How to treat fibrosis using PKM2 activators

Abstract

This provides a method for treating patients suffering from diseases caused by organ / tissue fibrosis. [Solution] The method includes administering a PKM2 activator. The PKM2 activator can reduce LOX protein expression and activate PKM2.

Description

Technical Field

[0001] This application relates to the prevention, treatment, and / or recovery of fibrosis. This application also relates to the use of an activator of pyruvate kinase M2 (PKM2) for the treatment, prevention, or improvement of fibrosis.

Background Art

[0002] Fibrosis is defined, for example, as the abnormal accumulation of fibrous tissue caused by tissue damage repair and / or immune responses. In humans, fibrosis (a fine movement often called atrial fibrosis) is seen in various organs and tissues such as the lungs, liver, pancreas, kidneys, bone marrow, and skin.

[0003] Collagen fibrils are overproduced during the progression of fibrosis as a result of the inflammatory response to tissue damage. Fibrosis affects tissues in almost all organs. Myofibroblasts are the main cell type involved in collagen synthesis and secretion. Collagen is an ECM protein mainly composed of triplet repeats of Gly-x-Pro and Gly-x-Hyp. Glycine is an amino acid that accounts for more than 30% of the collagen content. Myofibroblasts need to engage in excessive glycine production to meet the needs of the large amount of collagen production and secretion during the progression of fibrosis. Cells obtain glycine from two sources: (1) glycine from food breakdown, which can then lead to protein synthesis. (2) De novo biosynthesis.

[0004] The composition and stiffness of the extracellular matrix (ECM) are the main driving forces for the onset and persistence of fibrotic diseases. Lysyl oxidase (LOX) and LOX-like (LOXL) proteins play important roles in ECM remodeling due to their collagen cross-linking and intracellular functions.

[0005] Fibrosis is an essential process that plays a crucial role in tissue repair and wound healing. Excessive fibrosis is common in many rare and common diseases and is an important etiology. Diseases characterized by excessive fibrosis include, but are not limited to, systemic sclerosis, scleroderma, hypertrophic cardiomyopathy, dilated cardiomyopathy (DCM), atrial fibrosis, ventricular fibrosis, myocarditis, cirrhosis, kidney disease, eye disease, asthma, cystic fibrosis, arthritis, and idiopathic pulmonary fibrosis. Despite its significant impact on human health, therapeutic and diagnostic approaches to fibrosis remain an unmet medical need. [Overview of the project]

[0006] One aspect of this application includes the treatment, prevention, or mitigation of fibrosis in a person requiring treatment by administration of a PKM2 activator. A method for treating a patient suffering from a disease caused by organ / tissue fibrosis includes the step of administering a pharmaceutical composition containing an effective amount of a PKM2 activator to the patient in need.

[0007] Another embodiment includes a method in which the PKM2 activator is TEPP-46.

[0008] Another embodiment includes a method for treating a patient suffering from a disease caused by organ / tissue fibrosis, comprising the administration of a PKM2 activator. PKM2 activators can reduce collagen synthesis and LOX / L family expression in myofibroblasts. Treatment with PKM2 activators can reduce the apoptotic resistance of myofibroblasts. [Brief explanation of the drawing]

[0009] [Figure 1] This study demonstrates that TEPP-46 reverses hepatic and pulmonary fibrosis and reduces serine / glycine metabolism in fibrotic liver and lungs. [Figure 2] This study demonstrates that PKM2 dimers protect myofibroblasts from oxidative stress-induced apoptosis by upregulating NADPH metabolism. [Figure 3] This shows that PKM2 is expressed in myofibroblasts. [Figure 4] This study demonstrates that treatment of fibroblasts with the PKM2 activator (DASA-10) reduces metabolic enzymes involved in serine / glycine metabolism. [Figure 5] This study demonstrates that treatment with a PKM2 activator (TEPP-46) can reverse pancreatitis. [Figure 6] Treatment with a PKM2 activator (TEPP-46) reduces cardiac fibrosis after myocardial infarction. [Figure 7] This study demonstrates that treatment with a PKM2 activator (DASA-10) reduces the expression of LOX and LOXL2 in vivo in activated fibroblasts. [Figure 8] This study demonstrates that treatment with a PKM2 activator (TEPP-46) reduces LOX expression in vivo in mouse models of hepatic and pulmonary fibrosis. [Figure 9] This study demonstrates the effect of the PKM2 activator (Mitapivat) on collagen production in myofibroblasts. [Modes for carrying out the invention]

[0010] definition The “administration step” refers to the method of giving a patient a certain dosage of a pharmaceutical composition. The compositions described herein may be administered by routes selected from, for example, eye, inhalation, parenteral, skin, transdermal, buccal, rectal, vaginal, sublingual, perilingual, nasal, topical, and oral administration. Parenteral administrations include intravenous, intraperitoneal, subcutaneous, and intramuscular administration. The preferred method of administration may vary depending on various factors, such as the components of the composition being administered and the severity of the condition being treated.

[0011] The term "fibrosis" refers to a condition characterized by the development of excessive fibrous connective tissue, such as scar tissue, in a tissue or organ. Such scar tissue formation may occur in response to infection, inflammation, or injury to an organ resulting from disease, trauma, chemical toxicity, surgery, etc. Fibrosis can occur in a variety of different tissues and organs, including the liver, kidneys, intestines, lungs, and heart. These changes are sometimes called fibrocystic changes and were formerly known as fibrocystic diseases. The term "fibrosis" as used herein does not refer to cancerous fibrosis.

[0012] As used herein, the terms “inhibit” or “inhibit” refer to any detectable positive effect on the onset or progression of a disease or condition (e.g., fibrosis). Such positive effects may include delaying or preventing the onset of at least one symptom or sign of the disease or condition, alleviating or reversing a symptom or sign, and delaying or preventing further worsening of a symptom or sign.

[0013] As used herein, the term “effective dose” refers to the amount of a compound (e.g., a pyruvate kinase M2 (PKM2) activator) that, when administered in an appropriate dose, produces an acute or chronic therapeutic effect. This effect includes, to any detectable extent, prevention, correction, inhibition, or recovery of the symptoms, signs, and underlying pathology of a disease / condition (e.g., hepatic, renal, or intestinal fibrosis) and associated complications. The exact dose and dosage schedule will depend on the therapeutic objective and will be verifiable by those skilled in the art using known techniques.

[0014] The term "patient" means any animal, such as a mammal or a human.

[0015] The term "prodrug" refers to a compound that can be converted to the biologically active compound of this application under physiological conditions or by solvolysis. Therefore, the term "prodrug" refers to a pharmaceutically acceptable metabolic precursor of the compound of this application. A prodrug may be inactive when administered to a target requiring it, but is converted to the active compound of this application in vivo. Typically, prodrugs are rapidly converted in vivo, for example, by hydrolysis in the blood, to produce the parent compound of this application. Prodrug compounds often offer advantages in solubility, histocompatibility, or delayed release in mammalian organisms.

[0016] The term “pharmaceutically acceptable carrier, diluent or excipient” includes, but is not limited to, any adjuvants, carriers, excipients, glidants, sweeteners, diluents, preservatives, colorants, flavor enhancers, surfactants, humectants, dispersants, suspending agents, stabilizers, isotonic agents, solvents, or emulsifiers that are approved by the U.S. Food and Drug Administration as acceptable for use in humans or livestock.

[0017] Pharmaceutical compositions can be administered parenterally, for example, intravenously, subcutaneously, intradermally, or intramuscularly. Accordingly, this application provides compositions for parenteral administration, comprising a solution of a compound or salt dissolved or suspended in an acceptable carrier suitable for parenteral administration, including aqueous and non-aqueous isotonic sterile injection solutions.

[0018] The term "selective" means inhibition of PKM2 by at least 20%, 50%, 75%, 2x, 3x, 4x, 5x, 6x, or 10x greater than that of a second kinase, e.g., a second pyruvate kinase, e.g., a different isoform of PKM2. Therefore, in some embodiments, a drug is selective for PKM2 more than another isoform. For example, a drug is selective for PKM2 more than for PKM1. Selective regulation, e.g., inhibition or activation, or selective modulation is used interchangeably with a particular regulation or modulation.

[0019] The term "therapy" includes not only the prevention and treatment of fibrosis, but also maintenance for suppressing the progression of fibrosis in tissues, alleviation of inflammation, alleviation of symptoms associated with fibrosis, and prevention of recurrence. [Detailed Description]

[0020] This application features methods and kits that utilize an activator of pyruvate kinase M2 (PKM2) for the treatment, prevention, or amelioration of diseases caused by organ tissue fibrosis. This application provides methods for treating, ameliorating, or preventing fibrosis or fibrosis-related diseases, as well as methods for screening compounds and compositions useful in those methods. In certain embodiments, a method for treating a patient suffering from fibrosis, comprising administering to a patient in need thereof a pharmaceutical composition (i.e., a PKM2 activator) comprising an effective amount of a compound selected from the compositions disclosed herein.

[0021] Examples of fibrosis include pulmonary fibrosis, liver fibrosis, cardiac fibrosis, pancreatitis, renal fibrosis, prostatic hypertrophy due to fibrosis, myelofibrosis, and scleroderma. In these fibroses, symptoms associated with fibrosis such as inflammation and atrophy are observed depending on the organ with advanced fibrosis and the progression rate. Therefore, the treatment of symptoms associated with fibrosis is also included in this application.

[0022] Other examples include liver fibrosis; hepatitis, cirrhosis, NASH (non-alcoholic steatohepatitis), ASH / AH, primary biliary cholangitis, primary sclerosing cholangitis; pulmonary fibrosis; idiopathic pulmonary fibrosis, COPD, cystic fibrosis, scleroderma; chronic kidney disease; diabetic nephropathy, cystic fibrosis, glomerulonephritis / nephritis, hypertensive nephrosclerosis, allograft nephropathy; pancreatic fibrosis; pancreatitis; systemic sclerosis; cardiac fibrosis; cardiovascular fibrosis; vascular fibrosis; coronary artery disease and peripheral artery disease, arteriosclerosis, vasculitis; primary myelofibrosis; reduction of scars in reconstructive surgery; surgical complications due to fibrosis; and radiation-induced fibrosis.

[0023] Myocardial infarction is a significant complication of coronary artery disease, usually resulting from a critical reduction in coronary blood flow secondary to coronary thrombosis. Two important pathological changes in cardiac tissue after acute myocardial infarction are fibrosis and hypertrophic growth of the cardiac tissue. Both changes ("remodeling") contribute significantly to the pathogenesis of heart failure. Intravenous thrombolytic therapy is widely used to restore blood flow to the occluded coronary arteries. Thrombolytic agents are drugs that can dissolve fibrin-platelet thrombi, thereby allowing blood to flow again through the affected blood vessels. Such drugs include streptokinase, urokinase, prourokinase, leteplase, alteplase, and tissue plasminogen activators (t-PA). The mortality rate in patients with acute myocardial infarction remains high, even with treatment using hemolytic agents.

[0024] Patients treated using the methods described above may or may not have detectable fibrosis. In some embodiments, patients have a reduction of at least about 5%, 10%, 20%, 30%, 40%, or even more than 50% in the amount of fibrosis present in the patient after administration of a PKM2 activator, for example, 1 day, 2 days, 1 week, 1 month, or more than 6 months. Administration of such compounds may be, for example, at least daily. The delay in the clinical manifestations of fibrosis in patients as a result of administration of the compounds disclosed herein may be at least, for example, 6 months, 1 year, 18 months, or even more than 2 years, compared to patients who have not been administered compounds such as those disclosed herein.

[0025] In one example, a PKM2 activator reduced the expression of lysyl oxidase (LOX), a member of the LOX / L family of proteins. This protein family is thought to promote covalent crosslinking of ECM proteins such as collagen and elastin through the oxidation of lysylamine residues, leading to the establishment of ECM tensile strength. The LOX / L protein family is expressed in various tissues, and their dysregulation is associated with the pathology of all organ tissue fibrosis diseases. The expression of LOX, and different LOX family proteins including LOXL2 and LOXL4, differs depending on the disease. This may be due to several reasons, including differences in tissue distribution, processing, domains, regulation of activity, and other differences between proteins. Due to the functional role of LOX / L family proteins in the progression of organ tissue fibrosis diseases, inhibition of the LOX / L family has become an attractive strategy for developing treatments for fibrosis. Experiments have demonstrated that a PKM2 activator potently reduces the expression and secretion of LOX / L family proteins in myofibroblasts as well as in fibrous tissue and blood circulation in mouse models of hepatic and pulmonary fibrosis. The functional role of PKM2 in the decline of the LOX / L family suggests that PKM2 activators are excellent agents for the treatment of fibrosis.

[0026] The method of administering the therapeutic agents for fibrosis (e.g., PKM2 activators) and pharmaceutical compositions for treating fibrosis described in this application is appropriately determined, for example, according to the dosage form, the patient's age, sex, and other conditions, as well as the patient's symptoms. For example, tablets, pills, powders, granules, capsules, liquid formulations, suspensions, and emulsions are administered orally. Injectable preparations are administered intravenously, alone or in combination with common fluids such as glucose and amino acids, and, if necessary, injectable preparations are administered intra-arterial, intramuscular, intradermal, subcutaneous, or intraperitoneal. Suppositories are administered rectally. PKM2 activators may be administered in combination with other treatments or other PKM2 activators.

[0027] "Activator" means a drug that increases the level of pyruvate kinase activity of PKM2 from an inactive monomeric or dimeric state, or maintains or increases the activity of the active tetramer of PKM2 (e.g., in the presence of an endogenous inhibitor). Increased activity may include reducing the endogenous downregulation of PKM2 by an endogenous inhibitor (e.g., an endogenous phosphotyrosine peptide or protein). Binding of phosphotyrosine-containing peptides to activated PKM2 results in FBP dissociation and inactivation of PKM2. Autonomous proliferation signaling in proliferating cells or insulin-induced stimulation of adipocytes leads to the tyrosine phosphorylation cascade. Activators can exert their effects in several ways, including one or more of the following: they can make PKM2 resistant to inhibition by inhibitors, such as endogenous inhibitors; they can inhibit the release of activators, more specifically FBP; they can bind to PKM2 and prevent endogenous inhibitors from promoting the release of endogenous activators, more specifically FBP; or they can inhibit the dissolution of or promote the recombination of subunits constituting PKM2, for example, they can inhibit the oxidation of sulfhydryl moieties on such subunits, for example, they can inhibit the oxidation of cysteine ​​residues. Activators of the pyruvate kinase activity of PKM2 may be referred to as "PKM2 activators."

[0028] Activators can increase the pyruvate kinase activity of PKM2 to a level exceeding the activity level of PKM2 (e.g., the baseline level) (e.g., the level seen in the absence of endogenous or native activators / ligands, e.g., FBP). For example, activators may mimic the effects caused by endogenous or native ligands or activators (e.g., FBP). The activating effect caused by a drug may be the same as, greater than, or less than, the activating effect caused by endogenous or native ligands or activators, but the same type of effect may be produced. Peptides, nucleic acids, and small molecules can be activators. In certain embodiments, the activator has a molecular weight in the range of 10 or 20, 100 or 200 to 10,000, 100 or 200 to 5,000, 100 or 200 to 2,000, or more preferably 100 to 300, 200 to 500, 150 to 500, 200 to 500, 300 to 500, or 150 to 800 daltons or more.

[0029] The activator may be, for example, a peptide, nucleic acid, or small molecule. Peptides useful as activators in the methods, compositions, and kits described herein may include modifications, such as in vivo or in vitro chemical derivatization (e.g., acetylation or carboxylation) of polypeptides. Also included are modifications of glycosylation, such as those produced by modifying the glycosylation pattern of a polypeptide by exposing the peptide to an enzyme that affects glycosylation (e.g., a mammalian glycosylation enzyme) typically derived from cells providing such processing, during polypeptide synthesis and processing, or during further processing steps. Versions of the same primary amino acid sequence having phosphorylated amino acid residues, such as phosphotyrosine, phosphoserine, or phosphothreonine, are also included.

[0030] This application relates to the above-mentioned compounds or pharmaceutical compositions for use in combination with at least one therapeutic agent selected from STAT inhibitors, other anti-inflammatory agents, and / or immunosuppressants. The combination can be supplemented with one or more other active ingredients, such as anticoagulants, or by surgical methods such as angioplasty.

[0031] Peptides useful as activators may be synthetic peptides or purified peptides of natural origin. Peptides may be commercially available or produced in recombinant or non-recombinant cell lines. Characterization of isolated peptide activators can be achieved, for example, using solution assays, gel assays (e.g., SDS-PAGE), membrane conjugation methods, antibodies, enzyme-linked immunosorbent assays (ELISA), or liquid chromatography-electron spray ionization mass spectrometry (LCMS).

[0032] One exemplary activator of PKM2 is TEPP-46, which has the following formula: [ka]

[0033] One exemplary activator of PKM2 is mitapibat, which has the following formula / structure. [ka]

[0034] Other exemplary activators of PKM2 may be selected from the following candidates: 1,6-fructose-bisphosphate, dithiothreitol, 2,5-anhydro-D-mannitol 1,6-bisphosphate, AMP, phosphoenolpyruvic acid, and the following structures. [ka]

[0035] The pharmaceutical compositions of this application can be prepared by methodologies well known in the pharmaceutical field. For example, a pharmaceutical composition intended for administration by injection can be prepared by combining the compounds of this application with sterile distilled water to form a solution. A surfactant may be added to facilitate the formation of a homogeneous solution or suspension. The surfactant is a compound that interacts non-covalently with the compounds of this application to facilitate the dissolution or homogeneous suspension of the compounds in an aqueous delivery system.

[0036] The compounds of this application (e.g., PKM2 activators), or their pharmaceutically acceptable salts, are administered in therapeutically effective doses, which vary depending on various factors, including the activity of the specific compound used, the metabolic stability and duration of action of the compound, the patient's age, weight, overall health, sex, and diet, mode and timing of administration, excretion rate, drug combinations, the severity of a particular disorder or condition, and the patient receiving treatment.

[0037] In the case of topical administration or selective ingestion, the effective local concentration of the drug may not be related to the plasma concentration, and the correct dosage and interval can be determined using other procedures known in the art.

[0038] The amount of PKM2 activator administered depends, of course, on the patient being treated, the severity of their pain, the method of administration, and the prescribing physician's judgment. [Examples]

[0039] Figure 1 shows that TEPP-46 reverses hepatic and pulmonary fibrosis and reduces serine / glycine metabolism in fibrous liver and lung. Sections (A) and (E) of Figure 1 are schematic diagrams of the induction of TAA / alcoholic liver fibrosis (A) and bleomycin-induced pulmonary fibrosis (E) and the subsequent TEPP-46 treatment schedule. Section (B) of Figure 1 shows representative images of liver sections stained with Sirius Red (top) and a-SMA IF (bottom) from mice treated with the indicated drugs. Sections (C) and (D) of Figure 1 show the quantification of collagen levels using ImageJ software in Sirius Red (B) and Masson's Trichrome (F) staining of fibrous liver (B) and lung (F), as well as in a-SMA IF (B) and IHC (F) staining. Quantification was calculated from measurements of 10 mice. Four randomly selected tissue sections and three randomly selected fields of view from each section were quantified for each animal. Collagen levels in Sirius Red or Masson's trichrome staining and α-SMA IF or IHC staining are presented as a percentage of total area. Section (F) shows representative images of Masson's trichrome staining (top) and α-SMA IHC staining (bottom) of lung tissue sections from mice treated with the indicated drugs. Sections (G) and (H) show quantitative analysis of serine (G) and glycine (H) levels in liver and lung tissue extracts from mice treated with the indicated drugs by HPLC-ms. Serine and glycine levels are presented as relative abundances, with the vehicle treatment group defined as 100%. Error bars in sections C, D, G, and H represent mean ± SEM values.

[0040] Figure 2 shows that PKM2 dimers protect myofibroblasts from oxidative stress-induced apoptosis by upregulating NADPH metabolism. Representative images of IF co-staining of α-SMA with cleaved caspase 3 (CC3) from liver sections of mice treated with the indicated drugs are shown. Sections (B) and (C) Apoptosis of LX2 with or without (H2O2) treatment and with or without DASA-10 or DMSO treatment was measured by FACS (graph, B) and (quantitative, C). Sections (D) and (E) Cellular levels of GSH, GSSG, GSH / GSSG ratio (D), and NADPH (E) in LX2 were measured using a kit. Cells were treated with either DMSA or DASA-10. GSH and GSSG were measured in mM (1 × 10⁶ per 100 ml). 6 It is presented as a lysate of individual cells. NADPH is 10 6 The values ​​are presented as picomoles within individual cells. Error bars in C, D, and E represent the mean ± SEM value.

[0041] Figure 3 shows that PKM2 is expressed in myofibroblasts. Section (A) shows immunoblotting analysis of PKM2 (IB:PKM2) and α-SMA (IB:a-SMA) levels in LX-2 cells (LX2) and NLF with or without TGFβ treatment (+) or (-). The immunoblot for β-actin (IB:β-actin) is a loading control. Sections (B) and (C) show the quantification of α-SMA (B) and PKM2. Section (C) shows the levels in LX2 cells and NFL with or without TGFb treatment (TGFb, black bar) or without (TGFb-free, white bar). PKM2 and α-SMA levels are presented as magnification changes compared to the control. Error bars in sections B, C, D, and F represent mean ± SEM. Section (D) shows representative images of IF co-staining of a-SMA (red) and PKM2 (green) in liver sections from mice treated with TEPP-46 or vehicle after hepatic fibrosis was induced with TAA-alcohol. Co-staining (yellow) indicates PKM2 in myofibroblasts.

[0042] Figure 4 shows that the PKM2 activator (DASA-10) reduces metabolic enzymes involved in serine / glycine metabolism. Cellular levels of collagen (Col1A1), PHGDH, PSAT1, SHMT1, and SHMT2 mRNA in LX2 cells treated with DASA-10 (black bars) or DMSO (white bars) after TGFβ treatment were analyzed by qRT-PCR. Cellular mRNA levels are presented as magnification changes compared to the control. Error bars represent mean ± SEM.

[0043] Figure 5 shows that PKM2 activators reverse pancreatitis. Section (A) is a schematic diagram of the schedule for cerulein (red arrow) pancreatitis induction and subsequent TEPP-46 treatment (green arrow). Section (B) shows the quantification of collagen levels in Sirius Red staining in (C) using ImageJ software. Quantification was calculated from measurements of 6 mice. Four randomly selected tissue sections and three randomly selected fields of view in each section were quantified for each animal. The amount of collagen level in Sirius Red staining is presented as a percentage of the total area. Error bars in B mean ± SEM. Section (C) shows representative images of Sirius Red staining of collagen (top) and H&E staining of pancreatic sections from both vehicle and TEPP-46 treated animals.

[0044] Figure 6 shows that treatment with a PKM2 activator reduces post-infarction cardiofibrosis. C57BL / 6J mice (n=6 / group) had myocardial infarction (MI) induced by left ventricular ligation (LV). Sham mice were those that underwent the same surgical procedure without LV. After 70 minutes, the LV was released. All mice were immediately treated with TEPP-46 or vehicle by intraperitoneal injection. Sham mice were not treated. Section 2(A) shows images of Masson's trichrome staining of tissue sections from the infarct area of ​​mice treated with the indicated drugs. Blue indicates collagen staining. Scale bar, 100 mm. Section (B) shows the quantification of trichrome staining of collagen in tissue sections. Quantification means the mean of four fields randomly selected from each section. Four sections were randomly selected from each mouse. Quantification is presented as blue-stained collagen area (%). Error bars represent the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001, performed using an independent two-tailed Student's t-test.

[0045] Figure 7 shows that DASA-10 reduces LOX and LOXL2 expression. Section (A) is 1 × 10 6 Section (B) shows that LX2 and NLF cells were treated with either DMSO or 10 μM DASA-10 and cultured under hypoxic conditions for 24 hours. Relative band intensities for both LOX and LOXL2 were calculated using ImageJ. Section (C) shows isolated and cDNA-converted mRNA, and qPCR was performed on the expression of LOX and LOXL2 in LX2 cells. Section (D) shows isolated and cDNA-converted mRNA, and qPCR was performed on the expression of LOX and LOXL2 in NLF cells.

[0046] Figure 8 shows that TEPP-46 reduces LOX expression in vivo. Section (A) shows that fibrous lung (upper panel) and liver (lower panel) tissues from mice treated with the indicated drugs were thinned to 5 μm and probed for LOX via immunohistochemical staining. Section (B) shows that the LOX-positive area was quantified using ImageJ in both the lung (upper panel) and liver (lower panel). Section (C) shows that the LOX activity assay was measured using the LOX Activity Kit on lung lysate (left column) and serum (right column) in a bleomycin model of pulmonary fibrosis. Section (D) shows that the LOX activity assay was measured using the LOX Activity Kit on liver lysate (left column) and serum (right column) in a TAA / alcohol model of hepatic fibrosis.

[0047] Figure 9 shows the effect of the PKM2 activator (mitapibat) on collagen production in myofibroblasts. Sections (A) and (B) contain hydroxyproline in LX2, and sections (A) and lung fibroblasts (NFL), as well as section (B), were analyzed using a hydroxyproline kit with or without TGFβ treatment (black bars). After TGFβ treatment, cells were treated with mitapibat (gray bars) or vehicle (black bars). The hydroxyproline content in A and B was 1 x 10⁻⁶. 6 This is presented as μg of hydroxyproline in the lysate of individual cells.

Claims

1. A method for treating a patient suffering from a disease caused by organ / tissue fibrosis, comprising the step of administering a pharmaceutical composition containing an effective amount of a PKM2 activator to a patient in need thereof, wherein the PKM2 activator reduces LOX protein expression and activates PKM2.

2. The method according to claim 1, wherein the fibrosis is pulmonary fibrosis.

3. The method according to claim 1, wherein the fibrosis is pancreatic or pancreatic fibrosis.

4. The method according to claim 1, wherein the PKM2 activator is TEPP-46.

5. The method according to claim 1, wherein the fibrosis is fibrosis of the liver, kidneys, bone marrow, heart, pancreas, skin, intestines, vascular system, or joints.

6. The method according to claim 1, wherein the fibrosis is cholangitis or alcoholic hepatitis.

7. The method according to claim 1, wherein the fibrosis is non-alcoholic steatohepatitis (NASH).

8. The method according to claim 1, wherein the fibrosis is pulmonary fibrosis including IPF and COPD.

9. The method according to claim 1, wherein the fibrosis is idiopathic pulmonary fibrosis.

10. The method according to claim 1, wherein the fibrosis is cardiac fibrosis following cardiac surgery due to a heart attack.

11. The method according to claim 1, wherein the fibrosis is a surgical complication and scarring.

12. The method according to claim 1, wherein the PKM2 activator has the following structure. 【Chemistry 1】

13. The method according to claim 1, wherein the PKM2 activator has the following structure. 【Chemistry 2】

14. The method according to claim 1, wherein the PKM2 activator has the following structure. 【Transformation 3】

15. The method according to claim 1, wherein the PKM2 activator has the following structure. 【Chemistry 4】

16. The method according to claim 1, wherein the PKM2 activator has the following structure. 【Transformation 5】

17. The method according to claim 1, wherein the PKM2 activator has the following structure. 【Transformation 6】

18. The method according to claim 1, wherein the PKM2 activator has the following structure. 【Transformation 7】

19. The method according to claim 1, wherein the PKM2 activator has the following structure. 【Transformation 8】

20. A method for activating PKM2 in a mammal to reduce the synthesis of organ tissue fibrils and the expression of LOX protein in a subject, and to increase apoptosis of myofibroblasts that require these, the method comprising the step of administering an effective amount of a PKM2-activating compound to the mammal.

21. The method according to claim 20, wherein the PKM2 activator comprises a pharmaceutically acceptable carrier, diluent, or excipient.

22. The method according to claim 20, further comprising the step of administering an effective amount of one or more chemotherapeutic agents to the subject.

23. The method according to claim 20, wherein the therapeutically effective dose of the compound is about 0.5 mg to about 1000 mg to a human.

24. A method for treating a fibrous disease in a person requiring treatment for a fibrous disease, comprising the step of administering an effective amount of a PKM2 activator to the person, wherein the PKM2 activator is TEPP-46.

25. The method according to claim 24, wherein the fibrous disease is hepatic fibrosis, pulmonary fibrosis, cardiac fibrosis, cutaneous fibrosis, myelofibrosis, or intestinal fibrosis.

26. The method according to claim 25, wherein the PKM2 activator is administered as a pharmaceutical composition comprising an excipient, a dispersant, a solubilizer, a stabilizer, and / or a preservative.

27. The method according to claim 25, wherein the PKM2 activator is administered orally or parenterally.

28. The method according to claim 25, wherein the PKM2 activator is 1,6-fructose-bis-phosphate, dithiothreitol, 2,5-anhydro-D-mannitol 1,6-bis-phosphate, AMP, or phosphoenolpyruvic acid.

29. The method according to claim 25, wherein the PKM2 activator reduces the expression of LOX / L family members.

30. A method for inhibiting fibrosis in a patient who requires inhibition of fibrosis or is at risk of developing fibrosis, the method comprising the step of administering a therapeutically effective amount of a PKM2 activator to the patient, wherein the fibrosis is related to surgery.

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