Compounds and compositions for treating fibrosis

Abstract

To provide methods for treating or preventing MKP-5 modulated disease or disorder in mammals.SOLUTION: A method comprises the step for administering to a mammal a therapeutically effective amount of a compound of formula (la) or (1b). Examples of the compound include 3,3-dimethyl-1-((9-(methylthio)-5,6-dihydrothieno[3,4-h]quinazolin-2-yl)thio)butan-2-one.SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority under 35 USC § 119(e) to U.S. Provisional Application No. 62 / 607,110, filed December 18, 2017, which is incorporated by reference in its entirety.

[0002] STATEMENT REGARDING GOVERNMENT SPONSORED RESEARCH OR DEVELOPMENT This invention was made with Government support under AR066003 awarded by the National Institutes of Health. The Government has certain rights in this invention. [Background technology]

[0003] 2. Background of the Invention Accumulation of extracellular matrix components in response to tissue injury is a physiological process essential for tissue repair. Unfortunately, chronic injury exacerbating the injury promotes overproduction of extracellular matrix components by fibroblasts and myofibroblasts, thus causing excessive accumulation of fibrous connective tissue, a pathological condition known as fibrosis. Regardless of the etiology and intricate progressive complexity of many chronic diseases, fibrosis represents a common end stage of tissue death. The destructive sequelae of tissue fibrosis lead to organ dysfunction and ultimately failure. Thus, fibrosis is the single most common reason for organ transplantation.

[0004] Fibrosis can affect various organs such as the heart, liver, lungs, skeletal muscle, kidneys, vasculature and heart. Fibrosis is the final stage of several chronic tissue diseases that account for almost 50% of deaths worldwide: skeletal muscle tissue (dystrophic myopathies), cardiac and vascular tissue (myocardial infarction), liver tissue (non-alcoholic fatty liver disease / cirrhosis), lung tissue (idiopathic pulmonary fibrosis) and kidney tissue (chronic kidney disease / renal fibrosis).

[0005] For example, in dystrophic muscle diseases (such as Duchenne muscular dystrophy), dystrophic muscles gradually become fibrotic, reducing the patient's ability to walk and endangering the patient's respiratory and cardiovascular functions. There is no treatment or cure for dystrophic muscle diseases, and experimental gene replacement therapy has been unsuccessful.

[0006] In another example, idiopathic pulmonary fibrosis (commonly referred to as IDF or IPF) is a chronic fibrotic parenchymal lung disease of unknown origin that occurs in response to chronic lung injury. Thickening and scarring of lung tissue reduces the patient's ability to breathe. IDF is more common and deadly than many cancers, with a prevalence of 20-50 cases / 100,000. The disease is generally diagnosed in subjects aged 55 years or older, with a median survival of 3-4 years. There are approximately 200,000 patients in the United States at any given time, of which 30,000 die each year.

[0007] Despite increasing recognition that tissue fibrosis contributes to significant morbidity and mortality in many diseases, there are few FDA-approved treatments available for fibrosis.

[0008] Mitogen-activated protein (MAP) kinases (also known as MAPKs) are protein kinases specific for the amino acids serine, threonine, and tyrosine. MAPKs are involved in inducing cellular responses to a variety of stimuli, including mitogens, osmotic stress, heat shock, and proinflammatory cytokines. MAPKs regulate cellular functions, including proliferation, gene expression, differentiation, mitosis, cell survival, and apoptosis.

[0009] On the other hand, MAP kinase phosphatases (MKPs) dephosphorylate MAP kinases. Specifically, MKP-5 dephosphorylates p38 MAPK and JNK. MAP kinases have been implicated in the progression of tissue fibrosis. Based on multiple mouse models in which injurious tissue repair progresses to end-stage fibrosis, MKP-5 has been shown to be a key positive regulator of tissue fibrosis. Genetic inhibition of MKP-5 ameliorates fibrosis in lung and skeletal muscle and reduces early events occurring in vascular fibrosis and atherogenesis. At a mechanistic level, MKP-5-deficient cells are impaired in their ability to activate the TGF-β signaling pathway, which plays an established role in promoting fibrosis. These combined observations suggest MKP-5 as a target for tissue fibrosis therapy.

[0010] There is a need in the art for new compounds and compositions that can be used to treat and / or prevent fibrotic diseases. The present disclosure addresses this need. Summary of the Invention

[0011] BRIEF SUMMARY OF THE INVETION In one aspect, the present invention provides a method of treating or preventing an MKP-5 regulated disease or disorder in a mammal in need thereof. In various embodiments, the method comprises administering to the mammal a therapeutically effective amount of a compound of formula (1a) or (1b): TIFF0007676477000001.tif39128Where: Z is NR, NC(=O)R, CH 2 and O; TIFF0007676477000002.tif6128 is single; or Z is N, and TIFF0007676477000003.tif6128 is a duplicate;R 1 is C 1 -C 6 each occurrence of R is independently selected from the group consisting of H, C 1 -C 6A is selected from the group consisting of optionally substituted phenyl and optionally substituted heteroaryl.

[0012] In various embodiments, the compound is of formula (2): TIFF0007676477000004.tif35128Where: Z is CH 2 and O; R 1 is C 1 -C 6 is selected from the group consisting of alkyl, optionally substituted phenyl, and NRR, where each occurrence of R is independently selected from H, C 1 -C 3 R is selected from the group consisting of alkyl, and substituted or unsubstituted phenyl; 3 , H, C 1 -C 6 Alkyl, and C 1 -C 3 thioethers; and R 4 H and COOR 5 wherein R 5 H and C 1 -C 6 alkyl.

[0013] In various embodiments, the aryl or heteroaryl is 1 -C 6 Alkyl, C 1 -C 6 Thioalkyl, C(=O)OH, C(=O)OC 1 -C 6 Substituted with at least one substituent selected from the group consisting of alkyl, cyano, and halo.

[0014] In various embodiments, the compounds have a Ki for MKP-5 of ≦100 μM.

[0015] In various embodiments, the compound binds to an allosteric site of MKP-5.

[0016] In various embodiments, the compound is 3,3-dimethyl-1-((9-(methylthio)-5,6-dihydrothieno[3,4-h]quinazolin-2-yl)thio)butan-2-one; 1-((5,6-dihydrobenzo[h]quinazolin-2-yl)thio)-3,3-dimethylbutan-2-one; N,N-dimethyl-2-((9-(methylthio)-5,6-dihydrothieno[3,4-h ]quinazolin-2-yl)thio)acetamide;N-ethyl-N-methyl-2-((9-(methylthio)-5,6-dihydrothieno[3,4-h]quinazolin-2-yl)thio)acetamide;1-((9-fluoro-5,6-dihydrobenzo[h]quinazolin-2-yl)thio)-3,3-dimethylbutan-2-one;1-((6H-isochromeno[4,3-d]pyrimidine- 2-yl)thio)-3,3-dimethylbutan-2-one;1-(2,4-dichlorophenyl)-2-((4-(3,4-dimethylthieno[2,3-b]thiophen-2-yl)pyrimidin-2-yl)thio)ethan-1-one;1-((5,6-dihydrothieno[2,3-h]quinazolin-2-yl)thio)-3,3-dimethylbutan-2-one;3,3-Dimethyl-1-(pyrimidin-2-yl)thio) and 1-((6-acetyl-5,6-dihydropyrimido[5,4-c]quinolin-2-yl)thio)-3,3-dimethylbutan-2-one; and 3,3-dimethyl-1-((9-propyl-5,6-dihydrothieno[3,4-h]quinazolin-2-yl)thio)butan-2-one.

[0017] In various embodiments, the compound is selected from the group consisting of YU032149, HJ830, HJ845, HJ846, HJ858, and HJ862.

[0018] In various embodiments, the compound is administered as part of a pharmaceutical composition that further comprises at least one pharma- ceutically acceptable carrier.

[0019] In various embodiments, the MKP-5 modulated disease or disorder is a fibrotic disease or disorder.

[0020] In various embodiments, the MKP-5 modulated disease or disorder is selected from the group consisting of dystrophic muscle diseases, cardiac or vascular diseases, idiopathic pulmonary fibrosis, and any combination thereof.

[0021] In various embodiments, the mammal is a human.

[0022] In various embodiments, the MKP-5 inhibitor is administered to the mammal by at least one route selected from the group consisting of nasal, inhalation, topical, oral, buccal, rectal, pleural, peritoneal, vaginal, intramuscular, subcutaneous, transdermal, epidural, intratracheal, otic, intraocular, intrathecal, and intravenous routes.

[0023] In various embodiments, the method further comprises administering to the mammal at least one additional agent that treats or prevents an MKP-5 modulated disease or disorder in the mammal.

[0024] In various embodiments, the inhibitor and at least one additional agent are co-formulated.

[0025] In another aspect, the present invention provides a compound selected from the group consisting of: TIFF0007676477000005.tif114142.

[0026] In another aspect, the invention provides a kit for preventing or treating an MKP-5 regulated disease or disorder in a mammal, comprising an MKP-5 inhibitor, optionally an applicator, and instructions for its use, the instructions describing the amount of the MKP-5 inhibitor to be administered to a mammal to treat or prevent the MKP-5 regulated disease or disorder, and the frequency of administration.

[0027] In yet another aspect, the present invention provides a method for determining whether a test compound is an MKP-5 inhibitor. In certain embodiments, the method comprises contacting the test compound with (i) a substrate peptide comprising the amino acid sequence pThr-Gly-pTyr, and (ii) a catalytic polypeptide comprising and / or consisting of the catalytic domain of MKP-5 or an active fragment thereof, thus forming a composition. In other embodiments, the method comprises measuring MKP-5 activity in the composition. In yet other embodiments, the method comprises comparing MKP-5 activity in the composition to a control. In yet other embodiments, the substrate peptide comprises the amino acid sequence In yet other embodiments, the step of measuring the MKP-5 activity comprises measuring any change in inorganic free phosphate in the composition. [The present invention 1001] 1. A method of treating or preventing an MKP-5 regulated disease or disorder in a mammal in need thereof, comprising: Compounds of formula (1a) or (1b): TIFF0007676477000007.tif39128 (in the formula: Z is NR, NC(=O)R, CH 2 and O; TIFF0007676477000008.tif6128 is single; or Z is N, and TIFF0007676477000009.tif6128 is a duplicate; R 1 is C 1 -C 6 selected from the group consisting of alkyl, optionally substituted phenyl, and NRR; Each occurrence of R is independently H, C 1 -C 6 selected from the group consisting of alkyl, and optionally substituted phenyl; A is selected from the group consisting of optionally substituted phenyl and optionally substituted heteroaryl. Administering to a mammal a therapeutically effective amount of [The present invention 1002] The compound is a compound of formula (2): TIFF0007676477000010.tif35128 (in the formula: Z is CH 2 and O; R 1 is C 1 -C 6 is selected from the group consisting of alkyl, optionally substituted phenyl, and NRR, where each occurrence of R is independently selected from H, C 1 -C 3 selected from the group consisting of alkyl, and substituted or unsubstituted phenyl; R 3 , H, C 1 -C 6 Alkyl, and C 1 -C 3 thioethers; and R 4 H and COOR 5 wherein R 5 H and C 1 -C 6 alkyl) The method of the present invention 1001. [The present invention 1003] Aryl or heteroaryl is C 1 -C 6 Alkyl, C 1 -C 6 Thioalkyl, C(=O)OH, C(=O)OC 1 -C 6 The method of any one of claims 1001 to 1002, wherein said compound is substituted with at least one substituent selected from the group consisting of alkyl, cyano, and halo. [The present invention 1004] The method of any one of claims 1001 to 1002, wherein said compound has a Ki for MKP-5 of ≦100 μM. [The present invention 1005] The method of any one of claims 1001 to 1002, wherein said compound binds to an allosteric site of MKP-5. [The present invention 1006] The compound is 3,3-dimethyl-1-((9-(methylthio)-5,6-dihydrothieno[3,4-h]quinazolin-2-yl)thio)butan-2-one; 1-((5,6-dihydrobenzo[h]quinazolin-2-yl)thio)-3,3-dimethylbutan-2-one; N,N-dimethyl-2-((9-(methylthio)-5,6-dihydrothieno[3,4-h]quinazolin-2-yl )thio)acetamide;N-ethyl-N-methyl-2-((9-(methylthio)-5,6-dihydrothieno[3,4-h]quinazolin-2-yl)thio)acetamide;1-((9-fluoro-5,6-dihydrobenzo[h]quinazolin-2-yl)thio)-3,3-dimethylbutan-2-one;1-((6H-isochromeno[4,3-d]pyrimidin-2-yl)thio)-3,3- Dimethylbutan-2-one;1-(2,4-dichlorophenyl)-2-((4-(3,4-dimethylthieno[2,3-b]thiophen-2-yl)pyrimidin-2-yl)thio)ethan-1-one;1-((5,6-dihydrothieno[2,3-h]quinazolin-2-yl)thio)-3,3-dimethylbutan-2-one;3,3-Dimethyl-1-(pyrimido[5,4-c]quinoline-2 1-((6-acetyl-5,6-dihydropyrimido[5,4-c]quinolin-2-yl)thio)-3,3-dimethylbutan-2-one; and 3,3-dimethyl-1-((9-propyl-5,6-dihydrothieno[3,4-h]quinazolin-2-yl)thio)butan-2-one. [The present invention 1007] The method of any one of claims 1001 to 1002, wherein said compound is selected from the group consisting of YU032149, HJ830, HJ845, HJ846, HJ858, and HJ862. [The present invention 1008] The method of any one of claims 1001 to 1002, wherein said compound is administered as part of a pharmaceutical composition further comprising at least one pharma- ceutically acceptable carrier. [The present invention 1009] The method of any of claims 1001 to 1008, wherein the MKP-5 regulated disease or disorder is a fibrotic disease or disorder. [The present invention 1010] The method of claim 1009, wherein the MKP-5 regulated disease or disorder is selected from the group consisting of dystrophic muscle disease, cardiac or vascular disease, idiopathic pulmonary fibrosis, and any combination thereof. [The present invention 1011] The method according to any one of claims 1001 to 1010, wherein the mammal is a human. [The present invention 1012] The method of any of claims 1001 to 1011, wherein the MKP-5 inhibitor is administered to the mammal by at least one route selected from the group consisting of nasal, inhalation, topical, oral, buccal, rectal, pleural, peritoneal, vaginal, intramuscular, subcutaneous, transdermal, epidural, intratracheal, aural, intraocular, intrathecal, and intravenous routes. [The present invention 1013] The method of any of claims 1001 to 1012, further comprising the step of administering to the mammal at least one additional agent that treats or prevents an MKP-5 regulated disease or disorder in the mammal. [The present invention 1014] The method of claim 1013, wherein the inhibitor and the at least one additional agent are co-formulated. [The present invention 1015] A compound selected from the group consisting of: TIFF0007676477000011.tif114142. [The present invention 1016] A kit for preventing or treating an MKP-5 regulated disease or disorder in a mammal, comprising an MKP-5 inhibitor, optionally an applicator, and instructions for its use, the instructions describing the amount and frequency of administration of the MKP-5 inhibitor to be administered to the mammal to treat or prevent the MKP-5 regulated disease or disorder. [The present invention 1017] 1. A method for determining whether a test compound is an MKP-5 inhibitor, comprising: A test compound; (i) a substrate peptide comprising the amino acid sequence pThr-Gly-pTyr, and (ii) a catalytic polypeptide comprising and / or consisting of the catalytic domain of MKP-5 or an active fragment thereof; and thus forming a composition; measuring the MKP-5 activity in the composition; and Comparing the MKP-5 activity in the composition to a control. Including, Thereby, determining whether the test compound is an MKP-5 inhibitor. [The present invention 1018] The substrate peptide has the amino acid sequence The method of the present invention 1017, including TIFF0007676477000012.tif9163. [The present invention 1019] The method of claim 1017, wherein the step of measuring the MKP-5 activity comprises measuring any change in inorganic free phosphate in the composition. [Brief description of the drawings]

[0028] The following detailed description of exemplary embodiments of the invention will be better understood when read in conjunction with the accompanying drawings. For the purpose of illustrating the invention, certain exemplary embodiments are shown in the drawings. It should be understood, however, that the invention is not limited to the precise arrangements and instrumentalities of the embodiments shown in the drawings.

[0029] [Figure 1] FIG. 1 shows an exemplary role of MKP-5 in tissue fibrosis. [Figure 2A] 1 is a graph showing the dose response of MKP-5 to exemplary compounds of the invention. [Figure 2B] 1 is a graph showing the dose response of MKP-5 to an exemplary compound of the invention (YU032149). [Figure 2C] 1 is a graph showing the Kd determination of YU032149 with MKP-5. [Diagram 3]1 is a ribbon diagram based on the crystal structure of MKP-5 with bound ligand and inhibitor. [Figure 4A] Binding of YU032149 to MKP-5 is shown with residues that are constant. [Figure 4B] YU032149 relative to the allosteric site is shown with certain contacts between the inhibitor and the protein. [Diagram 5] Binding of YU032149 to the allosteric site of MKP-5 is shown, along with displacement of selected active site residues in response to binding. [Figure 6] FIG. 1 is a bar graph showing normalized turnover for selected analogs of YU032149 (50 μM inhibitor in each test). [Figure 7A] 1 is a series of bar graphs showing MAPK activity assays of selected compounds. [Figure 7B] 1 is a series of bar graphs showing MAPK activity assays of selected compounds. [Figure 7C] 1 is a series of bar graphs showing MAPK activity assays of selected compounds. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0030] Detailed Description of the Invention The present invention relates in part to the identification of novel MKP-5 inhibitors for use in the treatment and / or prevention of fibrosis. The present invention provides novel inhibitors of MKP-5, as well as compositions comprising the same, and their use in the treatment and / or prevention of fibrosis. For example, compound YU032149 has an IC of about 12 μM. 50 In certain embodiments, the compounds of the present invention inhibit MKP-5 through inhibition of an allosteric mode.

[0031] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. Although any method and material similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred method and material are described. Each of the following terms used herein has the meaning associated with it in this section.

[0032] Generally, the nomenclature and laboratory procedures in cell culture, molecular genetics, pharmacology and organic chemistry used herein are those well known and commonly used in the art.

[0033] Standard techniques are used for biochemical and / or biological manipulations. Techniques and procedures are generally carried out according to conventional methods in the art and various general references provided throughout this document (e.g., Sambrook and Russell, 2012, Molecular Cloning, A Laboratory Approach, Cold Spring Harbor Press, Cold Spring Harbor, NY, and Ausubel et al., 2002, Current Protocols in Molecular Biology, John Wiley & Sons, NY).

[0034] The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0035] As used herein, the term "about" when referring to a measurable value, such as an amount, duration, and the like, is meant to encompass variations of ±20% or ±10%, more preferably ±5%, even more preferably ±1%, and even more preferably ±0.1% from the specified value, as such variations are appropriate in the practice of the disclosed methods.

[0036] A disease or disorder is "alleviated" if the severity or frequency of at least one sign or symptom of the disease or disorder experienced by a patient is reduced.

[0037] As used herein, the terms "analog," "analogue," or "derivative" refer to a chemical compound or molecule made from a parent compound or molecule by one or more chemical reactions. Thus, an analog may be a structure having a similar structure to the small molecule inhibitors described herein, or may be based on the scaffold of the small molecule inhibitors described herein, but may differ therefrom with respect to certain components or structural components, and may have a similar or opposite metabolic action.

[0038] The term "binding" as used herein refers to the attachment of molecules to one another, such as, but not limited to, enzymes and substrates, antibodies and antigens, DNA strands and their complementary strands, etc. Binding occurs due to complementary shapes and chemical properties of portions of the molecular surfaces. A common metaphor is the "lock and key" used to describe how enzymes fit around their substrates.

[0039] The term "conservative variation" or "conservative substitution" as used herein refers to the replacement of an amino acid residue with another biologically similar residue. Conservative variation or substitution is unlikely to change the shape of the peptide chain. Examples of conservative variation or substitution include the replacement of one hydrophobic residue, such as isoleucine, valine, leucine or methionine, with another, or the replacement of one polar residue with another, such as the replacement of lysine with arginine, the replacement of aspartic acid with glutamic acid, or the replacement of asparagine with glutamine. Further examples include exchanges within groups, such as Gly / Ala; Val / Ile / Leu; Asp / Glu; Asn / Gln; Ser / Thr; Lys / Arg; and Phe / Tyr.

[0040] A "disease" is a state in the health of an animal in which the animal is unable to maintain homeostasis and where the animal's health continues to deteriorate if the disease is not ameliorated.

[0041] In contrast, a "disorder" in an animal is a health state in which the animal is able to maintain homeostasis, but in which the animal's health state is less favorable than it would be in the absence of the disorder. If left untreated, the disorder does not necessarily cause a further deterioration in the animal's health state.

[0042] An "effective amount" or "therapeutically effective amount" of a compound is an amount of the compound sufficient to provide a beneficial effect to the subject to which the compound is administered. An "effective amount" of a delivery vehicle is an amount sufficient to effectively bind or deliver the compound.

[0043] The term "inhibit" as used herein means to measurably reduce or completely prevent the expression, stability, function or activity of a molecule, reaction, interaction, gene, mRNA, and / or protein. Inhibitors are compounds, e.g., antagonists, that bind to, partially or completely block, reduce, prevent, delay activation, inactivate, desensitize, or downregulate protein, gene, and mRNA stability, expression, function, and activity.

[0044] The term "MKP-5" or "MKP5" or "DUSP10" as used herein refers to a protein known as dual specificity protein phosphatase 10 or mitogen-activated protein kinase phosphatase 5 (see, e.g., Tao & Tong, 2007, Protein Sci. 16(5):880-886, which is incorporated herein by reference in its entirety). This protein phosphatase is involved in the inactivation of MAP kinases and has specificity for the MAPK11-MAPK12-MAPK13-MAPK14 subfamily. In certain embodiments, MKP-5 dephosphorylates p38. Human MKP-5 corresponds to the polypeptide of the amino acid sequence of SEQ ID NO:4. In certain embodiments, the MAP kinase binding domain of human MKP-5 corresponds to residues 139-287 of SEQ ID NO:4, and the catalytic domain of human MKP-5 corresponds to residues 305-482 of SEQ ID NO:4.

[0045] SEQ ID NO:4 is provided herein. TIFF0007676477000013.tif96138

[0046] As used herein, the phrase "MKP-5 inhibitor" or "inhibitor of MKP-5" refers to a composition or compound that at least partially inhibits, either directly or indirectly, compared to a control system lacking inhibitor, MKP-5 activity, MKP-5 expression, and / or both, using any method known to one of skill in the art. MKP-5 inhibitors can be any type of compound, including, but not limited to, a nucleic acid, a peptide, an antibody, a small molecule, an antagonist, an aptamer, or a peptidomimetic.

[0047] As used herein, "MKP-5 regulated disease" or "MKP-5 dysregulation" refers to a disease associated with the pathological accumulation of excess extracellular matrix protein in an organ or tissue. Non-limiting examples of such diseases include, but are not limited to, cystic fibrosis or idiopathic pulmonary fibrosis.

[0048] "Natural" as applied to an object refers to the fact that the object can be found in nature. For example, a polypeptide or polynucleotide sequence present in an organism (including viruses) that can be isolated from a source in nature and has not been intentionally modified by humans is a naturally occurring sequence.

[0049] The terms "patient," "subject," "individual," and the like are used interchangeably herein and refer to any animal, or cells thereof, amenable to the methods described herein, whether in vitro or in situ. In certain non-limiting alternative embodiments, the patient, subject, or individual is a human.

[0050] The term "pharmaceutically acceptable carrier" as used herein means a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, stabilizer, dispersant, suspending agent, diluent, excipient, thickener, solvent or encapsulating material, involved in carrying or transporting a compound useful within the scope of the present invention into or to a patient so that it can perform its intended function. Typically, such a construct is carried or transported from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense of containing a compound useful within the scope of the present invention and not harmful to the patient, and being compatible with the other components of the formulation. Some examples of materials which can serve as pharma- ceutically acceptable carriers include: sugars such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository wax; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffers, such as magnesium hydroxide and aluminum hydroxide; surfactants; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances used in pharmaceutical preparations. As used herein, "pharmaceutically acceptable carriers" include any and all coatings, antibacterial and antifungal agents, absorption delaying agents, and the like, that are compatible with the activity of the compounds useful within the scope of the present invention and are physiologically acceptable to the patient. Supplementary active compounds may also be incorporated into the compositions. "Pharmaceutically acceptable carriers" may further include pharmaceutically acceptable salts of the compounds useful within the scope of the present invention.Other additional components that may be included in pharmaceutical compositions used in the practice of the invention are known in the art and described, for example, in Remington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, Pa.), which is incorporated herein by reference.

[0051] As used herein, the phrase "pharmaceutically acceptable salts" or "therapeutically acceptable salts" refers to salts of the compound to be administered prepared from pharma- ceutically acceptable non-toxic acids, including inorganic acids or bases, organic acids or bases, solvates, hydrates, or clathrates thereof.

[0052] The terms "pharmaceutical effective amount" and "effective amount" refer to a non-toxic but sufficient amount of an agent to provide a desired biological result. The result may be reduction and / or alleviation of the signs, symptoms, or causes of a disease or disorder, or any other desired change in a biological system. The appropriate effective amount in any individual case may be determined by one of ordinary skill in the art using routine experimentation.

[0053] As used herein, the terms "polypeptide," "protein," and "peptide" refer to polymers of amino acid residues linked through peptide bonds, related naturally occurring structural variants, and synthetic non-natural analogs thereof. Synthetic polypeptides can be synthesized, for example, using an automated polypeptide synthesizer.

[0054] As used herein, the term "specific binding" refers to a molecule, such as an antibody, that recognizes and binds to another molecule or feature in a sample, but does not substantially recognize or bind other molecules or features.

[0055] A "therapeutic" treatment is a treatment administered to a subject who exhibits symptoms of a pathology in order to reduce or eliminate those symptoms.

[0056] The term "therapeutically effective amount" as used herein is an amount of a compound of the present invention that, when administered to a patient, improves the symptoms of a disease or disorder. The amount of a compound of the present invention that constitutes a "therapeutically effective amount" will vary depending on the compound, the disease state and its severity, the age of the patient to be treated, etc. The therapeutically effective amount can be determined routinely by one of ordinary skill in the art having regard to his or her own knowledge and this disclosure.

[0057] As used herein, "treating a disease or disorder" means reducing the frequency with which a patient experiences symptoms of a disease or disorder. Disease and disorder are used interchangeably herein.

[0058] The term "treatment" or "treating" as used herein includes prevention and / or therapy. Thus, the compositions and methods of the present invention are not limited to therapeutic applications, but can also be used prophylactically. Thus, "treating" a condition, disorder or condition or their "treatment" includes: (i) preventing or delaying the appearance of clinical symptoms of a developing condition, disorder or condition in a subject who may be affected by or susceptible to a condition, disorder or condition, but who has not yet experienced or shown clinical or subclinical symptoms of the condition, disorder or condition; (ii) inhibiting a condition, disorder or condition, i.e., stopping or reducing the onset of the disease or at least one clinical or subclinical symptom thereof; or (iii) relieving the disease, i.e., causing the regression of the condition, disorder or condition, or at least one clinical or subclinical symptom thereof.

[0059] The term "wild-type" as used herein refers to the genotype and phenotype that occurs in nature and is characteristic of most members of a species, as opposed to a mutant genotype and phenotype.

[0060] As used herein, the term “alkyl,” by itself or as part of another substituent, means, unless otherwise stated, an alkyl group having the indicated number of carbon atoms (i.e., C 1 -C10 means a straight or branched chain hydrocarbon having from 1 to 10 carbon atoms, including straight, branched, or cyclic substituents. Examples include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, neopentyl, hexyl, and cyclopropylmethyl. Certain specific examples include (C 1 -C 6 ) Alkyl, including but not limited to, ethyl, methyl, isopropyl, isobutyl, n-pentyl, n-hexyl, and cyclopropylmethyl.

[0061] As used herein, the term “cycloalkyl,” by itself or as part of another substituent, means, unless otherwise stated, a cycloalkyl group having the indicated number of carbon atoms (i.e., C 3 -C 6 means a cyclic chain hydrocarbon having a ring group containing 3 to 6 carbon atoms, including straight chain, branched chain, or cyclic substituents. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Certain specific examples include (C 3 -C 6 ) Cycloalkyl, including but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0062] As used herein, the term "substituted alkyl" or "substituted cycloalkyl" refers to any group including halogen, -OH, alkoxy, tetrahydro-2-H-pyranyl, -NH 2 , -N(CH 3 ) 2 , (1-methyl-imidazol-2-yl), pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, -C(=O)OH, trifluoromethyl, -C≡N, -C(=O)O(C 1 -C 4 ) alkyl, -C(=O)NH 2 , -C(=O)NH(C 1 -C 4 ) alkyl, -C(=O)N((C 1 -C4 )Alkyl) 2 , -SO 2 NH 2 , -C(=NH)NH 2 , and -NO 2 and is substituted with 1, 2 or 3 substituents selected from the group consisting of halogen, -OH, alkoxy, -NH 2 , trifluoromethyl, -N(CH 3 ) 2 and -C(=O)OH, more conveniently containing one or two substituents selected from halogen, alkoxy and -OH. Examples of substituted alkyl include, but are not limited to, 2,2-difluoropropyl, 2-carboxycyclopentyl and 3-chloropropyl.

[0063] As used herein, the term "alkoxy", used alone or in combination with other terms, unless otherwise specified, refers to an alkyl group having the indicated number of carbon atoms as defined above, linked to the remainder of the molecule via an oxygen atom, such as, for example, methoxy, ethoxy, 1-propoxy, 2-propoxy (isopropoxy) and higher homologs and isomers. In certain embodiments, alkoxy includes alkoxy groups such as, but not limited to, ethoxy and methoxy (C 1 -C 3 ) alkoxy.

[0064] The terms "halo" or "halogen" as used herein, alone or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom, conveniently fluorine, chlorine, or bromine, more conveniently fluorine or chlorine.

[0065] The term "heteroalkyl" as used herein, by itself or in combination with another term, means, unless otherwise specified, a stable straight or branched chain alkyl group consisting of the indicated number of carbon atoms and one or two heteroatoms selected from the group consisting of O, N, and S, where the nitrogen and sulfur atoms may be optionally oxidized and the nitrogen heteroatom may be optionally quaternized. The heteroatom may be located at any position of the heteroalkyl group, including between the remainder of the heteroalkyl group and the fragment to which it is attached, as well as attached to the most distal carbon atom of the heteroalkyl group. Examples include: -O-CH 2 -CH 2 -CH 3 , -CH 2 -CH 2 -CH 2 -OH, -CH 2 -CH 2 -NH-CH 3 , -CH 2 -S-CH 2 -CH 3 , and -CH 2 CH 2 -S(=O)-CH 3 For example, -CH 2 -NH-OCH 3 , or -CH 2 -CH 2 -SS-CH 3 Up to two heteroatoms may be consecutive, such as:

[0066] The term "aromatic" as used herein refers to a carbocyclic or heterocyclic ring having one or more polyunsaturated rings and having aromatic character, i.e., having (4n+2) delocalized π (pi) electrons, where n is an integer.

[0067] As used herein, the term "aryl", used alone or in combination with other terms, unless otherwise specified, refers to a carbocyclic aromatic system containing one or more rings (typically 1, 2 or 3 rings), where such rings may be linked together in a pendant manner, such as biphenyl, or may be fused, such as naphthalene. Examples include phenyl, anthracyl, and naphthyl. In certain embodiments, aryl includes phenyl and naphthyl, especially phenyl.

[0068] The term "heterocycle" or "heterocyclyl" or "heterocyclic" as used herein, by itself or in combination with another term, means, unless otherwise specified, a stable monocyclic or polycyclic heterocyclic ring system, unsubstituted or substituted, consisting of carbon atoms and at least one heteroatom selected from the group consisting of N, O, and S, where the nitrogen and sulfur heteroatoms are optionally oxidized, and the nitrogen atom is optionally quaternized. The heterocyclic ring system may be linked at any heteroatom or carbon atom that provides a stable structure, unless otherwise specified. The heterocycle may be aromatic or non-aromatic in nature. In certain embodiments, the heterocycle is a heteroaryl.

[0069] The term "heteroaryl" or "heteroaromatic" as used herein refers to a heterocycle having aromatic character. Polycyclic heteroaryls may contain one or more rings that are partially saturated. Examples include tetrahydroquinoline and 2,3-dihydrobenzofuryl.

[0070] Examples of non-aromatic heterocycles include monocyclic groups such as aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, pyrroline, imidazoline, pyrazolidine, dioxolane, sulfolane, 2,3-dihydrofuran, 2,5-dihydrofuran, tetrahydrofuran, thiophane, piperidine, 1,2,3,6-tetrahydropyridine, 1,4-dihydropyridine, piperazine, morpholine, thiomorpholine, pyran, 2,3-dihydropyran, tetrahydropyran, 1,4-dioxane, 1,3-dioxane, homopiperazine, homopiperidine, 1,3-dioxepane, 4,7-dihydro-1,3-dioxepine, and hexamethylene oxide.

[0071] Examples of heteroaryl groups include pyridyl, pyrazinyl, pyrimidinyl (such as, but not limited to, 2- and 4-pyrimidinyl), pyridazinyl, thienyl, furyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,3,4-triazolyl, tetrazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl, 1,3,4-thiadiazolyl, and 1,3,4-oxadiazolyl.

[0072] Examples of polycyclic heterocycles include indolyl (such as, but not limited to, 3-, 4-, 5-, 6-, and 7-indolyl), indolinyl, quinolyl, tetrahydroquinolyl, isoquinolyl (such as, but not limited to, 1- and 5-isoquinolyl), 1,2,3,4-tetrahydroisoquinolyl, cinnolinyl, quinoxalinyl (such as, but not limited to, 2- and 5-quinoxalinyl), quinazolinyl, phthalazinyl, 1,8-naphthyridinyl, 1,4-benzodioxanyl, coumarin, dihydrocoumarin, 1,5-naphthyridinyl, benzofuryl (such as, but not limited to, Examples of benzothienyl include, but are not limited to, 3-, 4-, 5-, 6-, and 7-benzofuryl, 2,3-dihydrobenzofuryl, 1,2-benzisoxazolyl, benzothienyl (such as, but not limited to, 3-, 4-, 5-, 6-, and 7-benzothienyl), benzoxazolyl, benzothiazolyl (such as, but not limited to, 2-benzothiazolyl and 5-benzothiazolyl), purinyl, benzimidazolyl, benzotriazolyl, thioxanthinyl, carbazolyl, carbolinyl, acridinyl, pyrrolidinyl, and quinolidinyl.

[0073] The foregoing lists of heterocyclyl and heteroaryl moieties are intended to be representative and not limiting.

[0074] The term "substituted" as used herein means that an atom or group of atoms replaces a hydrogen as a substituent linked to another group.

[0075] For aryl and heterocyclyl groups, the term "substituted" as applied to the rings of these groups refers to any level of substitution where such substitution is permitted, i.e., mono-, di-, tri-, tetra-, or penta-substitution. The substituents are independently selected and the substitution may be at any chemically feasible position. In certain embodiments, the number of substituents varies between 1 and 4. In other embodiments, the number of substituents varies between 1 and 3. In yet another embodiment, the number of substituents varies between 1 and 2. In yet another embodiment, the substituents are independently selected from C1-6 Alkyl, -OH, C 1-6 It is selected from the group consisting of alkoxy, halo, amino, acetamido and nitro. As used herein, when a substituent is an alkyl or alkoxy group, the carbon chain may be branched, straight chain or cyclic, in particular straight chain.

[0076] Ranges: Throughout this disclosure, various aspects of the invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity, and should not be construed as an inflexible limitation on the scope of the invention. Thus, the description of a range should be considered to have specifically disclosed all possible subranges as well as individual numerical values ​​within that range. For example, the description of a range such as 1-6 should be considered to have specifically disclosed subranges such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, and individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

[0077] method In certain embodiments, the present invention includes treating an MKP-5 regulated disease by administering an MKP-5 inhibitor. In certain embodiments, the MKP-5 regulated disease is any disease that can be treated or prevented by inhibiting MKP-5. In other embodiments, the present invention includes a method of treating a fibrotic disease by administering an MKP-5 inhibitor to a patient. In yet other embodiments, the MKP-5 regulated disease is a dystrophic muscle disease, a heart or vascular disease, idiopathic pulmonary fibrosis, and any combination thereof.

[0078] Compounds and Compositions In one aspect, the present invention provides a compound of formula (1a) or (1b): TIFF0007676477000014.tif31128 formula: Z is NR, NC(=O)R, CH 2 and O; TIFF0007676477000015.tif2128 The bond is single; or Z is N, and TIFF0007676477000016.tif2128 The bond is double; R 1 is C 1 -C 6 selected from the group consisting of alkyl, optionally substituted phenyl, and NRR; Each occurrence of R is independently H, C 1 -C 6 selected from the group consisting of alkyl, and optionally substituted phenyl; A is selected from the group consisting of optionally substituted phenyl and optionally substituted heteroaryl.

[0079] In certain embodiments, the aryl or heteroaryl is 1 -C 6 Alkyl, C 1 -C 6 Thioalkyl, C(=O)OH, C(=O)OC 1 -C 6 Substituted with at least one substituent selected from the group consisting of alkyl, cyano, and halo.

[0080] In another aspect, the present invention provides a compound of formula (2): TIFF0007676477000017.tif35128 formula: Z is CH 2 and O; R 1 is C 1 -C 6 selected from the group consisting of alkyl, optionally substituted phenyl, and NRR; Each occurrence of R is independently H, C 1 -C 3 selected from the group consisting of alkyl, and substituted or unsubstituted phenyl; R 3 , H, C 1 -C 6 Alkyl, and C1 -C 3 thioethers; and R 4 H and COOR 5 where R 5 H and C 1 -C 6 alkyl.

[0081] In certain embodiments, the compound is selected from the group consisting of: TIFF0007676477000018.tif155149.

[0082] In certain embodiments, the compound is 3,3-dimethyl-1-((9-(methylthio)-5,6-dihydrothieno[3,4-h]quinazolin-2-yl)thio)butan-2-one; 1-((5,6-dihydrobenzo[h]quinazolin-2-yl)thio)-3,3-dimethylbutan-2-one; N,N-dimethyl-2-((9-(methylthio)-5,6-dihydrothieno[3,4-h ]quinazolin-2-yl)thio)acetamide;N-ethyl-N-methyl-2-((9-(methylthio)-5,6-dihydrothieno[3,4-h]quinazolin-2-yl)thio)acetamide;1-((9-fluoro-5,6-dihydrobenzo[h]quinazolin-2-yl)thio)-3,3-dimethylbutan-2-one;1-((6H-isochromeno[4,3-d]pyrimidine- 2-yl)thio)-3,3-dimethylbutan-2-one;1-(2,4-dichlorophenyl)-2-((4-(3,4-dimethylthieno[2,3-b]thiophen-2-yl)pyrimidin-2-yl)thio)ethan-1-one;1-((5,6-dihydrothieno[2,3-h]quinazolin-2-yl)thio)-3,3-dimethylbutan-2-one;3,3-Dimethyl-1-(pyrimidin-2-yl)thio) and 1-((6-acetyl-5,6-dihydropyrimido[5,4-c]quinolin-2-yl)thio)-3,3-dimethylbutan-2-one; and 3,3-dimethyl-1-((9-propyl-5,6-dihydrothieno[3,4-h]quinazolin-2-yl)thio)butan-2-one.

[0083] In certain embodiments, the compound is TIFF0007676477000019.tif32128. In certain embodiments, the compound is TIFF0007676477000020.tif32128. In certain embodiments, the compound is TIFF0007676477000021.tif33128. In certain embodiments, the compound is TIFF0007676477000022.tif32128. In certain embodiments, the compound is TIFF0007676477000023.tif35128. In certain embodiments, the compound is TIFF0007676477000024.tif32128. In certain embodiments, the compound is The file is TIFF0007676477000025.tif33128. In certain embodiments, the compound is TIFF0007676477000026.tif39128. In certain embodiments, the compound is The file is TIFF0007676477000027.tif33128.

[0084] Combination therapy In certain embodiments, the compounds of the invention are useful in the methods of the invention in combination with at least one additional agent useful for treating or preventing an MKP-5 regulated disease in a mammal in need thereof, which may include compounds identified herein or compounds known to treat, prevent or reduce symptoms of an MKP-5 regulated disease in a subject, e.g., commercially available compounds.

[0085] In certain embodiments, the at least one additional compound useful for treating or preventing an MKP-5 regulated disease includes an acetylcholinesterase inhibitor, such as, but not limited to, nintedanib (methyl(3Z)-3-{[(4-{methyl[(4-methylpiperazin-1-yl)acetyl]amino}phenyl)amino](phenyl)methylidene}-2-oxo-2,3-dihydro-1H-indole-6-carboxylate) or pirfenidone (5-methyl-1-phenylpyridin-2-one).

[0086] Synergistic effects may be calculated using any suitable method, such as, for example, the sigmoid-Emax formula (Holford & Scheiner, 1981, Clin. Pharmacokinet. 6:429-453), the Loewe additivity formula (Loewe & Muischnek, 1926, Arch. Exp. Pathol Pharmacol. 114:313-326) and the median effect formula (Chou & Talalay, 1984, Adv. Enzyme Regul. 22:27-55). Each of the above formulas may be applied to experimental data to generate corresponding graphs that aid in evaluating the effect of drug combinations. The corresponding graphs associated with the above formulas are the concentration-effect curve, the isobologram curve and the combination coefficient curve, respectively.

[0087] Method for screening compounds as MKP-5 inhibitors In one aspect, the present invention provides a method for identifying a compound that inhibits MKP-5. In certain embodiments, the method includes contacting a putative inhibitor compound with (i) a substrate peptide comprising the sequence pThr-Gly-pTyr, and (ii) a catalytic polypeptide comprising and / or consisting of the catalytic domain of MKP-5 (such as, but not limited to, residues 305-482 of SEQ ID NO:4) or an active fragment thereof, thus forming a composition. In other embodiments, the method includes measuring MKP-5 activity in the composition. In yet other embodiments, the method includes comparing MKP-5 activity in the composition with a control; thereby identifying the putative inhibitor compound as a compound that inhibits MKP-5.

[0088] In certain embodiments, the catalytic polypeptide has at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and / or 100% homology to human MKP-5 in an amino acid sequence corresponding to the catalytic domain of MKP-5. Conservative substitutions are contemplated within such homology.

[0089] In certain embodiments, the catalytic polypeptide has at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and / or 100% sequence identity to human MKP-5 in an amino acid sequence corresponding to the catalytic domain of MKP-5.

[0090] In certain embodiments, the method is performed as a high throughput screen in which a plurality of compounds (putative inhibitors) are contacted with a peptide comprising pThr-Gly-pTyr and a polypeptide comprising and / or consisting of the catalytic domain of MKP-5 or an active fragment thereof, and inhibitors are identified from among the plurality of compounds by comparing their individual activities with a control.

[0091] Without wishing to be limited by theory, it is believed that inclusion of peptides containing the pThr180-Gly-pTyr182 motif of p38α MAPK (which is present on the activation loop of p38α MAPK and is the primary MKP-5 substrate) in the assay reveals more selective and therefore more useful inhibitor compounds. TIFF0007676477000028.tif4128, where Nle is norleucine.

[0092] Those skilled in the art will understand that activity can be measured by combining a substrate with a polypeptide comprising and / or consisting of MKP-5 or any catalytically active fragment thereof, i.e., a polypeptide comprising and / or consisting of the MKP-5 catalytic domain or an active fragment thereof. Those skilled in the art will understand that various methods of measuring MKP-5 activity and controls are possible and will be familiar with them by analogy with activity assays. The control can be, as a non-limiting example, a predetermined standard or a reaction performed without an MKP-5 inhibitor and / or without a peptide comprising pThr-Gly-pTyr.

[0093] In a non-limiting example, the assay includes contacting a test compound with a peptide comprising the sequence pThr-Gly-pTyr, a polypeptide comprising and / or consisting of the catalytic domain of MKP-5 (such as, but not limited to, residues 305-482 of SEQ ID NO:4) or an active fragment thereof. If the test compound is an MKP-5 inhibitor, the compound will inhibit the dephosphorylation of the peptide comprising the sequence pThr-Gly-pTyr. If the test compound is not an MKP-5 inhibitor, the compound will not inhibit the dephosphorylation of the peptide comprising the sequence pThr-Gly-pTyr. The dephosphorylation of the peptide can be qualitatively and / or quantitatively assessed using any method known in the art, such as, but not limited to, the malachite green phosphate assay. This assay provides a rapid, reproducible colorimetric method for measuring inorganic free phosphate in aqueous solutions. The assay is based on the formation of a complex between malachite green molybdate and free orthophosphate, which absorbs at 620-640 nm. The assay is a reliable and suitable means of detecting and quantifying minimal amounts of inorganic free phosphate and is suitable for high throughput screening applications.

[0094] kit The present invention includes kits comprising at least one MKP-5 inhibitor, optionally an applicator, and instructions for its use.

[0095] The instructional material included in the kit includes instructions for preventing or treating an MKP-5 regulated disease in a mammal. The instructional material describes the amount of the MKP-5 inhibitor to be administered to the mammal, and the frequency of administration. In certain embodiments, the kit further includes at least one additional agent that prevents or treats an MKP-5 regulated disease in a mammal. In other embodiments, the kit further includes at least one additional agent that improves and / or prevents further cognitive loss in the mammal.

[0096] Administration / Dosage / Formulation The regimen of administration can affect what constitutes an effective amount. The therapeutic formulation can be administered to the subject either before or after the onset of the disease or disorder contemplated in the present invention. In addition, several divided doses, as well as staggered doses, can be administered daily or sequentially, or the dose can be continuously infused or bolus injected. Furthermore, the dose of the therapeutic formulation can be proportionally increased or decreased as indicated by the exigencies of the treatment or prophylactic situation.

[0097] Administration of the compositions of the present invention to a patient, preferably a mammal, more preferably a human, may be carried out using known procedures at a dosage and for a duration effective to treat the disease or disorder contemplated in the present invention. The effective amount of the therapeutic compound required to achieve a therapeutic effect may vary depending on factors such as the state of the disease or disorder in the patient; the age, sex, and weight of the patient; and the ability of the therapeutic compound to treat the disease or disorder contemplated in the present invention. The administration regimen may be adjusted to provide an optimal therapeutic response. For example, several divided doses may be administered daily, or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. A non-limiting example of an effective dosage range of the therapeutic compounds of the present invention is about 1 to 5,000 mg / kg body weight / day. Pharmaceutical compositions useful in carrying out the present invention may be administered to deliver a dose of ng / kg / day to 100 mg / kg / day. In certain embodiments, the present invention contemplates administration of a dosage that results in a concentration of the compound of the present invention in a mammal of 1 μM to 10 μM. One of ordinary skill in the art would be able to study the relevant factors and make a determination regarding the effective amount of the therapeutic compound without undue experimentation.

[0098] Actual dosage levels of the active ingredients in the pharmaceutical compositions of the present invention may be varied to obtain an amount of the active ingredient effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without being toxic to the patient.

[0099] In particular, the selected dosage level will depend on a variety of factors, including the activity of the particular compound used, the time of administration, the rate of excretion of the compound, the duration of treatment, other drugs, compounds or materials used in combination with the compound, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and similar factors known in the medical arts.

[0100] A medical practitioner, such as a physician or veterinarian, having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start the dosage of the compound of the present invention used in the pharmaceutical composition at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

[0101] In certain embodiments, it is particularly advantageous to formulate compound in unit dosage form for ease of administration and uniformity of dosage.Unit dosage form as used herein refers to a physically separate unit suitable as a unitary dose for the patient to be treated; each unit contains a predetermined amount of therapeutic compound calculated to produce desired therapeutic effect together with necessary pharmaceutical medium.The unit dosage form of the present invention is determined by and directly depends on (a) the inherent characteristics of therapeutic compound and the specific therapeutic effect to be achieved, and (b) the inherent limitations of the technical field of compounding / formulating such therapeutic compound for the treatment of disease or disorder contemplated in the present invention.

[0102] In certain embodiments, the compositions of the present invention are formulated with one or more pharma- ceutically acceptable excipients or carriers.In other embodiments, the pharmaceutical compositions of the present invention comprise a therapeutically effective amount of the compounds of the present invention and a pharma- ceutically acceptable carrier.

[0103] The carrier may be a solvent or dispersion medium, for example, containing water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oils. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. Prevention of microbial action can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it is preferable to include an isotonic agent, for example, sugar, sodium chloride, or a polyalcohol such as mannitol and sorbitol, in the composition. Prolonged absorption of an injectable composition can be brought about by including an agent that delays absorption, for example, aluminum monostearate or gelatin, in the composition.

[0104] In certain embodiments, the compositions of the present invention are administered to a patient in a dosage range ranging from 1 to 5 or more times per day. In other embodiments, the compositions of the present invention are administered to a patient in a dosage range including, but not limited to, once per day, once every 2 days, once every 3 days to once per week, and once every 2 weeks. It will be readily apparent to one of skill in the art that the frequency of administration of the various combination compositions of the present invention will vary from individual to individual depending on many factors including, but not limited to, age, disease or disorder being treated, sex, general health, and other factors. Thus, the present invention should not be construed as being limited to any particular dosing regimen, and the exact dose and composition to be administered to any patient will be determined by the attending physician taking into account all other factors related to the patient.

[0105] The compound of the present invention for administration may be administered in the following dosage amounts: about 1 μg to about 10,000 mg, about 20 μg to about 9,500 mg, about 40 μg to about 9,000 mg, about 75 μg to about 8,500 mg, about 150 μg to about 7,500 mg, about 200 μg to about 7,000 mg, about 3050 μg to about 6,000 mg, about 500 μg to about 5,000 mg, about 750 μg to about 4,000 mg, about 1 mg to about 3, The range may be from about 1,000 mg, about 10 mg to about 2,500 mg, about 20 mg to about 2,000 mg, about 25 mg to about 1,500 mg, about 30 mg to about 1,000 mg, about 40 mg to about 900 mg, about 50 mg to about 800 mg, about 60 mg to about 750 mg, about 70 mg to about 600 mg, about 80 mg to about 500 mg, and any and all whole or partial increments therebetween.

[0106] In some embodiments, the dosage of the compound of the present invention is about 1 mg to about 2,500 mg. In some embodiments, the dosage of the compound of the present invention used in the compositions described herein is less than about 10,000 mg, or less than about 8,000 mg, or less than about 6,000 mg, or less than about 5,000 mg, or less than about 3,000 mg, or less than about 2,000 mg, or less than about 1,000 mg, or less than about 500 mg, or less than about 200 mg, or less than about 50 mg. Similarly, in some embodiments, the dose of the second compound described herein is less than about 1,000 mg, or less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 400 mg, or less than about 300 mg, or less than about 200 mg, or less than about 100 mg, or less than about 50 mg, or less than about 40 mg, or less than about 30 mg, or less than about 25 mg, or less than about 20 mg, or less than about 15 mg, or less than about 10 mg, or less than about 5 mg, or less than about 2 mg, or less than about 1 mg, or less than about 0.5 mg, as well as any and all whole or partial increments thereof.

[0107] In certain embodiments, the present invention is directed to a packaged pharmaceutical composition comprising a container holding a therapeutically effective amount of a compound of the present invention, alone or in combination with a second pharmaceutical agent; and instructions for using the compound to treat, prevent, or reduce one or more symptoms of a disease or disorder contemplated herein.

[0108] The preparations may be used in admixture with conventional excipients, i.e., pharma- ceutically acceptable organic or inorganic carrier substances suitable for oral, parenteral, nasal, intravenous, subcutaneous, intestinal, or any other suitable administration mode known in the art.The pharmaceutical preparations may be sterilized and, if desired, adjuvants, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts that affect osmotic pressure, buffers, coloring, flavoring and / or aromatic substances, etc.They may be combined, if desired, with other active agents, such as antifibrotic agents.

[0109] The route of administration of any composition of the present invention includes oral, nasal, rectal, intravaginal, parenteral, buccal, sublingual or topical.The compounds used in the present invention may be formulated for administration by any suitable route, such as oral or parenteral, for example, transdermal, transmucosal (e.g. sublingual, lingual, (trans)buccal, (trans)urethral, ​​vaginal (e.g. vaginal and perivaginal), (trans)nasal and (trans)rectal), intravesical, intrapulmonary, intraduodenal, intragastric, intrathecal, subcutaneous, intramuscular, intradermal, intraarterial, intravenous, intrabronchial, inhalation and topical administration.

[0110] Suitable compositions and dosage forms include, for example, tablets, capsules, caplets, pills, gelcaps, troches, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pellets, magmas, lozenges, creams, pastes, plasters, lotions, discs, suppositories, liquid sprays for nasal or oral administration, dry powder or aerosolized formulations for inhalation, compositions and formulations for intravesical administration, etc. It should be understood that the formulations and compositions that would be useful in the present invention are not limited to the specific formulations and compositions described herein.

[0111] Oral route For oral application, tablets, dragees, liquids, drops, suppositories, or capsules, caplets and gelcaps are particularly suitable. Compositions intended for oral use may be prepared according to any method known in the art, and may contain one or more agents selected from the group consisting of inert and non-toxic pharmaceutical excipients suitable for the manufacture of tablets. Such excipients include, for example, inert diluents such as lactose; granulating and disintegrating agents such as cornstarch; binding agents such as starch; and lubricants such as magnesium stearate. Tablets may be uncoated or may be coated by known techniques for lubrication or to delay the release of active ingredients. Preparations for oral use may be presented as hard gelatin capsules in which the active ingredient is mixed with an inert diluent.

[0112] In certain embodiments, the tablet of the present invention comprises saracatinib difumaric acid salt, mannitol, anhydrous calcium hydrogen phosphate, crospovidone, hypromellose and magnesium stearate, together with a film coat comprising hypromellose, macrogol 400, red iron oxide, black iron oxide and titanium dioxide. In other embodiments, the tablet of the present invention comprises about 50 or 125 mg of saracatinib as the free base. In yet other embodiments, the tablet of the present invention comprises about 71.4 or 178.6 mg of saracatinib as the difumaric acid salt.

[0113] For oral administration, the compounds of the present invention may be in the form of tablets or capsules prepared by conventional means with pharma- ceutically acceptable excipients, such as binders (e.g., polyvinylpyrrolidone, hydroxypropylcellulose or hydroxypropylmethylcellulose); fillers (e.g., corn starch, lactose, microcrystalline cellulose or calcium phosphate); lubricants (e.g., magnesium stearate, talc, or silica); disintegrants (e.g., sodium starch glycolate); or wetting agents (e.g., sodium lauryl sulfate). If desired, tablets may be coated using a suitable method and coating material, such as OPADRY™ film coating system (e.g., OPADRY™ OY Type, OYC Type, Organic Enteric OY-P Type, Aqueous Enteric OY-A Type, OY-PM Type, and OPADRY™ White, 32K18400), available from Colorcon, West Point, Pa. Liquid preparations for oral administration may be in the form of solutions, syrups, or suspensions. Liquid preparations may be prepared by conventional means with pharma- ceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, methylcellulose, or hydrogenated edible fats); emulsifying agents (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters, or ethyl alcohol); and preservatives (e.g., methyl or propyl p-hydroxybenzoates or sorbic acid).

[0114] Granulation techniques are well known in the pharmaceutical arts for modifying starting powders of active ingredients or other granular materials.Powder is typically mixed with binder material to form larger permanently flowing aggregates or granules, which is called "granulation".For example, the "wet" granulation process using solvent is generally characterized by mixing powder with binder material, wetting with water or organic solvent under conditions that result in the formation of a wet granular mass, from which the solvent must then be evaporated.

[0115] Melt granulation generally uses materials that are solid or semi-solid at room temperature (i.e., have a relatively low softening point or melting point range) to facilitate the granulation of powders or other materials without the addition of water or other liquid solvents. When the low melting point solid is heated to a temperature within its melting point range, it liquefies and acts as a binder or granulation medium. The liquefied solid spreads over the surface of the powdered materials it contacts and, after cooling, forms a solid granular mass in which the initial materials are bound together. The resulting molten granules may then be provided to a tablet press to prepare an oral dosage form or may be encapsulated. Melt granulation improves the dissolution rate and bioavailability of the active substance (i.e., drug) by forming a solid dispersion or solid solution.

[0116] US Patent No. 5,169,645 discloses a directly compressible wax-containing granule with improved flow properties. The granule is obtained by mixing the wax with certain flow-improving additives in the melt, followed by cooling and granulating the mixture. In certain embodiments, only the wax itself melts during the molten combination of the wax and additives, and in other cases, both the wax and the additives melt.

[0117] The present invention also includes multi-layer tablets, comprising a layer providing delayed release of one or more compounds of the present invention, and a further layer providing immediate release of an agent for treating a disease or disorder contemplated in the present invention. A wax / pH-sensitive polymer mixture may be used to obtain a gastric insoluble composition in which the active ingredient is entrapped, ensuring its delayed release.

[0118] Parenteral Administration As used herein, "parenteral administration" of pharmaceutical compositions includes any route of administration characterized by physical breakthrough of the tissue of a subject and administration of pharmaceutical compositions through a gap in tissue.Thus, parenteral administration includes, but is not limited to, administration of pharmaceutical compositions by injection of the composition, application of the composition through a surgical incision, application of the composition through a tissue-permeable non-surgical wound, etc.In particular, parenteral administration is intended to include, but is not limited to, subcutaneous, intravenous, intraperitoneal, intramuscular, intrasternal injection, and kidney dialysis infusion techniques.

[0119] A formulation of a pharmaceutical composition suitable for parenteral administration comprises the active ingredient in combination with a pharma- ceutically acceptable carrier, such as sterile water or sterile isotonic saline. Such formulations may be prepared, packaged, or sold in a form suitable for bolus administration or continuous administration. Injectable formulations may be prepared, packaged, or sold in unit dosage form, such as in ampoules or in multi-dose containers containing a preservative. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous media, pastes, and implantable sustained-release or biodegradable formulations. Such formulations may further comprise one or more additional components, including, but not limited to, suspending agents, stabilizing agents, or dispersing agents. In certain embodiments of formulations for parenteral administration, the active ingredient is provided in a dry (i.e., powder or granules) form for reconstitution with a suitable vehicle (e.g., sterile pyrogen-free water) prior to parenteral administration of the reconstituted composition.

[0120] The pharmaceutical composition may be prepared, packaged, or sold in the form of a sterile injectable aqueous or oily suspension or solution. The suspension or solution may be formulated according to known techniques and may contain, in addition to the active ingredient, additional ingredients such as dispersants, wetting agents, or suspending agents described herein. Such sterile injectable preparations may be prepared using, for example, a non-toxic parenterally acceptable diluent or solvent, such as water or 1,3-butanediol. Other acceptable diluents and solvents include, but are not limited to, Ringer's solution, isotonic sodium chloride solution, and fixed oils such as synthetic mono- or diglycerides. Other parenterally administrable formulations that are useful include those that contain the active ingredient in microcrystalline form, in a liposomal preparation, or as a component of a biodegradable polymer system. Compositions for sustained release or implantation may include pharma-ceutically acceptable polymers or hydrophobic materials, such as emulsions, ion exchange resins, sparingly soluble polymers, or sparingly soluble salts.

[0121] Further dosage forms Additional dosage forms of the present invention include those described in U.S. Patent Nos. 6,340,475; 6,488,962; 6,451,808; 5,972,389; 5,582,837; and 5,007,790. Additional dosage forms of the present invention also include those described in U.S. Patent Application Nos. 20030147952; 20030104062; 20030104053; 20030044466; 20030039688; and 20020051820. Further dosage forms of the present invention include those described in PCT Publication Nos. WO 03 / 35041; WO 03 / 35040; WO 03 / 35029; WO 03 / 35177; WO 03 / 35039; WO 02 / 96404; WO 02 / 32416; WO 01 / 97783; WO 01 / 56544; WO 01 / 32217; WO 98 / 55107; WO 98 / 11879; WO 97 / 47285; WO 93 / 18755; and WO 90 / 11757.

[0122] Controlled Release Formulations and Drug Delivery Systems In certain embodiments, the formulations of the present invention may be short-term, rapid-offset, as well as controlled, including, but not limited to, sustained-release, delayed-release and pulsatile-release formulations.

[0123] The term sustained release is used in its ordinary sense to refer to a drug formulation that gradually releases drug over an extended period of time and can, but need not, result in substantially constant blood levels of drug over an extended period of time, which may be as long as a month or longer and should be a longer release than the same amount of drug administered in bolus form.

[0124] For sustained release, the compounds may be formulated with polymers or hydrophobic materials that provide sustained release properties to the compounds.Thus, the compounds for use in the methods of the present invention may be administered in the form of microparticles, for example by injection, or by implantation in the form of wafers or disks.

[0125] In certain embodiments, the compounds of the invention, alone or in combination with another agent, are administered to a patient in a sustained release formulation.

[0126] The term delayed release is used herein in its ordinary sense to refer to a drug formulation that provides an initial release of drug after some delay following drug administration, and which may, but need not, include a delay of from about 10 minutes up to about 12 hours.

[0127] The term pulsatile release is used herein in its ordinary sense to refer to a drug formulation that provides release of the drug in a manner that results in a pulsatile plasma profile of the drug following drug administration.

[0128] The term immediate release is used in its ordinary sense to refer to a drug formulation that provides release of the drug immediately after drug administration.

[0129] Short-term, as used herein, refers to any period of time up to and including about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 40 minutes, about 20 minutes, or about 10 minutes and any and all whole or partial increments thereof after drug administration.

[0130] As used herein, rapid clearance means any period of time up to and including about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 40 minutes, about 20 minutes, or about 10 minutes and any and all whole or partial increments therein after drug administration.

[0131] dosage The therapeutically effective amount or dose of the compound of the present invention depends on the age, sex and weight of the patient, the current medical condition of the patient, and the progression of the disease or disorder contemplated in the present invention. Those skilled in the art can determine the appropriate dose depending on these and other factors.

[0132] A suitable dose of the compound of the present invention may range from about 0.01 mg to about 5,000 mg per day, for example, from about 0.1 mg to about 1,000 mg per day, for example, from about 1 mg to about 500 mg, for example, from about 5 mg to about 250 mg. The dose may be administered in a single dose or multiple doses, for example, 1 to 4 or more doses per day. When multiple doses are used, the amount of each dose may be the same or different. For example, a dose of 1 mg per day may be administered as two 0.5 mg doses about 12 hours apart.

[0133] It is understood that the amount of compound administered daily can be, in non-limiting examples, administered every day, every other day, every 2nd day, every 3rd day, every 4th day, or every 5th day. For example, every other day administration can begin with a 5 mg daily dose on Monday, followed by a first 5 mg daily dose on Wednesday, followed by a second 5 mg daily dose on Friday, and so on.

[0134] If the patient's condition improves, at the physician's discretion, administration of the inhibitor of the present invention is optionally continued; or the dose of the drug being administered is temporarily reduced or temporarily stopped for a period of time (i.e., a "drug holiday"). The length of the drug holiday can optionally vary between 2 days and 1 year, including, by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or 365 days. Dose reductions during drug holidays include, by way of example only, 10% to 100%, including 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.

[0135] Once improvement of the patient's condition occurs, a maintenance dose is administered as necessary.Then, the dose or frequency of administration, or both, is reduced as a function of the disease or disorder to a level at which improvement of the disease is maintained.In certain embodiments, the patient requires long-term intermittent treatment upon any recurrence of symptoms and / or infection.

[0136] The compound for use in the method of the present invention may be formulated in a unit dosage form. The term "unit dosage form" refers to a physically discrete unit suitable as a unitary dose for a patient undergoing treatment, each unit containing a predetermined amount of active material calculated to produce a desired therapeutic effect, optionally together with a suitable pharmaceutical carrier. The unit dosage form may be for a single daily dose or for one of multiple daily doses (e.g., about 1-4 or more times per day). When multiple daily doses are used, the unit dosage form may be the same or different for each administration.

[0137] The toxicity and therapeutic efficacy of such treatment regimen are optionally determined in cell cultures or experimental animals, including but not limited to determining LD50 (the dose that is lethal to 50% of the population) and ED50 (the dose that is therapeutically effective in 50% of the population). The dose ratio between toxicity and therapeutic effect is the therapeutic index, which is expressed as the ratio of LD50 to ED50. The data obtained from cell culture assays and animal tests are optionally used to formulate a range of dosages for use in humans. The dosage of such compounds is preferably within a range of circulating concentrations that includes the ED50 with minimal toxicity. Dosage can optionally vary within this range, depending on the dosage form used and the route of administration used.

[0138] Those skilled in the art will recognize or be able to ascertain, using no more than routine experimentation, many equivalents to the specific procedures, embodiments, claims, and examples described herein. Such equivalents are considered to be within the scope of the present invention and the appended claims. For example, variations in reaction conditions, including but not limited to reaction times, reaction sizes / volumes, and experimental reagents such as solvents, catalysts, pressures, atmospheric conditions, and reducing / oxidizing agents, using art-recognized substitutions and no more than routine experimentation, should be understood to be within the scope of this application.

[0139] Whenever values ​​and ranges are provided herein, it is to be understood that all values ​​and ranges that fall within these values ​​and ranges are included within the scope of the present invention. Moreover, all values ​​that fall within these ranges, as well as the upper or lower limits of a range of values, are also contemplated by the application.

[0140] The following examples further illustrate aspects of the present invention, but do not in any way limit the present teachings or disclosure herein. EXAMPLES

[0141] Experimental Examples The present invention will be further described in conjunction with the following examples. These examples are provided for illustrative purposes only and are not intended to be limiting unless otherwise specified. Therefore, the present invention should not be construed as being limited to the following examples in any way, but rather as including any and all variations that become evident as a result of the teachings provided herein.

[0142] Example 1: Assay for MKP-5 inhibitors Compounds from Microsource GenPlus, Microsource Natural Product, NIH Clinical collections, ChemBridge DIVERSet, ChemBridge MicroFormats, Maybridge Diversity, and pilot collections consisting of ChemBridge Amines and Alcohols were screened. Assay conditions were 1.5 μM MKP-5 and 25 μM dually phosphorylated peptide DDE(Nle)(pT)G(pY)VATR (total volume 10 μL) incubated at 37 °C for 30 min followed by addition of 40 μL malachite green reagent. Z' values ​​were approximately 0.7-0.8. Malachite green reagent was made by mixing 30 mL of 0.045% malachite green (Sigma #M-9636) with 20 mL of 4.2% ammonium molybdate / 4M HCl, stirring for at least 30 min, and adding 0.01% Tween 20. Dose responses of the four most potent compounds (structures below) are shown in Figure 2 and Table 1.

[0143] The following % MKP-5 inhibition data were obtained at a compound concentration of 12.5 μM: YU252252, 37%; YU241658, 15%; YU241659, 12%.

[0144] [Table 1]

[0145] Example 2: Crystal structure of human MKP-5 Crystallization was achieved using the hanging drop vapor diffusion method with a protein concentration of 12 mg / mL and 5 mM inhibitor (YU032149, formula 3) incubated at 4 °C. The well solution contained 200 mM ammonium acetate, 100 mM HEPES (pH 7.5), 25% w / v PEG3350. Crystals were grown for 4-7 days. Mother liquor containing an additional 3% w / v PEG3350 was used as a cryoprotectant. A ribbon diagram of the structure is shown in Figure 3. Data statistics are shown in Table 2.

[0146] [Table 2]

[0147] Without wishing to be limited by theory, YU032149 appears to inhibit MKP-5 by binding to the allosteric site, based on the crystal structure, which causes a conformational change in the protein, ultimately reducing the amount of catalytic site by about 30%. Contacts between residues in the allosteric site and YU032149 are shown in Figure 4. Replacements of active site residues are shown in Figure 5.

[0148] Example 3: Other inhibitors of MKP-5 Commercially available derivatives of YU032149 were evaluated for activity against MKP-5. The results are shown in Figure 6. A series of compounds, referred to herein as the HJ series, was derived from YU032149 based on the co-crystal structure. The structures are shown above. Activity data for the HJ series is shown in Figure 7.

[0149] Example 4: Synthesis of HJ series Exemplary schemes applicable to certain compounds contemplated within the scope of the present invention are provided herein. TIFF0007676477000031.tif123148

[0150] The disclosures of each and every patent, patent application, and publication cited herein are incorporated herein by reference in their entirety. Although the present invention has been disclosed with respect to certain embodiments, it is clear that other embodiments and modifications of the present invention may be devised by those skilled in the art without departing from the true spirit and scope of the present invention. It is intended that the appended claims be construed to include all such embodiments and equivalent modifications.

[0151] Sequence information SEQUENCE LISTING <110> Yale University <120> Compounds and Compositions for Treating Fibrosis <150> US 62 / 607,110 <151> 2017-12-18 <160> 4 <170> PatentIn version 3.5 <210> 1 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Chemically synthesized <220> <221> MOD <222> (1)..(1) <223> N-terminus derivatived with FITC-AHA [5-(fluorescein-amido)hexylacrylic acid] <220> <221> MOD_RES <222> (6)..(6) <223> PHOSPHORYLATION <220> <221> MOD_RES <222> (10)..(10) <223> AMIDATION <400> 1 Asp Glu Leu Thr Gly Tyr Val Ala Thr Arg 1 5 10 <210> 2 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Chemically synthesized <220> <221> MOD_RES <222> (1)..(1) <223> N-terminus derivatived with FITC-AHA [5-(fluorescein-amido)hexylacrylic acid] <220> <221> MOD_RES <222> (4)..(4) <223> PHOSPHORYLATION <220> <221> MOD_RES <222> (10)..(10) <223> AMIDATION <400> 2 Asp Glu Leu Thr Gly Tyr Val Ala Thr Arg 1 5 10 <210> 3 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Chemically synthesized <220> <221> MOD_RES <222> (4)..(4) <223> Nle <220> <221> MOD_RES <222> (5)..(5) <223> PHOSPHORYLATION <220> <221> MOD_RES <222> (7)..(7) <223> PHOSPHORYLATION <400> 3 Asp Asp Glu Xaa Tyr Gly Tyr Val Ala Thr Arg 1 5 10 <210> 4 <211> 482 <212> PRT <213> Homo sapiens <400> 4 Met Pro Pro Ser Pro Leu Asp Asp Arg Val Val Val Ala Leu Ser Arg 1 5 10 15 Pro Val Arg Pro Gln Asp Leu Asn Leu Cys Leu Asp Ser Ser Tyr Leu 20 25 30 Gly Ser Ala Asn Pro Gly Ser Asn Ser His Pro Pro Val Ile Ala Thr 35 40 45 Thr Val Val Ser Leu Lys Ala Ala Asn Leu Thr Tyr Met Pro Ser Ser 50 55 60 Ser Gly Ser Ala Arg Ser Leu Asn Cys Gly Cys Ser Ser Ala Ser Cys 65 70 75 80 Cys Thr Val Ala Thr Tyr Asp Lys Asp Asn Gln Ala Gln Thr Gln Ala 85 90 95 Ile Ala Ala Gly Thr Thr Thr Thr Ala Ile Gly Thr Ser Thr Thr Cys 100 105 110 Pro Ala Asn Gln Met Val Asn Asn Asn Glu Asn Thr Gly Ser Leu Ser 115 120 125 Pro Ser Ser Gly Val Gly Ser Pro Val Ser Gly Thr Pro Lys Gln Leu 130 135 140 Ala Ser Ile Lys Ile Ile Tyr Pro Asn Asp Leu Ala Lys Lys Met Thr 145 150 155 160 Lys Cys Ser Lys Ser His Leu Pro Ser Gln Gly Pro Val Ile Ile Asp 165 170 175 Cys Arg Pro Phe Met Glu Tyr Asn Lys Ser His Ile Gln Gly Ala Val 180 185 190 His Ile Asn Cys Ala Asp Lys Ile Ser Arg Arg Arg Leu Gln Gln Gly 195 200 205 Lys Ile Thr Val Leu Asp Leu Ile Ser Cys Arg Glu Gly Lys Asp Ser 210 215 220 Phe Lys Arg Ile Phe Ser Lys Glu Ile Ile Val Tyr Asp Glu Asn Thr 225 230 235 240 Asn Glu Pro Ser Arg Val Met Pro Ser Gln Pro Leu His Ile Val Leu 245 250 255 Glu Ser Leu Lys Arg Glu Gly Lys Glu Pro Leu Val Leu Lys Gly Gly 260 265 270 Leu Ser Ser Phe Lys Gln Asn His Glu Asn Leu Cys Asp Asn Ser Leu 275 280 285 Gln Leu Gln Glu Cys Arg Glu Val Gly Gly Gly Ala Ser Ala Ala Ser 290 295 300 Ser Leu Leu Pro Gln Pro Ile Pro Thr Thr Pro Asp Ile Glu Asn Ala 305 310 315 320 Glu Leu Thr Pro Ile Leu Pro Phe Leu Phe Leu Gly Asn Glu Gln Asp 325 330 335 Ala Gln Asp Leu Asp Thr Met Gln Arg Leu Asn Ile Gly Tyr Val Ile 340 345 350 Asn Val Thr Thr His Leu Pro Leu Tyr His Tyr Glu Lys Gly Leu Phe 355 360 365 Asn Tyr Lys Arg Leu Pro Ala Thr Asp Ser Asn Lys Gln Asn Leu Arg 370 375 380 Gln Tyr Phe Glu Glu Ala Phe Glu Phe Ile Glu Glu Ala His Gln Cys 385 390 395 400 Gly Lys Gly Leu Leu Ile His Cys Gln Ala Gly Val Ser Arg Ser Ala 405 410 415 Thr Ile Val Ile Ala Tyr Leu Met Lys His Thr Arg Met Thr Met Thr 420 425 430 Asp Ala Tyr Lys Phe Val Lys Gly Lys Arg Pro Ile Ile Ser Pro Asn 435 440 445 Leu Asn Phe Met Gly Gln Leu Leu Glu Phe Glu Glu Asp Leu Asn Asn 450 455 460 Gly Val Thr Pro Arg Ile Leu Thr Pro Lys Leu Met Gly Val Glu Thr 465 470 475 480 Val Val

Claims

1. 1. A pharmaceutical composition for treating or preventing an MKP-5 regulated disease or disorder in a mammal in need thereof, comprising: The pharmaceutical composition comprises a compound selected from the group consisting of: 1-((5,6-dihydrothieno[2,3-h]quinazolin-2-yl)thio)-3,3-dimethylbutan-2-one: ； 3,3-Dimethyl-1-(pyrimido[5,4-c]quinolin-2-ylthio)butan-2-one: and 1-((6-acetyl-5,6-dihydropyrimido[5,4-c]quinolin-2-yl)thio)-3,3-dimethylbutan-2-one: A pharmaceutical composition comprising a therapeutically effective amount of

2. 2. The pharmaceutical composition of claim 1, wherein the compound has a Ki for MKP-5 of ≦100 μM.

3. The pharmaceutical composition of claim 1 , wherein the compound binds to an allosteric site of MKP-5.

4. 10. The pharmaceutical composition of claim 1, further comprising at least one pharma- ceutically acceptable carrier.

5. The pharmaceutical composition of any one of claims 1 to 4, wherein the MKP-5 regulated disease or disorder is a fibrotic disease or disorder.

6. 6. The pharmaceutical composition of claim 5, wherein the MKP-5 regulated disease or disorder is selected from the group consisting of dystrophic muscle diseases, cardiac or vascular diseases, idiopathic pulmonary fibrosis, and any combination thereof.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the mammal is a human.

8. 8. The pharmaceutical composition of any one of claims 1 to 7, wherein the pharmaceutical composition is administered to a mammal by at least one route selected from the group consisting of nasal, inhalation, topical, oral, buccal, rectal, pleural, peritoneal, vaginal, intramuscular, subcutaneous, transdermal, epidural, intratracheal, otic, intraocular, intrathecal, and intravenous routes.

9. The pharmaceutical composition of any one of claims 1 to 8, for use in combination with at least one additional agent for treating or preventing an MKP-5 regulated disease or disorder in a mammal.

10. 10. The pharmaceutical composition of claim 9, wherein the compound and the at least one additional agent are co-formulated.

11. 1. A kit for preventing or treating an MKP-5 regulated disease or disorder in a mammal, comprising an MKP-5 inhibitor, optionally an applicator, and instructions for its use, the instructions describing the amount and frequency of administration of the MKP-5 inhibitor to be administered to a mammal to treat or prevent the MKP-5 regulated disease or disorder, The MKP-5 inhibitor is a compound selected from the group consisting of: 1-((5,6-dihydrothieno[2,3-h]quinazolin-2-yl)thio)-3,3-dimethylbutan-2-one: ； 3,3-Dimethyl-1-(pyrimido[5,4-c]quinolin-2-ylthio)butan-2-one: and 1-((6-acetyl-5,6-dihydropyrimido[5,4-c]quinolin-2-yl)thio)-3,3-dimethylbutan-2-one: That's it, Kit.

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