Therapeutic compounds and methods

Abstract

The invention provides a compound of formula (I): or a salt thereof, wherein R1-R7 and X have any of the values described in the specification, as well as compositions comprising a compound of formula (I) or a salt thereof. The compounds and salts are useful for activating a focal adhesion kinase (FAK) and for treating epithelial disease or damage. The invention also provides methods to treat bladder disease and damage, interstitial cystitis and transplant corneas with compounds discussed in herein.

Description

[0001] THERAPEUTIC COMPOUNDS AND METHODS

[0002] CROSS REFERENCE TO RELATED APPLICATION

[0003] This application claims priority to United States Provisional Application Number 63 / 681,481 that was filed on August 9, 2024. The entire content of the application referenced above is hereby incorporated by reference herein.

[0004] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0005] This invention was made with government support under HL 152410 awarded by the National Institutes of Health. The government has certain rights in the invention.

[0006] BACKGROUND

[0007] Failure of mucosal healing is central to diseases as diverse as inflammatory bowel disease (IBD), peptic ulcer, and necrotizing enterocolitis (NEC). Failure to heal impairs quality of life, may require risky surgery, and can lead to death. Approximately 1 million people in the US are afflicted with IBD, and 51,200 died of IBD in the United States alone in 2013. In addition to the GI tract, mucosal injury and repair are critical issues for oral and bladder injury and repair, such as radiation injury, interstitial cystitis, or oral ulceration from infectious or toxic agents, while many patients with skin wounds also suffer from non-healing epithelial wounds. Many patients also suffer from non-healing corneal ulcerations or wounds.

[0008] Focal adhesion kinase (FAK), a nonreceptor protein tyrosine kinase, is expressed in most tissues and cell types and is highly conserved across mammalian and other eukaryotic species (Schaller MD (2010) J Cell Sci 123: 1007-1013). The phosphorylation of FAK's tyrosine and serine residues in response to integrin engagement, mitogenic neuropeptides, lysophosphatidic acid, platelet-derived growth factor, activated Rho, and selected oncogenes leads to the formation of docking sites for a variety of signaling molecules that may regulate cell morphology, locomotion, proliferation, differentiation, and apoptosis (Schaller MD; Parsons JT (2003) J Cell Sci 116: 1409-1416; Hanks SK, Polte TR (1997) Bioessays 19: 137- 145). FAK activation is a convergent target for many growth factors and inhibiting or reducing FAK inhibits migration of epithelial cells. However, activated FAK is decreased in migrating intestinal epithelial cells in vitro and at the edge of human mucosal ulcers, making FAK an attractive target to promote mucosal and other epithelial healing.

[0009] International Patent Application Numbers PCT / US2018 / 060765 and PCT / US2021 / 056595 relate to FAK activators that are reported to promote mucosal and other epithelial healing. In spite of these reports, there remains a need for FAK activators that are useful to promote mucosal and other epithelial healing. In particular, there is a need for FAK activators with improved properties, such as, for example, improved potency, toxicity, in vivo half-lives, and / or physical properties relating to stability or formulate-ability.

[0010] SUMMARY

[0011] Small molecule FAK activators are provided.

[0012] Accordingly, one embodiment provides a compound of formula (I): or a salt thereof, wherein:

[0013] X is O, NRa, or CRbRc;

[0014] R1is (Ci-C3)alkyl;

[0015] R2is H or (Ci-C3)alkyl;

[0016] R3is a 6-membered heteroaryl;

[0017] R5is OR4, CHF2, OCH3, or CF3;

[0018] R4is trifluoromethyl, methoxymethyl, or -(CH2CH2)n-Rd;

[0019] R6is H or (Ci-C3)alkyl;

[0020] R7is H or (Ci-C3)alkyl;

[0021] Rais H or (Ci-C3)alkyl;

[0022] Rbis H, F, Cl, Br, or (Ci-C3)alkyl;

[0023] Rcis H, F, Cl, Br, or (Ci-C3)alkyl; n is 0, 1, 2, 3, or 4; and Rdis H, (Ci-C3)alkyl, or (Ci-C3)alkanoyl;

[0024] A compound of formula (I) which is a compound of formula (la): or a salt thereof, wherein:

[0025] X is O, NRa, or CRbRc;

[0026] R1is (Ci-C3)alkyl;

[0027] R2is H or (Ci-C3)alkyl;

[0028] R3is a 6-membered heteroaryl;

[0029] R4is trifluoromethyl, methoxymethyl, or -(CH2CH2O)n-Rd;

[0030] Rais H or (Ci-C3)alkyl;

[0031] Rbis H, F, Cl, Br, or (Ci-C3)alkyl;

[0032] Rcis H, F, Cl, Br, or (Ci-C3)alkyl; n is 0, 1, 2, 3, or 4; and

[0033] Rdis H, (Ci-C3)alkyl, or (Ci-C3)alkanoyl is also provided.

[0034] A pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient is also provided.

[0035] A method for treating an epithelial disease or epithelial damage in an animal comprising, administering compound of formula (I) or a pharmaceutically acceptable salt thereof to the animal is provided. A method for activating a focal adhesion kinase comprising, contacting the kinase (in vitro or in vivo) with a compound of formula (I) or a salt thereof is provided.

[0036] A method for activating a focal adhesion kinase in an animal comprising, administering a compound of formula (I) or a pharmaceutically acceptable salt thereof to the animal is provided.

[0037] A method for promoting mucosal healing in an animal comprising, administering a compound of formula (I) or a pharmaceutically acceptable salt thereof to the animal is provided.

[0038] A method for promoting epithelial healing in an animal comprising, administering a compound of formula (I) or a pharmaceutically acceptable salt thereof to the animal is provided.

[0039] A compound of formula (I) or a pharmaceutically acceptable salt thereof for use in medical therapy is provided.

[0040] A compound of formula (I) or a pharmaceutically acceptable salt thereof for the prophylactic or therapeutic treatment of an epithelial disease or epithelial damage is provided.

[0041] A compound of formula (I) or a pharmaceutically acceptable salt thereof for activating a focal adhesion kinase is provided.

[0042] A compound of formula (I) or a pharmaceutically acceptable salt thereof for promoting epithelial healing is provided.

[0043] A compound of formula (I) or a pharmaceutically acceptable salt thereof for promoting mucosal healing is provided.

[0044] Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof to prepare a medicament for treating an epithelial disease or epithelial damage in an animal is provided.

[0045] Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof to prepare a medicament for activating a focal adhesion kinase in an animal is provided.

[0046] Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof to prepare a medicament for promoting mucosal healing in an animal is provided.

[0047] Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof to prepare a medicament for promoting epithelial healing in an animal is provided.

[0048] Processes and intermediates disclosed herein that are useful for preparing a compound of formula (I) or a salt thereof are also provided.

[0049] BRIEF DESCRIPTION OF THE FIGURES

[0050] Figure 1 shows that M64HC1 can be used to promote skin wound healing. Human keratinocytes were cultured on plastic dishes precoated with 50 ug / ml type I collagen, small wounds were made in the monolayers, and wound closure was assessed at 12 hours as the difference between the wound area at 12 hours and the original wound area, expressed as a percentage of the original wound area. Monolayer wound closure was substantially accelerated by M64HC1 treatment. Wound closure after dosing with 100 nM M64HC1 was accelerated by 20+6% compared to control (p<0.006) and by 30+6% after treatment with 1000 nM M64HC1.

[0051] Figure 2 shows that M64HC1 can be used to promote skin wound healing. The study of Figure 1 was repeated on dishes precoated with 10 ug / ml type I collagen to simulate impaired wound healing due to abnormalities in the extracellular matrix. Monolayer wound closure was substantially accelerated by M64HC1 treatment. Wound closure after dosing with 1000 nM M64HC1 was accelerated by 30+8% compared to control (p<0.004). This suggests that the healing of human skin (which is covered with keratinocytes) can be accelerated by M64HC1 treatment.

[0052] Figure 3 shows that M64HC1 can be used to (a) promote corneal wound healing and (b) improve the quality of corneal transplants prior to transplantation. HCE-T human corneal epithelial cell monolayers were wounded and then treated with 100 or 1000 nM M64HC1 over 9 hours. The wounds were imaged at 0, 3, 6, and 9 hours and wound closure was expressed as the difference between the original wound area and the new wound area as a percentage of the original wound area. A statistically significant effect at 1000 nM (green bars above, p<0.05) at both 3 and 6 hours was observed. By 9 hours the wounds were all healed.

[0053] Figure 4 shows that FAK was being activated in HCE-T human corneal epithelial cell by Western blotting for phosphorylated FAK and expressing it as a ratio to total FAK. The result confirmed that M64HC1 was activating FAK within these cells. (n=8, p<0.001). This suggested that corneal wound healing might be accelerated by the ability of M64HC1 to activate FAK.

[0054] Figure 5 shows wound healing in human corneas. Human corneas that had been donated for transplantation but which for various reasons had been rejected for transplantation were used in the study. Wounds were made by placing a small filter paper disk soaked in heptanol on the cornea for 15 seconds, and the wounds were imaged at 0 and 24 hours. Paired human corneas in which one was treated with 1000 nM M64HC1 and one was not, were used. Wound healing was measured as described in figure 3 (the difference between the original wound area and the new wound area as a percentage of the original wound area was determined). These results confirm the marked acceleration of corneal wound healing induced by M64HC1. One cornea of the study was entirely dipped in heptanol, injuring the entire cornea. After 4 days of incubation with M64HC1, the cornea had effectively completely recovered. This further suggests that the condition corneas being stored awaiting transplantation or even those rejected for transplantation because of corneal injury can be improved by incubation with M64HC1. This is especially noteworthy as corneas can generally be stored for up to two weeks prior to transplantation.

[0055] Figure 6 shows that M64HC1 can be used to heal the bladder, including bladder injury related to interstitial cystitis and cyclophosphamide therapy. Interstitial cystitis is a challenging disease associated with disabling pelvic pain, dysuria, and frequency of urination. Although it occurs spontaneously, it can also be induced by cyclophosphamide and is seen in human patients receiving this anti-cancer drug. Human primary bladder epithelial cells were treated with 0, 100, and 1000 nM M64HC1 for one hour. Western blotting was used to assess the ratio of phosphorylated to total FAK as a measure of FAK activation. These results confirm that M64HC1 can activate FAK in bladder epithelium.

[0056] Figures 7A -7C show bladder weight, bladder hemorrhaging scoring and histiological image scoring in mice treated with cyclophosphamide or cyclophosphamide and M64HC1. The bladders of mice in whom cystitis had been induced by intraperitoneal injection of cyclophosphamide (CYP, 200 mg / kg) were compared. Mice received either a saline vehicle control or M64HC1 via osmotic minipump for one week. Figure 7A shows that at autopsy, the bladder weight was substantially lower in the mice receiving M64HC1, suggesting less tissue edema. Figure 7B shows that macroscopic inspection and blinded scoring of the bladder wall indicated a statistically significant improvement. Hemorrhaging was scored using a scale from 1 to 10, where 1 indicates no hemorrhage and 10 represents the most severe. Histological examination of the bladder wall in a CYP -induced mouse shows mucosal erosion, ulceration, hemorrhage, edema, and inflammatory cell infiltrates. By contrast, histological examination from a CYP -induced mouse treated with M64HC1 showed reduced epithelial denudation, erosion, ulceration and hemorrhage. Figure 7C shows the scoring of histological images. Histological images were scored by a blinded observer on a predetermined scale, where a higher score indicates increased injury. A statistically significant improvement in the injury score after M64HC1 treatment was observed. These results demonstrate that M64HC1 can be used to heal bladder injuries, and specifically to treat interstitial cystitis and cyclophosphamide-induced bladder injury DETAILED DESCRIPTION

[0057] The following definitions are used, unless otherwise described: halo or halogen is fluoro, chloro, bromo, or iodo. Alkyl, etc. denote both straight and branched groups; but reference to an individual radical, such as propyl, embraces only the straight chain radical, a branched chain isomer, such as isopropyl, being specifically referred to.

[0058] The term "alkyl", by itself or as part of another substituent, means, unless otherwise stated, a straight or branched chain hydrocarbon radical, having the number of carbon atoms designated (i.e., Ci-8 means one to eight carbons). Examples include (Ci-Cs)alkyl, (C2-Cs)alkyl, Ci-Ce)alkyl, (C2-Ce)alkyl and (C3-Ce)alkyl. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, iso-butyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and higher homologs and isomers.

[0059] The term “heteroaryl” as used herein refers to a single aromatic ring that has at least one atom other than carbon in the ring, wherein the atom is selected from the group consisting of oxygen, nitrogen and sulfur. Thus, “heteroaryl” includes single aromatic rings of from about 1 to 5 carbon atoms and about 1-4 heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur. The sulfur and nitrogen atoms may also be present in an oxidized form provided the ring is aromatic. Exemplary heteroaryl ring systems include but are not limited to pyridyl and pyrimidinyl.

[0060] The term “alkanoyl” as used herein refers to a group (alkyl)-C(=O)-, wherein the term alkyl has the meaning defined herein.

[0061] As used herein, the term "heteroatom" is meant to include oxygen (O), nitrogen (N), sulfur (S) and silicon (Si).

[0062] As used herein, the term "protecting group" refers to a substituent that is commonly employed to block or protect a particular functional group on a compound. For example, an "amino-protecting group" is a substituent attached to an amino group that blocks or protects the amino functionality in the compound. Suitable amino-protecting groups include acetyl, trifluoroacetyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBZ) and 9- fluorenylmethylenoxycarbonyl (Fmoc). Similarly, a "hydroxy-protecting group" refers to a substituent of a hydroxy group that blocks or protects the hydroxy functionality. Suitable protecting groups include acetyl and silyl. A "carboxy-protecting group" refers to a substituent of the carboxy group that blocks or protects the carboxy functionality. Common carboxy-protecting groups include phenylsulfonylethyl, cyanoethyl, 2-(trimethylsilyl)ethyl, 2-(trimethylsilyl)ethoxymethyl, 2-(p- toluenesulfonyl)ethyl, 2-(p-nitrophenylsulfenyl)ethyl, 2-(diphenylphosphino)-ethyl, nitroethyl and the like. For a general description of protecting groups and their use, see P.G.M. Wuts and T.W. Greene, Greene's Protective Groups in Organic Synthesis 4thedition, Wiley-Interscience, New York, 2006.

[0063] As used herein a wavy line “ ” that intersects a bond in a chemical structure indicates the point of attachment of the bond that the wavy bond intersects in the chemical structure to the remainder of a molecule.

[0064] The terms “epithelial disease” and “epithelial damage” include conditions of the gastrointestinal tract (e.g., inflammatory bowel disease IBD) as well as oral mucosal injury and mucosal injury of the bladder, including radiation injury, interstitial cystitis, and oral ulceration from infectious or toxic agents. These terms also include non-healing epithelial wounds and non-healing corneal ulcerations or wounds.

[0065] The terms “treat”, “treatment”, or “treating” to the extent it relates to a disease or condition includes inhibiting the disease or condition, eliminating the disease or condition, and / or relieving one or more symptoms of the disease or condition. The terms “treat”, “treatment”, or “treating” also refer to both therapeutic treatment and / or prophylactic treatment or preventative measures, wherein the object is to prevent or slow down (lessen) an undesired physiological change or disorder, such as, for example, the development or spread of cancer. For example, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease or disorder, stabilized (i.e., not worsening) state of disease or disorder, delay or slowing of disease progression, amelioration or palliation of the disease state or disorder, and remission (whether partial or total), whether detectable or undetectable. “Treat”, “treatment”, or “treating,” can also mean prolonging survival as compared to expected survival if not receiving treatment. Those in need of treatment include those already with the disease or disorder as well as those prone to have the disease or disorder or those in which the disease or disorder is to be prevented. In one embodiment “treat”, “treatment”, or “treating” does not include preventing or prevention,

[0066] The phrase "therapeutically effective amount" or “effective amount” includes but is not limited to an amount of a compound of the that (i) treats or prevents the particular disease, condition, or disorder, (ii) attenuates, ameliorates, or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of the particular disease, condition, or disorder described herein.

[0067] The term “animal” includes mammals, fish, amphibians, reptiles, birds and invertebrates. The term “mammal” includes humans, higher non-human primates, rodents, domestic, cows, horses, pigs, sheep, dogs and cats. In one embodiment, the animal is a mammal. In one embodiment, the animal is a human. The term “patient” as used herein refers to any animal including mammals. In one embodiment, the patient is a mammalian patient. In one embodiment, the patient is a human patient.

[0068] The compounds disclosed herein can also exist as tautomeric isomers in certain cases. Although only one delocalized resonance structure may be depicted, all such forms are contemplated.

[0069] It is understood by one skilled in the art that this invention also includes any compound claimed that may be enriched at any or all atoms above naturally occurring isotopic ratios with one or more isotopes such as, but not limited to, deuterium (2H or D). As a non-limiting example, a - CH3 group may be substituted with -CD3.

[0070] The pharmaceutical compositions can comprise one or more excipients. When used in combination with the pharmaceutical compositions the term “excipients” refers generally to an additional ingredient that is combined with the compound of formula (I) or the pharmaceutically acceptable salt thereof to provide a corresponding composition. For example, when used in combination with the pharmaceutical compositions the term “excipients” includes, but is not limited to: carriers, binders, disintegrating agents, lubricants, sweetening agents, flavoring agents, coatings, preservatives, and dyes.

[0071] Stereochemical definitions and conventions used herein generally follow S. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. The compounds can contain asymmetric or chiral centers, and therefore exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds, including but not limited to, diastereomers, enantiomers and atropisomers, as well as mixtures thereof such as racemic mixtures, form part of the present invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D and L, or R and S, are used to denote the absolute configuration of the molecule about its chiral center(s). The prefixes d and 1 or (+) and (-) are employed to designate the sign of rotation of plane-polarized light by the compound, with (-) or 1 meaning that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of one another. A specific stereoisomer can also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture or a racemate, which can occur where there has been no stereoselection or stereospecificity in a chemical reaction or process. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomeric species, devoid of optical activity.

[0072] It will be appreciated by those skilled in the art that compounds having a chiral center may exist in and be isolated in optically active and racemic forms. Some compounds may exhibit polymorphism. It is to be understood that the present invention encompasses any racemic, optically-active, polymorphic, or stereoisomeric form, or mixtures thereof, of a compound, which possess the useful properties described herein, it being well known in the art how to prepare optically active forms (for example, by resolution of the racemic form by recrystallization techniques, by synthesis from optically-active starting materials, by chiral synthesis, or by chromatographic separation using a chiral stationary phase.

[0073] When a bond in a compound formula herein is drawn in a non-stereochemical manner (e.g. flat), the atom to which the bond is attached includes all stereochemical possibilities. When a bond in a compound formula herein is drawn in a defined stereochemical manner (e.g. bold, bold-wedge, dashed or dashed-wedge), it is to be understood that the atom to which the stereochemical bond is attached is enriched in the absolute stereoisomer depicted unless otherwise noted. In one embodiment, the compound may be at least 51% the absolute stereoisomer depicted. In another embodiment, the compound may be at least 60% the absolute stereoisomer depicted. In another embodiment, the compound may be at least 80% the absolute stereoisomer depicted. In another embodiment, the compound may be at least 90% the absolute stereoisomer depicted. In another embodiment, the compound may be at least 95 the absolute stereoisomer depicted. In another embodiment, the compound may be at least 99% the absolute stereoisomer depicted.

[0074] Specific values listed below for radicals, substituents, and ranges, are for illustration only; they do not exclude other defined values or other values within defined ranges for the radicals and substituents. It is to be understood that two or more values may be combined. It is also to be understood that the values listed herein below (or subsets thereof) can be excluded.

[0075] Specifically, (Ci-C3)alkyl can be methyl, ethyl, propyl, or isopropyl; and (Ci-C3)alkanoyl can be formyl, acetyl, or propanoyl.

[0076] A specific value for X is O.

[0077] A specific value for X is NRa.

[0078] A specific value for X is CRbRc.

[0079] A specific value for R1is methyl.

[0080] A specific value for R2is H.

[0081] A specific value for R2is methyl.

[0082] A specific value for R3is a 6-membered heteroaryl that comprises one or two nitrogens.

[0083] A specific value for R3is pyridyl.

[0084] A specific value for R3is 3-pyridyl.

[0085] A specific value for R3is 4-pyridyl.

[0086] A specific value for R4is trifluoromethyl or methoxymethyl.

[0087] A specific value for R4is -(CH2CH2O)n-Rd

[0088] A specific value for R5is OR4or CHF2.

[0089] A specific value for R5is OR4.

[0090] A specific value for R6is H or CH3.

[0091] A specific value for R7is H or CH3.

[0092] A specific value for R6is H.

[0093] A specific value for R7is H.

[0094] A specific value for n is 0.

[0095] A specific value for n is i.

[0096] A specific value for n is 2.

[0097] A specific value for n is 3.

[0098] A specific value for n is 4.

[0099] A specific value for Rais H.

[0100] A specific value for Rais (Ci-C3)alkyl.

[0101] A specific value for Rbis H.

[0102] A specific value for Rbis F, Cl, or Br. A specific value for Rbis (Ci-C3)alkyl.

[0103] A specific value for Rcis H.

[0104] A specific value for Rcis F, Cl, or Br.

[0105] A specific value for Rcis (Ci-C3)alkyl.

[0106] A specific compound or salt is selected from the group consisting of and salts thereof.

[0107] A specific compound or salt is selected from the group consisting of

[0108]

[0109] A specific compound or salt is selected from the group consisting of

[0110] and salts thereof. Processes for preparing compounds of formula (I) are provided as further embodiments and are illustrated by the following procedures in which the meanings of the generic radicals are as given above unless otherwise qualified.

[0111] The term “transplant cornea” as used herein refers to a cornea that will be or has been removed from a donor (e.g., an animal or human donor), and that is to be transplanted in a recipient (e.g., an animal or human recipient). The recipient can also be referred to as a patient (e.g., an animal or human patient). The transplant cornea can be contacted (e.g., treated) with a compound as described herein before or after removal of the transplant cornea from the donor. The transplant cornea can be treated for any period of time (e.g., 1, 3, 6, 9, 12, or more hours, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more days) prior to or after removal from the donor. The transplant cornea can also be contacted (e.g., treated) with a compound as described herein after the transplant cornea has been transplanted in the recipient. The transplant cornea can also be maintained or stored (e.g., after removal from donor but before transplant in recipient) in contact with a compound as described herein for any period of time (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more days, 2, 3, 4, 5, 6, or more weeks).

[0112] One embodiment provides a method of treating damage or disease of the bladder (e.g., epithelial disease or epithelial damage of the bladder) in an animal, comprising, administering a compound or a pharmaceutically acceptable salt thereof as described herein (e.g., a compound of formula I) or a compound of formula: or a pharmaceutically acceptable salt thereof, to the animal.

[0113] One embodiment provides a compound or a pharmaceutically acceptable salt thereof as described herein (e.g., a compound of formula I), or a compound of formula:

[0114] or a pharmaceutically acceptable salt thereof, for treating damage or disease of the bladder (e.g., epithelial disease or epithelial damage of the bladder).

[0115] One embodiment provides the use of a compound or a pharmaceutically acceptable salt thereof as described herein (e.g., a compound of formula I), or a compound of formula: or a pharmaceutically acceptable salt thereof, to prepare a medicament for treating damage or disease of the bladder (e.g., epithelial disease or epithelial damage of the bladder) in an animal.

[0116] One embodiment provides a method of treating interstitial cystitis in an animal, comprising, administering a compound or a pharmaceutically acceptable salt thereof as described herein (e.g., a compound of formula I), or a compound of formula: or a pharmaceutically acceptable salt thereof, to the animal.

[0117] One embodiment provides a compound or a pharmaceutically acceptable salt thereof as described herein (e.g., a compound of formula I), or a compound of formula: or a pharmaceutically acceptable salt thereof, for treating interstitial cystitis.

[0118] One embodiment provides the use of a compound or a pharmaceutically acceptable salt thereof as described herein (e.g., a compound of formula I), or a compound of formula: or a pharmaceutically acceptable salt thereof, to prepare a medicament for treating interstitial cystitis in an animal.

[0119] One embodiment provides a method of treating a transplant cornea, comprising contacting the transplant cornea with a compound or a pharmaceutically acceptable salt thereof as described herein (e.g., a compound of formula I), or a compound of formula: or a pharmaceutically acceptable salt thereof, to the animal.

[0120] One embodiment provides a compound or a pharmaceutically acceptable salt thereof as described herein (e.g., a compound of formula I), or a compound of formula: or a pharmaceutically acceptable salt thereof, for treating a transplant cornea.

[0121] One embodiment provides the use of a compound or a pharmaceutically acceptable salt thereof as described herein (e.g., a compound of formula I), or a compound of formula: or a pharmaceutically acceptable salt thereof, to prepare a medicament for treating a transplant cornea.

[0122] One embodiment provides a method of treating damage or disease of the bladder (e.g., epithelial disease or epithelial damage of the bladder) in an animal, comprising, administering a compound of formula II or a pharmaceutically acceptable salt thereof as described herein, to the animal.

[0123] One embodiment provides a compound of formula II or a pharmaceutically acceptable salt thereof as described herein, for treating damage or disease of the bladder (e.g., epithelial disease or epithelial damage of the bladder).

[0124] One embodiment provides a the use of a compound of formula II or a pharmaceutically acceptable salt thereof as described as described herein, to prepare a medicament for treating damage or disease of the bladder (e.g., epithelial disease or epithelial damage of the bladder) in an animal.

[0125] One embodiment provides a method of treating interstitial cystitis in an animal, comprising, administering a compound of formula II or a pharmaceutically acceptable salt thereof as described herein, to the animal.

[0126] One embodiment provides a compound of formula II or a pharmaceutically acceptable salt thereof as described herein, for treating interstitial cystitis.

[0127] One embodiment provides the use of a compound of formula II or a pharmaceutically acceptable salt thereof as described herein, to prepare a medicament for treating interstitial cystitis in an animal.

[0128] One embodiment provides a method of treating a transplant cornea, comprising contacting the transplant cornea with a compound of formula II or a pharmaceutically acceptable salt thereof as described herein.

[0129] One embodiment provides a compound of formula II or a pharmaceutically acceptable salt thereof as described herein for treating a transplant cornea.

[0130] One embodiment provides the use of a compound of formula II or a pharmaceutically acceptable salt thereof as described herein, to prepare a medicament for treating a transplant cornea.

[0131] The following aspects relate to compounds of formula II.

[0132] Aspect 1 provides a molecule used to treat an epithelial disease comprising a compound of Formula II or a salt (e.g., a pharmaceutically acceptable salt) thereof, wherein

[0133] X1is -C-, -N-, or -O-;

[0134] X2is -N- or -C-; and

[0135] R1and R2are independently selected from the group consisting of -H, -OH, and substituted or unsubstituted (Ci-C2o)hydrocarbyl, and combinations thereof.

[0136] Aspect 2 provides the molecule of Aspect 1, wherein the (Ci-C2o)hydrocarbyl is selected from the group consisting of (Ci-C2o)alkyl, (Ci-C2o)alkenyl, (Ci-C2o)alkynyl, (Ci-C2o)acyl, (Ci- C2o)cycloalkyl, (Ci-C2o)aryl, (Ci-C2o)alkoxy, (Ci-C2o)haloalkyl, and combinations thereof. Aspect 3 provides the molecule of Aspect 1, wherein the compound is represented by

[0137] Formula Ila: wherein

[0138] X1is -C-, -N-, or -O-;

[0139] X2is -N- or -C-; and

[0140] R1and R2are independently selected from the group consisting of -H, -F, -OH, substituted or unsubstituted (C1-C20) hydrocarbyl, and combinations thereof. Aspect 4 provides the molecule of Aspect 3, wherein the (Ci-C2o)hydrocarbyl is selected from the group consisting of (Ci-C2o)alkyl, (Ci-C2o)alkenyl, (Ci-C2o)alkynyl, (Ci-C2o)acyl, (Ci- C2o)cycloalkyl, (Ci-C2o)aryl, (Ci-C2o)alkoxy, (Ci-C2o)haloalkyl, and combinations thereof.

[0141] Aspect 5 provides the molecule of Aspect 1, wherein the compound is represented by Formula III:

[0142] wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, - CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof; and

[0143] Y1, Y2, and Y3are independently selected from the group consisting of -C-, -N-, -O-, and combinations thereof.

[0144] Aspect 6 provides the molecule of Aspect 1, wherein the compound is represented by Formula IV:

[0145]

[0146] Wherein at each occurrence, R1is independently selected from the group consisting of -H, -

[0147] F, -CF3, -CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof; and

[0148] Y1, Y2, and Y3are independently selected from the group consisting of -N-, -C-, -O-, and combinations thereof.

[0149] Aspect 7 provides the molecule of Aspect 1, wherein the compound is represented by

[0150] Formula V:

[0151]

[0152] Aspect 8 provides the molecule of Aspect 1, wherein the compound is represented by

[0153] Formula VI: wherein n is any positive integer.

[0154] Aspect 9 provides the molecule of Aspect 1, wherein the compound is represented by Formula VII: wherein at each occurrence, R1is independently selected from the group consisting of -H, -

[0155] F, -CF3, -CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof.

[0156] Aspect 10 provides the molecule of Aspect 1, wherein the compound is represented by Formula VIII: wherein at each occurrence, R1is independently selected from the group consisting of -H, - F, -CF3, -CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof.

[0157] The following term definitions are used only for compounds of formula II and sub-formulas thereof (i.e., formulas Ila, III, IV, V, VI„ VII, and VIII). The term “organic group” as used herein for compounds of formula II refers to any carbon- containing functional group. Examples can include an oxygen-containing group such as an alkoxy group, aralkyloxy group, a carboxyl group including a carboxylic acid, carboxylate, and a carboxylate ester; a sulfur-containing group such as an alkyl and aryl sulfide group; and other heteroatom-containing groups. Non-limiting examples of organic groups include OR, OOR, OC(O)N(R)2, CN, CF3, OCF3, R, C(O), methylenedi oxy, ethylenedioxy, N(R)2, SR, SOR, SO2R, SO2N(R)2, SO3R, C(O)R, C(O)C(O)R, C(O)CH2C(O)R, C(S)R, C(O)OR, OC(O)R, C(O)N(R)2, OC(O)N(R)2, C(S)N(R)2, (CH2)O-2N(R)C(0)R, (CH2)O-2N(R)N(R)2, N(R)N(R)C(O)R, N(R)N(R)C(O)OR, N(R)N(R)CON(R)2, N(R)SO2R, N(R)SO2N(R)2, N(R)C(O)OR, N(R)C(O)R, N(R)C(S)R, N(R)C(O)N(R)2, N(R)C(S)N(R)2, N(COR)COR, N(OR)R, C(=NH)N(R)2, C(O)N(OR)R, C(=NOR)R, and substituted or unsubstituted (Ci-Cioo)hydrocarbyl, wherein R can be hydrogen (in examples that include other carbon atoms) or a carbon-based moiety, and wherein the carbon-based moiety can be substituted or unsubstituted.

[0158] The term “substituted” as used herein as used herein for compounds of formula II in conjunction with a molecule or an organic group as defined herein refers to the state in which one or more hydrogen atoms contained therein are replaced by one or more non-hydrogen atoms. The term “functional group” or “substituent” as used herein refers to a group that can be or is substituted onto a molecule or onto an organic group. Examples of substituents or functional groups include, but are not limited to, a halogen (e.g., F, Cl, Br, and I); an oxygen atom in groups such as hydroxy groups, alkoxy groups, carboxyl groups including carboxylic acids, carboxylates, and carboxylate esters; a sulfur atom in groups such as thiol groups, alkyl and aryl sulfide groups, sulfoxide groups, sulfone groups, sulfonyl groups, and sulfonamide groups; a nitrogen atom in groups such as amines, hydroxyamines, nitriles, nitro groups, N-oxides, hydrazides, azides, and enamines; and other heteroatoms in various other groups. Non-limiting examples of substituents that can be bonded to a substituted carbon (or other) atom include F, Cl, Br, I, OR, OC(O)N(R)2, CN, NO, NO2, ONO2, azido, CF3, OCF3, R, O (oxo), S (thiono), C(O), S(O), methylenedioxy, ethylenedioxy, N(R)2, SR, SOR, SO2R, SO2N(R)2, SO3R, C(O)R, C(O)C(O)R, C(O)CH2C(O)R, C(S)R, C(O)OR, OC(O)R, C(O)N(R)2, OC(O)N(R)2, C(S)N(R)2, (CH2)o-2N(R)C(0)R, (CH2)o-2N(R)N(R)2, N(R)N(R)C(O)R, N(R)N(R)C(O)OR, N(R)N(R)CON(R)2, N(R)SO2R, N(R)SO2N(R)2, N(R)C(O)OR, N(R)C(O)R, N(R)C(S)R, N(R)C(O)N(R)2, N(R)C(S)N(R)2, N(COR)COR, N(OR)R, C(=NH)N(R)2, C(O)N(OR)R, and C(=NOR)R, wherein R can be hydrogen or a carbon-based moiety; for example, R can be hydrogen, (Ci-Cioo)hydrocarbyl, alkyl, acyl, cycloalkyl, aryl; or wherein two R groups bonded to a nitrogen atom or to adjacent nitrogen atoms can together with the nitrogen atom or atoms form a heterocyclyl.

[0159] The term “alkyl” as used herein as used herein for compounds of formula II refers to straight chain and branched alkyl groups and cycloalkyl groups having from 1 to 40 carbon atoms, 1 to about 20 carbon atoms, 1 to 12 carbons or, in some embodiments, from 1 to 8 carbon atoms. Examples of straight chain alkyl groups include those with from 1 to 8 carbon atoms such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups. Examples of branched alkyl groups include, but are not limited to, isopropyl, iso-butyl, sec-butyl, t-butyl, neopentyl, isopentyl, and 2,2-dimethylpropyl groups. As used herein, the term “alkyl” encompasses n-alkyl, isoalkyl, and anteisoalkyl groups as well as other branched chain forms of alkyl. Representative substituted alkyl groups can be substituted one or more times with any of the groups listed herein, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.

[0160] The term “alkenyl” as used herein as used herein for compounds of formula II refers to straight and branched chain and cyclic alkyl groups as defined herein, except that at least one double bond exists between two carbon atoms. Thus, alkenyl groups have from 2 to 40 carbon atoms, or 2 to about 20 carbon atoms, or 2 to 12 carbon atoms or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to vinyl, -CH=CH(CH3), -CH=C(CH3)2, - C(CH3)=CH2, -C(CH3)=CH(CH3), -C(CH2CH3)=CH2, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl among others.

[0161] The term “alkynyl” as used herein as used herein for compounds of formula II refers to straight and branched chain alkyl groups, except that at least one triple bond exists between two carbon atoms. Thus, alkynyl groups have from 2 to 40 carbon atoms, 2 to about 20 carbon atoms, or from 2 to 12 carbons or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to -OCH, -OC(CH3), -C =C(CH2CH3), -CH2C =CH, -CH2C =C(CH3), and -CH2CACXCH2CH3) among others.

[0162] The term “acyl” as used herein as used herein for compounds of formula II refers to a group containing a carbonyl moiety wherein the group is bonded via the carbonyl carbon atom. The carbonyl carbon atom is bonded to a hydrogen forming a “formyl” group or is bonded to another carbon atom, which can be part of an alkyl, aryl, aralkyl cycloalkyl, or cycloalkylalkyl. An acyl group can include 0 to about 12, 0 to about 20, or 0 to about 40 additional carbon atoms bonded to the carbonyl group. An acyl group can include double or triple bonds within the meaning herein. An acryloyl group is an example of an acyl group. An acyl group can also include heteroatoms within the meaning herein. A nicotinoyl group (pyridyl-3-carbonyl) is an example of an acyl group within the meaning herein. Other examples include acetyl, benzoyl, phenylacetyl, pyridylacetyl, cinnamoyl, and acryloyl groups and the like. When the group containing the carbon atom that is bonded to the carbonyl carbon atom contains a halogen, the group is termed a “haloacyl” group. An example is a trifluoroacetyl group.

[0163] The term “cycloalkyl” as used herein as used herein for compounds of formula II refers to cyclic alkyl groups such as, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In some embodiments, the cycloalkyl group can have 3 to about 8-12 ring members, whereas in other embodiments the number of ring carbon atoms range from 3 to 4, 5, 6, or 7. Cycloalkyl groups further include polycyclic cycloalkyl groups such as, but not limited to, norbornyl, adamantyl, bornyl, camphenyl, isocamphenyl, and carenyl groups, and fused rings such as, but not limited to, decalinyl, and the like. Cycloalkyl groups also include rings that are substituted with straight or branched chain alkyl groups as defined herein. Representative substituted cycloalkyl groups can be mono-substituted or substituted more than once, such as, but not limited to, 2,2-, 2,3-, 2,4- 2,5- or 2, 6-di substituted cyclohexyl groups or mono-, di- or trisubstituted norbornyl or cycloheptyl groups, which can be substituted with, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups. The term “cycloalkenyl” alone or in combination denotes a cyclic alkenyl group.

[0164] The term “aryl” as used herein as used herein for compounds of formula II refers to cyclic aromatic hydrocarbon groups that do not contain heteroatoms in the ring. Thus, aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chrysenyl, biphenylenyl, anthracenyl, and naphthyl groups. In some embodiments, aryl groups contain about 6 to about 14 carbons in the ring portions of the groups. Aryl groups can be unsubstituted or substituted, as defined herein. Representative substituted aryl groups can be mono-substituted or substituted more than once, such as, but not limited to, a phenyl group substituted at any one or more of 2-, 3-, 4-, 5-, or 6-positions of the phenyl ring, or a naphthyl group substituted at any one or more of 2- to 8-positions thereof.

[0165] The term “alkoxy” as used herein as used herein for compounds of formula II refers to an oxygen atom connected to an alkyl group, including a cycloalkyl group, as are defined herein. Examples of linear alkoxy groups include but are not limited to methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, and the like. Examples of branched alkoxy include but are not limited to isopropoxy, sec-butoxy, tert-butoxy, isopentyloxy, isohexyloxy, and the like. Examples of cyclic alkoxy include but are not limited to cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like. An alkoxy group can include about 1 to about 12, about 1 to about 20, or about 1 to about 40 carbon atoms bonded to the oxygen atom, and can further include double or triple bonds, and can also include heteroatoms. For example, an allyloxy group or a methoxyethoxy group is also an alkoxy group within the meaning herein, as is a methylenedioxy group in a context where two adjacent atoms of a structure are substituted therewith.

[0166] The term “amine” as used herein as used herein for compounds of formula II, refers to primary, secondary, and tertiary amines having, e.g., the formula N(group)s wherein each group can independently be H or non-H, such as alkyl, aryl, and the like. Amines include but are not limited to R-NFE, for example, alkylamines, arylamines, alkylarylamines; R2NH wherein each R is independently selected, such as dialkylamines, diarylamines, aralkylamines, and the like; and R3N wherein each R is independently selected, such as trialkylamines, dialkylarylamines, alkyldiarylamines, triarylamines, and the like. The term “amine” also includes ammonium ions as used herein.

[0167] The term “haloalkyl” group, as used herein as used herein for compounds of formula II, includes mono-halo alkyl groups, poly-halo alkyl groups wherein all halo atoms can be the same or different, and per-halo alkyl groups, wherein all hydrogen atoms are replaced by halogen atoms, such as fluoro. Examples of haloalkyl include trifluorom ethyl, 1,1 -di chloroethyl, 1,2-dichloroethyl, l,3-dibromo-3,3-difluoropropyl, perfluorobutyl, and the like.

[0168] As used herein for compounds of formula II, the term “hydrocarbyl” refers to a functional group derived from a straight chain, branched, or cyclic hydrocarbon, and can be alkyl, alkenyl, alkynyl, aryl, cycloalkyl, acyl, or any combination thereof. Hydrocarbyl groups can be shown as (Ca-Cb)hydrocarbyl, wherein a and b are integers and mean having any of a to b number of carbon atoms. For example, (Ci-C4)hydrocarbyl means the hydrocarbyl group can be methyl (Ci), ethyl (C2), propyl (C3), or butyl (C4), and (Co-Cb)hydrocarbyl means in certain embodiments there is no hydrocarbyl group.

[0169] In one embodiment a compound of formula II is:

[0170] or a salt (e.g., a pharmaceutically acceptable salt) thereof.

[0171] In one embodiment the compound of formula II is a compound described in PCT / US2021 / 056595, which document is hereby incorporated by reference herein. In cases where compounds are sufficiently basic or acidic, a salt of a compound of formula

[0172] (I) can be useful as an intermediate for isolating or purifying a compound of formula (I). Additionally, administration of a compound of formula (I) as a pharmaceutically acceptable acid or base salt may be appropriate. Examples of pharmaceutically acceptable salts are organic acid addition salts formed with acids which form a physiological acceptable anion, for example, tosylate, methanesulfonate, acetate, citrate, malonate, tartarate, succinate, benzoate, ascorbate, a- ketoglutarate, and a-glycerophosphate. Suitable inorganic salts may also be formed, including hydrochloride, sulfate, nitrate, bicarbonate, and carbonate salts.

[0173] Salts may be obtained using standard procedures well known in the art, for example by reacting a sufficiently basic compound such as an amine with a suitable acid affording a physiologically acceptable anion. Alkali metal (for example, sodium, potassium or lithium) or alkaline earth metal (for example calcium) salts of carboxylic acids can also be made.

[0174] The compounds of formula (I) can be formulated as pharmaceutical compositions and administered to a mammalian host, such as a human patient in a variety of forms adapted to the chosen route of administration, i.e., orally or parenterally, by intravenous, intramuscular, topical or subcutaneous routes.

[0175] Thus, the present compounds may be systemically administered, e.g., orally, in combination with a pharmaceutically acceptable vehicle such as an inert diluent or an assimilable edible carrier. They may be enclosed in hard or soft shell gelatin capsules, may be compressed into tablets, or may be incorporated directly with the food of the patient's diet. For oral therapeutic administration, the active compound may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and preparations should contain at least 0.1% of active compound. The percentage of the compositions and preparations may, of course, be varied and may conveniently be between about 2 to about 60% of the weight of a given unit dosage form. The amount of active compound in such therapeutically useful compositions is such that an effective dosage level will be obtained.

[0176] The tablets, troches, pills, capsules, and the like may also contain the following: binders such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, fructose, lactose or aspartame or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring may be added. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier, such as a vegetable oil or a polyethylene glycol. Various other materials may be present as coatings or to otherwise modify the physical form of the solid unit dosage form. For instance, tablets, pills, or capsules may be coated with gelatin, wax, shellac or sugar and the like. A syrup or elixir may contain the active compound, sucrose or fructose as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavoring such as cherry or orange flavor. Of course, any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed. In addition, the active compound may be incorporated into sustained-release preparations and devices.

[0177] The active compound may also be administered intravenously or intraperitoneally by infusion or injection. Solutions of the active compound or its salts can be prepared in water, optionally mixed with a nontoxic surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

[0178] The pharmaceutical dosage forms suitable for injection or infusion can include sterile aqueous solutions or dispersions or sterile powders comprising the active ingredient which are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. In all cases, the ultimate dosage form should be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions or by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0179] Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filter sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the previously sterile-filtered solutions.

[0180] For topical administration, the present compounds may be applied in pure form, i.e., when they are liquids. However, it will generally be desirable to administer them to the skin as compositions or formulations, in combination with a dermatologically acceptable carrier, which may be a solid or a liquid. For ophthalmic indications (e.g., promoting healing of corneal ulcers), the compounds can be formulated with an acceptable ophthalmic carrier and applied, for example, as eyedrops.

[0181] Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silica, alumina and the like. Useful liquid carriers include water, alcohols or glycols or water-alcohol / glycol blends, in which the present compounds can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants. Adjuvants such as fragrances and additional antimicrobial agents can be added to optimize the properties for a given use. The resultant liquid compositions can be applied from absorbent pads, used to impregnate bandages and other dressings, or sprayed onto the affected area using pump-type or aerosol sprayers.

[0182] Thickeners such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses or modified mineral materials can also be employed with liquid carriers to form spreadable pastes, gels, ointments, soaps, and the like, for application directly to the skin of the user.

[0183] Examples of useful dermatological compositions which can be used to deliver the compounds of formula (I) to the skin are known to the art; for example, see Jacquet et al. (U.S. Pat. No. 4,608,392), Geria (U.S. Pat. No. 4,992,478), Smith et al. (U.S. Pat. No. 4,559,157) and Wortzman (U.S. Pat. No. 4,820,508).

[0184] Useful dosages of the compounds of formula (I) can be determined by comparing their in vitro activity, and in vivo activity in animal models. Methods for the extrapolation of effective dosages in mice, and other animals, to humans are known to the art; for example, see U.S. Pat. No. 4,938,949.

[0185] The amount of the compound, or an active salt or derivative thereof, required for use in treatment will vary not only with the particular salt selected but also with the route of administration, the nature of the condition being treated and the age and condition of the patient and will be ultimately at the discretion of the attendant physician or clinician. The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations; such as multiple inhalations from an insufflator or by application of a plurality of drops into the eye.

[0186] Compounds of the invention can also be administered in combination with other therapeutic agents, for example, other agents that are useful for promoting epithelial healing or other therapeutic agents that reduce factors that injure the epithelium. Examples of such agents include, but are not limited to, growth factors (e.g., fibroblast growth factor FGF, platelet-derived growth factor PDGF, epidermal growth factor EGF, and transforming growth factor alpha TGF-alpha), proton-pump inhibitors and H2 blockers (e.g., omeprazole and famotidine), and anti-inflammatory agents (e.g., steroids and monoclonal antibodies). Accordingly, in one embodiment the invention also provides a composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, at least one other therapeutic agent, and a pharmaceutically acceptable diluent or carrier. The invention also provides a kit comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, at least one other therapeutic agent, packaging material, and instructions for administering the compound of formula (I) or the pharmaceutically acceptable salt thereof and the other therapeutic agent or agents to an animal to treat an epithelial disease or epithelial damage.

[0187] Compounds of the invention can also be administered in combination with other therapeutic agents, for example, other agents that have the undesirable effect of injuring the mucosa or impairing mucosal or epithelial healing. Examples of such agents include non-steroidal antiinflammatory agents. Accordingly, in one embodiment the invention also provides a composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, at least one other therapeutic agent that has the undesirable effect of injuring the mucosa or impairing mucosal or epithelial healing, and a pharmaceutically acceptable diluent or carrier. The invention also provides a kit comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, at least one other therapeutic agent that has the undesirable effect of injuring the mucosa or impairing mucosal or epithelial healing, packaging material, and instructions for administering the compound of formula (I) or the pharmaceutically acceptable salt thereof and the other therapeutic agent or agents to an animal to provide a beneficial therapeutic effect with reduced epithelial or mucosal damage.

[0188] The invention will now be illustrated by the following non-limiting Examples.

[0189] EXAMPLES

[0190] Chemistry. All reagents were purchased from commercial suppliers and used without further purification. All solvents used were reagent quality grade or anhydrous solvents. All reactions were performed under an inert atmosphere of nitrogen gas unless noted otherwise. 1H NMR spectra were recorded on a Bruker Advance II (400 MHz). 1H NMR data are reported as follows: chemical shift [multiplicity (s = singlet, d = doublet, t = triplet, q = quartet, dd = doublet of doublets, m = multiplet, s br = broad singlet), J = coupling constant(s) (Hz), integration]. Column chromatography was carried out employing Siliaflash silica gel (P60, 63-200 um). Precoated silica gel plates F-254 were used for thin-layer analytical chromatography. All final compounds 5-46 were purified to >95% purity as determined by LC / MS. Compounds were analyzed using a quadrupole / time of flight MS (G2S, Waters) coupled to UPLC binary pump system (Acquity, Waters). Compounds were resolved on ACQUITY UPLC HSS T3 column (1.8 pM, 100 A pore diameter, 2.1 x 150 mm; Waters) with a gradient of 0.1% formic acid in water (Solvent A) and acetonitrile with 0.1% formic acid (solvent B) at 0.3 mL / min. Percent B was increased from 1% to 75% over 2 min and then to 99% over 3 min. At 6 min, %B was returned back to 1 % over 1 min. Data were collected with MSE method, and MS was operated in sensitivity mode. Compounds were ionized with a positive electrospray ionization technique. All commercially available reagents and solvents were used without further purification. All yields reported were not optimized.

[0191] Example 1. Synthesis of: To a stirred solution of l-fluoro-2-nitro-4-(trifluoromethoxy)benzene (0.50 g, 2.22 mmol) in dry DMF (5 mL) was added morpholine (0.21 mL, 2.44 mmol), followed by K2CO3 (0.61 g, 4.44 mmol), The crude reaction was stirred under heating at 60 °C overnight. After cooling at room temperature, crude reaction was diluted with DCM and washed with brine several times. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. Product was purified by column chromatography using hexane: EtOAc (50:50) to afford a red-orange oil, 62% yield. 'H NMR (CDCI3, 400 MHZ): 3 7.69 (d, J= 4.0 Hz, 1H), 7.40-7.37 (m, 1H), 7.18 (d, J = 8.0 Hz, 1H), 3.85 (t, J = 4.0 Hz, 4H), 3.05 (t, J= 4.0 Hz, 4H). MS (ESI+) calculated for C11H11F3N2O4 [M+l]+: 293.2, observed: 293.2.

[0192] To a stirred solution of 4-(2-nitro-4-(trifluoromethoxy)phenyl)morpholine (0.40 g, 1.36 mmol) in EtOH (5 mL), purged under nitrogen, was added Pd / C (0.02 g) and hydrazine monohydrate (1.0 mL, 20.5 mmol) was added dropwise. The crude reaction was stirred at room temperature for 2 hours. Crude was filtered and concentrated in vacuo. Product was purified by column chromatography using hexane: EtOAc (30:70) to afford as a clear oil in 44% yield. 'H NMR (CDCI3, 400 MHZ): 3 7.01 (d, J= 8.0 Hz, 1H), 6.60 (s, 2H), 3.96 (s, 4H), 3.06 (t, J= 4.0 Hz, 4H). MS (ESI+) calculated for C11H13F3N2O2 [M+l]+: 263.2, observed: 263.2.

[0193] To a stirred solution of 2-morpholino-5-(trifluoromethoxy)aniline (0.08 g, 0.3 mmol) in dry DCM (5 mL), under N2 atmosphere at 0 °C, was added a solution of phenyl chloroformate (0.04 mL, 0.33 mmol), followed by pyridine (0.08 mL, 0.06 mmol). The crude reaction was allowed to warm up at room temperature and stirred overnight. Crude reaction was concentrated in vacuo and product was purified by column chromatography using hexane: EtOAc (70:30) to afford pure product as an orange oil in 85% yield. 'HNMR (CDCI3, 400 MHZ): 3 8.41 (s, 1H), 8.13 (s, 1H), 7.42 (t, J= 4.0 Hz, 2H), 7.27-7.25 (m, 2H), 7.22 (d, J= 8.0 Hz, 2H), 6.93 (dd, J = 4.0 Hz, 2.0 Hz, 1H), 3.93 (t, J= 4.0 Hz, 2H), 2.93 (t, J= 4.0 Hz, 4H). MS (ESI+) calculated for C18H17F3N2O4 [M+l]+: 383.3, observed: 383.3.

[0194] To a stirred solution of phenyl (2-morpholino-5-(trifluoromethoxy)phenyl)carbamate (0.025 g, 0.06 mmol) in dry pyridine (3 mL) was added N1,N1-dimethyl-l-(pyridin-4-yl)ethane-l,2- diamine (0.01 g, 0.06 mmol) and the solution was heated at 80 °C for 16 hours. Crude reaction was cooled down to room temperature and concentrated in vacuo. Product was purified by column chromatography using DCM:EtOH (95:5) to afford pure product as an orange solid in 60% yield. Obtained as a white solid in 25 % yield. 'H NMR (CDCI3, 400 MHZ): 3 8.78 (d, J= 4.0 Hz, 2H), 8.34 (s, br, 1H), 7.40 (d, J= 4.0 Hz, 2H), 7.14 (d, J= 8.0 Hz, 2H), 6.81 (dd, J= 8.0 Hz, 4.0 Hz, 1H), 4.34-4.30 (m, 1H), 4.02 (s, 4H), 3.93-3.90 (m, 2H), 2.86 (t, J= 4.0 Hz, 4H), 2.76 (s, br, 6H). MS (ESI+) calculated for C21H26F3N5O3 [M+l]+: 454.4, observed: 454.4.

[0195] Example 2. Synthesis of:

[0196] To a stirred solution of 4-fluoro-3 -nitrophenol (1.0 g, 6.36 mmol) in acetone (10 mL) was added K2CO3 (4.4 g, 31.8 mmol), followed M0MC1 (1.0 mL, 12.72 mmol). The crude reaction was stirred under heating at 50 °C overnight. After cooling at room temperature, crude reaction was diluted with DCM and filtered. The crude was concentrated in vacuo, redissolved in EtOAc (20 mL) and washed with IN NaOH (5 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to afford a red oil, 63% yield.JH NMR (CDCI3, 400 MHZ): 3 7.73-7.71 (m, 1H), 7.30-.7.28 (m, 1H), 7.20 (t, J= 8.0 Hz, 1H), 5.19 (s, 2H), 3.49 (s, 3H). MS (ESI+) calculated for C8H8FNO4[M+l]+: 202.1, observed: 202.1.

[0197] To a stirred solution of l-fluoro-4-(methoxymethoxy)-2-nitrobenzene (0.81 g, 4.0 mmol) in dry DMF (5 mL) was added morpholine (0.38 mL, 4.43 mmol), followed by K2CO3 (1.1 g, 8.0 mmol). The crude reaction was stirred under heating at 60 °C overnight. After cooling at room temperature, crude reaction was diluted with DCM and washed with brine several times. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. Product was purified by column chromatography using hexane: EtOAc (50:50) to afford a red oil 68 % yield. 'H NMR (CDCI3, 400 MHZ): 3 7.43 (d, J= 4.0 Hz, 1H), 7.19 (d, J= 4.0 Hz, 1H), 7.16 (d, J= 8.0 Hz, 1H), 5.16 (s, 2H), 3.81 (t, J= 4.0 Hz, 4H), 3.47 (s, 3H), 2.98 (t, J= 4.0 Hz, 4H). MS (ESI+) calculated for C12H16NO5 [M+l]+: 269.2, observed: 269.1.

[0198] To a stirred solution of 4-(4-(methoxymethoxy)-2-nitrophenyl)morpholine (0.74 g, 2.76 mmol) in MeOH (10 mL), purged under nitrogen, was added Pd / C (0.07 g) and then a balloon filled with H2. The crude reaction was stirred at room temperature for 1 hours. Crude was filtered and concentrated in vacuo. Product was purified by column chromatography using hexane: EtOAc (30:70) to afford a white solid in 90 % yield. 'H NMR (CDC13, 400 MHZ): 3 7.43 (d, J= 4.0 Hz, 1H), 7.20 (d, J= 4.0 Hz, 1H), 7.16 (d, J= 8.0 Hz, 1H), 6.95 (d, J= 4.0 Hz, 1H), 6.48 (d, J= 4.0 Hz, 1H), 5.16 (s, 2H), 3.82 (t, J= 4.0 Hz, 4H), 3.48 (s, 3H), 2.98 (t, J= 4.0 Hz, 4H). MS (ESI+) calculated for C12H18N2O3 [M+l]+: 239.1, observed: 239.1.

[0199] To a stirred solution of 5-(methoxymethoxy)-2-morpholinoaniline (0.6 g, 2.52 mmol) in dry DCM (10 mL), under N2 atmosphere at 0 °C, was added a solution of phenyl chloroformate (0.34 mL, 2.77 mmol), followed by EtsN (0.53 mL, 3.78 mmol). The crude reaction was allowed to warm up at room temperature and stirred overnight. Crude reaction was concentrated in vacuo and product was purified by column chromatography using hexane: EtOAc (70:30) to afford pure product as a cream solid in 60% yield.1H NMR (CDCI3, 400 MHZ): 3 8.44 (s, br, 1H), 7.92 (s, 1H), 7.40 (t, J= 4.0 Hz, 2H), 7.23 (d, J = 8.0 Hz, 2H), 7.14 (d, J= 8.0 Hz, 2H), 6.74 (dd, J= 8.0 Hz, 4.0 Hz, 1H), 5.16 (s, 2H), 3.81 (t, J= 4.0 Hz, 4H), 3.47 (s, 3H), 2.97 (t, J= 4.0 Hz, 4H). MS (ESI+) calculated for C19H22N2O5 [M+l]+: 359.3, observed: 359.3.

[0200] To a stirred solution of phenyl (5-(methoxymethoxy)-2-morpholinophenyl)carbamate (0.32 g, 0.89 mmol) in dry pyridine (5 mL) was added N1,N1-dimethyl-l-(pyridin-4-yl)ethane-l,2- diamine (0.13 g, 0.8 mmol) and the solution was heated at 80 °C for 16 hours. Crude reaction was cooled down to room temperature and concentrated in vacuo. Product was purified by column chromatography using DCM:EtOH (95:5) to afford pure product as a white solid in 45 % yield. 'H NMR (CDCI3, 400 MHZ): 3 8.67 (d, J= 4.0 Hz, 2H), 7.88 (s, 1H), 7.84 (d, J= 4.0 Hz, 1H), 7.30 (d, J= 8.0 Hz, 2H), 7.05 (d, J= 8.0 Hz, 1H), 6.64 (dd, J= 8.0 Hz, 4.0 Hz, 1H), 5.14 (s, 2H), 3.98- 3.94 (m, 2H), 3.88 (q, J= 4.0 Hz, 4H), 3.72-3.69 (m, 1H), 3.46 (s, 3H), 2.79 (s, br, 4H), 2.49 (s, 6H). MS (ESI+) calculated for C22H31N5O4 [M+l]+: 430.5, observed: 430.5.

[0201] Example s. Synthesis of: To a stirred solution of l-fluoro-4-(2-methoxyethoxy)-2-nitrobenzene (0.82 g, 3.81 mmol) in dry DMF (5 mL) was added morpholine (0.4 mL, 4.57 mmol), followed by K2CO3 (1.05 g, 7.62 mmol). The crude reaction was stirred under heating at 60 °C overnight. After cooling at room temperature, crude reaction was diluted with DCM and washed with brine several times. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. Product was purified by column chromatography using hexane: EtOAc (50:50) to afford a red oil, 68 % yield. 'HNMR (CDCI3, 400 MHZ): 3 7.30 (s, 1H), 7.19-7.13 (m, 2H), 4.12 (t, J= 4.0 Hz, 2H), 3.82 (t, J = 4.0 Hz, 4H), 3.75 (t, J= 4.0 Hz, 2H), 3.45 (s, 3H), 2.98 (t, J= 4.0 Hz, 4H). MS (ESI+) calculated for C13H18N2O5 [M+l]+: 283.3, observed: 283.3.

[0202] To a stirred solution of 4-(4-(2-methoxyethoxy)-2-nitrophenyl)morpholine (0.11 g, 0.41 mmol) in MeOH (5 mL), purged under nitrogen, was added Pd / C (0.05 g) and then a balloon filled with H2. The crude reaction was stirred at room temperature for 1 hour. Crude was filtered and concentrated in vacuo. Product was purified by column chromatography using hexane: EtOAc (30:70) to afford a white solid in 90 % yield. 'H NMR (CDCI3, 400 MHZ): 3 6.93 (d, J= 4.0 Hz, 1H), 6.35 (d, J= 4.0 Hz, 1H), 6.32 (dd, J= 4.0 Hz, 1H), 4.06 (t, J= 4.0 Hz, 2H), 3.86 (s, br, 4H), 3.70 (m, 2H), 3.49 (s, 3H), 2.84 (s, br, 4H). MS (ESI+) calculated for C13H20N2O3 [M+l]+: 253.3, observed: 253.3.

[0203] To a stirred solution of 5-(2-methoxyethoxy)-2-morpholinoaniline (0.6 g, 2.52 mmol) in dry DCM (10 mL), under N2 atmosphere at 0°C, was added a solution of phenyl chloroformate (0.34 mL, 2.77 mmol), followed by EtsN (0.53 mL, 3.78 mmol). The crude reaction was allowed to warm up at room temperature and stirred overnight. Crude reaction was concentrated in vacuo and product was purified by column chromatography using hexane: EtOAc (70:30) to afford pure product as a cream solid in 60% yield. 'HNMR (CDCI3, 400 MHZ): 3 8.58 (s, br, 1H), 7.83 (s, 1H), 7.43-7.39 (m, 3H), 7.23 (d, J= 8.0 Hz, 2H), 7.6 (d, J= 8.0 Hz, 1H), 6.68 (dd, J = 4.0 Hz, 2.0 Hz, 1H), 4.11 (t, J= 4.0 Hz, 2H), 3.92 (s, br, 4H), 3.71 (t, J= 4.0 Hz, 2H), 3.42 (s, 3H), 2.91 (s, br, 4H). MS (ESI+) calculated for C20H24N2O5 [M+l]+: 373.4, observed: 373.4.

[0204] To a stirred solution of phenyl (5-(2-methoxyethoxy)-2-morpholinophenyl)carbamate (0.32 g, 0.89 mmol) in dry pyridine (5 mL) was added N1,N1-dimethyl-l-(pyridin-4-yl)ethane-l,2- diamine (0.13 g, 0.8 mmol) and the solution was heated at 80 °C for 16 hours. Crude reaction was cooled down at room temperature and concentrated in vacuo. Product was purified by column chromatography using DCM:EtOH (95:5) to afford pure product as a white solid \in 25 % yield. 'H NMR (DMSO-d6, 400 MHZ): d 8.52 (d, J= 4.0 Hz, 2H), 7.87 (s, 1H), 7.77 (s, br, 1H), 7.61 (d, J = 4.0 Hz, 1H), 7.28 (d, J= 4.0 Hz, 2H), 7.16 (t, J = 8.0 Hz, 1H), 7.01 (d, J= 8.0 Hz, 1H), 6.44 (dd, J = 8.0 Hz, 4.0 Hz, 1H), 3.98-3.96 (m, 2H), 3.75 (t, J= 4.0 Hz, 4H), 3.64-3.59 (m, 2H), 3.46 (t, J= 4.0 Hz, 1H), 3.42-3.37 (m, 2H), 3.29 (s, 3H), 2.12 (s, 6H). MS (ESI+) calculated for C23H33N5O4 [M+l]+: 444.5, observed: 444.5.

[0205] Example 4. Synthesis of:

[0206] To a stirred solution of 4-(4-fluoro-2-nitrophenyl)morpholine (0.40 g, 1.76 mmol) in EtOH (5 mL), purged under nitrogen, was added Pd / C (0.1 g) and hydrazine monohydrate (1.5 mL, 30.4 mmol) was added dropwise. The crude reaction was stirred at room temperature for 2 h. Crude was filtered and concentrated in vacuo. The product was purified by column chromatography using hexane: EtOAc (30:70) to afford a white solid in 40% yield.JH NMR in accordance with the previously reported structure. MS (ESI+) calculated for C10H13FN2O [M+l]+: 197.2, observed: 197.2

[0207] To a stirred solution of 5-fhioro-2-morpholino aniline (0.61 g, 3.11 mmol) in dry DCM (25 mL), under an N2 atmosphere at 0 °C, was added a solution of phenyl chloroformate (0.4 mL, 3.42 mmol) in dry DCM (5 mL), followed by EtsN (0.5 mL, 6.22 mmol). The crude reaction was allowed to warm up at room temperature and stirred overnight. The crude reaction was concentrated in vacuo and the product was purified by column chromatography using hexane: EtOAc (70:30) to afford the pure product as a cream solid in 60% yield.JH NMR (CDCI3, 400 MHz): d 8.41 (s, 1H), 7.96 (d, J = 8.0 Hz, 1H), 7.42 (t, J= 8.0 Hz, 2H), 7.28-7.25 (m, 2H), 7.23-7.15 (m, 3H), 6.75 (td, J = 4.0 Hz, 2.0 Hz, 1H), 3.90 (t, J= 4.0 Hz, 4H), 2.88 (t, J= 4.0 Hz, 4H). MS (ESI+) calculated for C17H17FN2O3 [M+l]+: 317.3, observed: 317.3 To a stirred solution of phenyl (5-fluoro-2-morpholinophenyl)carbamate (0.39 g, 1.25 mmol) in dry pyridine (5 mL) was added N1,N1-dimethyl-l-(pyridin-4-yl)ethane-l,2-diamine (0.20 g, 1.25 mmol) and the solution was heated at 80 °C for 16 hrs. The crude reaction was cooled down to room temperature and concentrated in vacuo. The product was purified by column chromatography using DCM:EtOH (95:5) to afford the pure product as a white solid in 30 % yield. 'H NMR (DMSO-de, 400 MHz): 3 8.81 (d, J= 8.0 Hz, 2H), 8.01 (s, 1H), 7.85 (dd, J= 8.0 Hz, 4.0 Hz, 1H), 7.73 (d, J= 4.0 Hz, 2H), 7.59 (t, J= 4.0 Hz, 1H), 7.16 (d, J = 4.0 Hz, 1H), 6.75-6.70 (m, 1H), 4.64 (t, J= 4.0 Hz, 1H), 3.97-3.91 (m, 1H), 3.81-3.69 (m, 1H), 3.72 (t, J= 4.0 Hz, 4H), 2.80 (s, br, 6H), 2.73-2.62 (m, 4H). MS (ESI+) calculated for C20H26FN5O2 [M+l]+: 388.5, observed: 388.5.

[0208] Example 5. Synthesis of:

[0209] To a stirred solution of 4-(difluoromethyl)-l-fluoro-2-nitrobenzene (0.25 g, 1.3 mmol) in dry DMF (5 mL) was added morpholine (0.12 mL, 1.43 mmol), followed by K2CO3 (0.36 g, 2.6 mmol), The crude reaction was stirred under heating at 60 °C overnight. After cooling at room temperature, the crude reaction was diluted with DCM and washed with brine several times. The organic layer was dried over anhydrous ISfeSCU, filtered, and concentrated in vacuo. The product was purified by column chromatography using hexane: EtOAc (50:50) to afford a red-orange oil, 95% yield. 'H NMR (CDCI3, 400 MHz): 3 7.95 (s, 1H), 7.62 (d, J= 8.0 Hz, 1H), 7.18 (d, J= 8.0 Hz, 1H), 6.62 (t, J= 56.0 Hz, 1H), 3.85 (t, J= 4.0 Hz, 4H), 3.12 (t, J= 4.0 Hz, 4H). MS (ESI+) calculated for C11H12F2N2O3 [M+l]+: 259.2, observed: 259.2.

[0210] To a stirred solution of 4-(4-(difluoromethyl)-2-nitrophenyl)morpholine (0.33 g, 1.27 mmol) in MeOH (5 mL), purged under nitrogen, was added Pd / C (0.02 g) and a balloon filled with H2 was attached. The crude reaction was stirred at room temperature for 2 h. Crude was filtered and concentrated in vacuo. The product was purified by column chromatography using hexane: EtOAc (30:70) to afford a clear oil with a 44% yield. 'H NMR (CDC13, 400 MHz): 3 7.13 (d, J = 8.0 Hz, 1H), 6.94 (s, 2H), 6.54 (t, J = 56.0 Hz, 1H), 4.06 (s, 4H), 3.19 (s, 4H). MS (ESI+) calculated for C11H14F2N2O [M+l]+: 229.2, observed: 229.2.

[0211] To a stirred solution of 5-(difluoromethyl)-2-morpholinoaniline (0.13 g, 0.57 mmol) in dry DCM (15 mL), under an N2 atmosphere at 0°C, was added a solution of phenyl chloroformate (0.08 mL, 0.62 mmol) in dry DCM (5 mL), followed by EtsN (0.16 mL, 1.14 mmol). The crude reaction was allowed to warm up at room temperature and stirred overnight. The crude reaction was concentrated in vacuo and the product was purified by column chromatography using hexane: EtOAc (70:30) to afford product as a cream solid in 60% yield.1H NMR (CDCI3, 400 MHz): 3 8.35 (s, 2H), 7.7.44-7.40 (m, 2H), 7.28 (s, 2H), 7.22 (d, J= 8.0 Hz, 2H), 6.59 (t, J = 56.0 Hz, 1H), 3.95 (t, J= 4.0 Hz, 4H), 2.98 (t, J= 4.0 Hz, 4H). MS (ESI+) calculated for C18H18F2N2O3 [M+l]+: 349.3, observed: 349.3

[0212] To a stirred solution of phenyl (5-(difluoromethyl)-2-morpholinophenyl)carbamate (0.063 g, 0.18 mmol) in dry pyridine (5 mL) was added N1,N1-dimethyl-l-(pyridin-4-yl)ethane-l,2-diamine (0.03 g, 0.18 mmol) and the solution was heated at 80 °C for 16 hrs. The crude reaction was cooled down to room temperature and concentrated in vacuo. The product was purified by column chromatography using DCM:EtOH (95:5) and obtained as a white solid in 30% yield. 'H NMR (CDCI3, 400 MHz): 3 9.91 (s, 1H), 8.70 (d, J= 4.0 Hz, 1H), 8.57 (s, 1H), 7.70 (s, br, 1H), 7.55 (d, J = 4.0 Hz, 1H), 7.36 (d, J= 4.0 Hz, 1H), 7.22 (d, J= 40.0 Hz, 1H), 4.06-4.00 (m, 6H), 3.81-3.78 (m, 1H), 2.92 (s, 4H), 2.60 (s, br, 6H). MS (ESI+) calculated for C21H27F2N5O2 [M+l]+: 420.5, observed: 420.5.

[0213] Example 6. Synthesis of: To a stirred solution of 4-fluoro-3 -nitrophenol (1.0 g, 6.36 mmol) in acetone (10 mL) was added K2CO3 (4.4 g, 31.8 mmol), followed by CH3I (0.80 mL, 12.72 mmol), The crude reaction was stirred under heating at 50 °C overnight. After cooling at room temperature, the crude reaction was diluted with DCM and filtered. The crude was concentrated in vacuo, redissolved in EtOAc (20 mL), and washed with IN NaOH (5 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to afford a red oil with a 57% yield.JH NMR (CDCI3, 400 MHz): 3 7.54-7.51 (m, 1H), 7.23-7.15 (m, 2H), 3.86 (s, 3H). MS (ESI+) calculated for C7H6FNO3 [M+l]+: 172.2, observed: 172.2.

[0214] To a stirred solution of l-fluoro-4-methoxy-2-nitrobenzene (0.62 g, 3.62 mmol) in dry DMF (5 mL) was added morpholine (0.35 mL, 4.0 mmol), followed by K2CO3 (1.0 g, 7.2 mmol), The crude reaction was stirred under heating at 60 °C overnight. After cooling at room temperature, the crude reaction was diluted with DCM and washed with brine several times. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The product was purified by column chromatography using hexane: EtOAc (50:50) to afford a yellow solid 80% yield. 'H NMR (CDCI3, 400 MHz): 3 7.20 (s, 1H), 7.09 (d, J= 4.0 Hz, 1H), 7.07 (d, J= 2.0 Hz, 1H), 3.82 (s, 3H), 3.82 (t, J = 4.0 Hz, 4H), 2.98 (t, J = 4.0 Hz, 4H). MS (ESI+) calculated for C11H14N2O4 [M+l]+: 239.2, observed: 239.2.

[0215] To a stirred solution of 4-(4-methoxy-2-nitrophenyl)morpholine (0.69 g, 2.89 mmol) in MeOH (10 mL), purged under nitrogen, was added Pd / C (0.1 g) and then a balloon filled with H2was attached. The crude reaction was stirred at room temperature for 1 h. Crude was filtered and concentrated in vacuo. The product was purified by column chromatography using hexane: EtOAc (30:70) to afford a white solid with an 80% yield.1H NMR (CDCI3, 400 MHz): 3 7.20 (s, 1H), 7.09- 7.07 (dd, J= 4.0 Hz, 1H), 6.94 (d, J= 8.0 Hz, 1H), 6.32 (t, J= 4.0 Hz, 1H), 3.82 (s, 3H), 3.82 (t, J= 4.0 Hz, 4H), 2.87 (t, J= 4.0 Hz, 4H). MS (ESI+) calculated for C11H16N2O2 [M+l]+: 209.2, observed: 209.2.

[0216] To a stirred solution of 5 -methoxy -2-morpholinoaniline (0.49 g, 2.35 mmol) in dry DCM (15 mL), under an N2 atmosphere at 0°C, was added a solution of phenyl chloroformate (0.35 mL, 2.82 mmol) in dry DCM (5 mL), followed by EtsN (0.5 mL, 3.52 mmol). The crude reaction was allowed to warm up at room temperature and stirred overnight. The crude reaction was concentrated in vacuo and the product was purified by column chromatography using hexane: EtOAc (70:30) to afford product as a cream solid in 60% yield. 'H NMR (CDCI3, 400 MHz): 3 8.48 (s, 1H), 7.84 (m, 1H), 7.44-7.40 (m, 2H), 7.28-7.21 (m, 3H), 7.14 (d, J= 12.0 Hz, 1H), 6.61 (dd, J= 8.0 Hz, 4.0 Hz, 1H), 3.90 (t, J= 4.0 Hz, 4H), 3,78 (s, 3H), 2.87 (t, J= 4.0 Hz, 4H). MS (ESI+) calculated for C18H20N2O4 [M+l]+: 329.3, observed: 329.3

[0217] To a stirred solution of phenyl (5-methoxy-2-morpholinophenyl)carbamate (0.14 g, 0.42 mmol) in dry pyridine (5 mL) was added N1,N1-dimethyl-l-(pyridin-4-yl)ethane-l,2-diamine (0.07 g, 0.42 mmol) and the solution was heated at 80 °C for 16 hrs. The crude reaction was cooled down to room temperature and concentrated in vacuo. The product was purified by column chromatography using DCM:EtOH (95:5) and obtained as a white solid in 25% yield. 'H NMR (DMSO-d6, 400 MHz): 3 8.51 (d, J= 4.0 Hz, 2H), 7.87 (s, 1H), 7.61 (d, J= 4.0 Hz, 1H), 7.28 (d, J = 8.0 Hz, 2H), 7.13 (t, J= 4.0 Hz, 1H), 7.02 (d, J= 8.0 Hz, 1H), 6.44 (dd, J = 8.0 Hz, 4.0 Hz, 1H), 3.75 (t, J= 4.0 Hz, 4H), 3. 68 (s, 3H), 3.66-3.59 (m, 1H), 3.51-3.42 (m, 2H), 2.67-2.61 (m, 4H), 2.12 (s, 6H). MS (ESI+) calculated for C21H29N5O3 [M+l]+: 400.5, observed: 400.5.

[0218] Example 7. Synthesis of:

[0219] To a stirred solution of l-fluoro-2-nitro-4-(trifluoromethyl)benzene (0.50 g, 2.39 mmol) in dry DMF (5 mL) was added 3 -methylmorpholine (0.3 g, 2.87 mmol), followed by K2CO3 (0.66 g, 4.78 mmol). The crude reaction was stirred under heating at 60 °C overnight. After cooling at room temperature, the crude reaction was diluted with DCM and washed with brine several times. The organic layer was dried over anhydrous ISfeSCU, filtered, and concentrated in vacuo. The product was purified by column chromatography using hexane: EtOAc (50:50) to afford red-orange oil with a 95 % yield.1H NMR (CDCI3, 400 MHz): 3 7.97 (s, 1H), 7.12 (dd, J = 8.0 Hz, 4.0 Hz, 1H), 7.30 (d, J= 8.0 Hz, 1H), 3.82-3.77 (m, 3H), 3.50 (dd, J= 8.0 Hz, J = 4.0 Hz, 1H), 3.42-3.33 (m, 2H), 2.85- 2.83 (m, 1H), 1.02 (d, J = 8.0 Hz, 3H). MS (ESI+) calculated for C12H13F3N2O3 [M+l]+: 291.2, observed: 291.2.

[0220] To a stirred solution of 3-methyl-4-(2-nitro-4-(trifluorornethyl)phenyl)morpholine (0.68 g, 2.3 mmol) in EtOH (10 mL), purged under nitrogen, was added Pd / C (0.05 g) and hydrazine monohydrate (1.9 mL, 38.9 mmol) was added dropwise. The crude reaction was stirred at room temperature for 2 hrs. Crude was filtered and concentrated in vacuo. The product was purified by column chromatography using hexane: EtOAc (30:70) to afford the product as a white solid with an 87 % yield. 'HNMR (CDC13, 400 MHz): 3 7.80 (s, 1H), 7.50 (s, 1H), 7.19 (d, J= 4.0 Hz, 1H), 7.13 (d, J = 8.0 Hz, 1H), 6.97 (d, J = 8.0 Hz, 1H), 3.93-3.85 (m, 2H), 3.79-3.72 (m, 1H), 3.39-3.33 (m, 1H), 3.29-3.22 (m, 1H), 2.92-2.88 (m, 1H), 2.75-2.68 (m, 1H), 0.8 (d, J= 8.0 Hz, 3H). MS (ESI+) calculated for C12H15F3N2O [M+l]+: 261.2, observed: 261.2.

[0221] To a stirred solution of 2-(3-methylmorpholino)-5 (trifluoromethyl) aniline (0.53 g, 2.00 mmol) in dry DCM (15 mL), under an N2 atmosphere at 0°C, was added a solution of phenyl chloroformate (0.28 mL, 2.24 mmol) in dry DCM (5 mL), followed by EtsN (0.56 mL, 4.00 mmol). The crude reaction was allowed to warm up at room temperature and stirred overnight. The crude reaction was concentrated in vacuo and the product was purified by column chromatography using hexane: EtOAc (70:30) to afford a cream solid in 80% yield.1H NMR (CDCI3, 400 MHz): 3 8.55 (s, 1H), 7.43-7.41 (m, 2H), 7.35 (s, 1H), 7.25-7.21 (m, 4H), 6.83 (d, J= 8.0 Hz, 1H), 4.00-3.94 (m, 2H), 3.84 (t, J= 4.0 Hz, 1H), 3.45 (t, J= 8.0 Hz, 1H), 3.24-3.20 (m, 1H), 2.98-2.88 (m, 1H), 2.81 (d, J = 12.0 Hz, 1H), 0.8 (d, J = 4.0 Hz, 3H). MS (ESI+) calculated for C19H19F3N2O3 [M+l]+: 381.3, observed: 381.3

[0222] To a stirred solution of phenyl (2-(3 -methylmorpholino) 5-(trifluoromehty) phenyl)carbamate (0.091 g, 0.23 mmol) in dry pyridine (5 mL) was added N^N imethyl-l- (pyridin-4-yl)ethane-l,2-diamine (0.043 g, 0.26 mmol) and the solution was heated at 80°C for 16 hrs. The crude reaction was cooled down to room temperature and concentrated in vacuo. The product was purified by column chromatography using DCM:EtOH (95:5) and obtained as a white solid in 25% yield.XH NMR (DMSO-d6, 400 MHz): 3 8.54 (s, 2H), 8.43 (d, J= 8.0 Hz, 1H), 8.22 (d, J= 4.0 Hz, 1H), 7.39 (t, J= 4.0 Hz, 1H), 7.32 (s, br, 2H), 7.23 (d, J= 8.0 Hz, 1H), 3.84-3.77 (m, 4H), 3.68 (s, br, 1H), 3.39-3.34 (m, 1H), 3.13- 3.08 (m, 1H), 2.66 (s, br, 3H), 2.15 (s, br, 6H), 0.65 (t, J= 4.0 Hz, 3H). MS (ESI+) calculated for C22H28F3N5O2 [M+l]+: 452.5, observed: 452.5.

[0223] Example 8. Synthesis of:

[0224]

[0225] To a stirred solution of l-fluoro-2-nitro-4-(trifluoromethyl)benzene (0.73 g, 3.51 mmol) in dry DMF (10 mL) was added 3, 5 -dimethylmorpholine (0.71 g, 4.68 mmol), followed by K2CO3 (1.61 g, 11.6 mmol). The crude reaction was stirred under heating at 80 °C overnight. After cooling to room temperature, the crude reaction was diluted with DCM and washed with brine several times. The organic layer was dried over anhydrous ISfeSCU, filtered, and concentrated in vacuo. The product was purified by column chromatography using hexane: EtOAc (50:50) to afford a white solid with a 50% yield. 'H NMR (CDCI3, 400 MHz): 3 7.95 (s, 1H), 7.74 (dd, J= 8.0 Hz, 4.0 Hz, 1H), 7.33 (d, J = 8.0 Hz, 1H), 3.88 (dd, J = 8.0 Hz, 4.0 Hz, 2H), 3.68 (s, br, 2H), 3.31 (s, br, 2H), 1.10 (s, br, 3H), 0.82 (s, br, 3H). MS (ESI+) calculated for Ci3Hi5F3N2O3 [M+l]+: 304.2, observed: 304.2.

[0226] To a stirred solution of 3,5-dimethyl-4-(2-nitro-4-(trifluoromethyl)phenyl)morpholine (0.3 g, 1.0 mmol) in EtOH (10 mL), purged under nitrogen, was added Pd / C (0.05 g) and hydrazine monohydrate (0.8 mL, 16.3 mmol) was added dropwise. The crude reaction was stirred at room temperature for 2 hrs. Crude was filtered and concentrated in vacuo. The product was purified by column chromatography using hexane: EtOAc (30:70) to afford the product as a white solid with a 30% yield. 'H NMR (CDCI3, 400 MHz): 3 1.11 (s, 1H), 7.51 (d, J = 4.0 Hz, 1H), 7.17 (dd, J = 8.0 Hz, 4.0 Hz, 1H), 7.01 (d, J= 4.0 Hz, 1H), 3.92-3.85 (m, 2H), 3.68-3.64 (m, 2H), 3.24 (t, J = 8.0 Hz, 1H), 3.02 (dd, J= 8.0 Hz, 4.0 Hz, 1H), 1.06 (d, J= 8.0 Hz, 3H), 0.75 (d, J= 8.0 Hz, 3H). MS (ESI+) calculated for C13H17F3N2O [M+l]+: 275.3, observed: 274.3.

[0227] To a stirred solution of 2-(3,5-dimethylmorpholino)-5 (trifluoromethyl) aniline (0.076 g, 0.27 mmol) in dry DCM (15 mL), under an N2 atmosphere at 0°C, was added a solution of phenyl chloroformate (0.041 mL, 0.33 mmol) in dry DCM (5 mL), followed by EtsN (0.07 mL, 0.54 mmol). The crude reaction was allowed to warm up at room temperature and stirred overnight. The crude reaction was concentrated in vacuo and the product was purified by column chromatography using hexane: EtOAc (70:30), product was obtained as a cream solid in 60% yield.1H NMR (CDCI3, 400 MHz): 3 8.62 (s, 1H), 8.13 (s, 1H), 7.45-7.39 (m, 2H), 7.24-7.17 (m, 3H), 6.93 (d, J = 8.0 Hz, 1H), 6.82 (d, J = 8.0 Hz, 1H), 3.99-3.92 (m, 2H), 3.85 (d, J = 8.0 Hz, 1H), 3.71-3.63 (m, 1H), 3.35 (t, J= 8.0 Hz, 1H), 3.04 (d, J= 8.0 Hz, 1H), 1.18 (d, J= 4.0 Hz, 3H), 0.81 (d, J= 4.0 Hz, 3H). MS (ESI+) calculated for C20H21F3N2O3 [M+l]+: 395.3, observed: 395.3.

[0228] To a stirred solution of phenyl (2-(3,5-dimethylmorpholino)-5-(trifluoromehty) phenyl)carbamate (0.045 g, 0.11 mmol) in dry pyridine (5 mL) was added N^N imethyl-l- (pyridin-4-yl)ethane-l,2-diamine (0.018 g, 0.11 mmol) and the solution was heated at 80 °C for 16 hrs. The crude reaction was cooled down to room temperature and concentrated in vacuo. The product was purified by column chromatography using DCM:EtOH (95:5) and obtained as a white solid in 25% yield. 'H NMR (DMSO-d6, 400 MHz): 3 8.52 (d, J= 4.0 Hz, 2H), 8.30 (s, 1H), 8.03 (s, 1H), 7.29 (d, J = 4.0 Hz, 2H), 7.25-7.20 (m, 2H), 3.99-3.95 (m, 1H), 3.83-3.80 (dd, J = 8.0 Hz, 4.0 Hz, 1H), 3.69-3.355 (m, 4H), 3.44 (t, J= 4.0 Hz, 1H), 3.38-3.36 (m, 1H), 2.75-2.74 (m, 1H), 2.12 (s, 6H), 0.93 (d , J= 8.0 Hz, 3H), 0.65 (t, J = 4.0 Hz, 3H). MS (ESI+) calculated for C23H30F3N5O2 [M+l]+: 466.5, observed: 466.5.

[0229] Example 9. Synthesis of:

[0230] To a stirred solution of 4-fluoro-3 -nitrophenol (1.0 g, 6.36 mmol) in dry Acetonitrile (10 mL) was added K2CO3 (1.82 g, 13.22 mmol), followed by l-bromo-2-(2-methoxyethoxy)ethane (1.0 mL, 7.3 mmol). The crude reaction was stirred under heating at 80 °C overnight. After cooling at room temperature, crude reaction was diluted with DCM and filtered. The crude was concentrated in vacuo, redissolved in EtOAc (20 mL) and washed with IN NaOH (5 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to afford a red oil 63 % yield. 'HNMR (CDCI3, 400 MHZ): 3 7.58 (d, J= 4.0 Hz, 1H), 7.18 (d, J= 4.0 Hz, 2H), 4.17 (t, J = 4.0 Hz, 2H), 3.87 (t, J = 4.0 Hz, 2H), 3.71 (m, 2H), 3.59-3.58 (m, 2H), 3.39 (s, 3H). MS (ESI+) calculated for C11H14FN2O5 [M+l]+: 260.3, observed: 260.3. To a stirred solution of l-fluoro-4-(2-(2-methoxyethoxy)ethoxy)-2-nitrobenzene (0.56 g, 2.17 mmol) in dry DMF (5 mL) was added morpholine (0.21 mL, 2.39 mmol), followed by K2CO3 (0.6 g, 4.34 mmol). The crude reaction was stirred under heating at 60 °C overnight. After cooling at room temperature, crude reaction was diluted with DCM and washed with brine several times. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. Product was purified by column chromatography using hexane: EtOAc (50:50) to afford a red oil in a 68% yield. ^ NMR ^DCh, 400 MHZ): 3 7.38 (d, J= 4.0 Hz, 1H), 7.18 (d, J= 4.0 Hz, 1H), 7.11 (dd, , J= 8.0 Hz, 4.0 Hz, 1H), 4.14 (t, J= 4.0 Hz, 1H), 3.87-3.82 (m, 3H), 3.71 (m, 1H), 3.59-3.56 (m, 1H), 3.39 (s, 3H), 3.01-2.98 (m, 2H). MS (ESI+) calculated for C15H22N2O6 [M+l]+: 327.3, observed: 327.3.

[0231] To a stirred solution of 4-(4-(2-(2-methoxyethoxy)ethoxy)-2-nitrophenyl)morpholine (0.49 g, 1.52 mmol) in EtOH (10 mL), purged under nitrogen, was added Pd / C (0.05 g) and hydrazine monohydrate (1.25 mL, 25.7 mmol) was added dropwise. The crude reaction was stirred at room temperature for 2 hours. Crude was filtered and concentrated in vacuo. Product was purified by column chromatography using hexane: EtOAc (30:70) to afford a white solid in 81 % yield. 'H NMR (CDCI3, 400 MHZ): 3 6.93 (d, J= 4.0 Hz, 1H), 6.34 (d, J= 4.0 Hz, 1H), 6.30 (dd, J= 12.0 Hz, 4.0 Hz, 1H), 4.08 (t, J= 4.0 Hz, 2H), 3.89 (s, br, 4H), 3.82 (t, J= 4.0 Hz, 2H), 3.70 (t, J= 4.0 Hz, 2H), 3.57 (t, J = 4.0 Hz, 2H), 3.39 (s, 3H), 2.94 (s, br, 4H). MS (ESI+) calculated for C15H24N2O4 [M+l]+: 297.3, observed: 297.3.

[0232] To a stirred solution of 5-(2-(2-methoxyethoxy)ethoxy)-2-morpholinoaniline (0.36 g, 1.21 mmol) and K2CO3 (0.33 g, 2.42 mmol) in Acetone (10 mL), at 0 °C, was added dropwise phenyl chloroformate (0.16 mL, 1.33 mmol). The crude reaction was allowed to warm up at room temperature and stirred overnight. Crude reaction was concentrated in vacuo, dissolved in DCM (15 mL) and washed with water (10 mL). The organic layer was dried over anhydrous Na2SO4 and concentrated. Crude product was taken to the next reaction without further purification.1H NMR (CDCI3, 400 MHZ): 3 8.54 (s, br, 1H), 7.82 (s, 1H), 7.41 (t, J= 8.0 Hz, 2H), 7.23 (d, J= 8.0 Hz, 2H), 7.14 (d, J= 8.0 Hz, 2H), 6.65 (dd, J= 8.0 Hz, 4.0 Hz, 1H), 4.12 (t, J= 4.0 Hz, 2H), 3.92 (s, 4 H), 3.83 (t, J= 4.0 Hz, 2H), 3.70 (t, J = 4.0 Hz, 2H), 3.56 (t, J = 4.0 Hz, 2H), 3.38 (s, 3H), 2.91 (t, J= 4.0 Hz, 4H). MS (ESI+) calculated for C22H28N2O6 [M+l]+: 417.4, observed: 417.4.

[0233] To a stirred solution of phenyl (5-(2-(2-methoxyethoxy)ethoxy)-2-morpholinophenyl)- carbamate (0.062 g, 0.15 mmol) in dry pyridine (3 mL) was added N1,N1-dimethyl-l-(pyridin-4- yl)ethane-l,2-diamine (0.03 g, 0.17 mmol) and the solution was heated at 80 °C for 16 hours. Crude reaction was cooled down to room temperature and concentrated in vacuo. Product was purified by column chromatography using DCM:EtOH (95:5) to afford pure product as a white solid in 25% yield. 'HNMR (CDCI3, 400 MHZ): 3 8.58 (d, J= 4.0 Hz, 2H), 7.70 (s, br, 1H), 7.68 (d, J= 4.0 Hz, 1H), 7.23 (d, J= 4.0 Hz, 2H), 7.03 (d, J= 4.0 Hz, 1H), 6.54 (dd, J = 8.0 Hz, 4.0 Hz, 1H), 4.12 (t, J = 4.0 Hz, 2H), 3.86-3.80 (m, 3H), 3.77-3.76 (m, 4H), 3.72-3.70 (m, 3H), 3.58- 3.55 (m, 2H), 3.38 (s, 3H), 2.81-2.71 (m, 4H), 2.33 (s, br, 6H). MS (ESI+) calculated for C25H37N5O5 [M+l]+: 488.6, observed: 488.6.

[0234] Example 10. Synthesis of:

[0235] To a stirred solution of 4-fluoro-3 -nitrophenol (0.62 g, 4.0 mmol) in dry Acetonitrile (10 mL) was added K2CO3 (1.1 g, 8.0 mmol), followed by l-bromo-2-(2-(2-methoxyethoxy)- ethoxy)ethane (1.0 g, 4.4 mmol). The crude reaction was stirred under heating at 80 °C overnight. After cooling at room temperature, crude reaction was diluted with DCM and filtered. The crude was concentrated in vacuo, redissolved in EtOAc (20 mL) and washed with IN NaOH (5 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to afford a yellow-brown oil in 87% yield. 'H NMR (CDCI3, 400 MHZ): 3 7.56 (dd, J= 4.0 Hz, 1H), 7.19 (d, J = 4.0 Hz, 2H), 4.16 (t, J= 4.0 Hz, 1H), 3.87 (t, J= 4.0 Hz, 1H), 3.81 (t, J= 8.0 Hz, 1H), 3.75-3.72 (m, 1H), 3.70-3.64 (m, 5H), 3.57-3.54 (m, 2H), 3.47 (t, J= 8.0 Hz, 1H), 3.38 (s, 3H). MS (ESI+) calculated for C13H18FN2O6 [M+l]+: 304.3, observed: 304.3.

[0236] To a stirred solution of l-fluoro-4-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)-2-nitrobenzene (1.0 g, 3.43 mmol) in dry DMF (10 mL) was added morpholine (0.33 mL, 3.77 mmol), followed by K2CO3 (0.95 g, 6.86 mmol), The crude reaction was stirred under heating at 60°C overnight. After cooling at room temperature, crude reaction was diluted with DCM and washed with brine several times. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. Product was purified by column chromatography using hexane: EtOAc (50:50) to afford a red- orange oil in 68% yield. 'HNMR (CDC13, 400 MHZ): 3 7.30 (d, J= 4.0 Hz, 1H), 7.16 (d, J= 4.0 Hz, 1H), 7.11 (dd, , J= 8.0 Hz, 4.0 Hz, 1H), 4.13 (t, J= 4.0 Hz, 2H), 3.85 (t, J= 4.0 Hz, 2H), 3.82 (t, J= 4.0 Hz, 4H), 3.74-3.72 (m, 2H), 3.69-3.64 (m, 4H), 3.56-3.54(m, 2H), 3.38 (s, 3H), 2.97 (t, J = 4.0 Hz, 4H). MS (ESI+) calculated for C17H26N2O7 [M+l]+: 371.4, observed: 371.4.

[0237] To a stirred solution of 4-(4-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)-2-nitrophenyl)- morpholine (0.18 g, 0.48 mmol) in EtOH (10 mL), purged under nitrogen, was added Pd / C (0.05 g) and hydrazine monohydrate (0.4 mL, 8.22 mmol) was added dropwise.. The crude reaction was stirred at room temperature for 2 hours. Crude was filtered and concentrated in vacuo. Product was purified by column chromatography using hexane: EtOAc (30:70) to afford as a clear oil in 81 % yield. 'H NMR (CDCI3, 400 MHZ): 3 6.91 (d, J= 8.0 Hz, 1H), 6.34 (d, J= 4.0 Hz, 1H), 6.29 (dd, J = 12.0 Hz, 4.0 Hz, 1H), 4.07-4.05 (m, 2H), 3.82-3.80 (m, 6H), 3.74-3.71 (m, 2H), 3.69-3.64 (m, 4H), 3.56-3.54 (m, 2H), 3.38 (s, 3H), 2.85 (t, J = 4.0 Hz, 4H). MS (ESI+) calculated for C17H28N2O5 [M+l]+: 341.4, observed: 341.4.

[0238] To a stirred solution of 5-(2-(2-methoxyethoxy)ethoxy)-2-morpholinoaniline (0.09 g, 0.25 mmol) and K2CO3 (0.07 g, 0.5 mmol) in acetone (5 mL), at 0 °C, was added dropwise phenyl chloroformate (0.03 mL, 0.28 mmol). The crude reaction was allowed to warm up at room temperature and stirred overnight. Crude reaction was concentrated in vacuo, dissolved in DCM (15 mL) and washed with water (10 mL). The organic layer was dried over anhydrous Na2SO4 and concentrated. Crude product was taken to next reaction without further purification.1H NMR (CDCI3, 400 MHZ): 3 8.43 (s, 1H), 7.83 (s, 1H), 7.42 (t, J= 4.0 Hz, 2H), 7.28-7.26 (m,l H), 7.23 (d, J= 8.0 Hz, 2H), 713 (d, J= 8.0 Hz, 1H), 6.63 (dd, J = 4.0 Hz, 2.0 Hz, 1H), 4.13 (t, J= 4.0 Hz, 2H), 3.89 (t, J= 4.0 Hz, 4H), 3.82 (t, J= 4.0 Hz, 2H), 3.73-3.70 (m, 2H), 3.66-3.63 (m, 4H), 3.55.- 3.53 (m, 2H), 3.37 (s, 3H), 2.86 (t, J= 4.0 Hz, 4H). MS (ESI+) calculated for C24H32N2O7 [M+l]+: 461.5, observed: 461.5.

[0239] To a stirred solution of phenyl (5-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)-2- morpholinophenyl)carbamate (0.062 g, 0.15 mmol) in dry pyridine (3 mL) was added N^N1- dimethyl-l-(pyridin-4-yl)ethane-l,2-diamine (0.03 g, 0.17 mmol) and the solution was heated at 80°C for 16 hours. Crude reaction was cooled down to room temperature and concentrated in vacuo. Product was purified by column chromatography using DCM:EtOH (95:5) to afford pure product as a white solid in 25 % yield.1H NMR (CDCI3, 400 MHZ): 3 8.61 (d, J= 4.0 Hz, 2H), 7.80 (s, br, 1H), 7.69 (d, J= 4.0 Hz, 1H), 7.03 (d, J= 4.0 Hz, 2H), 6.54 (dd, J= 8.0 Hz, 4.0 Hz, 1H), 4.10 (t, J= 4.0 Hz, 2H), 3.86-3.78 (m, 6H), 3.74-3.71 (m, 2H), 3.68-3.62 (m, 7H), 3.56- 3.53 (m, 2H), 3.37 (s, 3H), 2.77 (d, J= 4.0 Hz, 4H), 2.41 (s, br, 6H). MS (ESI+) calculated for C27H41N5O6 [M+l]+: 532.6, observed: 532.6.

[0240] Example 11. Synthesis of:

[0241] To a stirred solution of 4-fluoro-3 -nitrophenol (0.77 g, 4.93 mmol) in dry Acetonitrile (10 mL) was added K2CO3 (1.3 g, 9.86 mmol), followed by 2-(2-bromoethoxy)ethan-l-ol (1.34 mL, 12.7 mmol). The crude reaction was stirred under heating at 80 °C overnight. After cooling at room temperature, crude reaction was diluted with DCM and filtered. The crude was concentrated in vacuo, redissolved in EtOAc (20 mL) and washed with IN NaOH (5 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to afford a red oil in 87 % yield. 'HNMR (CDCI3, 400 MHZ): 3 7.60 (dd, J= 4.0 Hz, 1H), 7.21-7.19 (d, J= 4.0 Hz, 2H), 4.18 (t, J = 4.0 Hz, 2H), 3.89 (t, J= 4.0 Hz, 2H), 3.78 (t, J= 4.0 Hz, 2H), 3.68 (t, J= 4.0 Hz, 2H), 1.97 (s, br, 1H). MS (ESI+) calculated for C10H12FNO5 [M+l]+: 246.2, observed: 246.2.

[0242] To a stirred solution of 2-(2-(4-fluoro-3 -nitrophen oxy)ethoxy)ethan-l-ol (0.48 g, 1.86 mmol) in dry DCM (10 mL) was added 2,6-Lutidine (0.47 mL, 4.09 mmol), followed by TBSOTf (0.51 mL, 2.23 mmol). The crude reaction was stirred at room temperature overnight. Crude reaction was diluted with DCM (20 mL) and washed with brine (15 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. Product was purified by column chromatography using hexane: EtOAc (80:20) to afford a clear oil in 71 % yield. 'H NMR (CDCI3, 400 MHZ): 3 7.55 (d, J= 4.0 Hz, 1H), 7.20-7.18 (m, 2H), 4.15 (t, J= 4.0 Hz, 2H), 3.86 (t, J= 4.0 Hz, 2H), 3.78 (t, J= 4.0 Hz, 2H), 3.63 (t, J= 4.0 Hz, 2H), 0.89 (s, 9H), 0.07 (s, 6H). MS (ESI+) calculated for C16H26FNO5 Si [M+l]+: 360.4, observed: 360.4. To a stirred solution of tert-butyl(2-(2-(4-fluoro-3-nitrophenoxy)ethoxy)ethoxy)- dimethylsilane (0.47 g, 1.33 mmol) in dry DMF (5.0 mL) was added morpholine (0.14 mL, 1.6 mmol), followed by K2CO3 (0.36 g, 2.66 mmol), The crude reaction was stirred under heating at 45 °C overnight. After cooling at room temperature, crude reaction was diluted with DCM and washed with brine several times. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. Product was purified by column chromatography using hexane: EtOAc (70:30) to afford a red-orange oil in 70 % yield.1H NMR (CDCI3, 400 MHZ): 3 7.32 (d, J= 4.0 Hz, 1H), 7.17 (d, J= 8.0 Hz, 1H), 7.11 (dd, J= 8.0 Hz, 4.0 Hz, 1H), 4.12 (t, J= 4.0 Hz, 2H), 3.86 (t, J = 4.0 Hz, 2H), 3.82 (t, J= 4.0 Hz, 4H), 3.79 (t, J = 4.0 Hz, 2H), 3.62 (t, J = 4.0 Hz, 2H), 2.97 (t, J= 4.0 Hz, 4H), 0.89 (s, 9H), 0.07 (s, 6H). MS (ESI+) calculated for C2oH34N206Si [M+l]+: 427.6, observed: 427.6.

[0243] To a stirred solution of 4-(4-(2-(2-((tert-butyldimethylsilyl)oxy)ethoxy)ethoxy)-2- nitrophenyl)morpholine (0.18 g, 0.48 mmol) in EtOH (10 mL), purged under nitrogen, was added Pd / C (0.02 g) and hydrazine monohydrate (0.4 mL, 8.17 mmol) was added dropwise. The crude reaction was stirred at room temperature for 2 hours. Crude was filtered and concentrated in vacuo. Product was purified by column chromatography using hexane: EtOAc (60:40) to afford the product as a clear oil in 81% yield. 'HNMR (CDCI3, 400 MHZ): 3 6.92 (d, J= 8.0 Hz, 1H), 6.34 (d, J= 4.0 Hz, 1H), 6.30 (dd, J= 8.0 Hz, 4.0 Hz, 1H), 4.05 (t, J= 4.0 Hz, 4H), 3.82 (t, J= 4.0 Hz, 4H), 3.78 (t, J= 4.0 Hz, 2H), 3.62 (t, J= 4.0 Hz, 2H), 2.85 (t, J= 4.0 Hz, 4H), 0.89 (s, 9H), 0.07 (s, 6H). MS (ESI+) calculated for C2oH36N204Si [M+l]+: 397.6, observed: 397.6.

[0244] To a stirred solution of 5-(2-(2-((tert-butyldimethylsilyl)oxy)ethoxy)ethoxy)-2- morpholinoaniline (0.06 g, 0.14 mmol) in dry DCM (5 mL), under N2 atmosphere at 0°C, was added a solution of phenyl chloroformate (0.02 mL, 0.16 mmol) in dry DCM (2 mL), followed by EtsN (0.03 mL, 0.21 mmol). The crude reaction was allowed to warm up at room temperature and stirred overnight. Crude reaction was concentrated in vacuo and product was purified by column chromatography using hexane: EtOAc (70:30) to afford pure product as a clear cream solid in 60% yield. 'H NMR (CDCI3, 400 MHZ): 3 8.44 (s, br, 1H), 7.84 (s, 1H), 7.42 (t, J= 4.0 Hz, 2H), 7.30- 7.27 (m, 2H), 7.12 (dd, J= 8.0 Hz, 4.0 Hz, 2H), 6.64 (dd, J = 4.0 Hz, 2.0 Hz, 1H), 4.09 (t, J= 4.0 Hz, 2H), 3.89 (t, J= 4.0 Hz, 4H), 3.82 (t, J= 4.0 Hz, 2H), 3.77 (t, J= 4.0 Hz, 2H), 3.60 (t, J= 4.0 Hz, 2H), 2.86 (t, J= 4.0 Hz, 4H), 0.88 (s, 9H), 0.06 (s, 6H) . MS (ESI+) calculated for C27H4oN206Si [M+l]+: 517.7, observed: 517.7. To a stirred solution of phenyl (5-(2-(2-((tert-butyldimethylsilyl)oxy)ethoxy)ethoxy)-2- morpholinophenyl)carbamate (0.08 g, 0.15 mmol) in dry pyridine (3 mL) was added N^N1- dimethyl-l-(pyridin-4-yl)ethane-l,2-diamine (0.03 g, 0.17 mmol) and the solution was heated at 80°C for 16 hours. Crude reaction was cooled down to room temperature and concentrated in vacuo. Crude product was stirred at room temperature in dry THF (2 mL) and 1.0 mL of 1 M TBAF solution in THF was added. The solution was stirred overnight and concentrated. Crude product was partitioned in MeOH: Water (1 : 1, 1 mL each) and filtered. Product was purified by HPLC using ACN:Water ( 10:90->95:5) to afford pure product as a white solid in 25 % yield. 'H NMR (CDCI3, 400 MHZ): 3 8.58 (d, J= 4.0 Hz, 2H), 7.80 (s, br, 1H), 7.23 (d, J= 4.0 Hz, 1H), 7.03 (d, J= 4.0 Hz, 2H), 6.54 (dd, J= 8.0 Hz, 4.0 Hz, 1H), 5.64 (s, br, 1 H), 4.13 (t, J= 4.0 Hz, 2H), 3.84-3.82 (m, 3H), 3.79-3.75 (m, 4H), 3.66-3.64 (m, 2H), 3.59- 3.52 (m, 1H), 3.38-3.34 (m, 1H), 2.77 (t, J= 4.0 Hz, 4H), 2.32 (s, br, 6H). MS (ESI+) calculated for C24H35N5O5 [M+l]+: 474.6, observed: 474.6.

[0245] Example 12. Synthesis of: l-(2-(Dimethylamino)-2-(pyridin-4-yl)ethyl)-3-(5-(methoxymethoxy)-2-morpholino- phenyl)urea (0.01 g) was stirred in 2 mL of a 2N solution of HC1 in diethyl ether at room temperature overnight. Crude was concentrated in vacuo and lyophilized to obtain pure product as a white solid in 95 % yield. 'HNMR (D2O, 400 MHZ): 3 9.00 (d, J= 8.0 Hz, 2H), 8.31 (d, J= 8.0 Hz, 2H), 7.56 (d, J= 4.0 Hz, 1H), 6.94 (dd, J= 8.0 HZ, 4.0 Hz, 1H), 6.74 (d, J= 4.0 Hz, 1H), 4.97 (t, J= 4.0 Hz, 1H), 4.11-4.09 (m, 7H), 3.66 (t, J = 4.0 Hz, 4H), 3.00 (s, 6H). MS (ESI+) calculated for C22H32CIN5O4 [M+l]+: 466.21, observed: 430.23 - HC1.

[0246] Example 13. Synthesis of:

[0247] l-(2-(Dimethylamino)-2-(pyridin-4-yl)ethyl)-3-(5-(2-(2-methoxyethoxy)ethoxy)-2- morpholinophenyl)urea (0.01 g) was stirred in 1 mL of a 2N solution of HC1 in diethyl ether at room temperature overnight. Crude was concentrated in vacuo and lyophilized to obtain pure product as an orange solid in 95 % yield. ‘H NMR (D2O, 400 MHZ): 3 8.74 (d, J= 8.0 Hz, 2H), 7.73 (d, J= 8.0 Hz, 2H), 7.21 (d, J= 4.0 Hz, 1H), 7.18 (d, J= 4.0 Hz, 1H), 6.77 (dd, J= 4.0 Hz, 1H), 4.69-4.65 (m, 1H), 4.17-4.14 (m, 2H), 4.07-3.94 (dq, J= 12.0 Hz, 8.0 Hz, 2H), 3.87-3.85 (m, 2H), 3.78 (t, J= 4.0 Hz, 4H), 3.73-3.72 (m, 2H), 3.62-3.60 (m, 2H), 3.35 (s, 3H), 2.92 (s, 6H), 2.80 (t, J = 4.0 Hz, 4H). MS (ESI+) calculated for C25H38CIN5O5 [M+l]+: 524.25, observed: 488.27 - HC1.

[0248] Example 14. Synthesis of: l-(2-(Dimethylamino)-2-(pyridin-4-yl)ethyl)-3-(2-morpholino-5-(trifluoromethoxy)- phenyl)urea (0.01 g) was stirred in 1 mL of a 2N solution of HC1 in diethyl ether at room temperature overnight. Crude was concentrated in vacuo and lyophilized to obtain pure product as a yellow solid in 95 % yield. 'HNMR (D2O, 400 MHZ): 3 8.68 (s, br, 2H), 7.57 (s, 2H), 7.49 (s, 1H), 7.20 (s, 1H), 7.04 (s, 1H), 4.60-4.59 (m, 1H), 4.02-3.96 (m, 2H), 3.75 (s, 4H), 2.89 (s, 6H), 2.73 (s, 4H). MS (ESI+) calculated for C21H27CIF3N5O3 [M+l]+: 490.17, observed: 454.20 - HC1.

[0249] Example 15. Synthesis of:

[0250] l-(2-(Dimethylamino)-2-(pyridin-4-yl)ethyl)-3-(5-fluoro-2-morpholinophenyl)urea (0.02 g) was stirred in 1 mL of a 2N solution of HC1 in diethyl ether at room temperature overnight. Crude was concentrated in vacuo and lyophilized to obtain pure product as a yellow solid in 95 % yield. 'H NMR (DMSO-de, 400 MHz): 3 8.99 (d, J= 8.0 Hz, 2H), 8.23 (s, 2H), 8.08 (s, 1H), 7.98 (s, br, 2H), 7.79 (dd, J= 8.0 Hz, 4.0 Hz, 1H), 7.15 (dd, J= 8.0 Hz, 4.0 HZ, 1H), 6.70 (td, J= 8.0 Hz, 4.0 Hz, 1H), 4.81 (t, J = 4.0 Hz, 1H), 4.06-4.00 (m, 1H), 3.85-3.82 (m, 1H), 3.78 (t, J = 4.0 Hz, 4H), 2.97 (s, br, 6 H), 2.66 (t, J = 4.0 Hz, 4H). MS (ESI+) calculated for C20H27CIFN5O2 [M+l]+: 424.18, observed: 388.20 - HC1.

[0251] Example 16. Synthesis of: l-(2-(Dimethylamino)-2-(pyridin-4-yl)ethyl)-3-(5-fluoro-2-morpholinophenyl)urea (0.02 g) was stirred in 1 mL of a 2N solution of HC1 in diethyl ether at room temperature overnight. Crude was concentrated in vacuo and lyophilized to obtain pure product as a white solid in 95 % yield.1H NMR (DMSO-d6, 400 MHz): 3 8.86 (d, J= 4.0 Hz, 2H), 8.34 (s, 1H), 8.03 (s, 1H), 7.96 (d, J= 4.0 Hz, 2H), 7.79 (s, br, 1H), 7.24 (s, 2H), 4.10-4.09 (m, 1H), 3.93 (d, J= 8.0 Hz, 1H), 3.79-3.68 (m, 2H), 3.59-3.46 (m, 3H), 2.93 (s, 2H), 2.60 (s, br, 6H), 0.93 (d, J= 4.0 Hz, 3H), 0.61 (d, J= 4.0 Hz, 3H). MS (ESI+) calculated for C23H31CIF3N5O2 [M+l]+: 502.21, observed: 466.23 - HC1.

[0252] Example 17. Biological Testing

[0253] Reagents'. Dulbecco's modified Eagle's medium (DMEM) (#25-500) was from Genesee Scientific (San Diego, CA, USA). 0.05% Trypsin-EDTA (#25-300) and Pierce bicinchoninic acid (BCA) Protein Assay Kit (#23225) were obtained from Thermo Fisher Scientific (Waltham, MA, USA). Human transferrin (#10652202001) was obtained from Roche Applied Science (Mannheim, Germany). IRDye conjugated secondary antibodies the anti-rabbit IRDye 680 (#925-68073) and anti-mouse IRDye 800 (#925-32213) were from LI-COR Biosciences (Lincoln, NE, USA). FAK- Tyr-397 antibody (#ab81298) was from Abeam (San Francisco, CA, USA). We also used antibodies to total FAK (Anti-FAK, clone 4.47, #05-537) from EMD Millipore (Temecula, CA, USA). Antibodies to Erkl / 2 T-202 / Y-204 (#4370), and ERK1 / 2 (#4696) were from Cell Signaling (Danvers, MA, USA). Collagen type-I (#C8919) was obtained from Sigma-Aldrich (St. Louis, MO, USA). The RhoA kinase inhibitor InsolutionTM Y-27632 (#688000) was from MilliporeSigma, and was used at working concentration 20pM. FAK inhibitor PF573328 (#s2013) was from Selleck Chem( Houston, TX, USA), and was used at working concentration 10 pM. ERK kinase (MEK) inhibitor PD98059 (#PHZ1164) was from Thermofisher Scientific, and was used at working concentration 10 pM. Rho kinase (ROCK) Activity Assay kit (STA-416) was from Cell Biolabs(San Diego, CA, USA).

[0254] The compound M64 (the first compound in the table below) has the formula:

[0255] The compound M64HC1 is the HC1 salt of M64.

[0256] Cell culture: Human Caco-2 cells were obtained from American Tissue Culture Collection

[0257] (ATCC) (Manassas, VA) and maintained at 37°C with 8% CO2 in Dulbecco’s modified Eagle medium (DMEM) supplemented with 4.5 g / L D-glucose, 4 mM glutamine, 1 mM sodium pyruvate, 100 U / ml penicillin, 100 pg / ml streptomycin, 10 pg / ml transferrin, 10 mM HEPES pH 7.4, 3.7 g / L NaHCCh and supplemented with 10% fetal bovine serum.

[0258] UPLC-MS analysis for M64HCI concentrations in cells and media: After a two-hour M64HC1 treatment, the M64HC1 -containing medium was removed, and cells were washed with PBS three times for one minute each, before adding normal cell culture medium. After one-hour incubation in a normal culture medium to permit equilibration, cells and media samples (500 pL of cells homogenate and 20 pL of media) were homogenized in methanol (80% final concentration) by sonication. Precipitated proteins were separated by centrifugation at 2,000 xg for 10 min, and 10 pL of supernatant was injected into the UPLC-MS / MS system for analysis. Precipitated proteins were incubated with 1 mL 0.2 M KOH per tube overnight in a water bath at 60 °C. Total protein concentrations were measured by using the Pierce BCA protein assay kit with a bovine serum albumin standard containing 0.2 M KOH. Then, M64HC1 concentrations within cells measured by MS were normalized to the total protein concentration measured by the Pierce BCA protein assay kit, making the assumption based upon the literature that the total protein concentration represents approximately 15% of wet cell weight. UPLC separation was achieved using Waters Acquity I Class UPLC system (Waters, Milford, MA, USA) on Waters ACQUITY UPLC HSS T3 column (1.8 pM, 100 A pore diameter, 2.1 x 150 mm; Waters) with an ACQUITY UPLC HSS T3 precolumn (1.8 pM, 100 A pore diameter, 2.1 x 5 mm; Waters) heated at 55 °C. We used a linear gradient of solvents A (0.1% formic acid in water) and B (0.1% formic acid in acetonitrile) at 0.3 mL / min. At 1 min of separation, initial %B was increased from 1% to 25% during 1 min, and at 2.1 min - to 90% during 1.9 min. At 5 min, solvent B was returned to 1% and allowed 4 min for equilibration between injections. MS / MS analysis was performed on Waters Xevo TQ-S triple quadrupole mass spectrometer (Waters, Milford, MA, USA) using multiple reaction monitoring mode. The MS was operated in a positive ESI mode. The following mass transitions (with collision energies (CE) indicated in parentheses, V) were used: 438.2 / 273.1 (25) for quantification; 438.2 / 192.1 (20), and 438.2 / 149.2 (24) for analyte confirmation. The UPLC-MS / MS system was controlled by MassLynx V4.1. Quantification was performed against an M64HC1 external standard using a generated response curve.

[0259] Wound closure: Caco-2 cells were seeded at 80% confluence into 6 well plates pre-coated with type-I collagen. When the cells reached 100% confluence (48 hours after seeding), they were wounded with 200 pl non-barrier autoclaved pipette tips. In the first experiment, Caco-2 cells were wounded, and 0-hour images were taken for each monolayer. Then, Caco-2 cells were treated with either M64HC1 (100 nM) for 1, 2, 3, 4, and 24 hours or with a water vehicle control. After the indicated incubation time with M64HC1, the M64HC1 -containing medium was removed, and cells were washed with PBS three times for one minute each, before adding normal cell culture medium. The cells were then incubated at 37 °C until images were taken of the monolayer wounds at 24 hours. In the second experiment, Caco-2 cells were wounded, and 0-hour images were taken for each monolayer. The first two groups of monolayers were treated with either M64HC1 (100 nM) or a water vehicle control for 24 hours. Group 3 and 4 were treated with either M64HC1 (100 nM) or a water vehicle control for 2 hours. After the two-hour incubation, the culture medium was removed, and cells were similarly washed with PBS three times before being incubated with a normal medium at 37 °C for 10 hours. Then, the cells were treated again for two more hours with either M64HC1 (100 nM) or a water vehicle control. After this second two-hour incubation, the medium was again removed and cells were similarly washed with PBS three times, before being incubated with normal media at 37 °C for 10 more hours. Then, 24-hour images were taken for all the conditions. Wound images were captured using an inverted light microscope (OLYMPUS CK2, Center Valley, PA) at 0 hours and 24 hours after wounding. Wound areas were measured with Fiji software, and the percentage of wound closure values was calculated in Excel.

[0260] Western Blotting: To study FAK activation and downstream signaling pathways in adherent migrating cells, Caco-2 cells were sparsely (3500 cells / cm2) seeded to create islands of migrating cells on type-I collagen pre-coated 150 mm bacteriologic plastic dishes using ELISA coating buffer. At 50-60 % confluence, cells were treated with 100 nM M64HC1 or water vehicle control for two hours at 37 °C in 8% CO2. After two hours of incubation, the treated medium was removed, and the cells were similarly washed with PBS three times. Then, cells were incubated with a normal medium at 37 °C until harvesting at the indicated time points after the initiation of the two-hour M64HC1 treatment. Caco-2 cells were lysed with 80 pL of protein lysis buffer (50 mM Tris, 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1% Triton-X-100, 1% deoxycholic acid, 0.1% SDS, 10% glycerol, and protease and phosphatase inhibitors) to extract protein. Protein concentration was estimated by bicinchoninic acid assay (Thermo Fisher, Waltham, MA, USA). 40 pg of proteins (20 pL of protein lysate) were loaded per lane onto 10% SDS-PAGE gels for resolution and transferred onto nitrocellulose membranes as previously described. Membranes were blotted with antibody to Y-397-phosphorylated FAK (1 : 1000) and T-202 / Y-204-phosphorylated ERK1 / 2 (1 :2000).

[0261] Antibody to total FAK (1 : 1000) and total ERK (1 :2000) served as a loading control after stripping and reprobing. Western blots were performed, and images were detected by LICOR -Odyssey -Fc imaging system (LI-COR Biosciences, Lincon, NE, USA). Densitometry was conducted on exposures within the linear range.

[0262] Data for representative compounds is provided in the following table.

[0263]

[0264] Example 18. The following illustrate representative pharmaceutical dosage forms, containing a compound of formula (I) ('Compound X'), for therapeutic or prophylactic use in humans. (i) Tablet 1 mg / tablet

[0265] Compound X= 100.0

[0266] Lactose 77.5

[0267] Povidone 15.0

[0268] Croscarmellose sodium 12.0 Microcrystalline cellulose 92.5

[0269] Magnesium stearate 3,0

[0270] 300.0

[0271] (ii) Tablet 2 mg / tablet Compound X= 20.0

[0272] Microcrystalline cellulose 410.0

[0273] Starch 50.0

[0274] Sodium starch glycolate 15.0 Magnesium stearate 5,0

[0275] 500.0

[0276] (iii) Capsule mg / capsule

[0277] Compound X= 10.0

[0278] Colloidal silicon dioxide 1.5

[0279] Lactose 465.5

[0280] Pregelatinized starch 120.0

[0281] Magnesium stearate 3,0

[0282] 600.0

[0283] (iv) Injection 1 (1 mg / ml) mg / ml

[0284] Compound X= (free acid form) 1.0

[0285] Dibasic sodium phosphate 12.0

[0286] Monobasic sodium phosphate 0.7

[0287] Sodium chloride 4.5

[0288] 1.0 N Sodium hydroxide solution

[0289] (pH adjustment to 7.0-7.5) q.s.

[0290] Water for injection q.s. ad 1 mL

[0291] (v) Injection 2 (10 mg / ml) mg / ml

[0292] Compound X= (free acid form) 10.0

[0293] Monobasic sodium phosphate 0.3

[0294] Dibasic sodium phosphate 1.1

[0295] Polyethylene glycol 400 200.0

[0296] 1.0 N Sodium hydroxide solution (pH adjustment to 7.0-7.5) q.s.

[0297] Water for injection q.s. ad 1 mL (vi) Aerosol mg / can

[0298] Compound X= 20.0

[0299] Oleic acid 10.0

[0300] Trichloromonofluoromethane 5,000.0 Dichlorodifluoromethane 10,000.0

[0301] Di chlorotetrafluoroethane 5,000.0

[0302] The above formulations may be obtained by conventional procedures well known in the pharmaceutical art. All publications, patents, and patent documents are incorporated by reference herein, as though individually incorporated by reference. The invention has been described with reference to various specific and preferred embodiments and techniques. However, it should be understood that many variations and modifications may be made while remaining within the spirit and scope of the invention.

Claims

CLAIMSWhat is claimed is:

1. A compound of formula (I):or a salt thereof, wherein:X is O, NRa, or CRbRc;R1is (Ci-C3)alkyl;R2is H or (Ci-C3)alkyl;R3is a 6-membered heteroaryl;R5is OR4, CHF2, OCH3, or CF3R4is trifluoromethyl, methoxymethyl, or -(CH2CH2O)n-Rd;R6is H or (Ci-C3)alkyl;R7is H or (Ci-C3)alkyl;Rais H or (Ci-C3)alkyl;Rbis H, F, Cl, Br, or (Ci-C3)alkyl;Rcis H, F, Cl, Br, or (Ci-C3)alkyl; n is 0, 1, 2, 3, or 4; andRdis H, (Ci-C3)alkyl, or (Ci-C3)alkanoyl; provided the compound is not;CLAIMSWhat is claimed is:

1. A compound of formula (I):or a salt thereof, wherein:X is O, NRa, or CRbRc;R1is (Ci-C3)alkyl;R2is H or (Ci-C3)alkyl;R3is a 6-membered heteroaryl;R5is OR4, CHF2, OCH3, or CF3R4is trifluoromethyl, methoxymethyl, or -(CH2CH2O)n-Rd;R6is H or (Ci-C3)alkyl;R7is H or (Ci-C3)alkyl;Rais H or (Ci-C3)alkyl;Rbis H, F, Cl, Br, or (Ci-C3)alkyl;Rcis H, F, Cl, Br, or (Ci-C3)alkyl; n is 0, 1, 2, 3, or 4; andRdis H, (Ci-C3)alkyl, or (Ci-C3)alkanoyl; provided the compound is not;2 The compound of claim 1 which is a compound of formula (la):or a salt thereof, wherein:X is O, NRa, or CRbRc;R1is (Ci-C3)alkyl;R2is H or (Ci-C3)alkyl;R3is a 6-membered heteroaryl;R4is trifluoromethyl, methoxymethyl, or -(CH2CH2O)n-Rd;Rais H or (Ci-C3)alkyl;Rbis H, F, Cl, Br, or (Ci-C3)alkyl;Rcis H, F, Cl, Br, or (Ci-C3)alkyl; n is 0, 1, 2, 3, or 4; andRdis H, (Ci-C3)alkyl, or (Ci-C3)alkanoyl.3 The compound or salt of claim 1, wherein R5is OR4or CHF2.4 The compound or salt of claim 1, wherein R5is OR4.

5. The compound or salt of any one of claims 1-4, wherein R6and R7are each independently is H or CH3.

6. The compound or salt of any one of claims 1-4, wherein R6and R7are each H.7 The compound or salt of any one of claims 1-6, wherein X is NRa.8 The compound or salt of any one of claims 1-7, wherein Rais H.9 The compound or salt of any one of claims 1-7, wherein Rais (Ci-C3)alkyl.10 The compound or salt of any one of claims 1-6, wherein X is CRbRc.11 The compound or salt of any one of claims 1-6 and 10, wherein Rbis H.12 The compound or salt of any one of claims 1-6 and 10, wherein Rbis F, Cl, or Br.The compound or salt of any one of claims 1-6 and 10, wherein Rbis (Ci-C3)alkyl.14 The compound or salt of any one of claims 1-6 and 10-13, wherein Rcis H.15 The compound or salt of any one of claims 1-6 and 10-13, wherein Rcis F, Cl, or Br.16 The compound or salt of any one of claims 1-6 and 10-13, wherein Rcis (Ci-C3)alkyl.17 The compound or salt of any one of claims 1-16, wherein R1is methyl.18 The compound or salt of any one of claims 1-17, wherein R2is H.19 The compound or salt of any one of claims 1-17, wherein R2is methyl.

20. The compound or salt of any one of claims 1-19, wherein R3is a 6-membered heteroaryl that comprises one or two nitrogens.

21. The compound or salt of any one of claims 1-19, wherein R3is pyridyl.

22. The compound or salt of any one of claims 1-19, wherein R3is 3-pyridyl.

23. The compound or salt of any one of claims 1-19, wherein R3is 4-pyridyl.

24. The compound or salt of any one of claims 1-23, wherein R4is -(CH2CH2O)n-Rd25. The compound or salt of any one of claims 1-24, wherein n is 0.

26. The compound or salt of any one of claims 1-24, wherein n is 1.

27. The compound or salt of any one of claims 1-24, wherein n is 2.

28. The compound or salt of any one of claims 1-24, wherein n is 3.

29. The compound or salt of any one of claims 1-24, wherein n is 4.

30. The compound or salt of any one of claims 1-29, wherein Rdis H.

31. The compound or salt of any one of claims 1-29, wherein Rdis (Ci-C3)alkyl.

32. The compound or salt of any one of claims 1-23, wherein R4is trifluoromethyl or methoxymethyl.

33. The compound or salt of claim 1, which is selected from the group consisting of:68and salts thereof.

34. The compound or salt of claim 1, which is selected from the group consisting of:

35. A compound as described in any one of claims 1-33.

36. A salt as described in any one of claims 1-33.

37. The salt of claim 36, which is a pharmaceutically acceptable salt.

38. The salt of claim 36, which is an HC1 salt.

39. A pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38 and a pharmaceutically acceptable excipient.

40. A method for treating an epithelial disease or epithelial damage in an animal comprising, administering a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38 to the animal.

41. A compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38 for use in medical therapy.

42. A compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38 for the prophylactic or therapeutic treatment of an epithelial disease or epithelial damage.

43. Use of a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38 to prepare a medicament for treating an epithelial disease or epithelial damage in an animal.

44. The method of claim 40, the compound of claim 42, or the use of claim 43, wherein the epithelial disease or epithelial damage is a disease or damage of the skin, gastrointestinal tract, cornea, or bladder.

45. A method for activating a focal adhesion kinase comprising, contacting the kinase (in vitro or in vivo) with a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38.

46. A method for activating a focal adhesion kinase in an animal comprising, administering a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38 to the animal.

47. A compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38 for activating a focal adhesion kinase.

48. Use of a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38 to prepare a medicament for activating a focal adhesion kinase in an animal.

49. A method for promoting mucosal healing in an animal comprising administering a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38 to the animal.

50. A compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38 for promoting mucosal healing.

51. Use of a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38 to prepare a medicament for promoting mucosal healing in an animal.

52. A method for promoting epithelial healing in an animal comprising, administering a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38 to the animal.

53. A compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38 for promoting epithelial healing.

54. Use of a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38 to prepare a medicament for promoting epithelial healing in an animal.

55. A method of treating damage or disease of the bladder (e.g., epithelial disease or epithelial72damage of the bladder) in an animal, comprising administering a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38 or a compound of formula:or a pharmaceutically acceptable salt thereof, to the animal.

56. A compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38, or a compound of formula:or a pharmaceutically acceptable salt thereof, for treating damage or disease of the bladder (e.g., epithelial disease or epithelial damage of the bladder).

57. Use of a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38, or a compound of formula:or a pharmaceutically acceptable salt thereof, to prepare a medicament for treating damage or disease of the bladder (e.g., epithelial disease or epithelial damage of the bladder) in an animal.

58. A method of treating interstitial cystitis in an animal, comprising administering a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38, or a compound of formula:or a pharmaceutically acceptable salt thereof, to the animal.

59. A compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38, or a compound of formula:or a pharmaceutically acceptable salt thereof, for treating interstitial cystitis.

60. Use of a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38, or a compound of formula:or a pharmaceutically acceptable salt thereof, to prepare a medicament for treating interstitial cystitis in an animal.

61. A method of treating a transplant cornea, comprising contacting the transplant cornea with a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38, or a compound of formula:or a pharmaceutically acceptable salt thereof.

62. A compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38, or a compound of formula:or a pharmaceutically acceptable salt thereof, for treating a transplant cornea.7563. Use of a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-38, or a compound of formula:or a pharmaceutically acceptable salt thereof, to prepare a medicament for treating a transplant cornea.76

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