure. The resultant brown residue was purified by column chromatography eluting with DCM/MeOH (0% MeOH→6% MeOH) to afford allyl 7-((6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)carbamoyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (183 mg, 1.0 mmol, 41%) as a tan amorphous solid.

Example 39: Synthesis of 2-allyl-N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)-1,2,3,4-tetrahydroisoquinoline-7-carboxamide

A solution of Pd(PPh3)4(2.7 mg, 2.4 μmol, 0.05 eq) in DCM (0.12 mL) was added to a solution of allyl 7-((6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)carbamoyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (21 mg, 0.05 mmol, 1.0 eq) in DCM (0.45 mL) and the reaction was stirred for 5 hrs. The reaction was loaded directly onto a column and purified by column chromatography eluting with DCM/MeOH (0% MeOH→15% MeOH) to afford 2-allyl-N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)-1,2,3,4-tetrahydroisoquinoline-7-carboxamide (12 mg, 0.03 mmol, 63%) as a colorless amorphous solid.

Example 74: Synthesis of benzyl 6-fluoro-7-((6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-yl)carbamoyl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

Step 1. Synthesis of 7-bromo-6-fluoro-1,2,3,4-tetrahydroisoquinoline

Sodium borohydride (1.5 g, 38.7 mmol) was added portion wise to a solution of 7-bromo-6-fluoroisoquinoline (2.5 g, 11.1 mmol) in AcOH (55.3 ml) and the reaction was stirred for 2 hr. The reaction was quenched carefully with sat. NaHCO3(dropwise, 850 mL) until the mixture was pH ˜8, then diluted with CH2Cl2(200 mL). The layers were separated and the aqueous layer was extracted with CH2Cl2(2×150 mL). The combined organic layers were washed with brine, dried (MgSO4), filtered, and concentrated under reduced pressure to give 2.29 g of crude 7-bromo-6-fluoro-1,2,3,4-tetrahydroisoquinoline as a yellow solid that was used without purification:1H NMR (400 MHz, Chloroform-d) δ 7.19 (d, J=7.0 Hz, 1H), 6.85 (dd, J=9.2, 2.5 Hz, 1H), 3.95 (s, 2H), 3.11 (t, J=6.0 Hz, 2H), 2.74 (t, J=6.0 Hz, 2H).

Step 2. Synthesis of 7-bromo-6-fluoro-3,4-dihydroisoquinoline

N-Bromosuccinimide (2.2 g, 12.1 mmol) was added portion wise over 20 minutes to a solution of 7-bromo-6-fluoro-1,2,3,4-tetrahydroisoquinoline (2.5 g, 11.0 mmol) in DCM (58.3 mL) and the reaction was stirred for 2 hr. 30% NaOH (15.3 ml) was added and the reaction was stirred for 2.5 hr. The layers were separated and the aqueous layer was extracted with DCM (2×). The organic layer was washed with H2O. The organic layer was extracted with 10% HCl (3×). The combined acidic extracts were washed with DCM, then made basic with concentrated ammonium hydroxide. This mixture was extracted with DCM (3×). The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure. The resultant clear gum was purified by column chromatography eluting with hexanes/EtOAc (50% EtOAc→100% EtOAc) to give 7-bromo-6-fluoro-3,4-dihydroisoquinoline (1.23 g, 5.4 mmol, 49% yield) over two steps as a yellow solid:1H NMR (400 MHz, Chloroform-d) δ 8.29 (t, J=2.3 Hz, 1H), 7.50 (d, J=6.7 Hz, 1H), 6.96 (dt, J=8.5, 0.9 Hz, 1H), 3.82-3.75 (m, 2H), 2.79-2.70 (m, 2H).

Step 3. Synthesis of benzyl 7-bromo-6-fluoro-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

TMSOTf (0.31 mL, 1.71 mmol) was added dropwise to a solution of 7-bromo-6-fluoro-3,4-dihydroisoquinoline (300 mg, 1.315 mmol) in 1,2-dimethoxyethane (1,2-DME) (6.6 mL) at −40° C. The reaction was stirred for 5 min at −40° C., then methylmagnesium chloride (0.88 mL, 2.63 mmol of a 3.0M solution in THF) was added dropwise and the reaction was stirred at −40° C. for 10 min. The cold bath was removed, and the reaction was stirred at rt for 30 min. The reaction was cooled to −40° C., and Cbz-Cl (0.56 mL, 3.95 mmol) was added dropwise. The cold bath was removed and the reaction was stirred for 80 min. The reaction was quenched with sat. NH4Cl and diluted with H2O and EtOAc. The layers were separated and the aqueous layer was extracted with EtOAc (2×). The combined organic layers were washed with sat. NaHCO3, dried (MgSO4), filtered, and concentrated under reduced pressure. The resultant yellow oil was purified by column chromatography eluting with hexanes/EtOAc (0% EtOAc→20% EtOAc) to give benzyl 7-bromo-6-fluoro-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate (442 mg, 1.17 mmol, 89% yield) as a colorless gum:1H NMR (400 MHz, Chloroform-d) δ 7.44-7.27 (comp, 6H), 6.88 (d, J=9.0 Hz, 1H), 5.29-5.13 (comp, 3H), 4.34-4.06 (m, 1H), 3.37-3.10 (m, 1H), 2.93-2.80 (m, 1H), 2.69 (d, J=16.5 Hz, 1H), 1.44 (d, J=6.8 Hz, 3H).

Step 4. Synthesis of 2-((benzyloxy)carbonyl)-6-fluoro-1-methyl-1,2,3,4-tetrahydroisoquinoline-7-carboxylic acid

Step 5. Synthesis of benzyl 6-fluoro-7-((6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-yl)carbamoyl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

Ghosez's Reagent (0.14 mL, 1.06 mmol) was added dropwise to a solution of 2-((benzyloxy)carbonyl)-6-fluoro-1-methyl-1,2,3,4-tetrahydroisoquinoline-7-carboxylic acid (180 mg, 0.52 mmol) in DCM (1.1 mL) and the reaction was stirred for 3 h. The reaction was concentrated under reduced pressure, chased with DCM, and dried under hi-vac. The resultant residue was dissolved in DCM (1.1 mL) and cooled to 0° C. 6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-amine (102 mg, 0.50 mmol) and pyridine (0.16 mL, 2.0 mmol) were added, and the reaction was stirred overnight, slowly warming to rt. The reaction was concentrated under reduced pressure. The resultant yellow gum was purified by column chromatography eluting with hexanes/EtOAc (0% EtOAc→45% EtOAc) to give benzyl 6-fluoro-7-((6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-yl)carbamoyl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate (233 mg, 0.44 mmol, 88% yield) as a colorless gum.

Example 68 was prepared according to the representative procedure for hydrogenolysis.

Example 69 was prepared according to the representative procedure for secondary urea formation.

Example 70 was prepared according to the representative procedure for sulfonamide formation.

Example 108: Synthesis of 6-fluoro-N-(6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-yl)-1-methyl-2-(2-oxo-2-(pyrrolidin-1-yl)ethyl)-1,2,3,4-tetrahydroisoquinoline-7-carboxamide

Example 98: Synthesis of 2-ethyl-6-fluoro-N-(6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-yl)-1-oxo-1,2,3,4-tetrahydroisoquinoline-7-carboxamide

Step 1. Synthesis of 7-bromo-6-fluoro-3,4-dihydroisoquinolin-1(2H)-one

Step 2. Synthesis of 7-bromo-2-ethyl-6-fluoro-3,4-dihydroisoquinolin-1(2H)-one

Step 3. Synthesis of 2-isocyanato-6-(1-isopropyl-1H-tetrazol-5-yl)pyridine

Triphosgene (19 mg, 0.063 mmol) was added portion wise to a solution of 6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-amine (36.0 mg, 0.176 mmol) in DCE (0.77 mL). Et3N (50 μL, 0.352 mmol) was added and the reaction was stirred at 50° C. for 2.5 h. The reaction was concentrated to give a mixture of 2-isocyanato-6-(1-isopropyl-1H-tetrazol-5-yl)pyridine and triethylammonium chloride. This mixture was used without purification.

Step 4. Synthesis of 2-ethyl-6-fluoro-N-(6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-yl)-1-oxo-1,2,3,4-tetrahydroisoquinoline-7-carboxamide

Isopropylmagnesium chloride (81 μL, 0.162 mmol) was added dropwise to a solution of 7-bromo-2-ethyl-6-fluoro-3,4-dihydroisoquinolin-1(2H)-one (40 mg, 0.147 mmol) in THF (0.3 mL) at −10° C. and the reaction was stirred at −10° C. for 1 h. This Grignard solution was added dropwise to a mixture of crude 2-isocyanato-6-(1-isopropyl-1H-tetrazol-5-yl)pyridine (41 mg, 0.176 mmol) in THF (0.7 mL) at 0° C. The cold bath was removed and the reaction was stirred overnight. The reaction was quenched carefully with sat. NH4Cl and diluted with EtOAc. The layers were separated and the aqueous layer was extracted with EtOAc (2×). The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure. The resultant yellow gum was purified by column chromatography eluting with hexanes/EtOAc (0% EtOAc→80% EtOAc) to give 2-ethyl-6-fluoro-N-(6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-yl)-1-oxo-1,2,3,4-tetrahydroisoquinoline-7-carboxamide (2.9 mg, 6.85 μmol, 5% yield) as an orange residue.

The synthesis of example 153 was completed using the representative procedure for amide formation with trimethylaluminum, reacting with 6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-amine.

Example 110: Synthesis of 6-(cyclopropylsulfonyl)-N-(6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide

Step 1. Synthesis of N-(6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide hydrochloride

The synthesis of example 110 was completed using the representative procedure for sulfonamide formation, with the following modification: 8 equivalents of Et3N were used.

Examples 138, 139, 140, 141, and 172 were prepared according to the representative procedure for sulfonamide formation, with the following modification: 8 equivalents of Et3N were used.

Example 111 was prepared according to the representative procedure for secondary sulfonyl urea formation, with the following modification: 8 equivalents of Et3N were used.

Examples 112-116, 137, and 173 were prepared according to the representative procedure for secondary urea formation, with the following modification: 8 equivalents of Et3N were used.

Example 117 was prepared according to the representative procedure for primary urea formation, with the following modification: 8 equivalents of Et3N were used.

Example 118 was prepared according to the representative procedure for carbamate, with the following modification: 8 equivalents of Et3N were used.

Example 119 was prepared according to the representative procedure 2 for amide formation, with the following modification: 8 equivalents of Et3N were used.

Example 143 was prepared according to the representative procedure for amide formation with trimethylaluminum, reacting with (R)-2-(5-(6-aminopyridin-2-yl)-1H-tetrazol-1-yl)propan-1-ol, with the following modification: 3 equivalents of trimethylaluminum were used.

Example 142: Synthesis of (R)-6-formyl-N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide

Step 1. Synthesis of (R)—N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide hydrochloride

Step 2. Synthesis of (R)—N-(6-(1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-6-formyl-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide

Step 3. Synthesis of (R)-6-formyl-N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide

Concentrated HCl (22.00 μl, 0.264 mmol) was added to a solution of (R)—N-(6-(1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-6-formyl-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide (23 mg, 0.044 mmol) in MeOH (147 μl) and the reaction was stirred for 1 h. The reaction was quenched carefully with sat. NaHCO3and diluted with EtOAc. The layers were separated and the aqueous layer was extracted with EtOAc (2×). The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure. The resultant white solid was purified by column chromatography eluting with CH2Cl2/MeOH (0% MeOH→8% MeOH) to give (R)-6-formyl-N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide (7 mg, 0.017 mmol, 39% yield) as a colorless solid.

Example 144: Synthesis of (R)—N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-6-(pyrrolidine-1-carbonyl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide

Step 1. Synthesis of (R)—N-(6-(1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide

Step 2. Synthesis of (R)—N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-6-(pyrrolidine-1-carbonyl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide

Pyrrolidine-1-carbonyl chloride (5.1 μL, 0.045 mmol) was added dropwise to a solution of (R)—N-(6-(1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide (15 mg, 0.030 mmol), Et3N (9.3 μL, 0.067 mmol), and DMAP (0.4 mg, 3.03 μmol) in DCM (303 μl) and the reaction was stirred for 1.5 hrs. Another 3 eq (10.2 μL) of pyrrolidine-1-carbonyl chloride and 2.2 eq (9.3 μL) of Et3N were added and the reaction was stirred another 1.5 hrs. TBAF (182 μl, 0.182 mmol of a 1.0M solution in THF) was added and the reaction was stirred overnight. The reaction was quenched with sat. NaHCO3and diluted with CH2Cl2. The layers were separated and the organic layer was washed with sat. NaHCO3. The organic layer was dried (MgSO4), filtered, and concentrated under reduced pressure. The resultant clear residue was purified by column chromatography eluting with CH2Cl2/MeOH (0% MeOH→5% MeOH, 4 g column) to give (R)—N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-6-(pyrrolidine-1-carbonyl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide (9.1 mg, 0.019 mmol, 63% yield) as a clear gum.

Example 146: Synthesis of (R)-6-(cyclopentanecarbonyl)-N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide

Cyclopentanecarbonyl chloride (5.5 μL, 0.045 mmol) was added dropwise to a solution of (R)—N-(6-(1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide (15 mg, 0.030 mmol), Et3N (9.3 μL, 0.067 mmol), and DMAP (0.4 mg, 3.03 μmol) in DCM (303 μl) and the reaction was stirred for 1.5 hrs. TBAF (182 μl, 0.182 mmol of a 1.0M solution in THF) was added and the reaction was stirred overnight. The reaction was quenched with sat. NaHCO3and diluted with CH2Cl2. The layers were separated and the organic layer was washed with sat. NaHCO3. The organic layer was dried (MgSO4), filtered, and concentrated under reduced pressure. The resultant clear residue was purified by column chromatography eluting with CH2Cl2/MeOH (0% MeOH 5% MeOH) to give (R)-6-(cyclopentanecarbonyl)-N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide (6.5 mg, 0.014 mmol, 45% yield) as a colorless solid.

Example 162: Synthesis of isopropyl (R)-7-((6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)carbamoyl)-3,4-dihydro-2,6-naphthyridine-2(1H)-carboxylate

Representative Procedure for N-Functionalization and TBS Deprotection

Isopropyl carbonochloridate (76 μl, 0.076 mmol) was added dropwise to a solution of (R)—N-(6-(1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide (25 mg, 0.051 mmol), Et3N (15.5 μl, 0.111 mmol), and DMAP (0.62 mg, 5.05 μmol) in DCM (505 μl) and the reaction was stirred for 3 hrs. Concentrated HCl (25.3 μl1, 0.303 mmol) was added and the reaction was stirred for 3 hrs. The reaction was quenched carefully with sat. NaHCO3and diluted with CH2Cl2. The layers were separated and the aqueous layer was extracted with CH2Cl2(2×). The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure. The resultant colorless gum was purified by column chromatography eluting with CH2Cl2/MeOH (0% MeOH→5% MeOH) to give (R)-7-((6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)carbamoyl)-3,4-dihydro-2,6-naphthyridine-2(1H)-carboxylate (14.1 mg, 0.030 mmol, 60% yield) as a colorless solid.

Examples 126-128, 145, 156-160, and 193-195 were prepared according to the representative procedure for N-functionalization and TB S deprotection, utilizing the corresponding chloroformate reagents.

Examples 120, 148-150, 161, and 200 were prepared according to the representative procedure for N-functionalization and TB S deprotection, utilizing the corresponding sulfonyl chloride reagents.

Examples 121-125, 151, 152, 155, and 191-192 were prepared according to the representative procedure for N-functionalization and TB S deprotection, utilizing the corresponding acid chloride reagents.

Examples 129-132, 169-170, and 196 were prepared according to the representative procedure for N-functionalization and TBS deprotection, utilizing the corresponding isocyanate reagents.

Examples 154, 166, and 199 were prepared according to the representative procedure for N-functionalization and TBS deprotection, utilizing the corresponding sulfamoyl chloride reagents.

Examples 167-168 and 197 were prepared according to the representative procedure for N-functionalization and TBS deprotection, utilizing the corresponding carbamoyl chloride reagents.

Example 163: Synthesis of (R)-6-(cyclopropylmethyl)-N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide

NaBH(OAc)3(38.6 mg, 0.8 mmol) was added to a solution of (R)—N-(6-(1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide (30 mg, 0.06 mmol), and cyclopropanecarbaldehyde (6.4 mg, 0.09 mmol) in DCE (5 mL) and the resulting mixture was stirred for 2 h at rt under a nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure. The residue was diluted with THF (5 mL), then concentrated HCl (0.2 mL) was added and the reaction was stirred for another 1 h. The reaction was neutralized with saturated NaHCO3and extracted with EtOAc (2×20 mL). The combined organic layers were dried (Na2SO4), filtered, and concentrated under reduced pressure. The crude residue was purified by reverse phase column chromatography eluting with H2O/MeCN (0% MeCN→55% MeCN) to afford (R)-6-(cyclopropylmethyl)-N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide (10.7 mg) as an off-white solid.

Examples 133-136, 164, 165, 171, 181, and 198 were prepared according to the procedure for the synthesis of example 163.

Step 1. Synthesis of methyl 5-bromo-4-fluoro-2-methylbenzoate

Step 2. Synthesis of methyl 5-bromo-2-(bromomethyl)-4-fluorobenzoate

A mixture of 5-bromo-4-fluoro-2-methylbenzoate (4.2 g, 17.0 mmol), benzoyl peroxide (41 mg, 0.17 mmol), and NBS (3.1 g, 17.34 mmol) in CHCl3(43.6 ml) was heated at reflux overnight. The reaction was quenched with H2O/brine and diluted with CH2Cl2. The layers were separated and the aqueous layer was extracted with CH2Cl2(2×). The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure to give 5.9 g of a mixture of three compounds with the major component being methyl 5-bromo-2-(bromomethyl)-4-fluorobenzoate (˜4.2 g, 76% yield). The mixture was used directly without purification.

A mixture of crude methyl 5-bromo-2-(bromomethyl)-4-fluorobenzoate (4.2 g, 12.89 mmol) and calcium carbonate (7.74 g, 77 mmol) in H2O (129 ml)/1,4-Dioxane (129 ml) was heated at reflux for 3.5 hrs. The reaction was cooled to rt and filtered to remove solids. The filtrate was concentrated under reduced pressure to remove dioxane. The resultant aqueous mixture was extracted DCM (4×). The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure. The resultant white solid was recrystallized from DCM/hexanes to give 6-bromo-5-fluoroisobenzofuran-1(3H)-one (2.3 g, 9.96 mmol, 58% yield over 2 steps) as a white solid:1H NMR (400 MHz, Chloroform-d) δ 8.16-8.12 (m, 1H), 7.26 (d, J=7.2 Hz, 1H), 5.27 (s, 2H).

Step 4. Synthesis of 5-bromo-4-fluoro-2-(hydroxymethyl)-N-isopropylbenzamide

Step 5. Synthesis of 6-bromo-5-fluoro-2-isopropylisoindolin-1-one

The synthesis of example 177 was completed according to the representative procedure for palladium catalyzed carbonylation and the representative procedure for amide formation with trimethylaluminum, reacting with 6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-amine.

Example 175: Synthesis of 2-isopropyl-N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)-6-methoxy-3-oxoisoindoline-5-carboxamide

K2CO3(25.2 mg, 0.182 mmol) was added to a solution of 6-fluoro-2-isopropyl-N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)-3-oxoisoindoline-5-carboxamide (15.4 mg, 0.036 mmol) in MeOH (0.608 mL) and the reaction was stirred overnight. The reaction was quenched with H2O and diluted with CH2Cl2. The layers were separated and the aqueous layer was extracted with CH2Cl2(2×). The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure to give 2-isopropyl-N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)-6-methoxy-3-oxoisoindoline-5-carboxamide (13.6 mg, 0.031 mmol, 86% yield) as a colorless solid.

Examples 176 and 174 were prepared in an analogous fashion utilizing the same representative procedures as examples 177 and 175, respectively, and performing the amide formation with 6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-amine.

Example 179: Synthesis of benzyl 6-((6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)carbamoyl)-1,3-dihydro-2H-pyrrolo[3,4-c]pyridine-2-carboxylate

Step 1. Synthesis of benzyl 6-chloro-1,3-dihydro-2H-pyrrolo[3,4-c]pyridine-2-carboxylate

The synthesis of example 179 was completed according to the representative procedure for palladium catalyzed carbonylation and the representative procedure for amide formation with trimethylaluminum, reacting with 6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-amine.

Examples 178 and 180 were prepared in an analogous fashion utilizing the same representative procedures as example 179, reacting with the appropriate amine during the amide coupling step.

Example 189: Synthesis of ethyl 6-((6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)carbamoyl)-1,3-dihydro-2H-pyrrolo[3,4-c]pyridine-2-carboxylate

Step 1. Synthesis of N-(6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine-6-carboxamide hydrochloride

The synthesis of example 189 was completed according to the representative procedure for carbamate formation, with the following modification: 3.3 equivalents of Et3N were used.

Example 189: Synthesis of 6-((1R,5S)-8-azabicyclo[3.2.1]octane-8-carbonyl)-N-(6-(1-((R)-1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide

4-Nitrophenyl carbonochloridate (242 mg, 1.2 mmol) was added to (R)—N-(6-(1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide (500 mg, 1.0 mmol), and Et3N (303 mg, 3 mmol) in DCM (20 mL) and the mixture was stirred at room temperature overnight. The reaction was diluted with DCM (30 mL) and washed sequentially with H2O (20 mL) and brine (20 mL). The organic phase was dried (Na2SO4), filtered, and concentrated under reduced pressure. The crude residue was purified by column chromatography eluting with DCM/MeOH (50:1) to afford 4-nitrophenyl (R)-7-((6-(1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)carbamoyl)-3,4-dihydro-2,6-naphthyridine-2(1H)-carboxylate (580 mg, 87%) as an off white solid.

Step 2. Synthesis of 6-((1R,5S)-8-azabicyclo[3.2.1]octane-8-carbonyl)-N-(6-(1-((R)-1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide

A mixture of 4-nitrophenyl (R)-7-((6-(1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)carbamoyl)-3,4-dihydro-2,6-naphthyridine-2(1H)-carboxylate (80 mg, 0.12 mmol), K2CO3(52.79 mg, 0.38 mmol) and 8-aza-bicyclo[3.2.1]octane hydrochloride (89.5 mg, 0.61 mmol) in acetonitrile (5 mL) was stirred for 3 h at 80° C. The reaction was diluted with EtOAc (40 mL) and washed sequentially with H2O (20 mL) and brine (20 mL). The organic layer was dried (Na2SO4), filtered, and concentrated under reduced pressure to give crude 6-((1R,5 S)-8-azabicyclo[3.2.1]octane-8-carbonyl)-N-(6-(1-((R)-1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide that was used directly in the next step.

Crude 6-((1R,5S)-8-azabicyclo[3.2.1]octane-8-carbonyl)-N-(6-(1-((R)-1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide was diluted with THF (3 mL) and concentrated HCl (0.2 mL) was added and the reaction was stirred for 1 h at rt. The reaction was neutralized with saturated NaHCO3, and extracted with EtOAc (2×20 mL). The combined organic layers were dried (Na2SO4), filtered, and concentrated under reduced pressure. The crude residue was purified by reverse phase column chromatography eluting with H2O/MeCN (0% MeCN→70% MeCN over 15 minutes) to afford 6-((1R,5 S)-8-azabicyclo[3.2.1]octane-8-carbonyl)-N-(6-(1-((R)-1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide (13.1 mg) as an off-white solid.

Example 184: Synthesis of (R)—N2-cyclopentyl-N7-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-N2-methyl-3,4-dihydro-2,6-naphthyridine-2,7(1H)-dicarboxamide

Step 1. Synthesis of (R)-7-((6-(1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)carbamoyl)-3,4-dihydro-2,6-naphthyridine-2(1H)-carbonyl chloride

To a solution of (R)—N-(6-(1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide (50 mg, 0.10 mmol) and TEA (30 mg, 0.30 mmol) in THF was added trichloromethyl carbonochloridate (9 mg, 0.05 mmol) at rt under nitrogen atmosphere. The reaction was stirred for 30 minutes and the solution was used directly in the next step.

Step 2. Synthesis of (R)—N2-cyclopentyl-N7-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-N2-methyl-3,4-dihydro-2,6-naphthyridine-2,7(1H)-dicarboxamide

To the above solution was added N-methylcyclopentanamine hydrochloride (21 mg, 0.16 mmol) and the reaction was stirred for 1.5 h at rt under a nitrogen atmosphere. Concentrated HCl (0.2 mL) was added, and the reaction was stirred for 1 h at rt. The reaction was neutralized with saturated NaHCO3, and extracted with EtOAc (2×20 mL). The organic phase was dried (Na2SO4), filtered, and concentrated under reduced pressure. The crude residue was purified by reverse phase column chromatography eluting with H2O/MeCN (0% MeCN→70% MeCN, 15 minute gradient) to afford (R)—N2-cyclopentyl-N7-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-N2-methyl-3,4-dihydro-2,6-naphthyridine-2,7(1H)-dicarboxamide (15 mg) as an off-white solid.

Examples 183, 186, and 187 were prepared according to the procedure for the synthesis of example 184.

Example 185: Synthesis of (R)-6-(azetidine-1-carbonyl)-N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide

To a solution of 4-nitrophenyl (R)-7-((6-(1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)carbamoyl)-3,4-dihydro-2,6-naphthyridine-2(1H)-carboxylate (50 mg, 0.08 mmol) in THF was added concentrated HCl (0.2 mL) and the reaction was stirred for 1 h at rt. The reaction was neutralized with saturated NaHCO3, and extracted with EtOAc (2×20 mL). The combined organic layers were dried (Na2SO4), filtered, and concentrated under reduced pressure to afford 4-nitrophenyl (R)-7-((6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)carbamoyl)-3,4-dihydro-2,6-naphthyridine-2(1H)-carboxylate (37.5 mg, 91%) as an off-white solid.

Step 2. Synthesis of (R)-6-(azetidine-1-carbonyl)-N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide

A mixture of 4-nitrophenyl (R)-7-((6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)carbamoyl)-3,4-dihydro-2,6-naphthyridine-2(1H)-carboxylate (37.5 mg, 0.07 mmol), azetidine (5.9 mg, 0.10 mmol) and K2CO3(28.5 mg, 0.21 mmol) in acetonitrile was stirred for 3 h at 75° C. The mixture was filtered and concentrated under reduced pressure. The crude product was purified column chromatography eluting with PE/EtOAc (1:1) to afford (R)-6-(azetidine-1-carbonyl)-N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide (10.2 mg) as an off-white solid.

NaH (12.1 mg, 0.51 mmol) was added to a solution of (1-ethyl-1H-pyrazol-4-yl)methanol (25.5 mg, 0.20 mmol) in THF (3 mL) and the reaction was stirred for 30 min at rt. This mixture was added to the crude solution of (R)-7-((6-(1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-1 H-tetrazol-5-yl)pyridin-2-yl)carbamoyl)-3,4-dihydro-2,6-naphthyridine-2(1H)-carbonyl chloride and the reaction was stirred overnight. The reaction was quenched with sat. NH4Cl, and the aqueous layer was extracted with CH2Cl2(2×50 mL). The combined organic phase was dried (Na2SO4), filtered, and concentrated under reduced pressure. The crude residue was diluted with THF (5 mL) and concentrated HCl (0.2 mL) was added. After stirring for 1 h, the mixture was neutralized with saturated NaHCO3and extracted with EtOAc (2×20 mL). The combined organic layers were dried (Na2SO4), filtered, and concentrated under reduced pressure. The crude residue was purified by reverse phase column chromatography eluting with H2O/MeCN (0% MeCN→55% MeCN) to afford (1-ethyl-1H-pyrazol-4-yl)methyl (R)-7-((6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)carbamoyl)-3,4-dihydro-2,6-naphthyridine-2(1H)-carboxylate (26 mg) as an off-white solid.

4-Nitrophenyl carbonochloridate (85.6 mg, 0.42 mmol) was added to 8-methyl-8-aza-bicyclo[3.2.1]octan-3-ol (50 mg, 0.35 mmol), and TEA (106 mg, 1.05 mmol) in DCM (5 mL). After stirring overnight at rt, the solution was diluted with EtOAc (20 mL), and washed sequentially with H2O (10 mL) and brine (10 mL). The organic layer was dried (Na2SO4), filtered, and concentrated under reduced pressure to give crude 8-methyl-8-azabicyclo[3.2.1]octan-3-yl (4-nitrophenyl) carbonate that was used without purification.

A solution of crude 8-methyl-8-azabicyclo[3.2.1]octan-3-yl (4-nitrophenyl) carbonate was dissolved in acetonitrile (2 mL) was added to a mixture of (R)—N-(6-(1-(1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide hydrochloride (20 mg, 0.05 mmol), and K2CO3(138 mg, 1 mmol) in acetonitrile (2 mL). The mixture was stirred at 50° C. for 2 hours. The reaction was cooled to rt, filtered and concentrated under reduced pressure. The crude residue was purified by reverse phase column chromatography eluting with H2O/MeCN (0% MeCN→80% MeCN) to afford 8-methyl-8-azabicyclo[3.2.1]octan-3-yl 7-((6-(1-((R)-1-hydroxypropan-2-yl)-1H-tetrazol-5-yl)pyridin-2-yl)carbamoyl)-3,4-dihydro-2,6-naphthyridine-2(1H)-carboxylate (8 mg) as an off-white solid.

Example 202: Synthesis of (R)-6-(4-fluorobenzoyl)-N-(6-(4-(1-hydroxypropan-2-yl)-4H-1,2,4-triazol-3-yl)pyridin-2-yl)-5,6,7,8-tetrahydro-2,6-naphthyridine-3-carboxamide

Step 1. Synthesis of 6-aminopicolinohydrazide

Hydrazine hydrate (32.9 g, 658.00 mmol) was added to a mixture of methyl 6-aminopicolinate (20 g, 131.45 mmol) in EtOH (200 mL), and the resulting solution was stirred for 2 h at 80° C. The reaction was cooled to rt and the solids were collected by filtration and dried in vacuo to give 6-aminopicolinohydrazide (18.2 g, 90.5%) as an off-white solid.

Step 2. Synthesis of (E)-N′-(6-aminopicolinoyl)-N,N-dimethylformohydrazonamide

DMF-DMA (21.5 g, 180.4 mmol) was added to a solution of 6-aminopicolinohydrazide (18.3 g, 120.27 mmol) in PhMe (200 mL), and the resulting solution was stirred overnight at 50° C. The reaction was cooled to rt and the solids were collected by filtration and dried in vacuo to give (E)-N′-(6-aminopicolinoyl)-N,N-dimethylformohydrazonamide (23.0 g, 92.3%) as a light yellow solid.

Step 3. Synthesis of (R)-2-(3-(6-aminopyridin-2-yl)-4H-1,2,4-triazol-4-yl)propan-1-ol

(R)-2-aminopropan-1-ol (25 g, 332.97 mmol) was added to a mixture of ((E)-N′-(6-aminopicolinoyl)-N,N-dimethylformohydrazonamide (23 g, 110.99 mmol) in acetic acid (24 mL) and toluene (120 mL). The resulting solution was stirred overnight at 80° C. The reaction was concentrated under reduced pressure. The crude residue was purified by column chromatography eluting with DCM/MeOH (0% MeOH→10% MeOH), then further purified by reverse phase prep HPLC eluting with H2O/CH3CN (0% CH3CN→20% CH3CN) to give (R)-2-(3-(6-aminopyridin-2-yl)-4H-1,2,4-triazol-4-yl)propan-1-ol (7.8 g, 32%) as an off-white solid.

The synthesis of example 202 was completed from (R)-2-(3-(6-aminopyridin-2-yl)-4H-1,2,4-triazol-4-yl)propan-1-ol and in an analogous fashion to example 162 utilizing the representative procedure for N-functionalization and TB S deprotection and the corresponding acid chloride.

Examples 204 and 205 were prepared according to the representative procedure for N-functionalization and TBS deprotection, utilizing the corresponding chloroformate reagents.

Examples 211 and 212 were prepared according to the representative procedure for N-functionalization and TBS deprotection, utilizing the corresponding sulfonyl chloride reagents.

Example 203 was prepared according to the representative procedure for N-functionalization and TBS deprotection, utilizing the corresponding acid chloride reagents.

Example 206 was prepared according to the representative procedure for N-functionalization and TBS deprotection, utilizing the corresponding isocyanate reagents.

Example 210 was prepared according to the representative procedure for N-functionalization and TBS deprotection, utilizing the corresponding sulfamoyl chloride reagents.

Example 207 was prepared according to the representative procedure for N-functionalization and TBS deprotection, utilizing the corresponding carbamoyl chloride reagents.

Examples 208 and 209 were prepared according to the procedure for the synthesis of example 163.

Example 201 was prepared according to the procedure for the synthesis of example 110, utilizing 6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-amine during the representative procedure for amide formation with trimethylaluminum.

Example 200 was prepared in analogous fashion to the sythesis of example 201.

Examples 191 and 192 were prepared according to the procedure for the synthesis of example 119, utilizing 6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-amine during the representative procedure for amide formation with trimethylaluminum.

Examples 193 and 194 were prepared according to the procedure for the synthesis of example 118, utilizing 6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-amine during the representative procedure for amide formation with trimethylaluminum.

Example 195 was prepared according to the procedure for the synthesis of example 117, utilizing 6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-amine during the representative procedure for amide formation with trimethylaluminum.

Example 196 was prepared according to the procedure for the synthesis of example 116, utilizing 6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-amine during the representative procedure for amide formation with trimethylaluminum.

Example 199 was prepared according to the procedure for the synthesis of example 111, utilizing 6-(4-isopropyl-4H-1,2,4-triazol-3-yl)pyridin-2-amine during the representative procedure for amide formation with trimethylaluminum.

Examples 197 and 198 were prepared according to the procedure for the synthesis of example 163.

Examples 203-205 were prepared according to the representative procedure for N-functionalization and TBS deprotection, utilizing the corresponding chloroformate reagents.

Example 210 was prepared according to the representative procedure for N-functionalization and TBS deprotection, utilizing the corresponding sulfonyl chloride reagents.

Example 202 was prepared according to the representative procedure for N-functionalization and TBS deprotection, utilizing the corresponding acid chloride reagents.

Example 206 was prepared according to the representative procedure for N-functionalization and TBS deprotection, utilizing the corresponding isocyanate reagents.

Example 209 was prepared according to the representative procedure for N-functionalization and TBS deprotection, utilizing the corresponding sulfamoyl chloride reagents.

Example 207 was prepared according to the representative procedure for N-functionalization and TBS deprotection, utilizing the corresponding carbamoyl chloride reagents.

Example 208 was prepared according to the procedure for the synthesis of example 163.

Examples 213-248 are prepared from 7-chloro-6-methoxy-1,2,3,4-tetrahydroisoquinoline utilizing the procedures herein described.

Examples 249-283 are prepared in analogous fashion to example 189.

Examples 284-319 are prepared from tert-butyl 5-chloro-6-methoxyisoindoline-2-carboxylate, which is prepared as in WO 2006082001, utilizing the procedures herein described.

Examples 320-355 are prepared from tert-butyl 5-bromo-6-fluoroisoindoline-2-carboxylate, which is prepared as in US 20150315198, utilizing the procedures herein described.

Examples 356-363 are prepared using procedures similar to those described above.

ASK1 was purchased from Thermofisher (Catalogue # PV4011), ATP was purchased from Sigma (Catalogue # A7699), HI-TRF® KinEASE™ Assay System was obtained from Cisbio (Bedford, Mass.). 1/2 Area plate was purchased from Perkin Elmer (Catalogue # #6005560). HTRF® KinEASE™-STK is a generic method for measuring serine/threonine kinase activities using a time-resolved fluorescence resonance energy transfer (TR-FRET) immunoassay. The IC50value for each compound was determined in the presence of compound (various concentration from 0 to 10 μM) and a fixed amount of ATP and peptide substrates. The test compound, 1 Um STK3 peptide substrate, and 5 Nm of ASK1 kinase are incubated with kinase reaction buffer containing 50 Mm HEPES Ph 7.5, 0.01% BRIJ-35, 10 Mm MgCl2, and 1 Mm EGTA for 30 minutes. 100 Um ATP is added to start kinase reaction and incubated for 3 hours. The STK3-antibody labeled with Eu3+-Cryptate and 125 Nm streptavidin-XL665 are mixed in a single addition with stop reagents provided by the Cisbio kit used to stop the kinase reaction. Fluorescence is detected using an Envision Multilabeled 2014 reader from PerkinElmer. The Fluorescence is measured at 615 nm (Cryptate) and 665 nm (XL665) and a ratio of 665 nm/615 nm is calculated for each well. The resulting TR-FRET is proportional to the phosphorylation level. Staurosporine was used as the positive control. IC50was determined by Xlfit 5.3.