DUAL-ACTION INHBITORS AND METHODS OF USING SAME

Provided are compounds, compositions, and methods for treating diseases and conditions wherein an inhibitor of a kinase, such as rho kinase (ROCK), and an inhibitor of one or more of the monoamine transporters, such as NET or SERT, act in concert to improve the condition.

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Description
CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No. 61/174,672, filed May 1, 2009, and incorporated herein by reference in its entirety.

FIELD OF THE INVENTION

This invention relates to compounds, compositions, and methods for treating diseases and conditions in mammals.

BACKGROUND OF THE INVENTION

A variety of hormones, neurotransmitters and biologically active substances control, regulate or adjust the functions of living bodies via specific receptors located in cell membranes. Many of these receptors mediate the transmission of intracellular signals by activating guanine nucleotide-binding proteins (G proteins) to which the receptor is coupled. Such receptors are generically referred to as G-protein coupled receptors (GPCRs) and include, among others, α-adrenergic receptors, β-adrenergic receptors, opioid receptors, cannabinoid receptors and prostaglandin receptors. The biological effects of activating these receptors is not direct but is mediated by a host of intracellular proteins. The importance of these secondary proteins has only recently been recognized and investigated as intervention points in disease states. One of the most important classes of these downstream effectors is the “kinase” class.

Kinases thus play important roles in the regulation of various physiological functions. For example, alterations in kinase activity have been implicated in a number of disease states, including, but not limited to: cardiac indications such as angina pectoris, essential hypertension, myocardial infarction, supraventricular and ventricular arrhythmias, congestive heart failure, atherosclerosis, renal failure, diabetes, respiratory indications such as asthma, chronic bronchitis, bronchospasm, emphysema, airway obstruction, upper respiratory indications such as rhinitis, seasonal allergies, inflammatory disease, inflammation in response to injury, and rheumatoid arthritis. The importance of p38 MAPK inhibitors in particular as new drugs for rheumatoid arthritis is reflected by the large number of compounds that has been developed over the last years (J. Westra and P. C. Limburg Mini-Reviews in Medicinal Chemistry Volume 6, Number 8, August 2006) Other conditions that involve changes in kinase activity include chronic inflammatory bowel disease, glaucoma, hypergastrinemia, gastrointestinal indications such as acid/peptic disorder, erosive esophagitis, gastrointestinal hypersecretion, mastocytosis, gastrointestinal reflux, peptic ulcer, Zollinger-Ellison syndrome, pain, obesity, bulimia nervosa, depression, obsessive-compulsive disorder, organ malformations (e.g., cardiac malformations), neurodegenerative diseases such as Parkinson's Disease and Alzheimer's Disease, multiple sclerosis, Epstein-Barr infection and cancer (Nature Reviews Drug Discovery 1, 493-502 2002). In other disease states, the roles of the various kinases are only now becoming clear. The retina is a complex tissue composed of multiple interconnected cell layers, highly specialized for transforming light and color into electrical signals that are perceived by the brain. Damage or death of the primary light-sensing cells, the photoreceptors, results in devastating effects on vision. Despite the identification of numerous mutations that cause inherited retinal degenerations, the cellular and molecular mechanisms leading from the primary mutations to photoreceptor apoptosis are not well understood, but may involve the Wnt pathway (A S Hackam The Wnt Signaling Pathway in Retinal Degeneration IUBMB Life Volume 57, Number 6/June 2005).

The success of the tyrosine-kinase inhibitor STI571 (Gleevec) in the treatment of chronic myelogenous leukaemia (Nature Reviews Drug Discovery 2, 296-313 2003) has spurred considerable efforts to develop other kinase inhibitors for the treatment of a wide range of other cancers (Nature Reviews Cancer 3, 650-665 2003). In view of the role that kinases have in many disease states, there is an urgent and continuing need for small molecule ligands which inhibit or modulate the activity of kinases.

Another class of proteins that have proven important in the development of new medicaments is the monoamine transporter class. Monoamine transporters (MAT) are proteins found in both neuronal and non-neuronal cell membranes that function to transport monoamine neurotransmitters into or out of cells. There are several distinct monoamine transporters, or MATs: the dopamine transporter, DAT, the norepinephrine transporter, NET and the serotonin transporter, SERT. DAT, NET and SERT are related to each other in amino acid sequence and protein structure, with each containing 12 trans-membrane helices. The discovery of NET and SERT has led to new drugs, mostly acting via central nervous system (CNS) pharmacology. Modern antidepressants typically work by enhancing serotonergic, noradrenergic or dopaminergic neurotransmission by binding to the corresponding transporter, and inhibiting neurotransmitter reuptake, thereby raising active levels of neurotransmitter in the synapse. Examples include fluoxetine, a selective serotonin reuptake inhibitor; reboxetine, a norepinephrine reuptake inhibitor and bupropion, which inhibits both the norepinephrine and dopamine transporter. Relevant references include He R, Kurome T, Giberson K M, Johnson K M, Kozikowski A P (2005). “Further structure-activity relationship studies of piperidine-based monoamine transporter inhibitors: effects of piperidine ring stereochemistry on potency. Identification of norepinephrine transporter selective ligands and broad-spectrum transporter inhibitors”. J. Med. Chem. 48 (25): 7970-9 and Blough B E, Keverline K I, Nie Z, Navarro H, Kuhar M J, Carroll F l (2002). “Synthesis and transporter binding properties of 3beta-[4′-(phenylalkyl, -phenylalkenyl, and -phenylalkynyl)phenyltropane]-2beta-carboxylic acid methyl esters: evidence of a remote phenyl binding domain on the dopamine transporter”. J. Med. Chem. 45 (18): 4029-37 and G. E. Torres, R. R. Gainetdinov and M. G. Caron (2003). “Plasma membrane monoamine transporters: structure, regulation and function”. Nat. Rev. Neurosci. 4 (1): 13-25.

SUMMARY

In certain embodiments, provided are compounds according to Formulas I, II, III, IV, V, VI, and VII, as described below.

In certain embodiments, provided are compositions comprising a compound according to Formulas I, II, III, IV, V, VI, or VII, as described below, and a carrier. The composition may further include an activity enhancer.

In certain embodiments, provided are methods for treating a disease or condition, the method comprising administering to a mammal in need thereof a therapeutically effective amount of a compound according to Formulas I, II, III, IV, V, VI, or VII, as described below. The compound may inhibit a kinase and monoamine transport in concert to alleviate the symptoms associated with the disease or condition.

In certain embodiments, provided are methods of reducing intraocular pressure comprising contacting a cell with an effective amount of a compound according to Formulas I, II, III, IV, V, VI, or VII, as described below.

In certain embodiments, provided are methods for treating cardiac indications comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to Formulas I, II, III, IV, V, VI, or VII, as described below.

In certain embodiments, provided are methods of treating a respiratory disorder comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to Formulas I, II, III, IV, V, VI, or VII, as described below.

In certain embodiments, provided are methods of treating a renal disease comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to Formulas I, II, III, IV, V, VI, or VII, as described below.

In certain embodiments, provided are methods of treating upper respiratory indications comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to Formulas I, II, III, IV, V, VI, or VII, as described below.

In certain embodiments, provided are methods of treating inflammatory disease, inflammation in response to injury, or rheumatoid arthritis comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to Formulas I, II, III, IV, V, VI, or VII, as described below.

In certain embodiments, provided are methods for modulating the action of a kinase and a monoamine transporter in a cell comprising contacting the cell with a compound according to Formulas I, II, III, IV, V, VI, or VII, as described below, in an amount effective to modulate the action of a kinase and a monoamine transporter in a cell. The compound may be administered in conjunction with one or more additional therapeutic agents. The additional therapeutic agent may be selected from the group consisting of beta blockers, alpha-agonists, carbonic anhydrase inhibitors, prostaglandin-like compounds, miotic or cholinergic agents, and epinephrine compounds.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a scheme for the synthesis of compounds, including E1-E8.

FIG. 2 is a scheme for the synthesis of compounds, including E457-E466.

FIG. 3 is a scheme for the synthesis of benzamidines, E467-E476.

FIG. 4 is a scheme for the synthesis of para-aminobenzamide precursors, E477-E478, for the synthesis scheme of FIG. 3.

FIG. 5 is a scheme for the synthesis of para-aminobenzamide precursors, E479-E481, for the synthesis scheme of FIG. 3.

FIG. 6 is a scheme for the synthesis of compounds, including E8-E12.

FIG. 7 is a scheme for the synthesis of compounds, including E132-E139.

FIG. 8 is a scheme for the synthesis of compounds, including E140-E143.

FIG. 9 is a scheme for the synthesis of compounds, including E145-E148.

FIG. 10 is a scheme for the synthesis of compounds, including E197-S and E197-R.

FIG. 11 is a scheme for the synthesis of compounds, including E199-E203.

FIG. 12 is a scheme for the synthesis of compounds, including E204-E206.

FIG. 13 is a scheme for the synthesis of compounds, including E231-E241.

FIG. 14 is a scheme for the synthesis of compounds, including E249-253.

FIG. 15 is a scheme for the synthesis of compounds, including E275-E278.

FIG. 16 is a scheme for the synthesis of compounds, including E289-E290.

FIG. 17 is a scheme for the synthesis of compounds, including E300-E308.

FIG. 18 is a scheme for the synthesis of compounds, including E319-E325.

FIG. 19 is a scheme for the synthesis of compounds, including E371-E377.

FIG. 20 is a scheme for the synthesis of compounds, including E398-E404.

FIG. 21 is a general scheme for the synthesis of compounds, including compounds E429-E433.

FIG. 22 shows structures of NET inhibitors.

 DETAILED DESCRIPTION OF THE INVENTION

This invention relates to compounds, compositions, and methods for treating diseases and conditions wherein an inhibitor of a kinase, such as rho kinase (ROCK), and an inhibitor of one or more of the monoamine transporters, such as NET or SERT, will act in concert to improve the condition. One such area is the treatment of glaucoma by producing, inter alia, a reduction in intraocular pressure (IOP), or anti-inflammatory effects or neuroprotection. The method comprises administering the composition comprising a first ligand that interacts strongly with a kinase, specifically and at a minimum rho kinase, and a second ligand that interacts with MAT proteins, specifically and at minimum NET proteins. These may be separate molecules, but ideally they are part of the same molecule. The same portions of that molecule may act on both systems, or separate but connected parts of the molecule may act more or less independently to bring about inhibition of both systems.

Publications and patents are referred to throughout this disclosure. All U.S. patents cited herein are hereby incorporated by reference.

All percentages, ratios, and proportions used herein are by weight unless otherwise specified.

In the description of the invention various embodiments and individual features are disclosed. As will be apparent to a person having ordinary skill in the art, all combinations of such embodiments and features are possible and can result in preferred embodiments of the invention.

DEFINITION AND USAGE OF TERMS

The following is a list of definitions for terms as used herein:

“Acyl group” means a monovalent group suitable for acylating a nitrogen atom to form an amide or carbamate or an oxygen atom to form an ester group. Preferred acyl groups include pivaloyl, benzoyl, substituted benzoyl, acetyl, tert-butyl acetyl, para-phenyl benzoyl, and trifluoroacetyl. More preferred acyl groups include acetyl pivaloyl, benzoyl, and substituted benzoyl. The most preferred acyl groups are pivaloyl, benzoyl, substituted benzoyl.

“Alkylene” means a divalent alkyl group.

“Aromatic group” means a monovalent group having a monocyclic ring structure or fused bicyclic ring structure. Monocyclic aromatic groups contain 5 to 10 carbon atoms, preferably 5 to 7 carbon atoms, and more preferably 5 to 6 carbon atoms in the ring. Bicyclic aromatic groups contain 8 to 12 carbon atoms, preferably 9 or 10 carbon atoms in the ring. Aromatic groups are unsubstituted. The most preferred aromatic groups are phenyl and naphthyl.

“Carbocyclic group” means a monovalent saturated or unsaturated hydrocarbon ring. Carbocyclic groups are monocyclic, or are fused, spiro, or bridged bicyclic ring systems. Monocyclic carbocyclic groups contain 4 to 10 carbon atoms, preferably 4 to 7 carbon atoms, and more preferably 5 to 6 carbon atoms in the ring. Bicyclic carbocyclic groups contain 8 to 12 carbon atoms, preferably 9 to 10 carbon atoms in the ring. Carbocyclic groups are unsubstituted. Preferred carbocyclic groups include cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl. More preferred carbocyclic groups include cyclohexyl, cycloheptyl, and cyclooctyl. The most preferred carbocyclic group is cycloheptyl. Carbocyclic groups are not aromatic.

“Halogen atom” means F, Cl, Br, or I. Preferably, the halogen atom is F, Cl, or Br; more preferably Cl or F; and most preferably F.

“Halogenated hydrocarbon group” means a substituted monovalent hydrocarbon group or a substituted carbocyclic group, wherein at least one substituent is a halogen atom. Halogenated hydrocarbon groups can have a straight, branched, or cyclic structure. Preferred halogenated hydrocarbon groups have 1 to 12 carbon atoms, more preferably 1 to 6 carbon atoms, and most preferably 1 to 3 carbon atoms. Preferred halogen atom substituents are Cl and F. The most preferred halogenated hydrocarbon group is trifluoromethyl.

“Heteroaromatic group” means an aromatic ring containing carbon and 1 to 4 heteroatoms in the ring. Heteroaromatic groups are monocyclic or fused bicyclic rings. Monocyclic heteroaromatic groups contain 5 to 10 member atoms (i.e., carbon and heteroatoms), preferably 5 to 7, and more preferably 5 to 6 in the ring. Bicyclic heteroaromatic rings contain 8 to 12 member atoms, preferably 9 or 10 in the ring. Heteroaromatic groups are unsubstituted. Preferred heteroaromatic groups include thienyl, thiazolo, purinyl, pyrimidyl, pyridyl, and furanyl. More preferred heteroaromatic groups include thienyl, furanyl, and pyridyl. The most preferred heteroaromatic group is thienyl.

“Heteroatom” means an atom other than carbon in the ring of a heterocyclic group or the chain of a heterogeneous group. Preferably, heteroatoms are selected from the group consisting of nitrogen, sulfur, and oxygen atoms. Groups containing more than one heteroatom may contain different heteroatoms.

“Heterocyclic group” means a saturated or unsaturated ring structure containing carbon and 1 to 4 heteroatoms in the ring. No two heteroatoms are adjacent in the ring. Heterocyclic groups are not aromatic. Heterocyclic groups are monocyclic, or are fused or bridged bicyclic ring systems. Monocyclic heterocyclic groups contain 4 to 10 member atoms (i.e., including both carbon atoms and at least 1 heteroatom), preferably 4 to 7, and more preferably 5 to 6 in the ring. Bicyclic heterocyclic groups contain 8 to 12 member atoms, preferably 9 or 10 in the ring. Heterocyclic groups are unsubstituted.

Preferred heterocyclic groups include piperzyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, and piperdyl.

“Heterogeneous group” means a saturated or unsaturated chain containing 1 to 18 member atoms (i.e., including both carbon and at least one heteroatom). No two heteroatoms are adjacent. Preferably, the chain contains 1 to 12 member atoms, more preferably 1 to 6, and most preferably 1 to 4. The chain may be straight or branched. Preferred branched heterogeneous groups have one or two branches, preferably one branch. Preferred heterogeneous groups are saturated. Unsaturated heterogeneous groups have one or more double bonds, one or more triple bonds, or both. Preferred unsaturated heterogeneous groups have one or two double bonds or one triple bond. More preferably, the unsaturated heterogeneous group has one double bond. Heterogeneous groups are unsubstituted.

“Lower alkylene” means an alkylene group having 1 to 6, preferably 1 to 4 carbon atoms.

“Lower monovalent hydrocarbon group” or “lower alkyl group” means a monovalent hydrocarbon group having 1 to 6, preferably 1 to 4 carbon atoms.

“Monovalent hydrocarbon group” or “alkyl group” means a chain of 1 to 18 carbon atoms, preferably 1 to 12, more preferably 1 to 6, and most preferably 1 to 4 carbon atoms. Monovalent hydrocarbon groups may have a straight chain or branched chain structure. Preferred monovalent hydrocarbon groups have one or two branches. Preferred monovalent hydrocarbon groups are saturated. Unsaturated monovalent hydrocarbon groups have one or more double bonds, one or more triple bonds, or combinations thereof. Preferred unsaturated monovalent hydrocarbon groups have one or two double bonds or one triple bond; more preferred unsaturated monovalent hydrocarbon groups have one double bond.

“Pharmaceutically acceptable” means suitable for use in a human or other mammal.

“Substituted aromatic group” means an aromatic group wherein 1 to 4 of the hydrogen atoms bonded to carbon atoms in the ring have been replaced with other substituents. Preferred substituents include: hydroxy, methoxy, alkyloxy, acetoxy, benzyloxy, halogen atoms, cyano groups, monovalent hydrocarbon groups, substituted monovalent hydrocarbon groups, heterogeneous groups, aromatic groups, substituted aromatic groups, or any combination thereof. More preferred substituents include hydroxy, methoxy, alkyloxy, acetoxy, benzyloxy halogen atoms, monovalent hydrocarbon groups, and substituted monovalent hydrocarbon groups. Preferred substituted aromatic groups include meta and para acyloxyphenyl, para-methoxyphenyl, para-alkoxyphenyl, methylenedioxyphenyl, para-chlorophenyl, beta-naphthyl. The substituents may be substituted at the ortho, meta, or para position on the ring, or any combination thereof. The preferred substitution pattern on the ring is para or meta, or both. The most preferred substitution pattern is para.

“Substituted carbocyclic group” means a carbocyclic group wherein 1 to 4 hydrogen atoms bonded to carbon atoms in the ring have been replaced with other substituents. Preferred substituents include: hydroxyl, halogen atoms, cyano groups, monovalent hydrocarbon groups, monovalent heterogeneous groups, substituted monovalent hydrocarbon groups, aromatic groups, substituted aromatic groups, or any combination thereof. More preferred substituents include hydroxyl, halogen atoms and substituted monovalent hydrocarbon groups. Carbocyclic group does not include aromatic rings.

“Substituted heteroaromatic group” means a heteroaromatic group wherein 1 to 4 hydrogen atoms bonded to carbon atoms in the ring have been replaced with other substituents. Preferred substituents include: halogen atoms, cyano groups, monovalent hydrocarbon groups, substituted monovalent hydrocarbon groups, heterogeneous groups, substituted heterogeneous groups, phenyl groups, phenoxy groups, or any combination thereof. More preferred substituents include halogen atoms, halogenated hydrocarbon groups, monovalent hydrocarbon groups, and phenyl groups.

“Substituted heterocyclic group” means a heterocyclic group wherein 1 to 4 hydrogen atoms bonded to carbon atoms in the ring have been replaced with other substituents. Preferred substituents include: halogen atoms, cyano groups, monovalent hydrocarbon groups, substituted monovalent hydrocarbon groups, heterogeneous groups, substituted heterogeneous groups, halogenated hydrocarbon groups, phenyl groups, phenoxy groups, or any combination thereof. More preferred substituents include halogen atoms and halogenated hydrocarbon groups. Substituted heterocyclic groups are not aromatic.

“Substituted heterogeneous group” means a heterogeneous group, wherein 1 to 4 of the hydrogen atoms bonded to carbon atoms in the chain have been replaced with other substituents. Preferred substituents include halogen atoms, hydroxy groups, alkoxy groups (e.g., methoxy, ethoxy, propoxy, butoxy, and pentoxy), aryloxy groups (e.g., phenoxy, chlorophenoxy, tolyloxy, methoxyphenoxy, benzyloxy, alkyloxycarbonylphenoxy, and acyloxyphenoxy), acyloxy groups (e.g., propionyloxy, benzoyloxy, and acetoxy), carbamoyloxy groups, carboxy groups, mercapto groups, alkylthio groups, acylthio groups, arylthio groups (e.g., phenylthio, chlorophenylthio, alkylphenylthio, alkoxyphenylthio, benzylthio, and alkyloxycarbonylphenylthio), aromatic groups (e.g., phenyl and tolyl), substituted aromatic groups (e.g., alkoxphenyl, alkoxycarbonylphenyl, and halophenyl), heterocyclic groups, heteroaromatic groups, and amino groups (e.g., amino, mono- and di-alkylamino having 1 to 3 carbon atoms, methylphenylamino, methylbenzylamino, alkanylamido groups of 1 to 3 carbon atoms, carbamamido, ureido, and guanidino).

“Substituted monovalent hydrocarbon group” means a monovalent hydrocarbon group wherein 1 to 4 of the hydrogen atoms bonded to carbon atoms in the chain have been replaced with other substituents. Preferred substituents include halogen atoms; halogenated hydrocarbon groups; alkyl groups (e.g., methyl, ethyl, propyl, and butyl); hydroxy groups; alkoxy groups (e.g., methoxy, ethoxy, propoxy, butoxy, and pentoxy); aryloxy groups (e.g., phenoxy, chlorophenoxy, tolyloxy, methoxyphenoxy, benzyloxy, alkyloxycarbonylphenoxy, and acyloxyphenoxy); acyloxy groups (e.g., propionyloxy, benzoyloxy, and acetoxy); carbamoyloxy groups; carboxy groups; mercapto groups; alkylthio groups; acylthio groups; arylthio groups (e.g., phenylthio, chlorophenylthio, alkylphenylthio, alkoxyphenylthio, benzylthio, and alkyloxycarbonylphenylthio); aryl groups (e.g., phenyl, tolyl, alkoxyphenyl, alkoxycarbonylphenyl, and halophenyl); heterocyclyl groups; heteroaryl groups; and amino groups (e.g., amino, mono- and di-alkanylamino groups of 1 to 3 carbon atoms, methylphenylamino, methylbenzylamino, alkanylamido groups of 1 to 3 carbon atoms, carbamamido, ureido, and guanidino).

Compounds of the Invention

Dual-action inhibitors according to the present invention that produce this dual action of kinase inhibition and monoamine transport inhibition are characterized by the following general structure:


Monoamine transport inhibitor-Linkage-Kinase inhibitor  (I)

In some embodiments, the dual-action inhibitor is selective for rho kinase. In other embodiments, the dual-action inhibitor is selective for NET or SERT.

One specific example of such an arrangement is shown below:

wherein each X is independently selected from hydrogen, amino, lower alkyl, halogen, carbonyl, nitrile, hydroxyl, and alkoxy;
wherein Z is selected from substituted monovalent hydrocarbon groups, substituted heterogeneous groups, substituted heterocyclic groups, substituted heteroaromatic groups, substituted carbocyclic groups, substituted aromatic groups, heterogeneous groups, heterocyclic groups, heteroaromatic groups, carbocyclic groups, and aromatic groups;
wherein R is selected from guanidino or —N(R5)2;
wherein each R5 is independently selected from H, Me or Et;
wherein B is C═O, C═S, or —CH2—;
wherein n and m are independently selected from the integers 0, 1, 2 or 3 and represent a independently variable number of substituted or unsubstituted methylene units in the alkyl chain;
and pharmaceutically acceptable salts, hydrates, and biohydrolyzable amides, esters, and imides.

In Formula (II), the left-hand portion (which excludes the isoquinoline ring) of the molecule contains the NET inhibitor pharmacophore and the entire molecule represents the rho kinase inhibitor pharmacophore.

Tautomers, optical isomers, diastereomers, and enantiomers of Formula (II) are also suitable as components of this invention. At all stereocenters where stereochemistry is not defined both epimers are envisioned.

Another example of a compound according to the present invention is shown below:

wherein each X is independently selected from hydrogen, amino, lower alkyl, halogen, carbonyl, nitrile, hydroxyl, and alkoxy;
wherein Z is selected from substituted monovalent hydrocarbon groups, substituted heterogeneous groups, substituted heterocyclic groups, substituted heteroaromatic groups, substituted carbocyclic groups, substituted aromatic groups, heterogeneous groups, heterocyclic groups, heteroaromatic groups, carbocyclic groups, and aromatic groups;
wherein R is selected from guanidino or —N(R5)2;
wherein each R5 is independently selected from H, Me or Et;
wherein B is C═O, C═S, or —CH2—;
wherein n and m are independently selected from the integers 0, 1, 2 or 3 and represent a independently variable number of substituted or unsubstituted methylene units in the alkyl chain;
and pharmaceutically acceptable salts, hydrates, and biohydrolyzable amides, esters, and imides.

Tautomers, optical isomers, diastereomers, and enantiomers of Formula (III) are also suitable as components of this invention. At all stereocenters where stereochemistry is not defined both epimers are envisioned.

Another example of a compound according to the present invention is shown below:

wherein each X is independently selected from hydrogen, amino, lower alkyl, halogen, carbonyl, nitrile, hydroxyl, and alkoxy;
wherein Z is selected from substituted monovalent hydrocarbon groups, substituted heterogeneous groups, substituted heterocyclic groups, substituted heteroaromatic groups, substituted carbocyclic groups, substituted aromatic groups, heterogeneous groups, heterocyclic groups, heteroaromatic groups, carbocyclic groups, and aromatic groups;
wherein R is selected from guanidino or —N(R5)2;
wherein each R5 is independently selected from H, Me or Et;
wherein B is C═O, C═S, or —CH2—;
wherein n and m are independently selected from the integers 0, 1, 2 or 3 and represent a independently variable number of substituted or unsubstituted methylene units in the alkyl chain;
and pharmaceutically acceptable salts, hydrates, and biohydrolyzable amides, esters, and imides.

Tautomers, optical isomers, diastereomers, and enantiomers of Formula (IV) are also suitable as components of this invention. At all stereocenters where stereochemistry is not defined both epimers are envisioned.

Another example of a compound according to the present invention is shown below:

wherein each X is independently selected from hydrogen, amino, lower alkyl, halogen, carbonyl, nitrile, hydroxyl;
wherein Z is selected from substituted monovalent hydrocarbon groups, substituted heterogeneous groups, substituted heterocyclic groups, substituted heteroaromatic groups, substituted carbocyclic groups, substituted aromatic groups, heterogeneous groups, heterocyclic groups, heteroaromatic groups, carbocyclic groups, and aromatic groups;
wherein R is selected from guanidino or —N(R5)2;
wherein each R5 is independently selected from H, Me or Et;
wherein R2 is H or Me;
wherein B is C═O, C═S, or —CH2—;
wherein n and m are independently selected from the integers 0, 1, 2 or 3 and represent a independently variable number of substituted or unsubstituted methylene units in the alkyl chain;

and pharmaceutically acceptable salts, hydrates, and biohydrolyzable amides, esters, and imides.

Tautomers, optical isomers, diastereomers, and enantiomers of Formula (V) are also suitable as components of this invention. At all stereocenters where stereochemistry is not defined both epimers are envisioned.

Another example of a compound according to the present invention is shown below:

wherein each X is independently selected from hydrogen, amino, hydroxy, alkoxy, lower alkyl, halogen, carbonyl and nitrile;
wherein R1 and R2 are each independently selected from substituted monovalent hydrocarbon groups, substituted heterogeneous groups, substituted heterocyclic groups, substituted heteroaromatic groups, substituted carbocyclic groups, substituted aromatic groups, heterogeneous groups, heterocyclic groups, heteroaromatic groups, carbocyclic groups, or aromatic groups;
wherein R is selected from guanidino or —N(R5)2;
wherein each R5 is selected from H, Me or Et;
wherein A is an —NR4—, —S(O)2—NH—, —NH—S(O)2—C═O, or lower alkylene;
wherein B is C═O, C═S, or —CH2—, or NR4;
wherein each R4 is independently H, Me, or Et;
wherein n1 and n2 are independently selected from the integers 0, 1 or 2 and represent a variable number of carbon units in the alkyl chain;
and pharmaceutically acceptable salts, hydrates, and biohydrolyzable amides, esters, and imides.

Tautomers, optical isomers, diastereomers, and enantiomers of Formula (VI) are also suitable as components of this invention. At all stereocenters where stereochemistry is not defined both epimers are envisioned.

Another example of a compound according to the present invention is shown below:

wherein X7 is a —O— or —NH—;
A is a NET inhibitor; and
B is a rho kinase inhibitor.

Tautomers, optical isomers, diastereomers, and enantiomers of Formula (VII) are also suitable as components of this invention. At all stereocenters where stereochemistry is not defined both epimers are envisioned.

Other examples of compounds according to the present invention are shown below:

Other useful compounds and methods may be found in International Patent Application No. PCT/US10/22246 filed on Jan. 27, 2010, and U.S. patent application Ser. No. 12/694,965 filed on Jan. 27, 2010, which are hereby incorporated fully by reference. Compounds according to the invention may further include compounds E589-E604 shown below:

The amino isoquinoline amide or substituted benzamide compounds may be synthesized according to the general schemes shown in FIGS. 1-5.

According to the synthesis scheme in FIG. 1, ester (1) may be protected with the TIPS group and alkylated with bromomethylphthalimide to give compound (3). The ester may be then hydrolyzed with LiOH*H2O to give diacid (4) and coupled with 6-aminoisoquinoline using EDC as the coupling agent. The amine (6) may be accomplished using hydrazine which may be then protected with Boc2O to give (7). Deprotection of the hydroxyl group may be carried out with TBAF, and coupling with the appropriate acid may be achieved with EDC or using the acid chloride. Deprotection of the amine may be accomplished with HCl to give the final amino isoquinoline amides.

As in FIG. 1, the synthesis scheme in FIG. 2 may begin by protecting 2-(4-(hydroxymethyl)phenyl)acetic acid as the methyl ester and the TIPS alcohol to give E457. This methyl ester may be then alkylated with bromomethylphthalimide to give compound E459. The ester may be hydrolyzed with LiOH*H2O to give diacid E460 and coupled with 6-aminoisoquinoline using EDC as the coupling agent giving compound E461. Formation of the amine E462 may be accomplished using hydrazine which may be then protected with Boc2O to give E463. Deprotection of the hydroxyl group may be carried out with TBAF, and coupling with the appropriate acid may be achieved with EDC or using the acid chloride. Deprotection of the amine may be accomplished with HCl to give the final amino isoquinoline amides.

Benzamides may be synthesized using the procedures outlined in FIG. 2, but by substituting the para-amino benzamide of choice for the amino isoquinoline, as shown in the synthesis scheme in FIG. 3.

The para-aminobenzamide precursors of the synthesis scheme in FIG. 3 may be commercially-available, or may be synthesized by the general synthesis schemes of FIGS. 4-5.

According to FIG. 4, the appropriate acid may be converted to its acid chloride with oxalyl chloride then reacted with ammonia gas or another amine to give the amide. The nitro group may be reduced to the aniline with hydrogen or another reducing agent. The aniline may be then coupled with an appropriate acid using standard coupling procedures such as EDC and DMAP in pyridine as shown in FIG. 3.

An alternative synthetic route is outlined in the synthesis scheme of FIG. 5. According to FIG. 5, the aniline may be coupled with an appropriate acid using standard coupling procedures such as EDC and DMAP in pyridine. The ester may be then converted to the corresponding primary amide using formamide and NaOMe in DMF or to a substituted amide by heating with the appropriate amine in a solvent such as MeOH.

The abbreviations used in the synthetic schemes shown in the figures have the following meanings: Boc2O is di-tert-butyl-dicarbonate, DMAP is dimethyl aminopyridine, DMSO is Dimethyl Sulfoxide, HATU is 2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate, LDA is lithium diisopropyl amide, DMF is dimethylformamide, THF is tetrahydrofuran, and EDC is N-(3-dimethylaminopropyl)-W-ethylcarbodiimide hydrochloride.

Compositions of the Invention

Additionally, this invention relates to a composition useful for treating various disease and conditions. The composition may comprise A) a dual-action inhibitor as described above, and B) a carrier. The composition may further comprise C) one or more optional activity enhancers. Standard pharmaceutical formulation techniques may be used, such as those disclosed in Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa. (1990).

In some embodiments, component A) can be any one of Formulae (I), (II), (III), (IV), (V), (VI), (VII) or combinations thereof.

The composition further comprises component B) a carrier. “Carrier” means one or more compatible substances that are suitable for administration to a mammal. Carrier includes solid or liquid fillers, diluents, hydrotopes, surface-active agents, and encapsulating substances. “Compatible” means that the components of the composition are capable of being commingled with the dual-action inhibitors, and with each other, in a manner such that there is no interaction which would substantially reduce the efficacy of the composition under ordinary use situations. Carriers must be of sufficiently high purity and sufficiently low toxicity to render them suitable for administration to the mammal being treated. The carrier can be inert, or it can possess pharmaceutical benefits, cosmetic benefits, or both.

The choice of carrier for component B) depends on the route by which A) the dual-action inhibitor will be administered and the form of the composition. The composition may be in a variety of forms, suitable, for example, for systemic administration (e.g., oral, rectal, nasal, sublingual, buccal, or parenteral) or topical administration (e.g., local application on the skin, ocular, liposome delivery systems, or iontophoresis). Topical administration is preferred.

Carriers for systemic administration typically comprise one or more ingredients selected from the group consisting of a) diluents, b) lubricants, c) binders, d) disintegrants, e) colorants, f) flavors, g) sweeteners, h) antioxidants, j) preservatives, k) glidants, m) solvents, n) suspending agents, o) wetting agents, p) surfactants, combinations thereof, and others.

Component a) is a diluent. Suitable diluents include sugars such as glucose, lactose, dextrose, and sucrose; polyols such as propylene glycol; calcium carbonate; sodium carbonate; cellulose; glycerin; mannitol; and sorbitol.

Component b) is a lubricant. Suitable lubricants are exemplified by solid lubricants including silica, talc, stearic acid and its magnesium salts and calcium salts, calcium sulfate; and liquid lubricants such as polyethylene glycol and vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and oil of theobroma.

Component c) is a binder. Suitable binders include polyvinylpyrilidone; magnesium aluminum silicate; starches such as corn starch and potato starch; gelatin; tragacanth; and cellulose and its derivatives, such as sodium carboxymethylcellulose, ethyl cellulose, methylcellulose, and sodium carboxymethylcellulose.

Component d) is a disintegrant. Suitable disintigrants include starches, agar, alginic acid and the sodium salt thereof, effervescent mixtures, and croscarmelose.

Component e) is a colorant such as an FD&C dye.

Component f) is a flavor such as menthol, peppermint, and fruit flavors.

Component g) is a sweetener such as aspartame and saccharin.

Component h) is an antioxidants such as BHA, BHT, and vitamin E.

Component j) is a preservative such as methyl paraben and sodium benzoate.

Component k) is a glidant such as silicon dioxide.

Component m) is a solvent, such as water, isotonic saline, ethyl oleate, alcohols such as ethanol, and phosphate buffer solutions.

Component n) is a suspending agent. Suitable suspending agents include cellulose and its derivatives, such as methyl cellulose and sodium carboxymethyl cellulose; AVICEL® RC-591 from FMC Corporation of Philadelphia, Pa.; tragacanth and sodium alginate.

Component o) is a wetting agent such as lecithin, polysorbate 80, and sodium lauryl sulfate.

Component p) is a surfactant such as the TWEENS® from Atlas Powder Company of Wilmington, Del.

Compositions for parenteral administration typically comprise A) 0.1 to 10% of a dual-action inhibitor and B) 90 to 99.9% of a carrier comprising a) a diluent, b) a lubricant, c) a binder, and m) a solvent. Preferably, component a) is propylene glycol, b) is sesame oil, c) is pyrrolidone, and m) is ethanol or ethyl oleate.

Compositions for oral administration can have various dosage forms. For example, solid forms include tablets, capsules, granules, and bulk powders. These oral dosage forms comprise a safe and effective amount, usually at least 5%, and preferably from 25% to 50%, of A) the dual-action inhibitor. The oral dosage compositions further comprise B) 50 to 95% of a carrier, preferably 50 to 75%.

Tablets can be compressed, tablet triturates, enteric-coated, sugar-coated, film-coated, or multiple-compressed. Tablets typically comprise A) the dual-action inhibitor, and B) a carrier comprising ingredients selected from the group consisting of a) diluents, b) lubricants, c) binders, d) disintigrants, e) colorants, f) flavors, g) sweeteners, k) glidants, and combinations thereof. Preferred diluents include calcium carbonate, sodium carbonate, mannitol, lactose and cellulose. Preferred binders include starch, gelatin, and sucrose. Preferred disintegrants include starch, alginic acid, and croscarmelose. Preferred lubricants include magnesium stearate, stearic acid, and talc. Preferred colorants are the FD&C dyes, which can be added for appearance. Chewable tablets preferably contain g) sweeteners such as aspartame and saccharin, or f) flavors such as menthol, peppermint, and fruit flavors.

Capsules (including time release and sustained release formulations) typically comprise A) the dual-action inhibitor, and B) a carrier comprising one or more a) diluents disclosed above in a capsule comprising gelatin. Granules typically comprise A) the dual-action inhibitor, and preferably further comprise k) glidants such as silicon dioxide to improve flow characteristics.

The selection of ingredients in the carrier for oral compositions depends on secondary considerations like taste, cost, and shelf stability, which are not critical for the purposes of this invention. One skilled in the art would know how to select appropriate ingredients without undue experimentation.

The solid compositions may also be coated by conventional methods, typically with pH or time-dependent coatings, such that A) the dual-action inhibitor is released in the gastrointestinal tract in the vicinity of the desired application, or at various times to extend the desired action. The coatings typically comprise one or more components selected from the group consisting of cellulose acetate phthalate, polyvinylacetate phthalate, hydroxypropyl methyl cellulose phthalate, ethyl cellulose, EUDRAGIT® coatings (available from Rohm & Haas G.M.B.H. of Darmstadt, Germany), waxes and shellac.

Compositions for oral administration can also have liquid forms. For example, suitable liquid forms include aqueous solutions, emulsions, suspensions, solutions reconstituted from non-effervescent granules, suspensions reconstituted from non-effervescent granules, effervescent preparations reconstituted from effervescent granules, elixirs, tinctures, syrups, and the like. Liquid orally administered compositions typically comprise A) the dual-action inhibitor and B) a carrier comprising ingredients selected from the group consisting of: a) diluents, e) colorants, and f) flavors, g) sweeteners, j) preservatives, m) solvents, n) suspending agents, and p) surfactants. Peroral liquid compositions preferably comprise one or more ingredients selected from the group consisting of e) colorants, f) flavors, and g) sweeteners.

Other compositions useful for attaining systemic delivery of the subject compounds include sublingual, buccal and nasal dosage forms. Such compositions typically comprise one or more of soluble filler substances such as a) diluents including sucrose, sorbitol and mannitol; and c) binders such as acacia, microcrystalline cellulose, carboxymethyl cellulose, and hydroxypropyl methyl cellulose. Such compositions may further comprise b) lubricants, e) colorants, f) flavors, g) sweeteners, h) antioxidants, and k) glidants.

The compositions may further comprise component C) an optional activity enhancer. Component C) is preferably selected from the group consisting of i) other medicaments for treating elevated IOP such as a FP series prostaglandin and ii) penetration enhancers.

Component i) is an optional IOP treatment. Component i) is exemplified by, but not limited to, prostaglandins, carbonic anhydrase inhibitors, alpha agonists, and beta blockers.

Component ii) is a penetration enhancer that can be added to all of the compositions for systemic administration except compositions for oral administration. The amount of component ii), when present in the composition, is typically 1 to 5%. Examples of penetration enhancers include 2-methyl propan-2-ol, propan-2-ol, ethyl-2-hydroxypropanoate, hexan-2,5-diol, POE(2) ethyl ether, di(2-hydroxypropyl)ether, pentan-2,4-diol, acetone, POE(2) methyl ether, 2-hydroxypropionic acid, 2-hydroxyoctanoic acid, propan-1-ol, 1,4-dioxane, tetrahydrofuran, butan-1,4-diol, propylene glycol dipelargonate, polyoxypropylene 15 stearyl ether, octyl alcohol, POE ester of oleyl alcohol, ° leyl alcohol, lauryl alcohol, dioctyl adipate, dicapryl adipate, di-isopropyl adipate, di-isopropyl sebacate, dibutyl sebacate, diethyl sebacate, dimethyl sebacate, dioctyl sebacate, dibutyl suberate, dioctyl azelate, dibenzyl sebacate, dibutyl phthalate, dibutyl azelate, ethyl myristate, dimethyl azelate, butyl myristate, dibutyl succinate, didecyl phthalate, decyl oleate, ethyl caproate, ethyl salicylate, iso-propyl palmitate, ethyl laurate, 2-ethyl-hexyl pelargonate, iso-propyl isostearate, butyl laurate, benzyl benzoate, butyl benzoate, hexyl laurate, ethyl caprate, ethyl caprylate, butyl stearate, benzyl salicylate, 2-hydroxypropanoic acid, 2-hyroxyoctanoic acid, dimethyl sulphoxide, N,N-dimethyl acetamide, N,N-dimethyl formamide, 2-pyrrolidone, 1-methyl-2-pyrrolidone, 5-methyl-2-pyrrolidone, 1,5-dimethyl-2-pyrrolidone, 1-ethyl-2-pyrrolidone, phosphine oxides, sugar esters, tetrahydrofurfural alcohol, urea, diethyl-m-toluamide, 1-dodecylazacyloheptan-2-one, and combinations thereof.

In a preferred embodiment of the invention, the medicaments are topically administered. Topical compositions that can be applied locally to the eye may be in solutions, suspensions, ointments, gels, sprays, skin patches, and the like. Topical compositions comprise: component A) the medicament described above and component B) a carrier. The carrier of the topical composition preferably aids penetration of the medicament into the eye. Component B) may further comprise one or more optional components. Topical compositions preferably further comprise C) one or more of the optional activity enhancers described above.

Topical compositions that can be applied locally to the skin may be in any form including solids, solutions, oils, creams, ointments, gels, lotions, shampoos, leave-on and rinse-out hair conditioners, milks, cleansers, moisturizers, sprays, skin patches, and the like. Topical compositions comprise: component A, the compounds described above, and component B, a carrier. The carrier of the topical composition preferably aids penetration of the compounds into the skin. Component B may further comprise one or more optional components.

The exact amounts of each component in the topical composition depend on various factors. The amount of component A) depends on the binding affinity (IC50) of the medicament selected. The amount of component A) added to the topical composition is up to 10% of the total, but more typically is from about 0.01% to about 1%.

The topical composition further comprises 1 to 20% component C), and a sufficient amount of component B) such that the amounts of components A), B), and C), combined equal 100%. The amount of B) the carrier employed in conjunction with the medicament is sufficient to provide a practical quantity of material for administration per unit dose of the compound. Techniques and compositions for making dosage forms useful in the methods of this invention are described in the following references: Modern Pharmaceutics, Chapters 9 and 10, Banker & Rhodes, eds. (1979); Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1981); and Ansel, Introduction to Pharmaceutical Dosage Forms, 2nd Ed., (1976).

Component B) the carrier may comprise a single component or a combination of two or more components. Typical carriers for component B) in the topical compositions include water, alcohols, aloe vera gel, allantoin, glycerin, vitamin A and E oils, mineral oil, propylene glycol, PPG-2 myristyl propionate, dimethyl isosorbide, combinations thereof, and the like. Preferred carriers include water.

The carrier of the topical composition may further comprise one or more ingredients selected from the group consisting of (q) emollients, (r) propellants, (s) solvents, (t) humectants, (u) thickeners, (v) powders, and (w) fragrances.

Ingredient (q) is an emollient. The amount of ingredient (q) in the topical composition is typically 5 to 95%. Suitable emollients include stearyl alcohol, glyceryl monoricinoleate, glyceryl monostearate, propane-1,2-diol, butane-1,3-diol, mink oil, cetyl alcohol, iso-propyl isostearate, stearic acid, iso-butyl palmitate, isocetyl stearate, ° leyl alcohol, isopropyl laurate, hexyl laurate, decyl oleate, octadecan-2-ol, isocetyl alcohol, cetyl palmitate, polydimethylsiloxane, di-n-butyl sebacate, iso-propyl myristate, iso-propyl palmitate, iso-propyl stearate, butyl stearate, polyethylene glycol, triethylene glycol, lanolin, sesame oil, coconut oil, arachis oil, castor oil, acetylated lanolin alcohols, petroleum, mineral oil, butyl myristate, isostearic acid, palmitic acid, isopropyl linoleate, lauryl lactate, myristyl lactate, decyl oleate, myristyl myristate, and combinations thereof. Preferred emollients include stearyl alcohol and polydimethylsiloxane.

Ingredient (r) is a propellant. The amount of ingredient (r) in the topical composition is typically 5 to 95%. Suitable propellants include propane, butane, iso-butane, dimethyl ether, carbon dioxide, nitrous oxide, and combinations thereof. However, in a topical eyedrop no propellant is used.

Ingredient (s) is a solvent. The amount of ingredient (s) in the topical composition is typically 5 to 95%. Suitable solvents include water.

Ingredient (t) is a humectant. The amount of ingredient (t) in the topical composition is typically 5 to 95%. Suitable humectants include glycerin, sorbitol, sodium 2-pyrrolidone-5-carboxylate, soluble collagen, dibutyl phthalate, gelatin, and combinations thereof. Preferred humectants include glycerin.

Ingredient (u) is a thickener. The amount of ingredient (u) in the topical composition is typically 0 to 95%.

Ingredient (v) is a powder. The amount of ingredient (v) in the topical composition is typically 0 to 95%. Suitable powders include chalk, talc, fullers earth, kaolin, starch, gums, colloidal silicon dioxide, sodium polyacrylate, tetra alkyl ammonium smectites, trialkyl aryl ammonium smectites, chemically modified magnesium aluminum silicate, organically modified montmorillonite clay, hydrated aluminum silicate, fumed silica, carboxyvinyl polymer, sodium carboxymethyl cellulose, ethylene glycol monostearate, and combinations thereof.

Ingredient (w) is a fragrance. The amount of ingredient (w) in the topical composition is typically 0.001 to 0.5%, preferably 0.001 to 0.1%.

Component C) the optional activity enhancer is as described above. Any of the i) activity enhancers and ii) penetration enhancers may be added to the topical compositions. Preferably, the topical composition comprises 0.01 to 15% of component i) the optional additional IOP lowering medicament. More preferably, the composition comprises 0.1 to 10%, and most preferably 0.5 to 5% of component i). Preferably, the topical composition comprises 1 to 5% of component ii).

In an alternative embodiment of the invention, topical pharmaceutical compositions for ocular administration are prepared by conventional methods. Topical pharmaceutical compositions for ocular administration typically comprise A) a dual-action inhibitor, B) a carrier, such as purified water, and one or more ingredients selected from the group consisting of (y) sugars such as dextrans, particularly dextran 70, (z) cellulose or a derivative thereof, (aa) a salt, (bb) disodium EDTA (Edetate disodium), and (cc) a pH adjusting additive.

Examples of (z) cellulose derivatives suitable for use in the topical pharmaceutical composition for ocular administration include sodium carboxymethyl cellulose, ethyl cellulose, methyl cellulose, and hydroxypropylmethylcellulose. Hydroxypropylmethylcellulose is preferred.

Examples of (aa) salts suitable for use in the for use in the topical pharmaceutical composition for ocular administration include sodium chloride, potassium chloride, and combinations thereof.

Examples of (cc) pH adjusting additives include HCl or NaOH in amounts sufficient to adjust the pH of the topical pharmaceutical composition for ocular administration to 7.2-7.5.

The dual-action inhibitors may also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines. A preferred formulation for topical delivery of the present compounds uses liposomes as described in Dowton et al., “Influence of Liposomal Composition on Topical Delivery of Encapsulated Cyclosporin A”, S.T.P. Pharma Sciences, Vol. 3, pp. 404-407 (1993); Wallach and Philippot, “New Type of Lipid Vesicle: Novasome®”, Liposome Technology, Vol. 1, pp. 141-156 (1993); Wallach, U.S. Pat. No. 4,911,928, assigned to Micro-Pak, Inc., issued Mar. 27, 1990; and Weiner et al.

An effective amount of a compound according to the present invention will vary with the particular condition being treated, the age and physical condition of the patient being treated, the severity of the condition, the duration of treatment, the nature of concurrent therapy, the route of administration, the particular pharmaceutically-acceptable carrier utilized, and like factors within the knowledge and expertise of the attending physician. For example, an effective amount of the compounds of the present invention for systemic administration is from about 0.01 to about 1000 μg/kg body weight, preferably from about 0.1 to about 100 μg/kg per body weight, most preferably form about 1 to about 50 μg/kg body weight per day. The transdermal dosages will be designed to attain similar serum or plasma levels, based upon techniques known to those skilled in the art of pharmacokinetics and transdermal formulations. Plasma levels for systemic administration are expected to be in the range of 0.01 to 100 ng/mL, more preferably from 0.05 to 50 ng/mL and most preferably from 0.1 to 10 ng/mL. While these dosages are based upon a daily administration rate, the compounds of the present invention may also be administered at other intervals, such as twice per day, twice weekly, once weekly, or once a month. One of ordinary skill in the art would be able to calculate suitable effective amounts for other intervals of administration.

The exact amounts of each component in the topical composition depend on various factors. The amount of component A added to the topical composition is dependent on the IC50 of component A, typically expressed in nanomolar (nM) units. For example, if the IC50 of the medicament is 1 nM, the amount of component A will be from about 0.001 to about 0.3%. If the IC50 of the medicament is 10 nM, the amount of component A) will be from about 0.01 to about 1%. If the IC50 of the medicament is 100 nM, the amount of component A will be from about 0.1 to about 10%. If the IC50 of the medicament is 1000 nM, the amount of component A will be 1 to 100%, preferably 5% to 50%. If the amount of component A is outside the ranges specified above (i.e., lower), efficacy of the treatment may be reduced. One skilled in the art understands how to calculate and understand an IC50. The remainder of the composition, up to 100%, is component B.

The dual-action inhibitors may be included in kits comprising a dual-action inhibitor, a systemic or topical composition described above, or both; and information, instructions, or both that use of the kit will provide treatment for glaucoma in mammals (particularly humans). The information and instructions may be in the form of words, pictures, or both, and the like. In addition or in the alternative, the kit may comprise a dual-action inhibitor, a composition, or both; and information, instructions, or both, regarding methods of application of the dual-action inhibitor or composition, preferably with the benefit of lowering IOP in mammals.

In all of the foregoing compositions, and for all routes of administration, the dual-action inhibitor can be used alone or in combinations of two or more dual-action inhibitors. The compositions may further comprise additional drugs or excipients as appropriate for the indication. “Excipient” as used herein includes physiologically compatible additives useful in preparation of a pharmaceutical composition. Examples of pharmaceutically acceptable carriers and excipients can for example be found in Remington Pharmaceutical Science, 16th Ed.

Methods of the Invention

In certain embodiments, provided are methods for treating a disease or condition, the method comprising administering to a mammal in need thereof a therapeutically effective amount of a compound as described above. This invention further relates to a method for treating various diseases and conditions in mammals wherein a dual-action inhibitor will inhibit a kinase and monoamine transport in concert to alleviate the symptoms associated with the disease or condition. The methods may comprise administering to a mammal (preferably a human) in need thereof a therapeutically effective amount of a dual-action inhibitor as described above. “Administering” as used herein refers to administration of the compounds as needed to achieve the desired effect. “Therapeutically effective amount” as used herein refers to a dosage of the dual-action inhibitor or compositions thereof effective for influencing, reducing or inhibiting the activity of or preventing activation of a kinase and monoamine transport. This term as used herein may also refer to an amount effective at bringing about a desired in vivo effect in an animal, preferably, a human. Disease or conditions may include, but are not limited to, eye disease, bone disorder, obesity, heart disease, hepatic disease, renal disease, pancreatitis, cancer, myocardial infarct, gastric disturbance, hypertension, fertility control, disorders of hair growth, nasal congestion, neurogenic bladder disorder, gastrointestinal disorder, and dermatological disorder.

In one aspect, the present invention provides a method for treating an eye disease comprising administering to a subject in need thereof a therapeutically effective amount of a dual-action inhibitor as described above. “Eye disease” as used herein includes, but is not limited to, glaucoma, allergy, cancers of the eye, neurodegenerative diseases of the eye, and dry eye.

For example, a mammal diagnosed with glaucoma can be treated by the methods of this invention. Preferably, a topical composition comprising A) the dual-action inhibitor and B) a carrier is administered to the mammal. More preferably, the composition is a topical composition comprising A) the dual-action inhibitor, B) the carrier, and C) an optional activity enhancer.

In another aspect, the present invention provides a method of reducing intraocular pressure comprising contacting a cell with an effective amount of a dual-action inhibitor as described above. The term “contacting a cell” is used to mean contacting a cell in vitro or in vivo (i.e. in a subject, such as a mammal, including humans, rabbits, cats and dogs).

In yet another aspect, the present invention provides a method for treating cardiac indications comprising administering to a subject in need thereof a therapeutically effective amount of a dual-action inhibitor as described above. “Cardiac indications” as used herein includes, but is not limited to, angina pectoris, essential hypertension, myocardial infarction, supraventricular and ventricular arrhythmias, congestive heart failure, and atherosclerosis.

In a further aspect, the invention provides a method of treating a respiratory disorder comprising administering to a subject in need thereof a therapeutically effective amount of a dual-action inhibitor as described above. “Respiratory disorder” as used herein includes, inter alia, asthma, chronic bronchitis, bronchospasm, emphysema, and airway obstruction.

In another aspect, the invention provides a method of treating a renal disease comprising administering to a subject in need thereof a therapeutically effective amount of a dual-action inhibitor as described above. In yet another aspect, the invention provides a method of treating a diabetes comprising administering to a subject in need thereof a therapeutically effective amount of a dual-action inhibitor as described above.

In another aspect, the invention provides a method of treating upper respiratory indications comprising administering to a subject in need thereof a therapeutically effective amount of a dual-action inhibitor as described above. As used herein, “upper respiratory indications” include rhinitis and seasonal allergies.

In a further aspect, the invention provides a method of treating inflammatory disease, inflammation in response to injury, or rheumatoid arthritis comprising administering to a subject in need thereof a therapeutically effective amount of a dual-action inhibitor as described above.

Compounds according to Formulae (I), (II), (III), (IV), (V), (VI) or (VII) are useful in methods of inhibiting kinases and monoamine transporters in a cell, a tissue or a subject such as a human comprising contacting the cell with an amount of one or more of the compounds of the present invention effective to inhibit the kinase and the monoamine transporter. In one embodiment, the compounds are administered in a pharmaceutically acceptable composition, such as in or with a pharmaceutically acceptable carrier.

In another embodiment, the compounds of the present invention are used in methods for modulating the action of a kinase and a monoamine transporter in a cell comprising contacting the cell with amount of one or more compounds according to Formulae (I), (II), (III), (IV), (V), (VI) or (VII) effective to modulate the action of a kinase and a monoamine transporter in a cell. In one embodiment, the compounds of the present invention are administered in a pharmaceutically acceptable composition, such as in or with a pharmaceutically acceptable carrier.

Treatment or prevention of diseases or conditions for which the compounds of the present invention may be useful includes any of the diseases or conditions associated with kinase activity and monoamine transporter activity or diseases or conditions affected by kinases and monoamine transporters. Examples of these types of diseases include retinal degradation, glaucoma, cardiovascular diseases and cancer.

The term “disease or condition associated with kinase activity” is used to mean a disease or condition treatable, in whole or in part, by inhibition of one or more kinases.

The term “disease or condition associated with monoamine transport activity” is used to mean a disease or condition treatable, in whole or in part, by inhibition of one or more monoamine transporters.

The term “controlling the disease or condition” is used to mean changing the activity of one or more kinases or monoamine transporters to affect the disease or condition.

In some embodiments, the compounds of the present invention will be administered in conjunction with one or more additional therapeutic agents. Suitable additional therapeutic agents include, but are not limited to, beta blockers, alpha-agonists, carbonic anhydrase inhibitors, prostaglandin-like compounds, miotic or cholinergic agents, or epinephrine compounds.

Beta blockers. These reduce the production of aqueous humor. Examples include levobunolol (Betagan), timolol (Betimol, Timoptic), betaxolol (Betoptic) and metipranolol (OptiPranolol).

Alpha-agonists. These reduce the production of aqueous humor and increase drainage. Examples include apraclonidine (Iopidine) and brimonidine (Alphagan).

Carbonic anhydrase inhibitors. These also reduce the production of aqueous humor. Examples include dorzolamide (Trusopt) and brinzolamide (Azopt).

Prostaglandin-like compounds. These eyedrops increase the outflow of aqueous humor. Examples include latanoprost (Xalatan), bimatoprost (Lumigan) and travoprost (Travatan).

Miotic or cholinergic agents. These also increase the outflow of aqueous humor. Examples include pilocarpine (Isopto Carpine, Pilopine) and carbachol (Isopto Carbachol).

Epinephrine compounds. These compounds, such as dipivefrin (Propine), also increase the outflow of aqueous humor.

The additional therapeutic agent or agents can be administered simultaneously or sequentially with the compounds of the present invention. Sequential administration includes administration before or after the compounds of the present invention. In some embodiments, the additional therapeutic agent or agents can be administered in the same composition as the compounds of the present invention. In other embodiments, there can be an interval of time between administration of the additional therapeutic agent and the compounds of the present invention.

In some embodiments, the administration of an additional therapeutic agent with a compound of the present invention will enable lower doses of the other therapeutic agents to be administered for a longer period of time.

The dosage of the dual-action inhibitor administered depends on the method of administration. For systemic administration, (e.g., oral, rectal, nasal, sublingual, buccal, or parenteral), typically, 0.5 mg to 300 mg, preferably 0.5 mg to 100 mg, more preferably 0.1 mg to 10 mg, of a dual-action inhibitor described above is administered per day. These dosage ranges are merely exemplary, and daily administration can be adjusted depending on various factors. The specific dosage of the dual-action inhibitor to be administered, as well as the duration of treatment, and whether the treatment is topical or systemic are interdependent. The dosage and treatment regimen will also depend upon such factors as the specific dual-action inhibitor used, the efficacy of the compound, the personal attributes of the subject (such as, for example, weight, age, sex, and medical condition of the subject), compliance with the treatment regimen, and the presence and severity of any side effects of the treatment.

For topical administration (e.g., ocular), the topical composition is typically administered from once per day up to four times per day. In general, 1-2 weeks is sufficient to observe a noticeable decrease in IOP.

EXAMPLES

These examples are intended to illustrate the invention to those skilled in the art and should not be interpreted as limiting the scope of the invention set forth in the claims.

Reference Example One The Cell-Based Porcine Trabecular Meshwork (PTM) Assay

The anterior section of porcine eyes was harvested within 4 hours post-mortem. The iris and ciliary body were removed and trabecular meshwork cells were harvested by blunt dissection. Finely minced trabecular meshwork tissue was plated into collagen-coated 6-well plates in Medium-199 containing 20% fetal bovine serum (FBS). After two passages at confluence, cells were transferred to low-glucose DMEM containing 10% FBS. Cells were used between passage 3 and passage 8.

Cells were plated into fibronectin-coated, glass multiwell plates the day before compounds were tested under standard culture conditions. Compounds were added to cells in the presence of 1% FBS-containing DMEM and 1% DMSO. When compounds were incubated with the cells for the duration determined to be optimal, the media and compound is removed and cells fixed for 20 minutes in 3% methanol-free paraformaldehyde. Cells were rinsed twice with phosphate buffered saline (PBS) and cells are permeabilized with 0.5% Triton X-100 for two minutes. Following an additional two washes with PBS, F-actin was stained with Alexa-fluor 488-labelled phalloidin and nuclei are stained with DAPI.

Data was reduced to the mean straight actin-fiber length and normalized to DMSO-treated control cells (100%) and 50 μM Y-27632 (0%). Y-27632 is a rho-kinase inhibitor known to cause disruption of the actin cytoskeleton in these cells.

Reference Example Two NET/SERT Assay

Norepinephrine Transporter (NET) Membrane Radioligand Binding Assays. Total cell membranes were prepared from MDCK cells expressing the recombinant human norepinehrine transporter (hNET) grown to confluence in 150 mm tissue culture dishes. Cells were scraped into standard medium and pelleted at 1600 g. The medium was discarded and the pellet resuspended in 5 ml per plate of ice-cold binding buffer (100 mM NaCl, 50 mM Tris, pH 7.4 at room temperature) by trituration, and the cells were repelleted at 20,000 g. Supernatant was discarded and cells were resuspended in binding buffer (50 mM Tris-HCl, pH 7.4, 100 mM NaCl, 1 μM leupeptin, 10 μM PMSF) and homogenized with a polytron (Brinkman) at 25,000 revs/min for 5 s. Centrifugation, resuspension and homogenization were repeated and a sample of suspension was used for Bradford protein determination (BioRad). Samples of membrane suspensions were frozen at −80° C. prior to use. Typical yields were about 100 μg membrane protein per 106 cells. Assays performed in duplicate were initiated with 0.2 nM [125 l]RTI-55. Non-specific binding was determined by the inclusion of 10 μM desipramine. Incubation was carried out for 3 hours at 4° C. Assays were terminated by rapid filtration over GF/B glass-fiber filters soaked in 0.5% polyethylineimine using an automated cell harvester (Brandel) followed by three rapid 5 ml washes in ice-cold binding buffer. Bound radioactivity was measured by gamma emission spectrometry.

Serotonin Transporter (SERT) Membrane Radioligand Binding Assays. Total cell membranes were prepared from HEK-293 cells expressing the recombinant human serotonin transporter (hSERT) grown to confluence in 150 mm tissue culture dishes. Cells were scraped into standard medium and pelleted at 1600 g. The medium was discarded and the pellet resuspended in 5 ml per plate of ice-cold binding buffer (100 mM NaCl, 50 mM Tris, pH 7.4 at room temperature) by trituration, and the cells were repelleted at 20 000 g. Supernatant was discarded and cells were resuspended in binding buffer (50 mM Tris-HCl, pH 7.4, 120 mM NaCl, 5 mM KCl) and homogenized with a polytron (Brinkman) at 25,000 revs/min for 5 s. Centrifugation, resuspension and homogenization were repeated and a sample of suspension was used for Bradford protein determination (BioRad). Samples of membrane suspensions were frozen at −80° C. prior to use. Typical yields were about 100 μg membrane protein per 106 cells. Assays performed in duplicate were initiated with 0.4 nM [3H]paroxetine. Non-specific binding was determined by the inclusion of 10 μM imipramine. Incubation was carried out for 60 minutes at 25° C. Assays were terminated by rapid filtration over GF/B glass-fiber filters soaked in 0.5% polyethylineimine using an automated cell harvester (Brandel) followed by three rapid 5 ml washes in ice-cold binding buffer. Bound radioactivity was measured by beta emission spectrometry.

Reference Example Three Pharmacological Activity for Glaucoma Assay

Pharmacological activity for glaucoma can be demonstrated using assays designed to test the ability of the subject compounds to decrease intraocular pressure. Examples of such assays are described in the following reference, incorporated herein by reference: C. Liljebris, G. Selen, B. Resul, J. Sternschantz, and U. Hacksell, “Derivatives of 17-phenyl-18,19,20-trinorprostaglandin F2alpha Isopropyl Ester: Potential Anti-glaucoma Agents”, Journal of Medicinal Chemistry 1995, 38 (2): 289-304.

Reference Example Four

All temperatures were in degrees Centigrade. Reagents and starting materials were purchased from commercial sources or prepared following published literature procedures.

Unless otherwise noted, HPLC purification, when appropriate, was performed by redissolving the compound in a small volume of DMSO and filtering through a 0.45 micron (nylon disc) syringe filter. The solution was then purified using, for example, a 50 mm Varian Dynamax HPLC 21.4 mm Microsorb Guard-8 C8 column. A typical initial eluting mixture of 40-80% MeOH:H2O was selected as appropriate for the target compound. This initial gradient was maintained for 0.5 min then increased to 100% MeOH:0% H2O over 5 min. 100% MeOH was maintained for 2 more min before re-equilibration back to the initial starting gradient. A typical total run time was 8 min. The resulting fractions were analyzed, combined as appropriate, and then evaporated to provide purified material.

Proton magnetic resonance (1H NMR) spectra were recorded on either a Varian INOVA 600 MHz (1H) NMR spectrometer, Varian INOVA 500 MHz (1H) NMR spectrometer, Varian Mercury 300 MHz (1H) NMR spectrometer, or a Varian Mercury 200 MHz (1H) NMR spectrometer. All spectra were determined in the solvents indicated. Although chemical shifts are reported in ppm downfield of tetramethylsilane, they are referenced to the residual proton peak of the respective solvent peak for 1H NMR. Interproton coupling constants are reported in Hertz (Hz).

Analytical LCMS spectra were obtained using a Waters ZQ MS ESI instrument with an Alliance 2695 HPLC and a 2487 dual wavelength UV detector. Spectra were analyzed at 254 and 230 nm. Samples were passed through a Waters Symmetry C18 4.6×75 mm 3.5μ column with or without a guard column (3.9×20 mm 5μ). Gradients were run with mobile phase A: 0.1% formic acid in H2O and mobile phase B: ACN with a flow rate of 0.8 mL/min. Two gradients will illustrate:

Gradient A Gradient B Time A % B % Time A % B % 0.00 80.0 20.0 0.00 95.0 20.0 1.00 80.0 20.0 1.00 9.0 25.0 6.00 25.0 75.0 6.00 40.0 75.0 7.00 5.0 95.0 7.00 5.0 95.0 8.00 5.0 95.0 8.00 5.0 95.0 9.00 80.0 20.0 9.00 95.0 20.0 12.00 80.0 20.0 12.00 95.0 20.0

The settings for the MS probe were a cone voltage at 38 mV and a desolvation temperature at 250° C. Any variations in these methods are noted below.

The following preparations illustrate procedures for the preparation of intermediates and methods for the preparation of an amino isoquinoline amide derivatives or substituted benzamide derivatives.

Examples 1-12

Compounds E1-E12 may be synthesized according to the scheme shown in FIG. 1 and FIG. 6. For example, methyl 2-(4-(triisopropylsilyloxy)phenyl)acetate (E2) was synthesized from E1 according to the below:

To methyl 2-(4-hydroxyphenyl)acetate (E1) in CH2Cl2 at 0° C. was added 2,6-lutidine and TIPS-OTf. The ice bath was removed and the solution was allowed to warm to room temperature and stirred. After 4 h the solution was poured into NH4Cl(sat) and CH2Cl2 and the organic layer was further extracted with NH4Cl(sat). The organics were dried (Na2SO4) filtered and evaporated. Column chromatography (0-15% EtOAc/Hexanes) gave pure methyl-2-(4-(triisopropylsilyloxy)phenyl)acetate (E2).

Methyl 3-(1,3-dioxoisoindolin-2-yl)-2-(4-(triisopropylsilyloxy)phenyl)propanoate (E3) was prepared from E2 according to the below:

To a solution of LiHMDS in THF cooled to −78° C. was added a cooled solution (approx −78° C.) of methyl-2-(4-(triisopropylsilyloxy)phenyl)acetate (E2) in THF via syringe. The solution was stirred at −78° C. for 30 min. Bromo-methyl phthalimide was added directly to the anion, and the solution was immediately removed from the −78° C. bath and placed in an ice bath and stirred for 2 h. The reaction was then poured into NH4Cl(sat) and extracted with EtOAc. The organics were dried (Na2SO4), filtered, and evaporated. Column chromatography 0-20% EtOAc/Hexanes gave pure methyl 3-(1,3-dioxoisoindolin-2-yl)-2-(4-(triisopropylsilyloxy)phenyl)propanoate (E3).

2-(2-Carboxy-2-(4-(triisopropylsilyloxy)phenyl)ethylcarbamoyl)benzoic acid (E4) was prepared from E3 according to the below:

To methyl 3-(1,3-dioxoisoindolin-2-yl)-2-(4-(triisopropylsilyloxy)phenyl) propanoate (E3) in THF/H2O was added LiOH.H2O, and the solution was stirred for 1.5 h or until conversion to product was visible by LC-MS. The solution was then poured into EtOAc/NH4Cl(sat)/1 N HCl (3:1), and the aqueous layer was further extracted with EtOAc. The organics were dried (Na2SO4), filtered, evaporated, and dried to give crude 2-(2-carboxy-2-(4-(triisopropylsilyloxy)phenyl)ethylcarbamoyl)benzoic acid (E4).

3-(1,3-Dioxoisoindolin-2-yl)-N-(isoquinolin-6-yl)-2-(4-(triisopropylsilyloxy phenyl)propanamide (E5) was prepared from E4 according to the below:

To 2-(2-carboxy-2-(4-(triisopropylsilyloxy)phenyl)ethylcarbamoyl)benzoic acid (E4) in pyridine was added EDC, DMAP, and 6-aminoisoquinoline, and the solution was flushed with N2, capped, and stirred overnight. The mixture was poured into EtOAc/NaHCO3(sat) and the aqueous layer was further extracted with EtOAc. The organics were dried (Na2SO4), filtered, and evaporated. Column chromatography 5% MeOH/CH2Cl2 gave pure 3-(1,3-dioxoisoindolin-2-yl)-N-(isoquinolin-6-yl)-2-(4-(triisopropylsilyloxy phenyl)propanamide (E5).

3-amino-N-(isoquinolin-6-yl)-2-(4-(triisopropylsilyloxy)phenyl)propanamide (E6) was prepared from E5 according to the below:

To 3-(1,3-dioxoisoindolin-2-yl)-N-(isoquinolin-6-yl)-2-(4-(triisopropylsilyloxy phenyl)propanamide (E5) in EtOH was added NH2—NH2, and the solution was refluxed for 1.2 hrs-2 hrs. The solids were filtered and the solvents were evaporated. Column chromatography 5% 2N NH3-MeOH/CH2Cl2 gave pure 3-amino-N-(isoquinolin-6-yl)-2-(4-(triisopropylsilyloxy)phenyl)propanamide (E6).

Tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(4-(triisopropylsilyloxy)phenyl)propyl carbamate (E7) was prepared from E6 according to the below:

To 3-amino-N-(isoquinolin-6-yl)-2-(4-(triisopropylsilyloxy)phenyl)propanamide (E6) in CH2Cl2 (7.3 mL) at 0° C. was added a solution of Boc2O in CH2Cl2 also cooled to 0° C. before addition. The solution stirred for 30 min at 0° C. and additional Boc2O was added, and the solution was stirred for 30 min more then poured into CH2Cl2/NaHCO3(sat). The aqueous layers were further extracted with CH2Cl2, dried (Na2SO4), filtered, and evaporated. Column chromatography (3% MeOH/CH2Cl2) gave pure tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(4-(triisopropylsilyloxy)phenyl)propylcarbamate (E7).

Tert-Butyl 2-(4-hydroxyphenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E8) was prepared from E7 according to the below:

To tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(4-(triisopropyl silyloxy)phenyl)propylcarbamate (E7) in THF at 0° C. was added TBAF, and the solution was stirred for 45 min at 0° C. The compound was poured into EtOAc and washed with NH4Cl(sat), dried (Na2SO4), filtered, and evaporated. Column chromatography 6% MeOH/CH2Cl2 gave pure tert-butyl 2-(4-hydroxyphenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E8).

4-(3-tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenylpivalate (E9) was prepared from E8 according to the below:

To tert-butyl 2-(4-hydroxyphenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E8) in pyridine was added pivaloyl chloride, and the solution was stirred for 2 h at room temperature. The mixture was poured into NaHCO3 and extracted with EtOAc. The organics were dried (Na2SO4), filtered, and evaporated. Column chromatography 5% MeOH/CH2Cl2 gave pure 4-(3-tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenylpivalate (E9).

4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl pivalate (E10) was prepared from E9 according to the below:

To 4-(3-tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenylpivalate (E9) in CH2Cl2 was added HCl (4N in dioxane) and the solution was stirred for 8-10 h. The solvents were evaporated to give pure 4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl pivalate (E10).

4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl 1-methylcyclopropanecarboxylate (E11) was prepared from E8 according to the below:

To tert-butyl 2-(4-hydroxyphenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E8) in pyridine was added EDC, DMAP, and 1-methylcyclopropanecarboxylic acid, and the solution was flushed with N2, capped, and stirred overnight. The mixture was poured into EtOAc/NaHCO3(sat) and the aqueous layer was further extracted with EtOAc. The organics were dried (Na2SO4), filtered, and evaporated. Column chromatography 5% MeOH/CH2Cl2 gave pure 4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl 1-methylcyclopropanecarboxylate (E11).

4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl 1-methylcyclopropanecarboxylate dihydrochloride (E12) was prepared from E11 according to the below:

To 4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl 1-methylcyclopropanecarboxylate (E11) in CH2Cl2 was added HCl (4N in dioxane) and the solution was stirred for 8-10 h. The solvents were evaporated to give pure 4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl 1-methylcyclopropanecarboxylate dihydrochloride (E12).

Examples 13-122

Using commercially available compounds and largely the procedures set forth in Examples 2-12 and substituting the appropriate starting materials, the compounds E13-E91 were made and E92-E122 could be synthesized, shown in Tables 1 and 2, respectively.

TABLE 1 Compounds E13-E91. Example R2 R1 10 H -i-Pr 12 H 13 H Me 14 H t-Bu 15 H —C(CH3)2CH2CH3 16 H —(CH2)6CH3 17 H 18 H 19 H 20 H 21 H 22 H Ph 23 H 2-MePh 24 H 3-MePh 25 H 4-MePh 26 H 2,3-diMePh 27 H 2,4-diMePh 28 H 2,5-diMePh 29 H 3,4-diMePh 30 H 3,5-diMePh 31 H 2-F-Ph 32 H 3-F-Ph 33 H 4-F-Ph 34 H 2-Me, 3-F-Ph 35 H 2-Me, 4-F-Ph 36 H 2-Me, 5-F-Ph 37 H 4-t-BuPh 38 H 2-MeOPh 39 H 4-MeOPh 40 H 2,4-diMeOPh 41 H 2-MeO, 4-MePh 42 H 2-MeO-5-MePh 43 H 3,4-O—CH2—O-Ph 44 H 3-PhOPh 45 H —CH2-2-MeOPh 46 H 2-NH2-Ph 47 H 3-NH2-Ph 48 H 4-NH2-Ph 49 H 3-N(Me2)-Ph 50 H 4-N(Me2)-Ph 51 H 2-CN-Ph 52 H 4-CN-Ph 53 H 4-(CH2NH2)-Ph 54 H 2-CF3-Ph 55 H 2-pyridyl 56 H 3-pyridyl 57 H 4-pyridyl 58 H 2-Me-3-pyridyl 59 H 2-Ph-Ph 60 H 3-(COPh)-Ph 61 H 62 H 63 H —CH2NH2 64 H —CH(Ph)CH2NH2 65 H 66 H 67 H 68 H 69 H —Bn 70 H 4-(CH2NMe2)-Ph 71 H 72 H —CH(Me)Ph 73 H —CH2-3,4-diMeOPh 74 H —CH2CH2Ph 75 H —CH2CH2CH2Ph 76 H —CH2-2-MePh 77 H —CH2-3-MePh 78 H —CH2-4-MePh 79 H 80 H 81 H 82 H —CH2-4-FPh 83 H —CH2CO2tBu 84 H —CHEtPh 85 H —(CH2)10CH3 86 H —(CH2)7(Z)CH═CH(CH2)7CH3 87 H 88 H —CH2CH2CO2Me 89 H -(E)CH═CHCO2Me 90 H 91 H -3-MeOPh

TABLE 2 Compounds E92-E122. Example R2 R1 92 Me Me 93 Me 94 Me 95 Me Ph 96 Me 2-MePh 97 Et 2,5-diMePh 98 Et 3,4-diMePh 99 Et 2-Me, 3-F-Ph 100 Et 2-Me, 4-F-Ph 101 Propyl 2-MeOPh 102 Et 2,4-diMeOPh 103 Me 3,4-O—CH2—O— 104 Allyl 2-NH2-Ph 105 Allyl 3-NH2-Ph 106 H —CH2NH2 107 Me —CH(Ph)CH2NH2 108 Propyl —CH(Ph)CH2NH2 109 Et —CH(Ph)CH2NH2 110 Me Bn 111 Et Bn 112 Allyl Bn 113 Me 114 Me —CH(Me)Ph 115 Et —CH(Me)Ph 116 Propyl —CH(Me)Ph 117 Me —CH2CH2Ph 118 Et —CH2CH2CH2Ph 119 Me —CH2-2-MePh 120 Me —CH2-3-MePh 121 Me —CH(Et)Ph 122 Me

Examples 123-131

Using commercially available compounds and largely the procedures set forth in Examples 2-12 and substituting the appropriate starting materials, the compounds E123-E131 were made, shown in Table 3.

TABLE 3 Compounds E123-E131.

Examples 132-139

Compounds E132-E139 were prepared according to the scheme in FIG. 7.

Methyl 2-(4-(hydroxymethyl)phenyl)acetate (E132) was prepared according to the below:

To 2-(4-(hydroxymethyl)phenyl)acetic acid in MeOH at 0° C. was added TMS-CHN2. The solution was stirred for 3 h then quenched with a few drops of AcOH. The solvents were evaporated. Column chromatography (SiO2, 3-15% EtOAc/Hex) gave pure methyl 2-(4-(hydroxymethyl)phenyl)acetate (E132).

Methyl 2-(4-((triisopropylsilyloxy)methyl)phenyl)acetate (E133) was prepared from E132 according to the below:

To methyl 2-(4-(hydroxymethyl)phenyl)acetate (E132) in CH2Cl2 at 0° C. was added 2,6-lutidine and TIPS-OTf. The ice bath was removed and the solution was allowed to warm to room temperature and stir. After 4 h the solution was poured into NH4Cl(sat) and CH2Cl2 and the organic layer was further extracted with NH4Cl(sat). The organics were dried (MgSO4) filtered and evaporated. Column chromatography (SiO2, 0-15% EtOAc/Hexanes) gave pure methyl 2-(4-((triisopropylsilyloxy)methyl)phenyl)acetate (E133).

Methyl 3-(1,3-dioxoisoindolin-2-yl)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoate (E134) was prepared from E133 according to the below:

To a solution of LiHMDS in THF cooled to −78° C. was added a cooled solution (−78° C.) of methyl 2-(4-((triisopropylsilyloxy)methyl)phenyl)acetate (E133) in THF via syringe. The solution was stirred at −78° C. for 30 min. Bromo-methyl phthalimide was added directly to the anion and the solution stirred for 2 h at −78° C. The reaction was then poured into NH4Cl(sat) and extracted with EtOAc. The organics were dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 0-20% EtOAc/Hexanes) gave pure methyl 3-(1,3-dioxoisoindolin-2-yl)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoate (E134).

2-(2-carboxy-2-(4-((triisopropylsilyloxy)methyl)phenyl)ethylcarbamoyl)benzoic acid (E135) was prepared from E134 according to the below:

To methyl 3-(1,3-dioxoisoindolin-2-yl)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoate (E134) in THF/H2O was added LiOH.H2O, and the solution was stirred for 1.5 h or until conversion to product was visible by LC-MS. The solution was then poured into EtOAc/NH4Cl(sat)/1 N HCl (3:1) and the aqueous layer was further extracted with EtOAc. The organics were dried (MgSO4), filtered, and evaporated to give crude 2-(2-carboxy-2-(4-((triisopropylsilyloxy)methyl)phenyl)ethylcarbamoyl)benzoic acid (E135).

3-(1,3-dioxoisoindolin-2-yl)-N-(isoquinolin-6-yl)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanamide (E136) was prepared from E135 according to the below:

To 2-(2-carboxy-2-(4-((triisopropylsilyloxy)methyl)phenyl)ethylcarbamoyl)benzoic acid (E135) in pyridine was added EDC, DMAP and 6-aminoisoquinoline and the solution was flushed with N2, capped, and stirred overnight. The mixture was poured into EtOAc/NaHCO3(sat) and the aqueous layer was further extracted with EtOAc. The organics were dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 5% MeOH/CH2Cl2) gave pure 3-(1,3-dioxoisoindolin-2-yl)-N-(isoquinolin-6-yl)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanamide (E136).

3-amino-N-(isoquinolin-6-yl)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanamide (E137) was prepared from E136 according to the below:

To 3-(1,3-dioxoisoindolin-2-yl)-N-(isoquinolin-6-yl)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanamide (E136) in EtOH was added NH2—NH2 and the solution was refluxed for 1.2-2 h. The solids were filtered, and the solvents were evaporated. Column chromatography (SiO2, 5% 2N NH3-MeOH/CH2Cl2) gave pure 3-amino-N-(isoquinolin-6-yl)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanamide (E137).

Tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(4-((triisopropylsilyloxy)methyl)phenyl)propylcarbamate (E138) was prepared from E137 according to the below:

To 3-amino-N-(isoquinolin-6-yl)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanamide (E137) in CH2Cl2 at 0° C. was added a solution of Boc2O in CH2Cl2 also cooled to 0° C. before addition. The solution was stirred for 30 min at 0° C. and additional Boc2O was added and the solution was stirred for 30 min more then poured into CH2Cl2/NaHCO3(sat). The aqueous layers were further extracted with CH2Cl2, dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 3% MeOH/CH2Cl2) gave tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(4-((triisopropylsilyloxy)methyl)phenyl)propylcarbamate (E138).

Tert-butyl 2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E139).

To tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(4-((triisopropylsilyloxy)methyl)phenyl)propylcarbamate (E138) in THF at 0° C. was added TBAF, and the solution was stirred for 45 min at 0° C. The compound was poured into EtOAc and washed with NH4Cl(sat), dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 6% MeOH/CH2Cl2) gave pure tert-butyl 2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E139).

Examples 140-143

Compounds E140-E143 were prepared according to the scheme in FIG. 8.

4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2-methylbenzoate (E140) was prepared from E139 according to the below:

To tert-butyl 2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E139) in pyridine was added 2-methylbenzoyl chloride and the solution was stirred for 2 h at room temperature. The mixture was poured into NaHCO3 and extracted with EtOAc. The organics were dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 5% MeOH/CH2Cl2) gave pure 4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2-methylbenzoate (E140).

4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2-methylbenzoate dihydrochloride (E141) was prepared from E140 according to the below:

To 4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2-methylbenzoate (E140) in CH2Cl2 was added HCl (4N in dioxane) and the solution was stirred for 8-10 h. The solvents were evaporated to give pure 4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2-methylbenzoate dihydrochloride (E141).

4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl cyclohexanecarboxylate (E142) was prepared from E139 according to the below:

To tert-butyl 2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E139) in pyridine was added EDC, DMAP, and cyclohexanecarboxylic acid, and the solution was flushed with N2, capped, and stirred overnight. The mixture was poured into EtOAc/NaHCO3(sat) and the aqueous layer was further extracted with EtOAc. The organics were dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 5% MeOH/CH2Cl2) gave pure 4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl cyclohexanecarboxylate (E142).

4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl cyclohexanecarboxylate dihydrochloride (E143) was prepared from E142 according to the below:

To 4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl cyclohexanecarboxylate (E142) in CH2Cl2 was added HCl (4 N in dioxane) and the solution was stirred for 8-10 h. The solvents were evaporated to give pure 4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl cyclohexanecarboxylate dihydrochloride (E143).

Example 144

3-amino-2-(4-(hydroxymethyl)phenyl)-N-(isoquinolin-6-yl)propanamide dihydrochloride (E144) was prepared from E139 according to the below:

To tert-butyl 2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E139) in THF and water and cooled to 0° C. was added HCl (1 N in Et2O). After 30 min the mixture was warmed to room temperature and the solution was stirred for 48 h. 2 M NH3 in MeOH was added. The solvents were evaporated and the mixture purified by column chromatography (SiO2, 0-5-10% (2 M NH3 in MeOH)/CH2Cl2). The compound was dissolved in DCM/MeOH and 1 N HCl in Et2O added. The solvents were evaporated to give pure 3-amino-2-(4-(hydroxymethyl)phenyl)-N-(isoquinolin-6-yl)propanamide dihydrochloride (E144).

Examples 145-148

Compounds E145-E148 were prepared according to the scheme in FIG. 9.

4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E145) was prepared E139 according to the below:

To 2,4-dimethylbenzoic acid in pyridine was added EDC, DMAP, and tert-butyl 2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E139), and the solution was capped and stirred overnight. The mixture was poured into EtOAc/NaHCO3(sat) and the aqueous layer was further extracted with EtOAc. The organics were dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 0-5% MeOH/CH2Cl2 gradient) gave pure 4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E145).

4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate dihydrochloride (E146) was prepared from E145 according to the below:

To 4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E145) in CH2Cl2 was added HCl (4 N in dioxane) and the solution was stirred for 8-10 h. The solvents were evaporated to give pure 4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate dihydrochloride (E146).

4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl butyrate (E148) was prepared from E139 according to the below:

To butyric acid in pyridine was added EDC, DMAP, and tert-butyl 2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E139), and the solution was capped and stirred overnight. The mixture was poured into EtOAc/NaHCO3(sat) and the aqueous layer was further extracted with EtOAc. The organics were dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 0-5% MeOH/CH2Cl2 gradient) gave pure 4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl butyrate (E148).

4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl butyrate dihydrochloride (E148) was prepared from E147 according to the below:

To 4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl butyrate (E147) in CH2Cl2 was added HCl (4 N in dioxane) and the solution was stirred for 8-10 h The solvents were evaporated to give pure 4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl butyrateate dihydrochloride (E148).

Examples 149-175

Using commercially available compounds and largely the procedures set forth in Examples 140-143 and substituting the appropriate starting materials, the compounds E149-E175 have been made, shown in Table 4.

TABLE 4 Compounds E149-E175. Example R 149 150 -iPr 151 -tBu 152 —(CH2)6CH3 153 154 155 156 -Ph 157 -Bn 158 —CH2CH2Ph 159 —CH2—OPh 160 161 3,5-diMePh 162 163 —(CH2)10CH3 164 165 3-MeOPh 166 4-MeOPh 167 2,4-diOMePh 168 3,4-O—CH2—O-Ph 169 170 —CHPh2 171 2-Ph-Ph 172 173 174 175

Examples 176-196

Using commercially available compounds and largely the procedures set forth in Examples 140-143 and substituting the appropriate starting materials, the compounds E176-E196, could be made, shown in Table 5.

TABLE 5 Compounds E176-E196. Example R 176 2-MeOPh 177 4-NHMePh 178 4-NMe2Ph 179 4-OEtPh 180 3-MePh 181 4-MePh 182 2,3-diMePh 183 2,6-MePh 184 3,4-MePh 185 186 2-ClPh 187 3-ClPh 188 4-ClPh 189 2-FPh 190 3-FPh 191 4-FPh 192 2,4-diClPh 193 2,4-diFPh 194 195 196

Example 197

(S)-4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E145-S) and (R)-4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E145-R) were prepared from E145 according to the scheme in FIG. 10. 4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate was dissolve in methanol and the R and S enantiomers separated by supercritical fluid chromatography (Chiralpak AS-H column, eluent: 18.8% MeOH, 0.2% dimethylethylamine, 80% CO2). The enantiomers were then each purified by column chromotagraphy (SiO2, 0-5% MeOH/CH2Cl2 gradient). The enantiomeric excess for each enantiomer was >98%.

(S)-4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate dihydrochloride (E197-S) was prepared from E145-S according to the scheme in FIGS. 6. To (S)-4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E145-S) in CH2Cl2 was added HCl (4 N in dioxane) and the solution was stirred for 8-10 h. The solvents were evaporated to give pure (S)-4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate dihydrochloride (E197-S). Analysis by chiral HPLC (Chiralpak AS-H, eluent: 90:10:0.1 EtOH:H2O:diethylamine) showed enantiomeric excess>98%.

(R)-4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate dihydrochloride (E197-R) was prepared from E145-R according to the scheme in FIGS. 6. To (S)-4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E145-R) in CH2Cl2 was added HCl (4 N in dioxane) and the solution was stirred for 8-10 h. The solvents were evaporated to give pure (R)-4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate dihydrochloride (E197-R). Analysis by chiral HPLC (Chiralpak AS-H, eluent: 90:10:0.1 EtOH:H2O:diethylamine) showed enantiomeric excess>98%.

Examples 198-203

Compounds E199-E203 were prepared according to the scheme in FIG. 11.

(4-iodobenzyloxy)triisopropylsilane (E199) was prepared from E198 according to the below:

To a solution of (4-iodophenyl)methanol (E198) and imidazole in CH2Cl2 at 0° C. was added dropwise TIPSCI. The reaction mixture was stirred overnight. The solution was quenched with H2O and the CH2Cl2 layer separated. The organic layer was further washed with 0.5N HCl and NaHCO3(sat). The combined organic layers were dried (MgSO4), filtered, and evaporated. The crude yellow oil, (4-iodobenzyloxy)triisopropylsilane (E199), was used directly in the next step.

Ethyl 2-cyano-2-(4-((triisopropylsilyloxy)methyl)phenyl)acetate (E200) was prepared from E199 according to the below:

To a solution of ethyl 2-cyanoacetate and (4-iodobenzyloxy)triisopropylsilane (E199) in dioxane were added Cs2CO3, CuI, and picolinic acid. The mixture was stirred overnight at 90° C. The solid was removed by filtration and the dioxane concentrated under reduced pressure. Column chromatography (SiO2, hexane:ethyl acetate 25:1) gave pure ethyl 2-cyano-2-(4-((triisopropylsilyloxy)methyl)phenyl)acetate (E200) as a yellow oil.

Ethyl 3-amino-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoate (E201) was prepared from E200 according to the below:

To a suspension of CoCl26H2O in THF was added ethyl 2-cyano-2-(4-((triisopropylsilyloxy)methyl)phenyl)acetate (E200). The mixture was cooled to 0° C. and NaBH4 was added to the mixture in several portions over 30 min. The mixture was stirred at room temperature for 4 h. The reaction was quenched with water. The mixture was filtered and the filtrate extracted twice with ether. The organics were dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, DCM:EtOH=50:1) gave pure ethyl 3-amino-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoate (E201) as a yellow oil.

Ethyl 3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoate (E202) was prepared from E201 according to the below:

To a solution of ethyl 3-amino-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoate (E201) in DCM was added (Boc)2O and triethylamine. The mixture was stirred for 2 h, then washed with 0.5 N HCl and NaHCO3(sat). The organic layer was dried (MgSO4) and concentrated to give ethyl 3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoate (E202).

3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoic acid (E203) was prepared from E202 according to the below:

To a solution of ethyl 3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoate (E202) in methanol was added dropwise 4 N NaOH. The mixture was stirred for 2 h, adjusted the pH to 7 with 2 N HCl, and extracted with ethyl acetate. The combined organic layers were washed with 0.5 N HCl and brine, dried (MgSO4), and concentrated in vacuo to afford 3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoic acid (E203) as white solid.

Examples 204-206

Compounds E204-E206 were prepared according to the scheme in FIG. 12, which is a modified procedure by Cheung, S. T. et al. Can. J. Chem. 1977, 55, 906-910.

3-(tert-butoxycarbonyl(methyl)amino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoic acid (E204) was prepared from E203 according to the below:

To 3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoic acid (E203) in THF under N2 and cooled to 0° C. was added CH3I followed by NaH and the solution was warmed and allowed to stir for 18 h. The mixture was taken up in EtOAc and extracted with NH4Cl(sat), dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 0-10% MeOH/CH2Cl2 gradient) gave pure 3-(tert-butoxycarbonyl(methyl)amino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoic acid (E204).

Tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(4-((triisopropylsilyloxy)methyl)phenyl)propyl(methyl)carbamate (E205) was prepared from E204 according to the below:

To 3-(tert-butoxycarbonyl(methyl)amino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoic acid (E204) in pyridine was added was added EDC, DMAP, and 6-aminoisoquinoline, and the solution was stirred overnight at room temperature. The mixture was poured into NaHCO3(sat) and extracted with EtOAc. The organics were dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 0-6% MeOH/CH2Cl2 gradient) gave pure tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(4-((triisopropylsilyloxy)methyl)phenyl)propyl(methyl)carbamate (E205).

Tert-butyl 2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropyl(methyl)carbamate (E205-1) was prepared from E205 according to the below:

To tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(4-((triisopropylsilyloxy)methyl)phenyl)propyl(methyl)carbamate (E205) in THF under N2 at 0° C. was added TBAF, and the solution was stirred for 30 min at 0° C. The reaction was warmed to room temperature and stirred another 4.5 h. The compound was poured into EtOAc and washed with NH4Cl(sat), dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 0-20% MeOH/CH2Cl2 gradient) gave pure tert-butyl 2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropyl(methyl)carbamate (E205-1)

4-(3-(tert-butoxycarbonyl(methyl)amino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E205-2) was prepared from E205-1 according to the below:

To tert-butyl 2-(4-(hydroxymethyl)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropyl(methyl)carbamate (E205-1) in pyridine was added was added EDC, DMAP, and 2,4-dimethylbenzoic acid, and the solution was stirred overnight at room temperature. The mixture was poured into NaHCO3(sat) and extracted with EtOAc. The organics were dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 0-5% MeOH/CH2Cl2gradient) gave pure 4-(3-(tert-butoxycarbonyl(methyl)amino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E205-2).

4-(1-(isoquinolin-6-ylamino)-3-(methylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E206) was prepared from E205-2 according to the below:

To 4-(3-(tert-butoxycarbonyl(methyl)amino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E205-2) in CH2Cl2 was added HCl (4 N in dioxane) and the solution was stirred for 8-10 h. The solvents were evaporated to give pure 4-(1-(isoquinolin-6-ylamino)-3-(methylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E206).

Examples 207-211

Using commercially available compounds and largely the procedures set forth in Examples 204-206 and substituting the appropriate starting materials, the compounds E206-E211 have been made, shown in Table 6.

TABLE 6 Compounds E206-E211.

Example 212

3-(tert-butoxycarbonylamino)-2-(3-((triisopropylsilyloxy)methyl)phenyl)propanoic acid (E212) was prepared.

Using commercially available compounds and largely the procedures set forth in Examples 198-203 and substituting the appropriate starting materials 3-(tert-butoxycarbonylamino)-2-(3-((triisopropylsilyloxy)methyl)phenyl)propanoic acid (E212) was made.

Examples 213-216

Using commercially available compounds and largely the procedures set forth in Examples 204-206 and substituting the appropriate starting materials, the compounds E213-E216 have been made, shown in Table 7.

TABLE 7 Compounds E213-E216

Examples 217-225

Using commercially available compounds and largely the procedures set forth in Examples 198-203 and Examples 204-206 and substituting the appropriate starting materials, the compounds E217-E225 could be made, shown in Table 8.

TABLE 8 Compounds E217-E225.   4-(3-amino-1-(isoquinolin-6-ylamino)-1- oxopropan-2-yl)-2-methoxybenzyl cyclopentanecarboxylate E217   4-(3-amino-1-(isoquinolin-6-ylamino)-1- oxopropan-2-yl)-3-fluorobenzyl benzoate E218   4-(3-amino-1-(isoquinolin-6-ylamino)-1- oxopropan-2-yl)-2-fluorobenzyl 2- phenylacetate E219   4-(3-amino-1-(isoquinolin-6-ylamino)-1- oxopropan-2-yl)-3-methylbenzyl benzoate E220   3-(3-amino-1-(isoquinolin-6-ylamino)-1- oxopropan-2-yl)-4-methylbenzyl pivalate E221   5-(3-amino-1-(isoquinolin-6-ylamino)-1- oxopropan-2-yl)-2-fluorobenzyl cyclohexanecarboxylate E222   5-(3-amino-1-(isoquinolin-6-ylamino)-1- oxopropan-2-yl)-2-methoxybenzyl 4- methylbenzoate E223   3-(3-amino-1-(isoquinolin-6-ylamino)-1- oxopropan-2-yl)-5-fluorobenzyl 2- phenoxyacetate E224   3-(3-amino-1-(isoquinolin-6-ylamino)-1- oxopropan-2-yl)-2-fluorobenzyl 3- fluorobenzoate E225

Example 226

3-(isopropylamino)-N-(isoquinolin-6-yl)-2-phenylpropanamide dihydrochloride (E226) was prepared as shown below:

To 3-amino-N-(isoquinolin-6-yl)-2-phenylpropanamide in MeOH/AcOH was added acetone and NaCNBH3. Then after 15 min the mixture was poured into NaHCO3(sat) and extracted with CH2Cl2. The organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 5% 2 N NH3-MeOH/CH2Cl2) gave pure 3-(isopropylamino)-N-(isoquinolin-6-yl)-2-phenylpropanamide. The compound was taken up in CH2Cl2 and HCl (1 M in Et2O) was added. The solution was evaporated to give 3-(isopropylamino)-N-(isoquinolin-6-yl)-2-phenylpropanamide dihydrochloride (E226).

Examples 227-230

Using commercially available compounds and largely the procedures set forth in Example 226 and substituting the appropriate starting materials, the compounds E227-E230 could be made, shown in Table 9.

TABLE 9 Compounds E227-E230.   o)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)be E227   4-(3-(benzylamino)-1-(isoquinolin-6-ylamino)- 1-oxopropan-2-yl)benzyl benzoate E228   4-(1-(isoquinolin-6-ylamino)-1-oxo-3- (propylamino)propan-2-yl)benzyl 2-phenylacetate E229   4-(3-(isopropylamino)-1-(isoquinolin-6-ylamino)-1- oxopropan-2-yl)benzyl cyclopentanecarboxylate E230 indicates data missing or illegible when filed

Examples 231-241

Compounds E231-E241 were prepared according to the scheme in FIG. 13.

Methyl 2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)acetate (E232) was prepared from E231 according to the below:

To 2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)acetic acid (E231) in MeOH at 0° C. was added TMS-CH2N2 until the solution persisted in a yellow color and TLC indicated completion of the reaction. The solution stirred for 30 min and then was quenched with a few drops of AcOH. The solvents were evaporated and column chromatography (SiO2, 0-15% EtOAc/Hexanes) gave pure methyl 2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)acetate (E232).

Methyl 2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(1,3-dioxoisoindolin-2-yl)propanoate (E233) was prepared from E232 according to the below:

To a solution of LiHMDS in THF cooled to −78° C. was added a cooled solution (approx −78° C.) of methyl 2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)acetate (E232) in THF via syringe. The solution was stirred at −78° C. for 30 min. Bromo-methylphthalimide was added directly to the anion, and the solution was immediately removed from the −78° C. bath and placed in an ice bath and stirred for 2 h. The reaction was then poured into NH4Cl(sat) and extracted with EtOAc. The organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 15-20% EtOAc/Hexanes) gave pure methyl 2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(1,3-dioxoisoindolin-2-yl)propanoate (E233).

2-(2-(5-(tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-2-carboxyethylcarbamoyl)benzoic acid (E234) was prepared from E233 according to the below:

To methyl 2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(1,3-dioxoisoindolin-2-yl)propanoate (E233) in THF/H2O was added LiOH*H2O, and the solution was stirred for 1.5 h or until complete conversion to product was visible by LC-MS. The solution was then poured into EtOAc/NH4Cl(sat)/1 N HCl (3:1) and the aqueous layer was further extracted with EtOAc. The organics were dried (Na2SO4), filtered, evaporated, and dried to give crude 2-(2-(5-(tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-2-carboxyethylcarbamoyl)benzoic acid (E234).

2-(2-(5-(tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(1,3-dioxoisoindolin-2-yl)-N-(isoquinolin-6-yl)propanamide (E235) was prepared from E234 according to the below:

To 2-(2-(5-(tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-2-carboxyethylcarbamoyl)benzoic acid (E234) in pyridine was added EDC, DMAP, and 6-aminoisoquinoline, and the solution was flushed with N2, capped, and stirred overnight. The mixture was poured into EtOAc/NaHCO3(sat) and the aqueous layer was further extracted with EtOAc. The organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 4% MeOH/CH2Cl2) gave pure 2-(2-(5-(tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(1,3-dioxoisoindolin-2-yl)-N-(isoquinolin-6-yl)propanamide (E235).

3-amino-2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-N-(isoquinolin-6-yl)propanamide (E236) was prepared from E235 according to the below:

To 2-(2-(5-(tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(1,3-dioxoisoindolin-2-yl)-N-(isoquinolin-6-yl)propanamide (E235) in EtOH was added NH2—NH2 and the solution was stirred for 7 h at room temperature then heated to 50° C. for 1 h. The solution was cooled, the solids were filtered, and the solvents were evaporated. Column chromatography (SiO2, 5-8% 2 N NH3-MeOH/CH2Cl2) gave pure 3-amino-2-(5-((tert-butyldimethylsilyloxy) methyl)thiophen-2-yl)-N-(isoquinolin-6-yl)propanamide (E236).

Tert-butyl 2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E237) was prepared from E236 according to the below:

To 3-amino-2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-N-(isoquinolin-6-yl)propanamide (E236) in CH2Cl2 at 0° C. was added a solution of Boc2O in CH2Cl2 (also cooled to 0° C. before addition). The solution was stirred at 0° C. for 2 h and then poured into CH2Cl2 and NaHCO3(sat). The solution was further extracted with CH2Cl2 and the combined organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 3% MeOH/CH2Cl2) gave pure tert-butyl 2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E237).

Tert-butyl 2-(5-(hydroxymethyl)thiophen-2-yl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E238) was prepared from E237 according to the below:

To tert-butyl 2-(5-((tert-butyldimethylsilyloxy)methyl)thiophen-2-yl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E237) in THF at 0° C. was added TBAF and the solution was stirred for 0° C. for 30 min then warmed to room temperature for 2 h. The compound was poured into EtOAc and washed with NH4Cl(sat), dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 6% MeOH/CH2Cl2) gave pure tert-butyl 2-(5-(hydroxymethyl)thiophen-2-yl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E238).

3-amino-2-(5-(hydroxymethyl)thiophen-2-yl)-N-(isoquinolin-6-yl)propanamide dihydrochloride (E239) was prepared from E238 according to the below:

To a solution of tert-butyl 2-(5-(hydroxymethyl)thiophen-2-yl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E238) in CH2Cl2 was added 4 N HCl-dioxane and the solution was stirred for 4 h. The solvents were evaporated to give 3-amino-2-(5-(hydroxymethyl)thiophen-2-yl)-N-(isoquinolin-6-yl)propanamide dihydrochloride (E239).

(5-(3-tert-butoxylcarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)methyl 2,4 dimethylbenzoate (E240) was prepared from E239 according to the below:

To tert-butyl 2-(5-(hydroxymethyl)thiophen-2-yl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E238) in pyridine was added EDC, DMAP, and 2,4-dimethyl benzoic acid, and the solution was flushed with N2, capped, and stirred overnight. The mixture was poured into EtOAc/NaHCO3(sat) and the aqueous layer was further extracted with EtOAc. The organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 4% MeOH/CH2Cl2) gave pure (5-(3-tert-butoxylcarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)methyl 2,4 dimethylbenzoate (E240).

(5-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)thiophen-2-yl)methyl 2,4-dimethylbenzoate dihydrochloride (E241) was prepared from E240 according to the below:

To (5-(3-tert-butoxylcarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)methyl 2,4 dimethylbenzoate (E240) in CH2Cl2 was added HCl (4 N in dioxane) and the solution was stirred overnight. The solvents were evaporated to give pure (5-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)thiophen-2-yl)methyl 2,4-dimethylbenzoate dihydrochloride (E241).

Examples 242-248

Using commercially available compounds and largely the procedures set forth in Examples 231-241 and substituting the appropriate starting materials, E242-E248 could be synthesized, shown in Table 10.

TABLE 10 Compounds E242-E248. Example R1 R2 242 —CH2Ph H 243 -3,5-diMePh H 244 Me 245 H 246 —(CH2)2CH3 Me 247 i-Pr H 248 -Ph Me

Examples 249-253

Methyl 2-(4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenoxy)acetate (E250) was prepared from E249 according to the scheme in FIG. 14. To tert-butyl 2-(4-hydroxyphenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E249) in DMF cooled to −35° C. was added NaH, and the solution was stirred at −35° C. for 30 min. Then, methyl bromoacetate was added and the solution was warmed and stirred at 0° C. for 1 h. The solution was poured into NaHCO3(sat)/EtOAc and further extracted with EtOAc. The combined organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 3-4% MeOH/CH2Cl2) gave pure methyl 2-(4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenoxy)acetate (E250).

2-(4-(3-tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenoxy)acetic acid (E251) was prepared from E250 according to the below:

To methyl 2-(4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenoxy)acetate (E250) in THF/H2O/MeOH at 0° C. was added LiOH*H2O, and the solution was stirred for 2 h at 0° C. The mixture was then quenched with HCl (1 N, Et2O) and evaporated. Column chromatography (SiO2, 20% MeOH/CH2Cl2) gave pure 2-(4-(3-tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenoxy)acetic acid (E251).

2,4-dimethylphenyl 2-(4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenoxy)acetate (E252) was prepared from E251 according to the below:

To 2-(4-(3-tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenoxy)acetic acid (E251) in pyridine was added EDC, DMAP, and 2,4-dimethylphenol, and the solution was stirred for 5 h. The mixture was then poured into EtOAc/NaHCO3(sat) and extracted with EtOAc. The combined organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 2-3% MeOH/CH2Cl2) gave 2,4-dimethylphenyl 2-(4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenoxy)acetate (E252).

2,4-dimethylphenyl 2-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenoxy)acetate dihydrochloride (E253) was prepared from E252 according to the below:

To 2,4-dimethylphenyl 2-(4-(3-(tert-butoxycarbonylamino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenoxy)acetate (E252) in CH2Cl2 was added HCl (4 N, dioxane) and the solution was stirred overnight. The solvents were evaporated to give 2,4-dimethylphenyl 2-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenoxy)acetate dihydrochloride (E253).

Examples 254-273

Using commercially available compounds and largely the procedures set forth in Examples 249-253 and substituting the appropriate starting materials E254-E261 (shown in Table 11) were made and E262-E273 (shown in Table 12) could be synthesized.

TABLE 11 Compounds E254-E261. Example X R 254 O Me 255 O H 256 O -2,4-diMePh 257 O -i-Pr 258 O —CH2Ph 259 O -3,5-diMePh 260 NH Ph 261 NH —(CH2)3CH3

TABLE 12 Compounds E262-E273. Example X R1 R2 262 O Ph H 263 O 4-MeOPh Me 264 O 2,4-di-F-Ph Me 265 O —CH2Ph H 266 O —CH2CH═CH2 H 267 O Me 268 NH 2,4-diMePh Me 269 NH 3,5-diMePh Me 270 NH 2-F-Ph H 271 NH —CH2-4-MeOPh Me 272 NH -2-MeOPh H 273 NH -3-pyridyl H

Example 274

Using commercially available compounds and largely the procedures set forth in Examples 249-253 and substituting the appropriate starting materials E274 was made.

Examples 275-278

Compounds E275-E278 were prepared according to the scheme presented in FIG. 15. Tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(4-(2-oxo-2-phenylethoxy)phenyl)propylcarbamate (E275) was prepared from E249 according to the below:

To tert-butyl 2-(4-hydroxyphenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E249) in DMF cooled to −35° C. was added NaH and the solution was stirred at −35° C. for 30 min. Then, 2-bromoacetophenone was added and the solution was warmed and stirred at 0° C. for 2 h. The solution was poured into NaHCO3(sat)/EtOAc and further extracted with EtOAc. The combined organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 3% MeOH/CH2Cl2) gave tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(4-(2-oxo-2-phenylethoxy)phenyl)propylcarbamate (E275).

3-amino-N-(isoquinolin-6-yl)-2-(4-(2-oxo-2-phenylethoxy)phenyl)propanamide dihydrochloride (E277) was prepared from E275 according to the below:

To tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(4-(2-oxo-2-phenylethoxy)phenyl)propylcarbamate (E275) in CH2Cl2 was added HCl (4 N, dioxane) and the solution was stirred overnight. The solvents were evaporated to give pure 3-amino-N-(isoquinolin-6-yl)-2-(4-(2-oxo-2-phenylethoxy)phenyl)propanamide dihydrochloride (E277).

Tert-butyl 2-(4-(2-hydroxy-2-phenylethoxy)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E276) was prepared from E275 according to the below:

To tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(4-(2-oxo-2-phenylethoxy)phenyl)propylcarbamate (E275) in EtOH was added NaBH4 and the solution was stirred for 20 min at room temperature. The mixture was then poured into NaHCO3(sat) and extracted with CH2Cl2. The combined organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 5% MeOH/CH2Cl2) gave pure tert-butyl 2-(4-(2-hydroxy-2-phenylethoxy)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E276).

3-amino-2-(4-(2-hydroxy-2-phenylethoxy)phenyl)-N-(isoquinolin-6-yl)propanamide dihydrochloride (E278) was prepared from E276 according to the below:

To tert-butyl 2-(4-(2-hydroxy-2-phenylethoxy)phenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E276) in CH2Cl2 was added HCl (4 N, dioxane) and the solution was stirred overnight. The solvents were evaporated to give 3-amino-2-(4-(2-hydroxy-2-phenylethoxy)phenyl)-N-(isoquinolin-6-yl)propanamide dihydrochloride (E278).

Examples 279-288

Using commercially available compounds and largely the procedures set forth in Examples 275-278 and substituting the appropriate starting materials E279-E282 (shown in Table 13) were made and E283-E288 (shown in Table 14) could be synthesized.

TABLE 13 Compounds E279-E282. Example X R 279 O PH 280 OH Ph 281 O -4-MeOPh 282 O -2-MeOPh

TABLE 14 Compounds E283-E288. Example X R1 R2 283 O -2-F-Ph Me 284 O -2,4 diCl-Ph H 285 O -3-MePh H 286 OH -4-MeOPh H 287 OH -2-MeOPh H 288 OH -3-MePh Me

Examples 289-290

Compounds E289 and E290 were prepared according to the scheme presented in FIG. 16.

Tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(phenethoxyphenyl)propylcarbamate (E289) was prepared from E249 according to the below:

To tert-butyl 2-(4-hydroxyphenyl)-3-(isoquinolin-6-ylamino)-3-oxopropylcarbamate (E249) in DMF cooled to −35° C. was added NaH and the solution was stirred at −40° C. for 30 min. Then, 2-bromoethylbenzene was added and the solution was warmed and stirred at room temperature for 2 h. The solution was poured into NaHCO3(sat)/EtOAc and further extracted with EtOAc. The combined organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 3-4% MeOH/CH2Cl2) gave pure tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(phenethoxyphenyl)propylcarbamate (E289).

3-amino-N-(isoquinolin-6-yl)-2-(4-phenethoxyphenyl)propanamide dihydrochloride (E290) was prepared from E289 according to the below:

To tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(phenethoxyphenyl)propylcarbamate (E289) in CH2Cl2 was added HCl (4 N, dioxane) and the solution was stirred overnight. The solvents were evaporated to give 3-amino-N-(isoquinolin-6-yl)-2-(4-phenethoxyphenyl)propanamide dihydrochloride (E290).

Examples 291-299

Using commercially available compounds and largely the procedures set forth in Examples 289-290 and substituting the appropriate starting materials E291-E292 (Table 15) were made and E294-E299 (Table 16) could be synthesized.

TABLE 15 Compounds E291-E292. Example R 291 —(CH2)2Ph 292 —CH2Ph** **E292 was synthesized from previous schemes carried out in which the benzyl was in place of the TIPS protecting group.

Using commercially available compounds and largely the procedures set forth in Examples 289-290 and substituting the appropriate starting materials E293 was made.

TABLE 16 Compounds E294-E299. Example R1 R2 294 —CH2-4-F-Ph H 295 —CH2-2-MePh Me 296 —CH2-2-CNPh Me 297 —(CH2)2-4-MePh H 298 —(CH2)2-2-FPh H 299 H

Examples 300-308

Compounds E300-E308 were prepared according to the scheme in FIG. 17.

Benzyl 2-(4-hydroxyphenyl)acetate (E301) was prepared from E300 according to the below:

To 2-(4-hydroxyphenyl)acetic acid in DMF cooled to 0° C. was added K2CO3 and the solution was stirred for 30 min. Then, benzyl bromide was added and the solution stirred at 0° C. and was allowed to slowly warm to 15-20° C. After all the ice was melted the solution was poured into NH4Cl(sat) and extracted with EtOAc. The combined organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 0-35% EtOAc/Hex) gave pure benzyl 2-(4-hydroxyphenyl)acetate (E301).

Benzyl 2-(4-(triisopropylsiloxy)phenyl)acetate (E302) was prepared from E301 according to the below:

To benzyl 2-(4-hydroxyphenyl)acetate (E301) in CH2Cl2 at 0° C. was added 2,6-lutidine and TIPS-OTf and the solution stirred for 2.5 h at 0° C. The mixture was poured into NH4Cl(sat) and extracted with CH2Cl2. The combined organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 0-15% EtOAc/Hex) gave pure benzyl 2-(4-(triisopropylsiloxy)phenyl)acetate (E302).

Benzyl 3-cyano-2-(triisopropylsilyloxy)phenyl)propanoate (E303) was prepared from E302 according to the below:

To a solution of LiHMDS in THF at −78° C. was added a solution of benzyl 2-(4-(triisopropylsiloxy)phenyl)acetate (E302) in THF also cooled to approx −78° C., and this mixture was allowed to stir at −78° C. for 30 min. Iodoacetonitrile was then added and the mixture was warmed to 0° C. and stirred for 2 h. The mixture was poured into NH4Cl(sat) and extracted with EtOAc. The combined organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 0-25% EtOAc/Hex) gave pure benzyl 3-cyano-2-(triisopropylsilyloxy)phenyl)propanoate (E303).

Benzyl 4-(tertbutoxycarbonylamino)-2-(4-(triisopropylsilyloxy)phenyl)butanoate (E304) was prepared from E303 according to the below:

To a solution of benzyl 3-cyano-2-(triisopropylsilyloxy)phenyl)propanoate (E303) in MeOH cooled to 0° C. was added CoCl2*6H2O and NaBH4 and the solution was allowed to stir for 20 min. Then, HCl (1.25 N in MeOH) was added and the solution stirred an additional 20 min at 0° C. The solvents were evaporated and the mixture was taken up in CH2Cl2 and cooled to 0° C. Boc2O and NEt3 were added and the solution stirred at 0° C. for 1.5 h. The mixture was poured into NH4Cl(sat) and extracted with CH2Cl2. The combined organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 10-20% EtOAc/Hexanes) gave pure benzyl 4-(tertbutoxycarbonylamino)-2-(4-(triisopropylsilyloxy)phenyl)butanoate (E304).

Preparation of 4-(tertbutoxycarbonylamino)-2-(4-(triisopropylsilyloxy)phenyl)butanoic acid (E305) was prepared from E304 according to the below:

To benzyl 4-(tert-butoxycarbonylamino)-2-(4-(triisopropylsilyloxy)phenyl)butanoate (E304) in EtOAc was added Pd/C (10%) and the solution was kept under a H2 atmosphere for 2 h. The mixture was filtered over Celite and the solvent was evaporated to give 4-(tert-butoxycarbonylamino)-2-(4-(triisopropylsilyloxy)phenyl)butanoic acid (E305).

Tert-butyl 4-(isoquinolin-6-ylamino)-4-oxo-3-(4-(triisopropylsilyloxy)phenyl)butylcarbamate (E306) was prepared from E305 according to the below:

To 4-(tert-butoxycarbonylamino)-2-(4-(triisopropylsilyloxy)phenyl)butanoic acid (E305) in pyridine was added EDC, DMAP, and 6-AIQ, and the solution was stirred at room temperature overnight. The mixture was poured into NaHCO3(sat) and extracted with EtOAc. The combined organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 4% MeOH/CH2Cl2) gave pure tert-butyl 4-(isoquinolin-6-ylamino)-4-oxo-3-(4-(triisopropylsilyloxy)phenyl)butylcarbamate (E306).

Preparation of tert-butyl 3-(4-hydroxyphenyl)-4-(isoquinolin-6-ylamino)-4-oxobutylcarbamate (E307) was prepared from E306 according to the following:

To tert-butyl 4-(isoquinolin-6-ylamino)-4-oxo-3-(4-(triisopropylsilyloxy)phenylbutylcarbamate (E306) in THF at 0° C. was added TBAF and the solution was stirred at 0° C. for 30 min. The solution was poured into NH4Cl(sat) and extracted with EtOAc. The combined organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 5-8% MeOH/CH2Cl2) gave pure tert-butyl 3-(4-hydroxyphenyl)-4-(isoquinolin-6-ylamino)-4-oxobutylcarbamate (E307).

Preparation of 4-amino-2-(4-hydroxyphenyl)-N-(isoquinolin-6-yl)butanamide dihydrochloride (E308) was prepared from E307 according to the below:

To tert-butyl 3-(4-hydroxyphenyl)-4-(isoquinolin-6-ylamino)-4-oxobutylcarbamate (E307) in CH2Cl2 was added HCl (4 N in dioxane) and 2 drops of H2O and the solution was stirred overnight at room temperature. The solvents were evaporated to give 4-amino-2-(4-hydroxyphenyl)-N-(isoquinolin-6-yl)butanamide dihydrochloride (E308).

Examples 309-318

Using commercially available compounds and largely the procedures set forth in this application and substituting the appropriate starting materials E309-E318 could be synthesized.

TABLE 17 Compounds E309-E318. Example R1 R2 309 H H 310 —CO-Ph Me 311 CO-2,4-diMePh H 312 —COCH2Ph H 313 —CO(CH2)3CH3 H 314 —CH2COPh Me 315 —CH2CO-4-MeOPh Me 316 —CH2—CH(OH)-Ph H 317 —CH2-3-MeOPh H 318 —(CH2)2Ph Me

Examples 319-325

Compounds E319-E325 were prepared according to the scheme in FIG. 18.

Preparation of methyl 3-(diphenylmethyleneamino)-2-phenylpropanoate (E320) was prepared from E319 according to the below:

To methyl 3-amino-2-phenylpropanoate hydrochloride in CH2Cl2 was added benzophenone imine, and the solution was stirred overnight at room temperature. The mixture was then washed with H2O and the organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 5-20% EtOAc/Hexanes) gave pure methyl 3-(diphenylmethyleneamino)-2-phenylpropanoate (E320).

Methyl 3-(diphenylmethyleneamino)-2-methyl-2-phenylpropanoate (E321) was prepared from E320 according to the below:

To a solution of LiHMDS in THF cooled to −78° C. was added a solution of methyl 3-(diphenylmethyleneamino)-2-phenylpropanoate (E320) in THF also cooled to approximately −78° C. This solution stirred for 30 min at −78° C., then methyl iodide was added directly and the solution was warmed to 0° C. After 3 h the solution was poured into NH4Cl(sat) and extracted with EtOAc. The combined organics were dried (Na2SO4) filtered, and evaporated. Column chromatography (SiO2, 0-15% EtOAc/Hexanes) gave pure methyl 3-(diphenylmethyleneamino)-2-methyl-2-phenylpropanoate (E321).

3-amino-2-methyl-2-phenylpropanoic acid hydrochloride (E322) was prepared from E321 according to the below:

A mixture of methyl 3-(diphenylmethyleneamino)-2-methyl-2-phenylpropanoate (E321) and 6 N HCl was refluxed overnight. The solution was cooled and evaporated to give 3-amino-2-methyl-2-phenylpropanoic acid hydrochloride (E322).

3-(tert-butoxycarbonylamino)-2-methyl-2-phenylpropanoic acid (E323) was prepared from E322 according to the below:

To a solution of Boc2O in dioxane cooled to 0° C. was added a solution of 3-amino-2-methyl-2-phenylpropanoic acid hydrochloride (E322) in 1 N NaOH and this solution stirred 3 h and the solution was then washed with NaHCO3(sat)/CH2Cl2. The aqueous layer was acidified with HCl (1 N) and extracted with CH2Cl2. These combined organics were dried (Na2SO4), filtered, and evaporated to give 3-(tert-butoxycarbonylamino)-2-methyl-2-phenylpropanoic acid (E323).

Tert-butyl 3-isoquinolin-6-yl)-2-methyl-3-oxo-2-phenylpropylcarbamate (E324) was prepared from E323 according to the below:

To 3-(tert-butoxycarbonylamino)-2-methyl-2-phenylpropanoic acid (E323) in pyridine was added EDC, DMAP, and 6-AIQ, and solution was stirred at room temperature for 48 h. The mixture was poured into NaHCO3(sat) and extracted with EtOAc. The combined organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 3% MeOH/CH2Cl2) gave pure tert-butyl 3-isoquinolin-6-yl)-2-methyl-3-oxo-2-phenylpropylcarbamate (E324).

3-amino-N-(isoquinolin-6-yl)-2-methyl-2-phenylpropanamide dihydrochloride (E325) was prepared from E324 according to the below:

To tert-butyl 3-isoquinolin-6-yl)-2-methyl-3-oxo-2-phenylpropylcarbamate (E324) in CH2Cl2 was added HCl (4 N in dioxane) and the solution was stirred overnight at room temperature. The solvents were evaporated to give 3-amino-N-(isoquinolin-6-yl)-2-methyl-2-phenylpropanamide dihydrochloride (E325).

Examples 326-334

Using commercially available compounds and largely the procedures set forth in the Examples above and substituting the appropriate starting materials, E326-E334 could be synthesized, shown in Table 18.

TABLE 18 Compounds E326-E334. Example X n R1 R2 326 —OH 1 Me Me 327 —CH2OH 1 Me H 328 —OCOPh 2 Me H 329 —OCO-2,4-diMePh 1 —CH2Ph H 330 —OCOCH2Ph 1 —CH2Ph H 331 —CH2OCO-3,5-diMePh 1 Me H 332 —CH2OCO-2,4-diMePh 1 —CH2-4-MeOPh Me 333 —CH2OCO—(CH2)2CH3 1 —CH2-2-MeOPh H 334 —CH2OCO-2,4-diMePh 2 Me H

Examples 335-338

2-fluoro-4-nitrobenzamide (E335) was prepared according to the below:

To 2-fluoro-4-nitrobenzoic acid suspended in CH2Cl2 under Ar was added DMF then oxalyl chloride. The reaction was stirred at room temperature 1.5 h then the solvent was evaporated. The residue was dissolved in THF and ammonia gas was bubbled through the reaction for 15 min. The solvent was evaporated and the residue partitioned between EtOAc and water. The aqueous layer was extracted with EtOAc. The extracts were dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 0-100% EtOAc/Hex) gave pure 2-fluoro-4-nitrobenzamide (E335).

4-amino-2-fluorobenzamide (E336) was prepared from E335 according to the below:

2-fluoro-4-nitrobenzamide (E335) was dissolved in EtOH under Ar and 10% Pd/C added. The reaction was pump-purged with H2 and left stirring at room temperature overnight. The catalyst was removed by filtration and the reaction concentrated to give pure 4-amino-2-fluorobenzamide (E336).

Tert-butyl 3-(4-carbamoyl-3-fluorophenylamino)-3-oxo-2-(4-((triisopropylsilyloxy)methyl)phenyl)propylcarbamate (E337) was prepared from E336 according to the below:

To 3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoic acid in pyridine was added EDC, DMAP, and 4-amino-2-fluorobenzamide (E336), and the solution was stirred overnight at room temperature. The mixture was poured into NaHCO3(sat) and extracted with EtOAc. The extracts were dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 0-6% MeOH/CH2Cl2 gradient) gave pure tert-butyl 3-(4-carbamoyl-3-fluorophenylamino)-3-oxo-2-(4-((triisopropylsilyloxy)methyl)phenyl)propylcarbamate (E337).

Tert-butyl 3-(4-carbamoyl-3-fluorophenylamino)-2-(4-(hydroxymethyl)phenyl)-3-oxopropylcarbamate (E337-1) was prepared from E337 according to the below:

To tert-butyl 3-(4-carbamoyl-3-fluorophenylamino)-3-oxo-2-(4-((triisopropylsilyloxy)methyl)phenyl)propylcarbamate (E337) in THF under N2 at 0° C. was added TBAF, and the solution was stirred for 30 min at 0° C. The reaction was warmed to room temperature and stirred another 3.5 h. The compound was poured into EtOAc and washed with NH4Cl(sat), dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 0-20% MeOH/CH2Cl2 gradient) gave pure tert-butyl 3-(4-carbamoyl-3-fluorophenylamino)-2-(4-(hydroxymethyl)phenyl)-3-oxopropylcarbamate (E337-1)

4-(3-(tert-butoxycarbonylamino)-1-(4-carbamoyl-3-fluorophenylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E337-2) was prepared from E337-1 according to the below:

To tert-butyl 3-(4-carbamoyl-3-fluorophenylamino)-2-(4-(hydroxymethyl)phenyl)-3-oxopropylcarbamate (E337-1) in pyridine was added was added EDC, DMAP, and 2,4-dimethylbenzoic acid, and the solution was stirred overnight at room temperature. The mixture was poured into NaHCO3(sat) and extracted with EtOAc. The organics were dried (MgSO4), filtered, and evaporated. Column chromatography (SiO2, 0-5% MeOH/CH2Cl2 gradient) gave pure 4-(3-(tert-butoxycarbonylamino)-1-(4-carbamoyl-3-fluorophenylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E337-2).

4-(3-amino-1-(4-carbamoyl-3-fluorophenylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E338) was prepared from E337-2 according to the below:

To 4-(3-(tert-butoxycarbonylamino)-1-(4-carbamoyl-3-fluorophenylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E337-2) in CH2Cl2 was added HCl (4 N in dioxane) and the solution was stirred overnight. The solvents were evaporated to give pure 4-(3-amino-1-(4-carbamoyl-3-fluorophenylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E338).

Examples 339-370

Using commercially available compounds and largely the procedures set forth in Examples 335-338 and substituting the appropriate starting materials, the compounds E339-E354 (Table 19) and E355-E370 (Table 20) could be made.

TABLE 19 Compounds E339-E354. Example R1 R2 R3 R4 R5 339 H Bu H H F 340 Me Bu H H F 341 H Ph H H H 342 Me Ph H H H 343 H 3,5-diMePh F H H 344 H 2,4-diMePh H F H 345 H Bn H H F 346 H cyclohexyl Me H H 347 Me cyclopentyl H Me H 348 Me 3-MePh H H Me 349 H 4-MePh H H H 350 H 3-thienyl H H H 351 Me 2,4-diFPh H H H 352 H 3,5-diClPh H H H 353 Me 2-thienyl H H H 354 H 4-MeOPh H H H

TABLE 20 Compounds E355-E370. Example R1 R2 R3 R4 R5 355 H Bu H H F 356 Me Bu H H F 357 H Ph H H H 358 Me Ph H H H 359 H 3,5-diMePh F H H 360 H 2,4-diMePh H F H 361 H Bn H H F 362 H cyclohexyl Me H H 363 Me cyclopentyl H Me H 364 Me 3-MePh H H Me 365 H 4-MePh H H H 366 H 3-thienyl H H H 367 Me 2,4-diFPh H H H 368 H 3,5-diClPh H H H 369 Me 2-thienyl H H H 370 H 4-MeOPh H H H

Examples 371-377

Compounds E371-E377 were prepared according to the scheme in FIG. 19.

For the preparation of methyl 3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoate (E372), to a 0° C. solution of methyl 3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoic acid (E371) in MeOH was added a 2.0 M solution of trimethylsilyldiazomethane in hexanes. The solution was stirred for 20 min at room temperature and then quenched by the addition of a few drops of AcOH. The solution was concentrated and the residue, methyl 3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoate (E372), was used without purification.

For the preparation of methyl 3-(tert-butoxycarbonylamino)-2-(4-(hydroxymethyl)phenyl)propanoate (E373), to a 0° C. solution of methyl 3-(tert-butoxycarbonylamino)-2-(4-((triisopropylsilyloxy)methyl)phenyl)propanoate (E372) in THF was added a 1 M solution of tetrabutylammonium fluoride in THF, and the reaction was stirred overnight at room temperature. The reaction was quenched with saturated aqueous NH4Cl, and extracted with EtOAc (3×). The combined organics were washed with brine, dried over Na2SO4, and concentrated. The residue was purified by silica gel column chromatography (eluting with 0% to 50% EtOAc/hexanes) to yield methyl 3-(tert-butoxycarbonylamino)-2-(4-(hydroxymethyl)phenyl)propanoate (E373).

For the preparation of 4-(3-(tert-butoxycarbonylamino)-1-methoxy-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E374, R=2,4-Me2Ph), to a solution of methyl 3-(tert-butoxycarbonylamino)-2-(4-(hydroxymethyl)phenyl)propanoate (E373) in pyridine was added 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (EDCI), 4-(dimethylamino)pyridine (DMAP), and 2,4-dimethylbenzoic acid. The reaction was stirred overnight at room temperature. After addition of EtOAc and saturated aqueous NaHCO3, the mixture was extracted with EtOAc (3×). The combined organics were washed with brine, dried over Na2SO4, and concentrated. The residue was purified by silica gel column chromatography (eluting with 20% to 80% EtOAc/hexanes) to yield 4-(3-(tert-butoxycarbonylamino)-1-methoxy-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E374, R=2,4-Me2Ph).

For the preparation of 3-(tert-butoxycarbonylamino)-2-(4-((2,4-dimethylbenzoyloxy)methyl)phenyl)propanoic acid (E375, R=2,4-Me2Ph), to a solution of 4-(3-(tert-butoxycarbonylamino)-1-methoxy-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E374, R=2,4-Me2Ph) in 2:1 THF/H2O was added LiOH.H2O and the solution was stirred at room temperature for 3 h. After addition of 1 N HCl (until the pH was acidic), the mixture was extracted with EtOAc (3×). The combined organics were washed with brine, dried over Na2SO4, filtered, and concentrated to yield 3-(tert-butoxycarbonylamino)-2-(4-((2,4-dimethylbenzoyloxy)methyl)phenyl)propanoic acid (E375, R=2,4-Me2Ph).

For the preparation of 4-(3-(tert-butoxycarbonylamino)-1-(1-methoxyisoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E376, R=2,4-Me2Ph, X=1-OMe), to a solution of 3-(tert-butoxycarbonylamino)-2-(4-((2,4-dimethylbenzoyloxy)methyl)phenyl)propanoic acid (E375, R=2,4-Me2Ph) in pyridine was added EDCI, DMAP, and 6-amino-1-methoxyisoquinoline. The solution was stirred overnight at room temperature. The mixture was diluted with EtOAc and saturated aq. NaHCO3 solution. The mixture was extracted with EtOAc (3×). The combined organics were washed with brine, dried over Na2SO4, and concentrated. The residue was purified by silica gel column chromatography (eluting with 0% to 80% EtOAc/hexanes) to yield 4-(3-(tert-butoxycarbonylamino)-1-(1-methoxyisoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E376, R=2,4-Me2Ph, X=1-OMe).

For the preparation of 4-(3-amino-1-(1-methoxyisoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E377, R=2,4-Me2Ph, X=1-OMe), to a solution of 4-(3-(tert-butoxycarbonylamino)-1-(1-methoxyisoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E376, R=2,4-Me2Ph, X=3-Me) in CH2Cl2 was added 4N HCl in dioxane and the solution was stirred overnight at room temperature. The solution was concentrated. The residue was diluted with dichloromethane and concentrated again to yield 4-(3-amino-1-(1-methoxyisoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (E377, R=2,4-Me2Ph, X=2-OMe) as the hydrochloride salt.

Examples 378-380

Using largely the procedures shown above, the following compounds E378-E380 were synthesized, shown in Table 21.

TABLE 21 Compounds E378-E380. Example R 378 379 380

Examples 381-397

Using largely the procedures shown above, the following compounds E381-E397 could be synthesized, shown in Table 22.

TABLE 22 Compounds E381-E397. Example R 381 382 383 384 385 386 387 388 389 390 391 392 393 394 395 396 397

Examples 398-404

Compounds E399-E404 were prepared according to the scheme in FIG. 20.

Methyl 2-phenyl-3-(triisopropylsilyloxy)propanoate (E399) was prepared from E398 according to the below:

To methyl 3-hydroxy-2-phenylpropanolate in CH2Cl2 was at 0° C. was added 2,6-lutidine and TIPS-OTf, and this solution was stirred for 2 h at room temperature. The mixture was poured into NH4Cl(sat) and extracted with CH2Cl2. The organics were dried (Na2SO4), filtered, and evaporated. Column chromatography 0-10% EtOAc/Hex gave pure methyl 2-phenyl-3-(triisopropylsilyloxy)propanoate (E399).

2-phenyl-3-(triisopropylsilyloxy)propanoic acid (E400) was prepared from E399 according to the below:

To methyl 2-phenyl-3-(triisopropylsilyloxy)propanoate (E399) in THF/H2O/MeOH was added LiOH*H2O and the solution was stir at room temperature overnight. The solution was poured into NH4Cl(sat)/HCl (1 N) (3:1) and extracted with EtOAc. The combined organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (0%-4% MeOH/CH2Cl2) gave pure 2-phenyl-3-(triisopropylsilyloxy)propanoic acid (E400).

N-(isoquinolin-6-yl)-2-phenyl-3-(triisopropylsilyloxy)propanamide (E401) was prepared from E400 according to the below:

To 2-phenyl-3-(triisopropylsilyloxy)propanoic acid (E400) in pyridine was added EDC, DMAP, and 6-aminoisoquinoline, and the solution was flushed with N2, capped, and stirred overnight. The mixture was poured into NaHCO3(sat) and extracted with EtOAC. The combined organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (3-4% MeOH/CH2Cl2) gave pure N-(isoquinolin-6-yl)-2-phenyl-3-(triisopropylsilyloxy)propanamide (E401).

3-hydroxy-N-(isoquinolin-6-yl)-2-phenylpropanamide (E402) was prepared from E401 according to the below:

To N-(isoquinolin-6-yl)-2-phenyl-3-(triisopropylsilyloxy)propanamide (E401) in THF cooled to 0° C. was added TBAF and this solution was stirred for 3 h at 0° C. The mixture was poured into EtOAc/NH4Cl(sat) and washed with NH4Cl(sat). The organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (0-10% MeOH/CH2Cl2) gave pure 3-hydroxy-N-(isoquinolin-6-yl)-2-phenylpropanamide (E402).

3-(isoquinolin-6-ylamino)-3-oxo-2-phenylpropyl methanesulfonate (E403) was prepared from E402 according to the below:

To 3-hydroxy-N-(isoquinolin-6-yl)-2-phenylpropanamide (E402) in pyridine at 0° C. was added MsCl, and this solution was stirred at 0° C. for 2.5 h. The mixture was poured into NaHCO3(sat) and extracted with EtOAc. The combined organics were dried (Na2SO4), filtered, and evaporated to give 3-(isoquinolin-6-ylamino)-3-oxo-2-phenylpropyl methanesulfonate (E403).

N-(isoquinolin-6-yl)-3-(4-methylpiperazin-1-yl)-2-phenylpropanamide (E404) was prepared from E403 according to the below:

To 3-(isoquinolin-6-ylamino)-3-oxo-2-phenylpropyl methanesulfonate (E403) in methanol was added 1-methypiperazine, and the solution was stirred overnight at 50° C. The solvents were evaporated and column chromatography 10-20% 2 N NH3-MeOH/CH2Cl2 gave N-(isoquinolin-6-yl)-3-(4-methylpiperazin-1-yl)-2-phenylpropanamide (E404).

Examples 405-428

Using commercially available compounds and largely the procedures set forth in the previous examples and substituting the appropriate starting materials, the compounds E405-E410 (Table 23) and E411-E428 (Table 24 and Table 25) could be synthesized.

TABLE 23 Compounds E405-E410.   4-(1-isoquinolin-6-ylamino)-1-oxo-3-(4-phenylpiperazin-1- yl)propan-2-yl)phenyl 2-phenylacetate 405   4-(3-(4-acetylpiperazin-1-yl)-1-(isoquinolin-6-ylamino)-1- oxopropan-2-yl)phenyl benzoate 406   4-(3-(4-(cyclopropylmethyl)piperazin-1-yl)-1-(isoquinolin-6-ylamino)- 1-oxopropan-2-yl)phenyl 3,5-dimethylbenzoate 407   3-(1,4-diazepan-1-yl)-N-(isoquinolin-6-yl)-2-(4-(2- oxo-2-phenylethoxy)phenyl)propanamide 408   4-(3-(4-benzyl-1,4-diazepan-1-yl)-1-(isoquinolin-6-ylamino)- 1-oxopropan-2-yl)phenyl 2-phenylacetate 409   4-(1-(isoquinolin-6-ylamino)-1-oxo-3-(piperidin-1-yl)propan-2- yl)phenyl 2-phenylpropanoate 410

TABLE 24 Compounds E411-E419. Example R1 R2 R3 n 411 Me Me CO-2,4diMePh 1 412 Me Me CO—CH2Ph 1 413 Me CH2-4-HOPh CO—(CH2)2CH3 1 414 Me CH2-2-HOPh CH2—COPh 1 415 Me CH2-4-FPh CH2CO-4-MeOPh 1 416 Et CH2—Ph CH2C(OH)-2-MeOPh 2 417 Et Me CH2CH2Ph 2 418 Me CH2-3-pyridyl COBn 2 419 Me CH2-4-pyridyl COPh 2

TABLE 25 Compounds E420-E428. Example R1 R2 R3 n 420 Me Me CO-2,4diMePh 1 421 Me Me 1 422 Me CH2-4-MeOPh CO—(CH2)2CH3 2 423 Me CH2-2-HOPh CO-4-MePh 1 424 Me CH2-3-FPh COPh 1 425 Et CH2—Ph CO-3,5-diMePh 2 426 Et Me CO-Bn 1 427 Me CO(CH2)2CH3 2 428 Me CH2-4-pyridyl CO(CH2)2Ph 1

Examples 429-433

Compounds E429-E433 were prepared according to the scheme in FIG. 21, which is a modified procedure of Calmes et al., Eur. J. Org. Chem. 2000, 2459-2466.

Methyl 3-(1,3-dioxoisoindolin-2-yl)-2-(thiophen-3-yl)propanoate (E429) was prepared according to the below:

To pure methyl 2-(thiophen-3-yl)acetate in THF cooled to −78° C. was added LiHMDS and the solution stirred at −78° C. for 30 min. Then N-(bromomethyl)phthalimide was added directly and the solution was allowed to warm to 0° C. The mixture was poured into NaHCO3(sat) extracted with EtOAc, dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 0-40% EtOAc/Hex) gave pure methyl 3-(1,3-dioxoisoindolin-2-yl)-2-(thiophen-3-yl)propanoate (E429).

3-amino-2-(thiophen-3-yl)propanoic acid hydrochloride (E430) was prepared from E429 according to the below:

To methyl 3-(1,3-dioxoisoindolin-2-yl)-2-(thiophen-3-yl)propanoate (E429) was added 6 N HCl and the solution was refluxed for 4 h. The solvents were evaporated to give 3-amino-2-(thiophen-3-yl)propanoic acid (E430).

3-(tert-butoxycarbonylamino)-2-(thiophen-3-yl)propanoic acid (E431) was prepared from E430 according to the below:

To Boc2O in dioxane at 0° C. was added a cooled solution (0° C.) of 3-amino-2-(thiophen-3-yl)propanoic acid hydrochloride (E430) in 1 N NaOH. The solution was stirred at 0° C. for 30 min, then at room temperature for 4 h. The mixture was acidified with HCl and extracted with EtOAc and NH4Cl(sat). The organics were dried (Na2SO4), filtered, and evaporated to give pure of 3-(tert-butoxycarbonylamino)-2-(thiophen-3-yl)propanoic acid (E431).

Tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(thiophen-3-yl)propylcarbamate (E432) was prepared from E431 according to the below:

To 3-(tert-butoxycarbonylamino)-2-(thiophen-3-yl)propanoic acid (E431) in pyridine was added was added EDC, DMAP, and 6-aminoisoquinoline, and the solution was stirred overnight at room temperature. The mixture was poured into NaHCO3(sat) and extracted with EtOAc. The organics were dried (Na2SO4), filtered, and evaporated. Column chromatography (SiO2, 3% MeOH/CH2Cl2) gave pure tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(thiophen-3-yl)propylcarbamate (E432).

3-amino-N-(isoquinolin-6-yl)-2-(thiophen-3-yl)propanamide dihydrochloride (E433) was prepared from E432 according to the below:

To tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(thiophen-3-yl)propylcarbamate (E432) in CH2Cl2 was added HCl (4 N in dioxane) and the solution was stirred for 8-10 h. The solvents were evaporated to give pure 3-amino-N-(isoquinolin-6-yl)-2-(thiophen-3-yl)propanamide dihydrochloride (E433).

Examples 434-456

Using commercially available compounds and largely the procedures set forth in Examples 429-433 and substituting the appropriate starting materials, the following compounds E434-E441 (Table 26) and E442-E456 (Table 27) were made.

TABLE 26 Compounds E434-E441. Example X R4 R2 R1 434 H (±)-3-thienyl Me Me 435 H (±)-3-thienyl H H 436 H C6H5 H H 437 H C6H5 Me Me 438 F C6H5 H H 439 F C6H5 Me Me 440 H (±)-2-thienyl H H 441 Cl (±)-2-thienyl Me Me

TABLE 27 Compounds E442-E456. Example X R4 R2 R1 442 H (R)-C6H5 H H 443 H (S)-C6H5 H H 444 OH p-fluoro-C6H4 Me Me 445 H p-fluoro-C6H4 benzyl H 446 H Benzyl Me H 447 H p-fluoro benzyl Me H 448 OH 3-pyridyl H H 449 H 4-pyridyl Me Me 450 OH 3-furyl H H 451 H cyclopropyl Me Me 452 H cyclopentyl Me Me 453 OH cyclohexyl H H 454 H 3-benzo[b]thiophene Me Me 455 H H H 456 OH 2-oxazole H H

Reference Example Five Synthesis of Compounds According to Formula (II)

Compounds according to Formula (II) may be synthesized according to the scheme below:

Example 457

Preparation of methyl 3-(1,3-dioxoisoindolin-2-yl)-2-(thiophen-3-yl)propanoate (E1)

To pure methyl 2-(thiophen-3-yl)acetate in THF cooled to −78° C. was added LiHMDS and the solution stirred at −78° C. for 30 min. Then N-(bromomethyl)phthalimide was added directly and the solution was allowed to warm to 0° C. The mixture was poured into NaHCO3(sat) extracted with EtOAc, dried (Na2SO4), filtered and evaporated. Column chromatography (SiO2, 0-40% EtOAc/Hex) gave pure methyl 3-(1,3-dioxoisoindolin-2-yl)-2-(thiophen-3-yl)propanoate (E457).

Example 458 Preparation of 3-amino-2-(thiophen-3-yl)propanoic acid hydrochloride (E458)

To methyl 3-(1,3-dioxoisoindolin-2-yl)-2-(thiophen-3-yl)propanoate (E457) was added 6 N HCl and the solution was refluxed for 4 hours. The solvents were evaporated to give 3-amino-2-(thiophen-3-yl)propanoic acid (E458).

Example 459 Preparation of 3-(tert-butoxycarbonylamino)-2-(thiophen-3-yl)propanoic acid (E459)

To Boc2O in dioxane at 0° C. was added a cooled solution (0° C.) of 3-amino-2-(thiophen-3-yl)propanoic acid hydrochloride (E458) in 1 N NaOH. The solution was stirred at 0° C. for 30 min, then at room temperature for 4 hours. The mixture was acidified with HCl and extracted with EtOAc and NH4Cl(sat). The organics were dried (Na2SO4), filtered and evaporated to give pure of 3-(tert-butoxycarbonylamino)-2-(thiophen-3-yl)propanoic acid (E459).

Example 460 Preparation of tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(thiophen-3-yl)propylcarbamate (E460)

To 3-(tert-butoxycarbonylamino)-2-(thiophen-3-yl)propanoic acid (E459) in pyridine was added was added EDC, DMAP and 6-aminoisoquinoline and the solution was stirred overnight at room temperature. The mixture was poured into NaHCO3(sat) and extracted with EtOAc. The organics were dried (Na2SO4), filtered and evaporated. Column chromatography (SiO2, 3% MeOH/CH2Cl2) gave pure tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(thiophen-3-yl)propylcarbamate (E460).

Example 461 Preparation of 3-amino-N-(isoquinolin-6-yl)-2-(thiophen-3-yl)propanamide dihydrochloride (E461)

To tert-butyl 3-(isoquinolin-6-ylamino)-3-oxo-2-(thiophen-3-yl)propylcarbamate (E460) in CH2Cl2 was added HCl (4N in dioxane) and the solution was stirred for 8-10 hours The solvents were evaporated to give pure 3-amino-N-(isoquinolin-6-yl)-2-(thiophen-3-yl)propanamide dihydrochloride (E461).

Using commercially available compounds and largely the procedures set forth in Examples 457-461 and substituting the appropriate starting materials, the following compounds were made.

Example X R4 R2 R1 462 H (±)-3-thienyl Me Me 463 H (±)-3-thienyl H H 464 H C6H5 H H 465 H C6H5 Me Me 466 F C6H5 H H 467 F C6H5 Me Me 468 H (±)-2-thienyl H H 469 Cl (±)-2-thienyl Me Me

Example X R4 R2 R1 470 H (R)-C6H5 H H 471 H (S)-C6H5 H H 472 OH p-fluoro-C6H4 Me Me 473 H p-fluoro-C6H4 benzyl H 474 H Benzyl Me H 475 H p-fluoro benzyl Me H 476 OH 3-pyridyl H H 477 H 4-pyridyl Me Me 478 OH 3-furyl H H 479 H cyclopropyl Me Me 481 H cyclopentyl Me Me 482 OH cyclohexyl H H 483 H 3-benzo[b]thiophene Me Me 484 H H H 485 OH 2-oxazole H H

Example 486

Compounds according to Formula (III) may be synthesized according to the Scheme Two below:

To a solution of amino acid (1.0 eq.) in pyridine was added EDCI (1.1 eq.), DMAP (0.05 eq.), and 6-aminocinnoline (0.95 eq.). The solution was stirred at room temperature overnight. Saturated sodium bicarbonate solution was added and the mixture was extracted 3 times with ethyl acetate. Combined organics were washed with brine and dried over Na2SO4. The organics were concentrated and the residue was purified by column chromatography (eluting with 0 to 10% MeOH/CH2Cl2). (In some instances, it may be better to use ethyl acetate/hexanes as the eluent).

The BOC groups can be deprotected in two different ways:

(1) The protected amine (˜100 mg) was mixed with 4 mL of CH2Cl2 and 0.5 mL of 4N HCl in dioxane was added. The reaction was allowed to stir overnight. The reaction was concentrated, then methanol was added and the reaction was concentrated. This was repeated twice. Ether was added and the solution was concentrated one last time to obtain the product as a solid.

(2) The protected amine (˜100 mg) was mixed with 5 mL of CH2Cl2 and 0.5 mL of trifluoroacetic acid was added. The reaction was stirred at room temperature overnight. After the addition of saturated aqueous NaHCO3, the mixture was extracted 3 times with ethyl acetate. The combined organics were washed with brine, dried over Na2SO4 and concentrated. After analysis, the residue was dissolved in ethyl acetate or dichloromethane (sometimes a minimum volume of methanol is added to dissolve the product) and 1N HCl in ether is added to make the HCl salt if necessary.

Using largely the procedures above in Example 486 and substituting the appropriate starting materials, the following compounds were synthesized:

Example A B Y Z R 487 Direct bond C═O H Direct bond H 488 Direct bond C═O 4-Cl CH2 H

Using largely the procedures above in Example 486 and substituting the appropriate starting materials, the following compounds could be synthesized:

Example A B Y Z R 489 Direct bond C═O H CH2 H 490 CH2 C═O H Direct bond H 491 Direct bond C═O 4-Cl Direct bond H 492 Direct bond C═O 4-OH Direct bond H

Using largely the procedures above in Example 486 and substituting the appropriate starting materials, the following compounds could be synthesized:

Example 496

Compounds according to Formula (IV) may be synthesized according to Scheme Three shown below:

To a solution of amino acid (1.0 eq.) in pyridine is added EDCI (1.1 eq.), DMAP (0.05 eq.), and 5-aminoindazole (0.95 eq.). Stir at room temperature overnight. Saturated sodium bicarbonate solution is added and extract 3 times with ethyl acetate. Combined organics are washed with brine and dry over Na2SO4. The organics are concentrated and the residue is purified by column chromatography (eluting with 0 to 10% MeOH/CH2Cl2). (In some instances, it may be better to use ethyl acetate/hexanes as the eluent).

The compounds can be deprotected in two ways.

The protected amine (˜100 mg) is mixed with 4 mL of CH2Cl2 and 0.5 mL of 4N HCl in dioxane is added. The reaction is allowed to stir overnight. The reaction is concentrated, then methanol is added and the reaction is concentrated. This is repeated twice. Ether is added and the solution is concentrated one last time to obtain the product as a solid.

The protected amine (˜100 mg) is mixed with 5 mL of CH2Cl2 and 0.5 mL of trifluoroacetic acid is added. The reaction is stirred at room temperature overnight. After the addition of saturated aqueous NaHCO3, the mixture is extracted 3 times with ethyl acetate. The combined organics are washed with brine, dried over Na2SO4 and concentrated. After analysis, the residue is dissolved in ethyl acetate or dichloromethane (sometimes a minimum volume of methanol may be added to dissolve the product) and 1N HCl in ether is added to make the HCl salt if necessary.

Using largely the procedures above, the following compounds could be synthesized:

Example A B X1 X2 Y Z R 497 Direct C═O H H H CH2 H bond 498 CH2 C═O H H H Direct H bond 499 Direct C═O H H 4-Cl Direct H bond bond 500 Direct C═O H H 4-OH Direct H bond bond 501 Direct C═O H H H Direct H bond bond 502 Direct C═O H H 4-Cl CH2 H bond

Using largely the procedures above, the following compounds could be synthesized:

Synthesis of Compounds According to Formula (V)

Example 506

Compounds according to Formula (V) may be synthesized according to Scheme Four shown below:

The appropriate acid was converted to its acid chloride with oxalyl chloride then reacted with ammonia gas or another amine to give the amide. The nitro group was reduced to the aniline with hydrogen or another reducing agent. The aniline was coupled with the appropriate acid using standard coupling procedures such as EDC and DMAP in pyridine.

Example 507 Preparation of 2-fluoro-4-nitrobenzamide (E507)

To 2-fluoro-4-nitrobenzoic acid suspended in CH2Cl2 under Ar was added DMF then oxalyl chloride. The reaction was stirred at room temperature 1.5 hours then the solvent was evaporated. The residue was dissolved in THF and ammonia gas was bubbled through the reaction for 45 minutes. The solvent was evaporated and the residue partitioned between EtOAc and water. The aqueous layer was extracted with EtOAc. The extracts were dried (MgSO4), filtered and evaporated. Column chromatography (SiO2, 0-100% EtOAc/Hex) gave pure 2-fluoro-4-nitrobenzamide (E507).

Example 508 Preparation of 4-amino-2-fluorobenzamide (E508)

2-fluoro-4-nitrobenzamide (E507) was dissolved in EtOH under Ar and 10% Pd/C added. The reaction was pump-purged with H2 and left stirring at room temperature overnight. The catalyst was removed by filtration and the reaction concentrated to give pure 4-amino-2-fluorobenzamide (E508).

Example 509 Preparation of 4-(4-tert-butoxycarbonylamino-3-(4-chlorophenyl)butanamido)-2-fluorobenzamide (E509)

To 4-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)butanoic acid in pyridine was added EDC, DMAP and 4-amino-2-fluorobenzamide (E508) and the solution was stirred at room temperature. The mixture was poured into NaHCO3(sat) which resulted in a suspension. The suspension was separated from the aqueous layer which was then extracted with EtOAc. The suspension and extracts were combined and concentrated. Column chromatography (SiO2, 3:1 EtOAc/Hex then 100% EtOAc) gave pure 4-(4-tert-butoxycarbonylamino-3-(4-chlorophenyl)butanamido)-2-fluorobenzamide (E509).

Example 510 Preparation of 4-(4-amino-3-(4-chlorophenyl)butanamido)-2-fluorobenzamide (E510)

To 4-(4-tert-butoxycarbonylamino-3-(4-chlorophenyl)butanamido)-2-fluorobenzamide (E509) suspended in dichloromethane was added 4N HCl in dioxane. All of the starting material dissolved then a precipitate began to form. The reaction was stirred at room temperature. The mixture was poured into NaHCO3(sat) and extracted with EtOAc. The combined extracts were dried (MgSO4), filtered and evaporated. Column chromatography (SiO2, 5%-7.5% (2M NH3 in MeOH)/dichloromethane) gave pure 4-(4-amino-3-(4-chlorophenyl)butanamido)-2-fluorobenzamide (E510).

Using commercially available compounds and largely the procedures set forth in Examples 506 to 510 and substituting the appropriate starting materials, the following compounds were made:

Example A B X1 X2 Y Z R R4 511 Direct C═O H H H Direct H H bond bond 512 Direct C═O 3-F H H Direct H H bond bond 513 Direct C═O 3-F H H CH2 H H bond 514 (R) CH2 C═O 3-F H 4-Cl Direct H H bond 514 (S) CH2 C═O 3-F H 4-Cl Direct H H bond

Using commercially available compounds and largely the procedures set forth in Examples 506 to 510 and substituting the appropriate starting materials, the following compounds can be made:

Example A B X1 X2 Y Z R R4 516 CH2 C═O 3-OMe H H Direct H H bond 517 CH2 C═O 2-F H 3-Me Direct Et H bond 518 Direct C═O H H H Direct Me H bond bond 519 Direct C═O 3-F H H Direct H Me bond bond 520 Direct C═O 2-F H 4-Cl CH2 H H bond 521 Direct C═O H H 4- Direct H H bond OCOPh bond 522 Direct C═O 3-Cl H H O H H bond 523 Direct C═O H H 4-OMe Direct H H bond bond 524 Direct C═O 2-Me H 2-Cl, 4- Direct H H bond Cl bond

Using commercially available compounds and largely the procedures set forth in Examples 506 to 510 and substituting the appropriate starting materials, the following compounds can be made:

Example 528-532 Preparation of tert-Butyl 2-amino-1-phenylethylcarbamate (E532)

Tert-Butyl 2-hydroxy-1-phenylethylcarbamate (E529). To 2-(tert-butoxycarbonylamino)-2-phenylacetic acid (E528) in THF is added BH3-THE at 0° C. After stirring a few hours the solution is washed with NH4Cl (sat) and EtOAc, dried, filtered and evaporated to give crude E529. Column chromatography EtOAc/Hex gives pure tert-Butyl 2-hydroxy-1-phenylethylcarbamate (E529).

2-(tert-butoxycarbonylamino)-2-phenylethyl-4-methylbenzenesulfonate (E530). To E529 in CH2Cl2 is added NEt3 and TsCl. After stirring for 5 hours the solution is poured into NH4Cl (sat) and washed with CH2Cl2, dried and evaporated to give crude E530. Column chromatography EtOAc/Hex gives pure 2-(tert-butoxycarbonylamino)-2-phenylethyl-4-methylbenzenesulfonate (E530).

Tert-butyl 2-azido-1-phenylethylcarbamate (E531). To E530 in DMF is added NaN3 and the solution is stirred at 50° C. The reaction is then cooled and poured into NH4Cl (sat) and washed with EtOAc, dried, filtered and evaporated to give crude E531. Column chromatography EtOAc/Hex gives pure tert-butyl 2-azido-1-phenylethylcarbamate (E531).

Tert-Butyl 2-amino-1-phenylethylcarbamate (E532). To E531 in THF is added triphenylphosphine and the solution stirred 3-4 hours. Then H2O is added and the solution is stirred an additional 3-4 hours.

The solution is poured into NH4Cl (sat) and washed with EtOAc (to remove the triphenylphosphine oxide). Then the aqueous solution is made basic by addition of NaCO3(sat) and reextracted with CH2Cl2, dried and evaporated to give tert-butyl 2-amino-1-phenylethylcarbamate (E532).

Alternatively, to the reaction is added 1N HCl in Et2O and the mixture is triturated with benzene (to remove triphenylphosphine oxide) to give tert-butyl 2-amino-1-phenylethylcarbamate (E532).

Example 533-536 Preparation of N-(2-amino-2-phenylethyl)isoquinoline-5-sulfonamide (E536)

Isoquinoline-5-sulfonyl chloride (E534). To isoquinoline-5-sulfonic acid (E533) in DMF is added thionyl chloride and the solution is refluxed for 2 hours and the solution is evaporated. The residue is suspended with CHCl3, filtered and washed with more CHCl3 to give isoquinoline-5-sulfonyl chloride (E534).

Tert-Butyl 2-(isoquinoline-5-sulfonamido)-1-phenylethylcarbamate (E535). To a solution of E533 in CH2Cl2 at 0° C. is added NEt3 followed by a solution of E534 in CH2Cl2. After warming to room temperature and stirring for 2 h the mixture is poured into NaHCO3I(sat) and extracted with CH2Cl2, dried and evaporated to give crude E535. Column chromatography MeOH/CH2Cl2 gives tert-butyl 2-(isoquinoline-5-sulfonamido)-1-phenylethylcarbamate (E535).

N-(2-amino-2-phenylethyl)isoquinoline-5-sulfonamide dihydrochloride (E536). To E535 in CH2Cl2 is added 4 NHCl-dioxane and the solution is stirred for 12 hours at room temperature. The solvents are evaporated to give N-(2-amino-2-phenylethyl)isoquinoline-5-sulfonamide (E536).

Using largely the procedure set forth in Examples 528-536 and substituting the appropriate starting materials the following compounds can be made.

Example R X1 X2 537 H Me 538 Cl H 539 OH Me 540 H Me 541 OH H 542 H H 543 H Me 544 OH H 545 H H

Example 546 Preparation of N-(2-amino-2-phenylethyl)isoquinoline-5-carboxamide (E548)

Tert-butyl 2-(isoquinoline-5-carboxamido)-1-phenylethylcarbamate E547. To isoquinoline-5-carboxylic acid (lit. Loge, C; Siomboing, X et al. J. of Enzy Inhib & Med Chem, 2003, 18, 127-128), E546 in pyridine is added EDC, DMAP and E532 dissolved in pyridine. The solution is stirred overnight then poured into NaHCO3 and extracted with EtOAc, dried, evaporated to give crude E547. Column chromatography (MeOH/CH2Cl2) gives pure tert-butyl 2-(isoquinoline-5-carboxamido)-1-phenylethylcarbamate E547.

N-(2-amino-2-phenylethyl)isoquinoline-5-carboxamide dihydrochloride (E548). To E547 was added CH2Cl2 and 4 NHCl-dioxane and the solution is stirred at room temperature of 12 hours. The solvents are evaporated to give N-(2-amino-2-phenylethyl)isoquinoline-5-carboxamide dihydrochloride (E548).

Using largely the procedure set forth in Examples 528-532 and 546-548 and substituting the appropriate starting materials the following compounds can be made.

Example R X 549 H 550 H 551 OH 552 H Example No R X 553 H 554 OH 555 H 556 OH

Example 557-559 Preparation of 3-amino-N-(isoquinolin-5-yl)-2-phenylpropanamide (E24)

Tert-Butyl 3-(isoquinolin-5-ylamino)-3-oxo-2-phenylpropylcarbamate (E558). To 3-(tert-butoxycarbonylamino)-2-phenylpropanoic acid in pyridine is added EDC, DMAP and isoquinolin-5-amine (E557) and the solution is stirred for 12 hours at room temperature. The mixture is poured into NaHCO3(sat) and extracted with EtOAc, dried, filtered and evaporated to give crude E558. Column chromatography (MeOH/CH2Cl2) gave tert-butyl 3-(isoquinolin-5-ylamino)-3-oxo-2-phenylpropylcarbamate (E558).

3-amino-N-(isoquinolin-5-yl)-2-phenylpropanamide dihydrochloride (E559). To E558 in CH2Cl2 is added 4N HCl in dioxane and the solution is stirred at room temperature for 12 hours. The solvents are evaporated to give 3-amino-N-(isoquinolin-5-yl)-2-phenylpropanamide dihydrochloride (E559).

Using largely the procedure set forth in Examples 557-559 and substituting the appropriate starting materials the following compounds can be made.

Example No R X 560 H 561 OH 562 H 563 H 564 OH 565 H 566 H 567 OH

Example 568 Synthesis of Compounds According to Formula (VII)

Using commercially available compounds and procedures an ester link is created between venlafaxine (NET inhibitor) and fasudil (rho kinase inhibitor) to create 568, which metabolizes in the body to recreate the two independent moieties. Other suitable NET inhibitors are shown in FIG. 22.

Example 569

Topical pharmaceutical compositions for lowering intraocular pressure are prepared by conventional methods and formulated as follows:

Ingredient Amount (wt %) Dual-action inhibitor 0.50 Dextran 70 0.1 Hydroxypropyl methylcellulose 0.3 Sodium Chloride 0.77 Potassium chloride 0.12 Disodium EDTA 0.05 Benzalkonium chloride 0.01 HCl and/or NaOH pH 5.5-6.5 Purified water q.s. to 100%

Example 570

A compound according to this invention is used as the dual-action inhibitor in the composition according to Example 569. When the composition is topically administered to the eyes once daily, the above composition decreases intraocular pressure in a subject suffering from glaucoma.

Example 571

A compound according to this invention is used as the dual-action inhibitor in the composition according to Example 569. When administered as a drop 2 times per day, the above composition decreases intraocular pressure and serves as a neuroprotective agent.

Example 572

A compound according to this invention is used as the dual-action inhibitor in the composition according to Example 569. When administered as a drop twice per day, the above composition decreases intraocular pressure.

Example 573

A compound according to this invention is used as the dual-action inhibitor in the composition according to Example 569. When administered as a drop twice per day, the above composition substantially decreases allergic symptoms and relieves dry eye syndrome.

Example 574

A compound according to this invention is used as the dual-action inhibitor in the composition according to Example 569. When administered as a drop as needed, the above composition decreases hyperemia, redness and ocular irritation.

Example 575

A compound according to this invention is used as the dual-action inhibitor in the composition according to Example 569. When administered as a drop 4 times per day, the above composition decreases intraocular pressure and serves as a neuroprotective agent.

Example 576

A compound according to this invention is used as the dual-action inhibitor in the composition according to Example 569. When administered as a drop twice per day, the above composition decreases ocular pressure, allergic symptoms and relieves dry eye syndrome.

Claims

1. A compound comprising a monoamine transport inhibitor and a kinase inhibitor.

2. A compound of Formula II:

wherein each X is independently selected from hydrogen, amino, lower alkyl, halogen, carbonyl, nitrile, hydroxyl, and alkoxy;
Z is selected from substituted monovalent hydrocarbon groups, substituted heterogeneous groups, substituted heterocyclic groups, substituted heteroaromatic groups, substituted carbocyclic groups, substituted aromatic groups, heterogeneous groups, heterocyclic groups, heteroaromatic groups, carbocyclic groups, and aromatic groups;
R is selected from guanidino or —N(R5)2;
R5 is independently selected from H, methyl, or ethyl groups;
B is C═O, C═S, or —CH2—;
n and m are independently selected from the integers 0, 1, 2 or 3 and represent an independently variable number of substituted or unsubstituted methylene units in the alkyl chain; and
pharmaceutically acceptable salts, hydrates, and biohydrolyzable amides, esters, and imides thereof.

3. (canceled)

4. (canceled)

5. A compound of Formula III:

wherein X1 and X2 are independently selected from hydrogen, amino, lower alkyl, halogen, carbonyl, nitrile, hydroxyl, and alkoxy;
Z is selected from substituted monovalent hydrocarbon groups, substituted heterogeneous groups, substituted heterocyclic groups, substituted heteroaromatic groups, substituted carbocyclic groups, substituted aromatic groups, heterogeneous groups, heterocyclic groups, heteroaromatic groups, carbocyclic groups, and aromatic groups;
R is selected from guanidino or —N(R5)2;
R5 is independently selected from H, methyl, or ethyl groups;
B is C═O, C═S, or —CH2—;
n and m are independently selected from the integers 0, 1, 2 or 3 and represent an independently variable number of substituted or unsubstituted methylene units in the alkyl chain; and
pharmaceutically acceptable salts, hydrates, and biohydrolyzable amides, esters, and imides thereof.

6. (canceled)

7. A compound of Formula IV:

wherein each X1 and X2 are independently selected from hydrogen, amino, lower alkyl, halogen, carbonyl, nitrile, hydroxyl, and alkoxy;
Z is selected from substituted monovalent hydrocarbon groups, substituted heterogeneous groups, substituted heterocyclic groups, substituted heteroaromatic groups, substituted carbocyclic groups, substituted aromatic groups, heterogeneous groups, heterocyclic groups, heteroaromatic groups, carbocyclic groups, and aromatic groups;
R is selected from guanidino or —N(R5)2;
R5 is independently selected from H, methyl, and ethyl groups;
B is C═O, C═S, or —CH2—;
n and m are independently selected from the integers 0, 1, 2 or 3 and represent an independently variable number of substituted or unsubstituted methylene units in the alkyl chain; and
pharmaceutically acceptable salts, hydrates, and biohydrolyzable amides, esters, and imides thereof.

8. (canceled)

9. (canceled)

10. A compound of Formula V:

wherein X1 and X2 are independently selected from hydrogen, amino, lower alkyl, halogen, carbonyl, nitrile, and hydroxyl;
Z is selected from substituted monovalent hydrocarbon groups, substituted heterogeneous groups, substituted heterocyclic groups, substituted heteroaromatic groups, substituted carbocyclic groups, substituted aromatic groups, heterogeneous groups, heterocyclic groups, heteroaromatic groups, carbocyclic groups, and aromatic groups;
R is selected from guanidino or —N(R5)2;
R5 is independently selected from H, methyl, or ethyl groups;
R2 is H or methyl group;
B is C═O, C═S, or —CH2—;
n and m are independently selected from the integers 0, 1, 2 or 3 and represent an independently variable number of substituted or unsubstituted methylene units in the alkyl chain; and
pharmaceutically acceptable salts, hydrates, and biohydrolyzable amides, esters, and imides thereof.

11. (canceled)

12. (canceled)

13. A compound of Formula VI:

wherein each X is independently selected from hydrogen, amino, hydroxy, alkoxy, lower alkyl, halogen, carbonyl, and nitrile;
R1 and R2 are each independently selected from substituted monovalent hydrocarbon groups, substituted heterogeneous groups, substituted heterocyclic groups, substituted heteroaromatic groups, substituted carbocyclic groups, substituted aromatic groups, heterogeneous groups, heterocyclic groups, heteroaromatic groups, carbocyclic groups, and aromatic groups;
R is selected from guanidino and —N(R5)2;
R5 is selected from H, methyl group, and ethyl group;
A is —NR4—, —S(O)2—NH—, —NH—S(O)2—, C═O, or lower alkylene;
B is C═O, C═S, —CH2—, or NR4;
R4 is H, methyl group, or ethyl group;
n1 and n2 are independently selected from the integers 0, 1, and 2 and represent a variable number of carbon units in the alkyl chain; and
pharmaceutically acceptable salts, hydrates, and biohydrolyzable amides, esters, and imides thereof.

14. The compound of claim 13, wherein each X is independently selected from hydrogen, amino, hydroxy, and alkoxy;

R is —N(R5)2;
R5 is selected from H and methyl group; and
R4 is H or methyl group.

15. A compound of Formula VII:

wherein X7 is a —O— or —NH—;
A is a NET inhibitor; and
B is a rho kinase inhibitor.

16. (canceled)

17. A composition comprising:

(a) a compound according claim 13; and
(b) a carrier.

18. The composition of claim 17, further comprising (c) one or more activity enhancers.

19. A method for treating a disease or condition, the method comprising administering to a mammal in need thereof a therapeutically effective amount of a compound according to claim 13.

20. The method of claim 19, wherein the compound inhibits a kinase and monoamine transport in concert to alleviate the symptoms associated with the disease or condition.

21. The method of claim 19, wherein the disease or condition comprises at least one of eye disease, bone disorder, obesity, heart disease, hepatic disease, renal disease, pancreatitis, cancer, myocardial infarct, gastric disturbance, hypertension, fertility control, disorders of hair growth, nasal congestion, neurogenic bladder disorder, gastrointestinal disorder, and dermatological disorder.

22. The method of claim 21, wherein the disease or condition comprises an eye disease.

23. The method of claim 22, wherein the eye disease comprises glaucoma.

24. A method of reducing intraocular pressure comprising contacting a cell with an effective amount of a compound according to claim 13.

25. A method for treating cardiac indications comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to claim 13.

26. A method of treating a respiratory disorder comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to claim 13.

27. A method of treating a renal disease comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to claim 13.

28. A method of treating upper respiratory indications comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to claim 13.

29. A method of treating inflammatory disease, inflammation in response to injury, or rheumatoid arthritis comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to claim 13.

30. A method for modulating the action of a kinase and a monoamine transporter in a cell comprising contacting the cell with a compound according to claim 13 in an amount effective to modulate the action of a kinase and a monoamine transporter in a cell.

31. The method of claim 19, wherein the compound is administered in conjunction with one or more additional therapeutic agents.

32. The method of claim 31, wherein the additional therapeutic agent is selected from the group consisting of beta blockers, alpha-agonists, carbonic anhydrase inhibitors, prostaglandin-like compounds, miotic or cholinergic agents, and epinephrine compounds.

Patent History
Publication number: 20120135984
Type: Application
Filed: May 1, 2010
Publication Date: May 31, 2012
Inventors: Mitchell A. deLong (Chapel Hill, NC), Jill Marie Sturdivant (Chapel Hill, NC), Susan M. Royalty (Davis, CA), Geoffrey Richard Heintzelman (Durham, NC), Jeffrey D. Yingling (Apex, NC), Casey Kopczynski (Chapel Hill, NC)
Application Number: 13/318,457
Classifications
Current U.S. Class: Hetero Ring Is Seven-membered Consisting Of Two Nitrogens And Five Carbon Atoms (514/218); Nitrogen, Other Than As Nitro Or Nitroso, Attached Directly To The Isoquinoline Ring System By Nonionic Bonding (546/143); Nitrogen, Other Than As Nitro Or Nitroso, Attached Directly To The Isoquinoline Ring System By Nonionic Bonding (514/310); Amino Nitrogen, Not Bonded Directly To Carbonyl, Containing (564/157); Plural Carboxamide Groups Or Plural C=o Groups Bonded Directly To The Same Nitrogen (514/616); Ring In Alcohol Moiety (560/106); Z Or Y Radical Contains A Nitrogen Atom (514/534); Cyclopentyl-coor, -c-coor Or -c-c-coor (560/122); Z Contains A Cyclopentyl Or Cyclopentene Ring (514/530); The -c(=x)- Is Part Of A -c(=x)x- Group, Wherein The X's Are The Same Or Diverse Chalcogens (549/71); C=o Bonded Directly To The Hetero Ring (x Is Chalcogen) (514/448); Chalcogen Attached Directly To The Six-membered Hetero Ring By Nonionic Bonding (546/141); Chalcogen Attached Directly To The Six-membered Hetero Ring By Nonionic Bonding (514/309); Quinoline Or Isoquinoline (including Hydrogenated) (544/363); Isoquinolines (including Hydrogenated) (514/253.05); The Nitrogens Are In The 1,4-positions Of The Hetero Ring (540/575); Bicyclo Ring System Having The 1,2-diazine Ring As One Of The Cyclos (544/235); Polycyclo Ring System Having A 1,2- Or 1,4-diazine As One Of The Cyclos (514/248); The Other Ring Is A Benzene Ring (548/361.1); Pyrazoles (514/406); The Nitrogen Is Bonded Directly To A -c(=x)- Group, Wherein X Is Chalcogen (549/441); Carboxylic Acid Esters (560/1); Nitrogen Containing (514/466); Enzyme Inactivation By Chemical Treatment (435/184)
International Classification: A61K 31/4725 (20060101); C07D 217/02 (20060101); A61K 31/472 (20060101); C07D 409/12 (20060101); C07D 417/12 (20060101); C07D 409/14 (20060101); C07C 237/42 (20060101); A61K 31/167 (20060101); A61K 31/235 (20060101); C07C 69/74 (20060101); A61K 31/215 (20060101); C07D 333/38 (20060101); A61K 31/381 (20060101); A61K 31/496 (20060101); A61K 31/551 (20060101); C07D 237/28 (20060101); A61K 31/502 (20060101); C07D 231/56 (20060101); A61K 31/416 (20060101); C07D 317/60 (20060101); A61P 27/02 (20060101); A61P 27/06 (20060101); A61P 9/00 (20060101); A61P 11/00 (20060101); A61P 13/12 (20060101); A61P 29/00 (20060101); A61K 31/36 (20060101); C12N 9/99 (20060101); C07D 401/12 (20060101);