Flavonoid compounds and methods and materials for using flavonoid compounds to treat fibrotic conditions

Abstract

This document relates to flavonoid compounds and methods and materials for using flavonoid compounds to treat one or more fibrotic and / or aberrant wound healing / scarring conditions (e.g., idiopathic pulmonary fibrosis (IPF) and primary sclerosing cholangitis (PSC)). For example, one or more flavonoid compounds having the structure of Formula (I) can be administered to a mammal (e.g., a human) having one or more fibrotic and / or aberrant wound healing / scarring conditions (e.g., IPF, NASH, and PSC) to treat the mammal.

Description

[0001] 07039-2361WO1 / 2025-008

[0002] FLAVONOID COMPOUNDS AND METHODS AND MATERIALS FOR USING FLAVONOID COMPOUNDS TO TREAT FIBROTIC CONDITIONS

[0003] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U. S. Application Serial No. 63 / 737,017, filed on December 20, 2024. The disclosure of the prior application is considered part of the disclosure of this application, and is incorporated in its entirety into this application.

[0004] TECHNICAL FIELD

[0005] This document relates to flavonoid compounds and methods and materials for using flavonoid compounds to treat fibrotic and / or aberrant wound healing / scarring conditions.

[0006] BACKGROUND INFORMATION

[0007] Fibrotic disease is a leading cause of morbidity and mortality, and can affect nearly all tissues and organ systems. The United States government estimates that 45% of deaths in the United States can be attributed to fibrotic diseases. Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal disease with undefined etiology and minimally effective therapies. The greatest risk factor for developing IPF is aging. The central paradigm to developing antifibrotic drugs for the last half century has focused on directly targeting proliferative lung fibroblasts. However, recent high-resolution analyses of IPF patient lungs suggests disease unique populations of resident lung cells are enriched for markers of senescence. Multiple naturally derived flavonoids can selectively induce apoptosis in senescent cells (e.g., senolytic) and improve endpoints in models of lung fibrosis; however, these natural phytochemicals are not structurally optimized to maximize their translational potential.

[0008] There is therefore a need to prepare novel flavonoids with ~50X greater senolytic potency compared to naturally derived compounds such as fisetin or quercetin. There is also a need to prepare novel flavonoids that, in a preclinical model of lung fibrosis, promote clearance of senescence burden and resolution of chronic lung fibrosis, and enhance markers of mature alveolar epithelium. 07039-2361WO1 / 2025-008

[0009] SUMMARY

[0010] This document provides flavonoid compounds and methods and materials for using flavonoid compounds to treat mammals (e.g., humans) having one or more fibrotic and / or aberrant wound healing / s carring conditions disclosed herein. For example, this document provides flavonoid compounds having the structure of Formula (I) as well as methods and materials for using one or more flavonoid compounds having the structure of Formula (1). In some cases, one or more flavonoid compounds having the structure of Formula (I) can be administered to a mammal (e.g, a human) having one or more fibrotic and / or aberrant wound healing / scarring conditions to treat that condition within the mammal. As demonstrated herein, one or more flavonoid compounds having the structure of Formula (I) can induce apoptosis in senescent cells (e.g, senescent fibroblasts), and can be used to treat a fibrotic and / or aberrant wound healing / scarring condition within a mammal (e.g., a human).

[0011] In general, one aspect of this document features compositions including a flavonoid compound having a structure of Formula (I):

[0012]

[0013] ° (I),

[0014] or a pharmaceutically acceptable salt thereof, wherein R1, R2and A-G are as disclosed herein.

[0015] In one aspect this document features a method of treating a fibrotic and / or aberrant wound healing / scarring condition, comprising administering a compound of claim 1, or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0016] In one aspect this document features pharmaceutical compositions comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0017] The fibrotic and / or aberrant wound healing / scarring condition can be IPF, PSC, or NASH. The fibrotic and / or aberrant wound healing / scarring condition can be IPF, and the method can include administering an agent used to treat IPF to the mammal. The agent used to treat IPF can be pirfenidone, nintedanib, N-acetylcysteine, sildenafil, vardenafil, tadalafil, avanafil, promethazine, FTY720, AM 152, BMS-986020, VPC 12249, AM966, AM095, taribavirin, BI-2545, GLPG1690, BBT 877, SARI 00842, BMS-986,020, 07039-2361WO1 / 2025-008

[0018] minaprine, dopamine, levodopa, apomorphine, fenoldopam, pergolide, bromocriptine, cabergoline, dasatinib, hydroxyfasudil, ripasudil, netarsudil, belumosudil, lebrikizumab, tralokinumab, dupilumab, or pamrevlumab. The fibrotic and / or aberrant wound healing / scarring condition can be PSC, and the method can include administering an agent used to treat PSC to the mammal. The agent used to treat said PSC can be ursodeoxycholic acid (UDCA), a corticosteroid, a bile acid sequestrant. an antibiotic, or an antihistamine.

[0019] In another aspect, this document features methods for reducing fibrosis in a mammal having a fibrotic and / or aberrant wound healing / scarring condition. The methods can include, or consist essentially of, administering a composition including a flavonoid compound having a structure of Formula (I):

[0020]

[0021] O (I),

[0022] or a pharmaceutically acceptable salt thereof, to a mammal having a fibrotic and / or aberrant wound healing / scarring condition. The mammal can be a human. The method can include identifying the mammal as having the fibrotic and / or aberrant wound healing / scarring condition. The fibrotic and / or aberrant wound healing / scarring condition can be IPF, PSC, or NASH.

[0023] In another aspect, this document features methods for reducing a number of senescent cells in a mammal having a fibrotic and / or aberrant wound healing / scarring condition. The methods can include, or consist essentially of. administering a composition including a flavonoid compound having a structure of Formula (I):

[0024]

[0025] ° (I),

[0026] or a pharmaceutically acceptable salt thereof, to a mammal having a fibrotic and / or aberrant wound healing / scarring condition. 07039-2361WO1 / 2025-008

[0027] The mammal can be a human. The method can include identifying the mammal as having the fibrotic and / or aberrant wound healing / scarring condition. The

[0028] fibroti c / aberrant wound healing / scarring condition can be pulmonary interstitial lung diseases including: sarcoidosis, nonspecific interstitial pneumonitis, IPF, hypersensitivity pneumonitis, autoimmune disease-related ILD, and / or inherited / genetic / familial ILD. The condition can be liver associated including: NASH / MASH / NAFLD, autoimmune and viral hepatitis, alcoholic liver disease, and cirrhosis, and / or cholestatic liver / biliary diseases, including PSC and PBC.. The condition can be kidney and / or heart associated including: PKD, CKD, diabetes-associated kidney dysfunction, and myocardial infarction. The condition can be dermal associated including: scleroderma / systemic sclerosis, diabetes-associated non-healing wounds, hypertrophic scars, and bums. The condition can be skeletal muscle associated including muscular dystrophy. The condition can be ocular associated including: retinal fibrosis associated with macular degeneration, proliferative diabetic retinopathy, and proliferative retinopathy, or comeal fibrosis due to injury’ or post-surgical scarring. The condition can be GI related including: inflammatory bowel disease (IBD), such as Crohn’s disease and ulcerative colitis. The condition can also be kidney disease, polycystic kidney disease (PKD), diabetes, bums, wound healing, and muscular dystrophy and other degenerative diseases. The senescent cell can be a mesenchymal or of epithelial lineage, sometimes specific to that organ, for example, but not limited to, alveolar epithelial cells in the lung and cholangiocytes in the liver.

[0029] In another aspect, this document features methods for inhibiting a serine / threonine kinase 17 (STK17) polypeptide in a mammal. The methods can include, or consist essentially of, administering a composition including a flavonoid compound having a structure of Formula (I):

[0030]

[0031] O (I),

[0032] or a pharmaceutically acceptable salt thereof, to a mammal. The mammal can be a human. The STK17 polypeptide can be a STK17A (DRAK1) polypeptide or a STK17B (DRAK2) polypeptide. 07039-2361WO1 / 2025-008

[0033] In another aspect, this document features methods for reducing fibrosis in a mammal having a fibrotic and / or aberrant wound healing / scarring condition. The mammal can be a human. The method can include identifying the mammal as having the fibrotic and / or aberrant wound healing / scarring condition. The fibrotic and / or aberrant wound healing / scarring condition can be IPF, PSC, or NASH.

[0034] In another aspect, this document features methods for reducing a number of senescent cells in a mammal having a fibrotic and / or aberrant wound healing / scarring condition.

[0035] The mammal can be a human. The method can include identifying the mammal as having the fibrotic and / or aberrant wound healing / scarring condition. The

[0036] fibroti c / aberrant wound healing / scarring condition can be pulmonary interstitial lung diseases including: sarcoidosis, nonspecific interstitial pneumonitis, IPF, hypersensitivity pneumonitis, autoimmune disease-related ILD, and / or inherited / genetic / familial ILD. The condition can be liver associated including: NASH / MASH / NAFLD, autoimmune and viral hepatitis, alcoholic liver disease, and cirrhosis, and / or cholestatic liver / biliary diseases, including PSC and PBC.. The condition can be kidney and / or heart associated including: PKD, CKD, diabetes-associated kidney dysfunction, and myocardial infarction. The condition can be dermal associated including: scleroderma / systemic sclerosis, diabetes-associated non-healing wounds, hypertrophic scars, and bums. The condition can be skeletal muscle associated including muscular dystrophy. The condition can be ocular associated including: retinal fibrosis associated with macular degeneration, proliferative diabetic retinopathy, and proliferative retinopathy, or comeal fibrosis due to injury or post-surgical scarring. The condition can be GI related including: inflammatory bowel disease (IBD). such as Crohn’s disease and ulcerative colitis. The condition can also be kidney disease, polycystic kidney disease (PKD), diabetes, bums, wound healing, and muscular dystrophy and other degenerative diseases. The senescent cell can be a mesenchymal or of epithelial lineage, sometimes specific to that organ, for example, but not limited to, alveolar epithelial cells in the lung and cholangiocytes in the liver.

[0037] In another aspect, this document features methods for inhibiting a serine / threonine kinase 17 (STK17) polypeptide in a mammal. The mammal can be a human. The STK17 polypeptide can be a STK17A(DRAK1) polypeptide or a STK17B (DRAK2) polypeptide.

[0038] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this 07039-2361WO1 / 2025-008

[0039] invention pertains. Although methods and materials similar or equivalent to those described herein can be used to practice the invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0040] International PCT Patent Application Publication No. WO 2023 / 244619 is incorporated by reference herein in its entirety7. In case of conflict between WO ‘619 and the present specification, the latter will control.

[0041] The details of one or more embodiments of this document are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

[0042] DESCRIPTION OF THE DRAWINGS

[0043] Figure 1. Synthetic flavonols containing a para-ethoxy substitution on the B ring potently and selectively induces apoptosis in senescent lung fibroblasts. Human lung fibroblasts were passaged in culture (P= 18-20) until they presented with markers associated with cellular senescence, referred to as “Sen” and compared against the same cell lines at passage 3 “Pro” in assays measuring: A. cellular proliferation, B. qPCR analysis for transcript expression of cyclin dependent kinase inhibitors- CDKN2A and CDKN1A. and C senescence associated ^-galactosidase staining. D. Dose-response curves comparing natural product flavonols quercetin and fisetin to induce cleaved caspase-3 staining in Sen vs. Pro fibroblasts, measured by immunocytochemistry staining after cells were treated for 72 hours with the indicated concentration of flavonol. E. Structural diversity of commercially available flavonols. 2-dimensional structural overlay with each flavonol shown at 90% transparency and A, B, and C ring nomenclature identified. Below, a Tanimoto metrics similarity index was calculated, relative to quercetin for each flavonol to highlight the greater chemical diversity opportunity in synthetic flavonols compared to naturally derived. F. Senolytic cleaved caspase-3 staining for each of the most potent synthetic flavonols identified in the screen. All data are plotted as the mean + / - the SEM from three biologically independent experiments.

[0044] Comparisons made using t-test, *** p < 0.001, **** p < 0.0001 vs. the indicated group. 07039-2361WO1 / 2025-008

[0045] Figure 2. Synthetic flavonol F-19 supports clearance of senescence associated markers ex vivo and in vivo. A. Aged (18-month-old) mice were administered intratracheal ’‘sham” or bleomycin (Bleo) and 14 days after exposure the left lobe was harvested to generate 300 pm precision-cut lung slices that were cultured for 72 hours with F-19 (10 pM) or Nintedanib (1.0 pM) ex vivo, prior to homogenization, RNA isolation and qPCR analysis. N=4 sham or bleomycin mice. Comparisons made using ANOVA, * p < 0.05, ** p < 0.01 vs. the indicated group. B. Young (2-month-old) Colla2-mTmG, fibroblast lineage tracing mice were prelabeled with tamoxifen, X-days prior to intratracheal bleomycin administration. Starting on day 10 after Bleo injury, one group of mice was treated with F-19 (daily, 10 mg / kg, i.p.) for four days. C. On day 14 post Bleo the lungs were harvested for flow sorting (FACS) to collect populations of fibroblasts and epithelial cells for RNA isolation and qPCR analysis of Cdkn2a and Cdknla. All data are plotted as the mean + / - the SEM from three mice. Comparisons made using t-test, * p < 0.05 vs. the indicated group. D and E. Radar plots of additional transcripts analyzed from the fibroblasts and epithelial cell populations including senescence associated inflammatory cytokines, profibrotic ECM genes, markers of mature alveolar epithelial cells, and markers of aberrant epithelial cells associated with IPF. All data are plotted as the mean, F-19 treated mice (purple), relative to the mean, vehicle treated mice (red, set to 1). Data are also provided as individual plots in Figure 7.

[0046] Figure 3. Discovery of F-4N, a potent and highly selective senolytic flavonol. A.

[0047] Dose-response curves comparing F-4N capacity to induce cleaved caspase-3 staining in replicative Sen vs. Pro fibroblasts, measured by immunocytochemistry staining after cells were treated for 72 hours with the indicated concentration. B. Representative images of proliferating or replicative senescent lung fibroblasts treated for 72 hours + / -F-4N (3.0 pM) then stained for Cleaved-Caspase-3 and DAPI. Scale bar represents 100 pm. C. Caspase 3 / 7 activity measured by a luminescent substrate after 72 hours + / -F-4N (3.0 pM). D. Flow cytometry-based analysis of early and late apoptosis measured by Annexin V and Propidium Iodide staining. Shown is a representative contour plot from an experiment with senescent fibroblasts treated with F-4N for 72 hours and quantification of early and late apoptosis + / -F-4N (3.0 pM) in proliferating and senescent cells. N=3 independent experiments (***p<0.001, ****p<0.0001 vs. the indicated group).

[0048] Figure 4. F-4N efficacy in a bleomycin-induced model of lung fibrosis in aged mice A. Survival and study design. Bleomycin was administered one time intratracheally, on day 14 mice began receiving F-4N (10 mg / kg, daily i.p.) or vehicle and lungs collected 07039-2361WO1 / 2025-008

[0049] on day 28. B. Representative Masson’s trichrome stained histological sections and hydroxyproline content measured from whole left lung. Scale bar = 100 pm C. RNA Expression of senescence associated markers, ECM genes, and mature alveolar epithelial markers from whole lung at the completion of the study. D. Plasma IL-6 levels. N= 7-9 mice / group, male and female mice (*p<0.05, **p<0.01, ***p<0.001 vs. the indicated group).

[0050] Figure 5. F-4N efficacy in a repeated bleomycin-induced model of lung fibrosis.

[0051] A. Body mass and study design. Bleomycin was administered three times intratracheally, starting on day 0 and repeated on days 14 and 28. Three weeks after the final bleomycin administration, mice began receiving F-4N (10 mg / kg, daily i.p.) or vehicle for two weeks prior to lung collection. B. Representative Masson’s tri chrome stained histological sections and hydroxyproline content measured from whole left lung. Scale bar = 100 pm C. RNA Expression of senescence associated markers, ECM genes, mature alveolar epithelial markers, and aberrant epithelial cells markers from whole lung at the completion of the study. N= 3-7 mice / group, male and female mice (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 vs. the indicated group).

[0052] Figure 6. Synthetic flavonols containing a para-ethoxy substitution on the B ring reduce cell number in senescent lung fibroblasts. Human lung fibroblasts were passaged in culture (P= 18-20) then treated for 72 hours with the indicated flavonol (1, 10, and 100 pM). After incubation cells were fixed and stained with DAPI. Cells / Field of view were calculated using automated software. N=3 biologically independent experiments.

[0053] Figure 7. Individual datapoints from radar plots shown in Figure 2.

[0054] Figure 8. F-4N induces cleaved caspase-3 expression in senescent fibroblasts derived from multiple stimuli. Senescence was induced by replication, radiation (20 Gy of X-ray), etoposide (20 pM), or bleomycin (10 p). Cells were then incubated for 72 hours + / - F-4N (3 pM) prior to fixation and staining for cleaved caspase-3 and DAPI. N=3 independent experiments (**** p < 0.0001, vs. the indicated group).

[0055] Figure 9. Synthetic flavonol F-4N supports clearance of senescence associated markers ex vivo and in vivo. Aged (18-month-old) mice were administered intratracheal “sham’' or bleomycin (Bleo) and 14 days after exposure the left lobe was harvested to generate 300 pm precision-cut lung slices that were cultured for 72 hours with F-4N (3 pM), ABT-263 (5.0 pM), or ABT-199 (3 pM) ex vivo, prior to homogenization, RNA 07039-2361WO1 / 2025-008

[0056] isolation and qPCR analysis. N=6 sham or bleomycin mice. Comparisons made using ANOVA, * p < 0.05. ** p < 0.01 vs. the indicated group.

[0057] Figure 10 shows that administration of F-4N reverses fibrosis in a murine model of steatohepatitis (MASH / NASH) with metabolic syndrome. Male C57B1 / 6J mice were maintained on either a standard chow diet or high-fat, high-fructose, high-cholesterol (FFC) diet to induce steatohepatitis and fibrosis. After 24 weeks of feeding, FFC-fed mice were randomized to receive either vehicle (com oil) or F-4N (10 mg / kg of body weight). F-4N or vehicle was administered via intraperitoneal injection five times per week (Monday through Friday) for 6 weeks while continuing chow7or FFC diet. (A) Body weight throughout the study. (B) Body weight at the endpoint. (C) Liver weight to body¬ weight ratio. (D) Epididymal white adipose tissue (eWAT) weight normalized to body weight. (E) Plasma alanine aminotransferase (ALT) activity7as a marker of liver injury. (F) Plasma cholesterol. (G) Liver fibrosis was visualized by Sirius Red staining and polarized light microscopy. Scale bar = 50 pm (H) Quantification of collagen deposition using Sirius Red staining and polarized light microscopy. Statistical test: ANOVA followed by Tukey’s post hoc test. Sample sizes are indicated in the graphs. *p < 0.05, **p < 0.01, ***p < 0.001, ns, non-significant.

[0058] DETAILED DESCRIPTION

[0059] This document provides flavonoid compounds and methods and materials for using flavonoid compounds to treat mammals (e.g., humans) having one or more fibrotic and / or aberrant wound healing / s carring conditions (e.g., IPF, NASH, and PSC). For example, this document provides flavonoid compounds having the structure of Formula (I):

[0060]

[0061] ° (I),

[0062] or a pharmaceutically acceptable salt thereof,

[0063] wherein 07039-2361WO1 / 2025-008

[0064] R1is selected from the group consisting ofH, alkyl, substituted alkyl, cyclo-alkyl, substituted cyclo-alkyl, and OH;

[0065] R2is selected from the group consisting of H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, and optionally substituted heterocyclyl;

[0066] A, B, C are each independently N or CR3;

[0067] each R3is independently H or halogen;

[0068] D is N or CR4;

[0069] R4is H, halogen, OR5, NR6R7, optionally substituted alkyl, or optionally substituted cycloalkyl;

[0070] R5, R6, R7are each independently hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl;

[0071] E and F are each independently N or CR8;

[0072] each R8is independently H, halogen, OR9, NR10R11, optionally substituted alkyl, or optionally substituted cycloalkyl;

[0073] R9, R10, R11are each independently hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl;

[0074] G is O, NR12, S, SO, SO2;

[0075] R12is hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl and optionally substituted heterocyclyl;

[0076] and

[0077] each ring bond in the ring containing A, B, C and D is a single bond or a double bond, provided that no more than three ring bonds are double bonds, and provided that if all ring bonds are single bonds, then at least one of A. B, C or D is N.

[0078] In some embodiments, R1is OH.

[0079] In some embodiments, R2is optionally substituted alkyl.

[0080] In some embodiments, R2is substituted alkyl.

[0081] In some embodiments, R2is alkyl substituted with C1-C6 alkoxy, such as ethoxy. In some embodiments, R2is alkyl substituted with carbocyclyl, such as alkyl substituted with phenyl.

[0082] In some embodiments, R2is alkyl substituted with heterocyclyl, such as alkyl substituted with heteroaryl, such as alkyl substituted with pyridinyl, such as alkyl substituted with 2-pyridinyl. or as alkyl substituted with 3-pyridinyl, or as alkyl substituted with 4-pyridinyl. 07039-2361WO1 / 2025-008

[0083] In some embodiments, A is N.

[0084] In some embodiments, B is N.

[0085] In some embodiments, C is N.

[0086] In some embodiments, D is N.

[0087] In some embodiments, one A, B, C or D is N, and the remaining three are not N. In some embodiments, E is CR8.

[0088] In some embodiments, F is CR8.

[0089] In some embodiments, each of E and F is CR8

[0090] In some embodiments, G is O.

[0091] In some embodiments, provided herein is a compound as disclosed in Table 1 herein, or a pharmaceutically acceptable salt thereof.

[0092] In one embodiment provided herein is a compound as disclosed in Figure 6 herein, or a pharmaceutically acceptable salt thereof.

[0093] In one embodiment provided herein is a compound selected from the group consisting of the following compounds, or a pharmaceutically acceptable salt thereof

[0094]

[0095] 07039-2361WO1 / 2025-008

[0096]

[0097] 07039-2361WO1 / 2025-008

[0098] (“F-CB ’), and

[0099]

[0100] (“F-L132").

[0101] Salts

[0102] In some cases, a flavonoid compound provided herein (e.g., a flavonoid compound having the structure of Formula (I)) can be in the form of a salt (e.g., pharmaceutically acceptable salt). A salt of a compound provided herein can be formed between an acid and a basic group of the compound, such as an amino functional group, or a base and an acidic group of the compound, such as a carboxyl functional group. When a flavonoid compound having the structure of Formula (I) is in the form of a salt, the salt can include any appropriate acid (e.g., an organic acid or an inorganic acid).

[0103] Examples of acids that can be used to form a pharmaceutically acceptable salt of a compound described herein include, without limitation, inorganic acids such as hydrogen bisulfide, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid and phosphoric acid, as well as organic acids such as para-toluenesulfonic acid, salicylic acid, tartaric acid, bitartaric acid, ascorbic acid, maleic acid, besylic acid, fumaric acid, gluconic acid, glucuronic acid, formic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, lactic acid, oxalic acid, parabromophenylsulfonic acid, carbonic acid, succinic acid, citric acid, benzoic acid and acetic acid, as well as related inorganic and organic acids. Such pharmaceutically acceptable salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, 07039-2361WO1 / 2025-008

[0104] isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne- 1.4-dioate, hexyne-l,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxy benzoate, phthalate, terephthalate, sulfonate, xylene sulfonate, phenyl acetate, phenylpropionate, phenylbutyrate, citrate, lactate, 0-hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene- 1 -sulfonate, naphthalene-2- sulfonate, mandelate and other salts. In some embodiments, pharmaceutically acceptable acid addition salts can be used including, without limitation, those formed with mineral acids such as hydrochloric acid and hydrobromic acid, and those formed with organic acids such as maleic acid. Examples of bases that can be used to form a pharmaceutically acceptable salt of a compound described herein include, without limitation, hydroxides of alkali metals, including sodium, potassium, and lithium; hydroxides of alkaline earth metals such as calcium and magnesium; hydroxides of other metals, such as aluminum and zinc; ammonia, organic amines such as unsubstituted or hydroxyl-substituted mono-, di-, or tri-alkylamines, dicyclohexylamine; tributyl amine; pyridine; N-methyl, N-ethylamine; diethylamine; triethylamine; mono-, bis-, ortris-(2-OH-(Cl-C6)-alkylamine), such as N, N-dimethyl-N-(2-hydroxyethyl)amine or tri-(2-hydroxyethyl)amine; N-methyl-D-glucamine; morpholine; thiomorpholine; piperidine; pyrrolidine; and amino acids such as arginine, lysine, and the like. In some cases, a compound described herein, or a pharmaceutically acceptable salt thereof, can be substantially isolated.

[0105] Definitions

[0106] At various places herein, substituents of compounds described herein are described in groups or in ranges. It is specifically intended that this document include and describe each and every individual member or subcombination of the members of such groups and ranges. For example, the term “Ci-6 alkyl” is specifically intended to individually disclose methyl, ethyl, C3 alkyl, C4 alky l, C5 alky l, and Ce alkyl.

[0107] At various places herein, various aryl, heteroaryl, cycloalkyl, and heterocycloalkyd rings are described. Unless otherwise specified, these rings can be attached to the rest of the molecule at any ring member as permitted by valency. For example, the term “a pyridine ring” or “pyridinyl” may refer to a pyridin-2-yl, pyridin-3-yl, or pyridin-4-yl ring.

[0108] It is further appreciated that certain features herein, which are, for clarity, described in the context of separate embodiments, can also be provided in combination in 07039-2361WO1 / 2025-008

[0109] a single embodiment Conversely, various features herein which are, for brevity', described in the context of a single embodiment, can also be provided separately or in any suitable subcombination.

[0110] The term “compound” as used herein is meant to include all stereoisomers, geometric isomers, tautomers, and isotopes of the structures depicted. Compounds herein identified by name or structure as one particular tautomeric form are intended to include other tautomeric forms unless otherwise specified.

[0111] The term “n-membered” where n is an integer ty pically describes the number of ring-forming atoms in a moiety' where the number of ring-forming atoms is n. For example, piperidinyl is an example of a 6-membered heterocy cloalkyl ring, pyrazolyl is an example of a 5-membered heteroaryl ring, pyridyl is an example of a 6-membered heteroaryl ring, and 1,2,3,4-tetrahydro-naphthalene is an example of a 10-membered cycloalkyl group.

[0112] As used herein, the phrase “optionally substituted” means unsubstituted or substituted. The substituents are independently selected, and substitution may be at any chemically accessible position. As used herein, the term “substituted” means that a hydrogen atom is removed and replaced by a substituent. A single divalent substituent, e.g., oxo, can replace two hydrogen atoms. It is to be understood that substitution at a given atom is limited by valency.

[0113] As used herein, “substituted”, when used to describe an alkyd, alkylene, alkoxy, acyl, aryl, aroyl, cycloalkyl, and heterocyclyl, and / or heterocycloalkyl group, refers to a group substituted by one or more substituents. For example, — (C 1-9 alkyl) may be optionally substituted with one or more of hydroxyl, — NH2, — NH(CI-3 alkyl), and — N(CI-3 alkyl)2; — (C 1-9 haloalky 1); halide; hydroxyl; a carbonyl [such as — C(O)OR, and — C(O)R]; a thiocarbonyl [such as — C(S)OR, — C(O)SR, and — C(S)R]; — (C1-9 alkoxy) optionally substituted with one or more of halide, hydroxyl, — NH2, — NH(Ci-s alkyl), and — N(CI-3 alkyl)2; — OPO(OH)2; a phosphonate [such as — PO(OH)2 and — PO(OR')2]; — OPO(OR')R"; — NRR'; — C(0)NRR'; — C(NR)NR'R"; — C(NR')R"; a cyano; a nitro; an azido; — SH; — S — R; — OSChCOR); a sulfonate [such as — SCh OH) and — SCh(OR)]; — SChNR'R"; and — SO2R; in which each occurrence of R, R' and R" are independently selected from H; — (C1-9 alkyd); Ce-io ar l optionally substituted with from 1-3R'"; 5-10 membered heteroaryl having from 1-4 heteroatoms independently selected from N, O, and S and optionally substituted with from 1-3 R'"; C3-7 carbocyclyl optionally substituted with from 1-3 R”'; and 3-10 membered heterocyclyl having from 1- 07039-2361WO1 / 2025-008

[0114] 4 heteroatoms independently selected from N, O, and S and optionally substituted with from 1-3 R"; wherein each R”' is independently selected from — (Ci-6 alkyl). — (Ci-ehaloalkyl), a halide (e.g., F), a hydroxyl, — C(O)OR, — C(O)R, — (Ci-6 alkoxyl), — NRR', — C(O)NRR', and a cyano, in which each occurrence of R and R' is independently selected from H and — (Ci-6 alkyl). In some embodiments, the substituent is selected from — (C i-6 alkyl), — (C i-6 haloalky 1), a halide (e.g., F), a hydroxyl, — C(O)OR, — C(O)R, — (Ci-6 alkoxyl), — NRR', — C(O)NRR', and a cyano, in which each occurrence of R and R' is independently selected from H and — (Ci-6 alkyl). Examples of substituted — (Ci-9 alky l) include — (C1-9 alkyl) substituted with carbocyclyl, such as — (C1-9 alkyl) substituted with phenyl. Examples of — (C1-9 alkyl) substituted with — (C1-9 alkyl) substituted with heterocyclyl, such as (C1-9 alkyl) substituted with heteroaryl, such as (Ci-9 alkyl) substituted with pyridinyl, such as (C1-9 alkyl) substituted with 2-pyridinyl, or (Ci-9 alky l) substituted with 3-pyridinyl, or as (C1-9 alkyl) substituted with 4-pyridinyl.

[0115] Throughout the definitions, the term “Cn-m” indicates a range which includes the endpoints, wherein n and m are integers and indicate the number of carbons. Examples include Ci-4, C1-6, and the like. As used herein, the term ‘‘Cn-m alkyl”, used alone or in combination with other terms, refers to a saturated hydrocarbon group that may be straight-chain or branched, having n to m carbons. Examples of alkyd moieties include, without limitation, chemical groups such as methy l, ethyl, n-propyl, isopropyl, / i-butyl, / e / 7-butyl. isobutyl, sec-butyl; higher homologs such as 2-methyl-l -butyl, w-pentyl, 3-pentyl, w-hexyl, 1,2,2-trimethylpropyl, and the like. In some embodiments, the alkyl group contains from 1 to 20 carbon atoms, such as from 1 to 8 carbon atoms, from 1 to 6 carbon atoms, from 1 to 4 carbon atoms, from 1 to 3 carbon atoms, or 1 to 2 carbon atoms.

[0116] As used herein, the term "‘Cn-m alkydene”, used alone or in combination with other terms, refers to a divalent alkyd linking group having n to m carbons. Examples of alkylene groups include, without limitation, ethan- 1,1 -diyl, ethan-l,2-diyd, propan- 1,1,-diyl, propan-1, 3-diyl, propan- 1,2-diyd, butan-l,4-diyl, butan-l,3-diyl, butan-l,2-diyl, 2-methyl-propan-l,3-diyl, and the like. In some embodiments, the alkylene moiety contains 2 to 6, 2 to 4, 2 to 3, 1 to 6, 1 to 4, or 1 to 2 carbon atoms.

[0117] As used herein, the term “Cn-m alkoxy”, used alone or in combination with other terms, refers to a group of formula -O-alkyl, wherein the alkyl group has n to m carbons. Example alkoxy groups include, without limitation, methoxy, ethoxy, propoxy (e.g., n- 07039-2361WO1 / 2025-008

[0118] propoxy and isopropoxy), butoxy (e.g., n-butoxy and tert-butoxy), and the like. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0119] As used herein, “aryl’’ means a mono-, bi-, tri- or polycyclic group with only carbon atoms present in the ring backbone having 5 to 14 ring atoms, alternatively 5, 6, 9, or 10 ring atoms; and having 6, 10, or 14 pi electrons shared in a cyclic array; wherein at least one ring in the system is aromatic. Aryl groups can either be unsubstituted or substituted with one or more substituents. Examples of aryl include phenyl, naphthyl, tetrahydronaphthyl, 2,3-dihydro-lH-indenyl, and others. In some embodiments, the aryl is phenyl.

[0120] As used herein, “cycloalkyl” refers to non-aromatic cyclic hydrocarbons including cyclized alkyl and / or alkenyl groups. Cycloalkyl groups can include mono- or polycyclic (e.g., having 2, 3 or 4 fused rings) groups and spirocycles. Ring-forming carbon atoms of a cycloalkyl group can be optionally substituted by 1 or 2 independently selected oxo or sulfide groups (e.g., C(O) or C(S)). Also included in the definition of cycloalkyl are moi eties that have one or more aromatic rings fused (i. e., having a bond in common with) to the cycloalkyl ring, for example, benzo or thienyl derivatives of cyclopentane, cyclohexane, and the like. A cycloalkyl group containing a fused aromatic ring can be attached through any ring-forming atom including a ring-forming atom of the fused aromatic ring. Cycloalkyl groups can have 3, 4, 5, 6, 7, 8, 9, or 10 ring-forming carbons (C3-10). In some embodiments, the cycloalkyl is a C3-10 monocyclic or bicyclic cyclocalkyl. In some embodiments, the cycloalkyl is a C3-7 monocyclic cyclocalkyl. Example cycloalkyl groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cycloheptatrienyl, norbomyl, norpinyl, norcamyl, adamantyl, and the like. In some embodiments, cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0121] As used herein, “heterocycle”, “heterocyclic” and “heterocyclyl” refer to an optionally substituted, fully saturated or unsaturated, aromatic or nonaromatic cyclic group, for example, which is a 3 to 7 membered monocyclic, 7 to 11 membered bicyclic, or 10 to 15 membered tricyclic ring system, which has at least one heteroatom in at least one carbon atom-containing ring. Each ring of the heterocyclic group containing a heteroatom may have 1, 2 or 3 heteroatoms selected from nitrogen atoms, oxygen atoms and sulfur atoms, where the nitrogen and sulfur heteroatoms may also optionally be oxidized and the nitrogen heteroatoms may also optionally be quaternized. The heterocyclic group may be attached at any heteroatom or carbon atom 07039-2361WO1 / 2025-008

[0122] Exemplary monocyclic heterocyclic groups include pyrrolidiny 1, pyrrolyl, indolyl, pyrazolyl, oxetanyl, pyrazolinyl. imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl, oxazolidinyl, isoxazolinyi, isoxazolyl, thiazolyl, thiadiazolyl, thiazolidinyi, isothiazolyl, isothiazolidinyl, furyl, tetrahydrofuryl, thienyl, oxadiazolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxazepinyl, azepinyl, 4- piperidonyl, pyridyl, N-oxo-pyridyl, pyrazinyl, pyritnidinyl, pyridazinyl, tetrahydropyranyl, morpholinyL thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, 1,3-dioxolane and tetrahydro-1, 1 -dioxothieny 1, dioxanyl, isothiazolidinyl, thietanyl, thiiranyl, tnazinyl, triazolyl, oxiranyl, azirldmyl, and the like.

[0123] Exemplary bicyclic hetrocyclic groups include 2,3-dihydro-2-oxo-lH-indoly1, benzothiazolyl, benzoxazolyl, benzothienyl, quinuclidinyl, quinolinyl, quinoIinyl-N- oxide, tetrahydroisoquinolmyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl, benzofuryl, chromonyl, coumarinyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl, furopyridinyl (such as furo[2,3-c]pyridinyl, furo[3,l-b]pyridinyl] or furo[2,3-b]pyridinyl), dihydroisoindolyl, dihydroquinazolinyl (such as 3,4-dihydro-4-oxo-quinazolinyl), benzisothiazolyl, benzisoxazolyl, benzodiazinyl, benzofurazanyL benzothiopyranyl, benzotriazolyl, benzpyrazolyl, dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl, dihydrobenzothiopyranyl sulfone, dihydrobenzopyranyl, indolmyl, isochromanyl, isoindolinyl, naphthyridinyl, phthalazinyl, piperonyl, purinyl, pyridopyridyl, quinazolinyl, tetrahydroquinolinyl, thienofuryl, thienopyridyl, thienothienyl. and the like.

[0124] As used herein, the term “heteroaryl” means a mono-, bi-, tri- or polycyclic group having 5 to 14 ring atoms, alternatively 5, 6, 9, or 10 ring atoms; and having 6, 10, or 14 pi electrons shared in a cyclic array; wherein at least one ring in the system is aromatic, and at least one ring in the system contains one or more heteroatoms independently selected from the group consisting of N, O, and S. Heteroaryl groups can either be unsubstituted or substituted with one or more substituents. Examples of heteroaryl include thienyl, pyridinyl, furyl, oxazolyl, oxadiazolyl, pyrrolyl, imidazolyl, triazolyl, thiodiazolyl, pyrazolyl, isoxazolyl, thiadiazolyl, pyranyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, thiazolyl benzothienyl, benzoxadiazolyl, benzofuranyl, benzimidazolyl, benzotriazolyl, cinnolinyl, indazolyl, indolyl, isoquinolinyl, isothiazolyl, naphthyridinyl. purinyl, thienopyridinyl, pyrido[2,3-d]pyrimidinyl, pyrrolo[2,3-b]pyridinyl, quinazolinyl, quinolinyl, thieno[2,3-c]pyridinyl, pyrazolo[3,4-b]pyridinyl. pyrazolo[3,4-c]pyridinyl, pyrazolo[4,3-c]pyridine, pyrazolo[4,3-b]pyridinyl, tetrazolyl, 07039-2361WO1 / 2025-008

[0125] chromane, 2,3-dihydrobenzo[b][l,4]dioxine, benzo[d][ 1,3] di oxole, 2,3-dihydrobenzofuran, tetrahydroquinoline, 2,3-dihydrobenzo[b][l,4]oxathiine, isoindoline, and others. In some embodiments, the heteroaryl is selected from thienyl, pyridinyl, furyl, pyrazolyl, imidazolyl, isoindolinyl, pyranyl, pyrazinyl, and pyrimidinyl.

[0126] As used herein, “heterocycloalkyl” means a nonaromatic cyclic ring system comprising at least one heteroatom in the ring system backbone. Heterocycloalkyls may include multiple fused rings. Heterocycloalkyls may be substituted or unsubstituted with one or more substituents. In some embodiments, heterocycles have 3-11 members. In six membered monocyclic heterocycles, the heteroatom(s) are selected from one to three of O, N or S, and wherein when the heterocycle is five membered, it can have one or two heteroatoms selected from O, N. or S. Examples of heterocycloalkyls include azirinyl, aziridinyl, azetidinyl, oxetanyl, thietanyl, 1,4,2-dithiazolyl, dihydropyridinyl, 1,3-dioxanyl, 1,4-dioxanyl, 1,3-dioxolanyl, morpholinyl, thiomorpholinyl, piperazinyl, pyranyl, pyrrolidinyl, tetrahydrofuryl, tetrahydropyridinyl, oxazinyl, thiazinyl, thiinyl, thiazolidinyl. isothiazolidinyl, oxazolidinyl, isoxazolidinyl, piperidinyl. pyrazolidinyl imidazolidinyl, thiomorpholinyl, and others. In some embodiments, the heterocycloalkyl is selected from azetidinyl, morpholinyl, piperazinyl, pyrrolidinyl, and tetrahydropyridinyl.

[0127] Compounds provided herein also include tautomeric forms. Tautomeric forms result from the swapping of a single bond with an adjacent double bond together with the concomitant migration of a proton. Tautomeric forms include prototropic tautomers which are isomeric protonation states having the same empirical formula and total charge. Example prototropic tautomers include, without limitation, ketone - enol pairs, amide -imidic acid pairs, lactam - lactim pairs, enamine - imine pairs, and annular forms where a proton can occupy two or more positions of a heterocyclic system, for example, 1H- and 3H-imidazole, 1H-, 2H- and 4H- 1,2,4-triazole, 1H- and 2H- isoindole, and 1H- and 2H-pyrazole. Tautomeric forms can be in equilibrium or sterically locked into one form by appropriate substitution.

[0128] In some cases, one or more flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can lack chirality.

[0129] In some cases, one or more flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be a neutral molecule (e.g.. can lack any charged moiety). 07039-2361WO1 / 2025-008

[0130] In some cases, one or more flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can lack any catechol moiety.

[0131] In some cases, one or more flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be formulated into a composition (e.g., a pharmaceutically acceptable composition) for administration to a mammal (e.g, a human) having one or more fibrotic and / or aberrant wound healing / scarring conditions (e.g., IPF, NASH, and PSC). For example, one or more flavonoid compounds having the structure of Formula (I) can be formulated together with one or more pharmaceutically acceptable carriers (additives), excipients, and / or diluents. Examples of pharmaceutically acceptable carriers, excipients, and diluents that can be used in a composition described herein include, without limitation, cyclodextrins (e.g., beta-cyclodextrins such as KLEPTOSE"). dimethylsulfoxide (DMSO), sucrose, lactose, starch (e.g., starch glycolate), cellulose, cellulose derivatives (e.g., modified celluloses such as microcrystalline cellulose, and cellulose ethers like hydroxypropyl cellulose (HPC) and cellulose ether hydroxypropyl methylcellulose (HPMC)), xylitol, sorbitol, mannitol, gelatin, polymers (e.g., polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), crosslinked polyvinylpyrrolidone (crospovidone), carboxymethyl cellulose, polyethylene-polyoxypropylene-block polymers, and crosslinked sodium carboxymethyl cellulose (croscarmellose sodium)), titanium oxide, azo dyes, silica gel. fumed silica, talc, magnesium carbonate, vegetable stearin, magnesium stearate, aluminum stearate, stearic acid, antioxidants (e.g., vitamin A, vitamin E, vitamin C, retinyl palmitate, and selenium), citric acid, sodium citrate, parabens (e.g., methyl paraben and propyl paraben), petrolatum, dimethyl sulfoxide, mineral oil, serum proteins (e.g, human serum albumin), glycine, sorbic acid, potassium sorbate, water, salts or electrolytes (e.g., saline, protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, and zinc salts), colloidal silica, magnesium trisilicate, polyacrylates, waxes, wool fat, lecithin, and com oil.

[0132] In some cases, a composition containing one or more flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be designed for oral or parenteral (including, without limitation, a subcutaneous, intramuscular, intravenous, intradermal, intra-cerebral, intrathecal, or intraperitoneal (i.p.) injection) administration to a mammal. Compositions suitable for oral administration include, without limitation, liquids, tablets, capsules, pills, powders, gels, and granules. In 07039-2361WO1 / 2025-008

[0133] some cases, compositions suitable for oral administration can be in the form of a food supplement. In some cases, compositions suitable for oral administration can be in the form of a drink supplement. Compositions suitable for parenteral administration include, without limitation, aqueous and non-aqueous sterile injection solutions that can contain anti-oxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient.

[0134] This document also provides methods for making one or more flavonoid compounds described herein (e.g., one or more flavonoid compounds having the structure of Formula (I). Any appropriate method can be used to make one or more flavonoid compounds provided herein. In some cases, a flavonoid compound having the structure of Formula (I) can be made as show in Figure 1. In some cases, a flavonoid compound having the structure of Formula (I) can be made as described in Example 1.

[0135] This document also provides methods for using one or more flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)). For example, one or more flavonoid compounds having the structure of Formula (I) can be administered to a mammal (e.g., a human) having one or more fibrotic and / or aberrant wound healing / s carring conditions (e.g., IPF, NASH, and PSC) to treat the mammal. In some cases, a mammal (e.g., a human) having one or more fibrotic and / or aberrant wound healing / s carring conditions (e.g., IPF, NASH, and PSC) can be administered or instructed to self-administer one or more flavonoid compounds having the structure of Formula (I).

[0136] In some cases, one or more (e.g, one, two, three, four, or more) flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be used to reduce or eliminate of one or more symptoms of one or more fibrotic and / or aberrant wound healing / scarring conditions. For example, a composition including one or more flavonoid compounds having the structure of Formula (I) can be administered to a mammal (e.g., a human) in need thereof (e.g., a human having one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF, NASH, and PSC) to reduce or eliminate one or more symptoms of the fibrotic and / or aberrant wound healing / scarring condition (e.g, IPF, NASH, and PSC). Examples of symptoms of IPF disease include, without limitation, shortness of breath (dyspnea), persistent dry cough, tiredness, loss of appetite and weight loss, aching muscles and joints, and clubbing, which is widening and rounding of the tips of the fingers or toes. Examples of symptoms of PSC disease include, without limitation, feeling 07039-2361WO1 / 2025-008

[0137] tired or weak, itchy skin, pain in the abdomen, losing weight without try ing, poor appetite, fever, enlarged liver, enlarged spleen, yellow eyes and skin (jaundice), symptoms of cirrhosis and liver failure such as bloating, bruising and bleeding easily, confusion, difficulty thinking or memory loss, redness in the palms of hands, and swelling in legs, ankles or feet. In some cases, one or more (e.g, one, two, three, four, or more) flavonoid compounds provided herein (e.g, one or more flavonoid compounds having the structure of Formula (I)) can be used to reduce one or more symptoms of fibrotic and / or aberrant wound healing / scarring condition in a mammal having one or more fibrotic and / or aberrant wound healing / scarring conditions by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent.

[0138] In some cases, one or more (e.g.. one, two. three, four, or more) flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be used to reduce or eliminate one or more complications associated with a fibrotic and / or aberrant wound healing / scarring condition. For example, a composition including one or more flavonoid compounds having the structure of Formula (I) can be administered to a mammal (e.g, a human) in need thereof (e.g, a human having one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF, NASH, and PSC) to reduce or eliminate one or more complications associated with the fibrotic and / or aberrant wound healing / scarring condition. Examples of complications associated with IPF include, without limitation, pulmonary hypertension, acute exacerbation of pulmonary fibrosis, respiratory infection, acute coronary syndrome, thromboembolic disease, adverse medication effects, and lung cancer. Examples of complications associated with PSC include, without limitation, low levels of fat-soluble vitamins, osteoporosis, bile duct infection, portal hypertension, cirrhosis, liver failure, bile duct cancer, gall bladder cancer, colon cancer, and hepatocellular carcinoma. In some cases, one or more (e.g., one, two, three, four, or more) flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be used to reduce one or more complications associated with one or more fibrotic and / or aberrant wound healing / scarring conditions in a mammal having one or more fibrotic and / or aberrant wound healing / scarring conditions by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent.

[0139] In some cases, one or more (e.g, one, two. three, four, or more) flavonoid compounds provided herein (e.g, one or more flavonoid compounds having the structure of Formula (I)) can be used as an anti-fibrotic agent. For example, one or more flavonoid 07039-2361WO1 / 2025-008

[0140] compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be used to reduce or eliminate fibrotic scarring in a mammal (e.g. in one or more tissues within a mammal). For example, one or more flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be used to slow the progression of fibrosis in mammal (e.g., in one or more tissues within a mammal).

[0141] In some cases, one or more (e.g, one, two. three, four, or more) flavonoid compounds provided herein (e.g, one or more flavonoid compounds having the structure of Formula (I)) can be used to reduce or eliminate fibrotic scarring in a mammal. For example, a composition including one or more (e.g., one, two, three, four, or more) flavonoid compounds having the structure of Formula (I) can be administered to a mammal (e.g., a human) in need thereof (e.g., a human having one or more fibrotic and / or aberrant wound healing / s carring conditions such as IPF, NASH, and PSC) to reduce or eliminate fibrotic scarring in one or more tissues within the mammal. One or more flavonoid compounds provided herein can be used to reduce or eliminate fibrotic scarring in any appropriate tissue within a mammal. Examples of tissues that can have fibrotic scars and that one or more flavonoid compounds provided herein can be used to reduce or eliminate fibrotic scarring in include, without limitation, lung, liver, bile ducts, kidney, heart, and skin. In some cases, one or more (e.g., one, two, three, four, or more) flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be used to reduce fibrotic scarring in one or more tissues within a mammal having fibrotic scarring by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent.

[0142] In some cases, one or more (e.g. one, two. three, four, or more) flavonoid compounds provided herein (e.g, one or more flavonoid compounds having the structure of Formula (I)) can be used to slow the progression of fibrosis in a mammal. For example, a composition including one or more flavonoid compounds having the structure of Formula (I) can be administered to a mammal (e.g., a human) in need thereof (e.g, a human having one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF, NASH, and PSC) to slow the progression of fibrosis in the mammal. One or more flavonoid compounds provided herein can be used to slow the progression of fibrosis in any appropriate tissue within a mammal. Examples of tissues that can be fibrotic and that one or more flavonoid compounds provided herein can be used to slow the progression of fibrosis in include, without limitation, lung, liver, bile ducts, kidney, 07039-2361WO1 / 2025-008

[0143] heart and skin. In some cases, one or more (e.g., one, two, three, four, or more) flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be effective to slow the progression of fibrosis in a mammal having fibrosis by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent. In some cases, one or more (e.g., one, two, three, four, or more) flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be used to slow the progression of fibrosis in a mammal having fibrosis by, for example, at least 6 months (e.g., about 6 months, about 8 months, about 10 months, about 1 year, about 1.5 years, about 2 years, about 2.5 years, about 3 years, about 4 years, about 5 years, or more).

[0144] In some cases, one or more (e.g.. one, two. three, four, or more) flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be used as a senolytic agent. For example, one or more flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be used to induce apoptosis in one or more senescent cells within a mammal. In some cases, one or more flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can exhibit little, or no, ability to induce apoptosis in proliferating cells within a mammal (e.g., a human).

[0145] In some cases, one or more (e.g, one, two. three, four, or more) flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be used to inhibit one or more serine / threonine kinase 17 (STK17) polypeptides. For example, a flavonoid compound having the structure of Formula (I) can bind to a STK17 polypeptide to inhibit polypeptide function of the STK17 polypeptide], A flavonoid compound having the structure of Formula (I) can inhibit any appropriate STK17 polypeptide. Examples of STK17 polypeptides that be inhibited by one or more flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) include, without limitation, STK17A (DRAK1) polypeptides, and STK17B (DRAK2) polypeptides. In some cases, a flavonoid compound having the structure of Formula (I) can inhibit a STK17A (DRAK1) polypeptide set forth in any one of National Center for Biotechnology Information (NCBI) GenBank® or GenPept® Accession Nos. 9263 and Q9UEE5. In some cases, a flavonoid compound having the structure of Formula (I) can inhibit a STK17B (DRAK2) polypeptide set forth in any one of NCBI GenBank® or GenPept® Accession Nos. 9262 and 094768. 07039-2361WO1 / 2025-008

[0146] In some cases, one or more (e.g., one, two. three, four, or more) flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be used to inhibit fibroblast activation. A flavonoid compound having the structure of Formula (I) can inhibit any appropriate fibroblast activation. For example, a flavonoid compound having the structure of Formula (I) can inhibit TGFfi-induced fibroblast activation.

[0147] In some cases, one or more (e.g., one, two. three, four, or more) flavonoid compounds provided herein (e.g, one or more flavonoid compounds having the structure of Formula (I)) can be used to induce apoptosis of cells (e.g., senescent cells) within a mammal. For example, a composition including one or more flavonoid compounds having the structure of Formula (I) can be administered to a mammal (e.g, a human) in need thereof (e.g, a human having one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF, NASH, and PSC) to induce apoptosis in senescent cells within the mammal. One or more flavonoid compounds provided herein can be used to induce apoptosis in any appropriate type of senescent cell within a mammal. Examples of types of cells that can be senescent and that one or more flavonoid compounds provided herein can be used to induce apoptosis in include, without limitation, fibroblasts (e.g, lung fibroblasts) and epithelial cells (e.g, cholangiocytes). For example, one or more flavonoid compounds provided herein (e.g, one or more flavonoid compounds having the structure of Formula (I)) can be used to reduce the number of senescent cells within a mammal. In some cases, a composition including one or more flavonoid compounds having the structure of Formula (I) can be administered to a mammal (e g., a human) in need thereof (e.g, a human having one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF, NASH, and PSC) to reduce the number senescent cells within the mammal. In some cases, one or more (e.g., one, two, three, four, or more) flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be effective to reduce the number of senescent cells within a mammal having one or more fibrotic and / or aberrant wound healing / scarring conditions by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent.

[0148] In some cases, one or more (e.g, one, two, three, four, or more) flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be used to increase the survival of a mammal (e.g., a human). For example, a composition containing one or more flavonoid compounds having the 07039-2361WO1 / 2025-008

[0149] structure of Formula (I) can be administered to a mammal (e.g, a human) in need thereof (e.g, a human having one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF, NASH, and PSC) to increase the survival of the mammal. In some cases, one or more (e.g., one, two, three, four, or more) flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be used to increase the survival of a mammal having one or more fibrotic and / or aberrant wound healing / scarring conditions by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent.

[0150] In some cases, one or more (e., one, two, three, four, or more) flavonoid compounds provided herein (e.g, one or more flavonoid compounds having the structure of Formula (I)) can be used to reduce or eliminate inflammation in one or more tissues within a mammal. For example, a composition including one or more flavonoid compounds having the structure of Formula (I) can be administered to a mammal (e.g, a human) in need thereof (e.g, a human having one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF, NASH, and PSC) to reduce or eliminate inflammation in one or more tissues within the mammal. One or more flavonoid compounds provided herein can be used to reduce inflammation in any appropriate tissue within a mammal. Examples of tissues that can be inflamed and that one or more flavonoid compounds provided herein can be used to reduce inflammation in include, without limitation, lung, liver, bile ducts, kidney, heart and skin. In some cases, one or more (e.g., one, two, three, four, or more) flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be used to reduce inflammation in one or more tissues within a mammal having a fibrotic and / or aberrant wound healing / scarring condition (e.g., IPF) by. for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent.

[0151] Any appropriate mammal having one or more fibrotic and / or aberrant wound healing / scarring conditions (e.g, IPF, NASH, and PSC) can be treated as described herein (e.g, by administering one or more flavonoid compounds having the structure of Formula (I)). Examples of mammals that can have one or more fibrotic and / or aberrant wound healing / scarring conditions (e.g, IPF, NASH, and PSC) and can be treated as described herein include, without limitation, humans, non-human primates such as monkeys, dogs, cats, horses, cows, pigs, sheep, mice, and rats. In some cases, a human having one or more fibrotic and / or aberrant wound healing / scarring conditions can be 07039-2361WO1 / 2025-008

[0152] treated by administering one or more flavonoid compounds having the structure of Formula (I) as described herein.

[0153] When treating a mammal (e.g., a human) having one or more fibrotic and / or aberrant wound healing / s carring conditions, the mammal can have any type of fibrotic and / or aberrant wound healing / s carring condition(s). Examples of fibrotic and / or aberrant wound healing / scarring conditions that can be treated as described herein (e.g., by administering one or more flavonoid compounds having the structure of Formula (1)) include, without limitation, IPF, PSC, and NASH.

[0154] In some cases, the methods described herein can include identifying a mammal (e.g., a human) as having one or more fibrotic and / or aberrant wound healing / scarring conditions (e.g., IPF. NASH, and PSC). Any appropriate method can be used to identify a mammal as having one or more fibrotic and / or aberrant wound healing / scarring conditions (e.g., IPF, NASH, and PSC). For example, chest scans including X-ray and high-resolution computed tremography, breathing tests, pulse oximetry, blood test for oxygen and CO2. exercise capacity, and / or lung biopsy (e.g., to observe signs of scarring) can be used to identify mammals (e.g., humans) having an IPF disease. For example, imaging techniques (e.g., magnetic resonance imaging (MRI), magnetic resonance cholangiopancreatography (MRCP), and endoscopic retrograde cholangiopancreatography (ERCP)), a cholestatic biochemical profile, and / or liver biopsy can be used to identify mammals (e.g., humans) having, a PSC disease.

[0155] In some cases, one or more flavonoid compounds provided herein (e. g, one or more flavonoid compounds having the structure of Formula (I)) can be used to treat a mammal (e.g., a human) having an age-related disease. For example, one or more flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be administered to a mammal (e.g, ahuman) having an age-related disease (e.g., to treat the mammal). Examples of age-related diseases that can be treated as described herein (e.g., by administering one or more flavonoid compounds provided herein) include, without limitation, osteoporosis, frailty, cardiovascular diseases, osteoarthritis, pulmonary fibrosis, renal diseases, neurodegenerative diseases, hepatic steatosis, and metabolic dysfunction. In some cases, an age-related disease that can be treated as described herein can be as described elsewhere (see, e.g., Kaur et al., Transl. Res., 226:96-104 (2020)). A composition containing one or more flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) can be administered to a mammal (e.g., ahuman) having one or 07039-2361WO1 / 2025-008

[0156] more fibrotic and / or aberrant wound healing / scarring conditions (e.g., IPF, NASH, and PSC) in any appropriate amount (e.g, any appropriate dose). An effective amount of a composition containing one or more flavonoid compounds having the structure of Formula (I) can be any amount that can treat a mammal (e.g., a mammal having one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF, NASH, and PSC) as described herein without producing significant toxicity to the mammal. In some cases, an effective amount of one or more flavonoid compounds having the structure of Formula (I) can be from about 0.1 µM to about 100 µM (e.g., from about 0.1 µM to about 75 µM, from about 0.1 µM to about 60 µM, from about 0.1 µM to about 50 µM, from about 0.1 µM to about 40 µM, from about 0.1 µM to about 30 µM, from about 0.1 µM to about 20 µM, from about 0.1 µM to about 10 µM, from about 0.1 µM to about 1 µM, from about 10 µM to about 100 µM, from about 20 µM to about 100 µM, from about 30 µM to about 100 µM, from about 40 µM to about 100 µM, from about 50 µM to about 100 µM, from about 60 µM to about 100 µM, from about 75 µM to about 100 µM, from about 1 µM to about 75 µM, from about 10 µM to about 50 µM, from about 20 µM to about 40 µM, from about 1 µM to about 30 µM, from about 30 µM to about 50 µM, from about 40 µM to about 60 µM, from about 50 µM to about 70 µM, from about 60 µM to about 80 µM, or about 30 µM). In some cases, an effective amount of one or more flavonoid compounds having the structure of Formula (I) can be from about 200 nM IC50 to about 800 nM IC50 (e.g., from about 200 nM IC50 to about 700 nM IC50, from about 200 nM IC50 to about 600 nM IC50, from about 200 nM IC50 to about 500 nM IC50, from about 200 nM IC50 to about 400 nM IC50, from about 200 nM IC50 to about 300 nM IC50, from about 300 nM IC50 to about 800 nM IC50, from about 400 nM IC50 to about 800 nM IC50, from about 500 nM IC50 to about 800 nM IC50, from about 600 nM IC50 to about 800 nM IC50, from about 700 nM IC50 to about 800 nM IC50, from about 300 nM IC50 to about 700 nM IC50, from about 400 nM IC50 to about 600 nM IC50, from about 300 nM IC50 to about 400 nM IC50, from about 400 nM IC50 to about 500 nM IC50, from about 500 nM IC50 to about 600 nM IC50, from about 600 nM IC50 to about 700 nM IC50, about 200 nM IC50, or about 800 nM IC50). In some cases, an effective amount of one or more flavonoid compounds having the structure of Formula (I) can include from about 0.1 milligrams per kilogram body weight (mg / kg) to about 200 mg / kg (e.g., from about 0.1 mg / kg to about 175 mg / kg, from about 0.1 mg / kg to about 150 mg / kg, from about 0.1 mg / kg to about 125 mg / kg, from about 0.1 mg / kg to about 100 mg / kg, from about 0.1 mg / kg to about 75 mg / kg, from about 0.1 mg / kg to about 50 mg / kg, from about 0.1 07039-2361WO1 / 2025-008

[0157] mg / kg to about 25 mg / kg, from about 0.1 mg / kg to about 10 mg / kg, from about 0.1 mg / kg to about 5 mg / kg, from about 1 mg / kg to about 200 mg / kg, from about 10 mg / kg to about 200 mg / kg, from about 25 mg / kg to about 200 mg / kg, from about 50 mg / kg to about 200 mg / kg, from about 75 mg / kg to about 200 mg / kg, from about 100 mg / kg to about 200 mg / kg, from about 150 mg / kg to about 200 mg / kg, from about 1 mg / kg to about 150 mg / kg, from about 10 mg / kg to about 100 mg / kg, from about 25 mg / kg to about 75 mg / kg, from about 1 mg / kg to about 5 mg / kg, from about 5 mg / kg to about 10 mg / kg, from about 10 mg / kg to about 25 mg / kg, from about 25 mg / kg to about 50 mg / kg, from about 50 mg / kg to about 100 mg / kg, from about 100 mg / kg to about 150 mg / kg, or about 2 mg / kg) F-4N. The effective amount can remain constant or can be adjusted as a sliding scale or variable dose depending on the mammal’s response to treatment. Various factors can influence the actual effective amount used for a particular application. For example, the frequency of administration, duration of treatment, use of multiple treatment agents, route of administration, and / or severity of the fibrotic and / or aberrant wound healing / scarring condition (e.g, IPF, NASH, and PSC) in the mammal being treated may require an increase or decrease in the actual effective amount administered.

[0158] A composition containing one or more flavonoid compounds provided herein (e.g, one or more flavonoid compounds having the structure of Formula (I)) can be administered to a mammal (e.g, a mammal having one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF, NASH, and PSC) in any appropriate frequency. The frequency of administration can be any frequency that can treat a mammal (e.g., a mammal having one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF, NASH, and PSC) without producing significant toxicity to the mammal (e.g., human). For example, the frequency of administration can be from about once a day to about once a week, from about once a week to about once a month, or from about twice a month to about once a month. The frequency of administration can remain constant or can be variable during the duration of treatment. As with the effective amount, various factors can influence the actual frequency of administration used for a particular application. For example, the effective amount, duration of treatment, use of multiple treatment agents, and / or route of administration may require an increase or decrease in administration frequency.

[0159] A composition containing one or more flavonoid compounds provided herein (e.g, one or more flavonoid compounds having the structure of Formula (I)) can be administered to a mammal (e.g, a mammal having one or more fibrotic and / or aberrant 07039-2361WO1 / 2025-008

[0160] wound healing / scarring conditions such as IPF, NASH, and PSC) for any appropriate duration. An effective duration for administering or using a composition containing one or more flavonoid compounds having the structure of Formula (I) can be any duration that can treat a mammal (e.g., a mammal having one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF, NASH, and PSC) without producing significant toxicity to the mammal (e.g., a human). For example, the effective duration can vary from several weeks to several months, from several months to several years, or from several years to a lifetime. Multiple factors can influence the actual effective duration used for a particular treatment. For example, an effective duration can vary with the frequency of administration, effective amount, use of multiple treatment agents, and / or route of administration.

[0161] In some cases, the methods for treating a mammal (e.g., a mammal such as a human having one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF, NASH, and PSC) as described herein (e.g., by administering one or more flavonoid compounds having the structure of Formula (I)) can include administering to the mammal one or more flavonoid compounds having the structure of Formula (I) as the sole active ingredient(s) to treat the mammal. For example, a composition containing one or more flavonoid compounds having the structure of Formula (I) can include the flavonoid compound(s) having the structure of Formula (I) as the sole active ingredient in the composition that is effective to treat a mammal (e.g., a mammal having one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF, NASH, and PSC).

[0162] In some cases, the methods for treating a mammal (e.g., a mammal such as a human having one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF, NASH, and PSC) as described herein (e.g., by administering one or more flavonoid compounds having the structure of Formula (I)) also can include administering to the mammal one or more (e.g., one, two, three, four, five or more) additional agents / therapies used to treat a condition (e.g., one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF and / or PSC). For example, a combination therapy used to treat one or more fibrotic and / or aberrant wound healing / scarring conditions (e.g., IPF, NASH, and PSC) can include administering to the mammal (e.g., a human) one or more flavonoid compounds having the structure of Formula (I) described herein and one or more (e.g.. one, two, three, four, five or more) agents used to treat one or more fibrotic and / or aberrant wound healing / scarring conditions (e.g., IPF, NASH, and 07039-2361WO1 / 2025-008

[0163] PSC). Examples of agents that can be administered to a mammal to treat IPF disease include, without limitation, pirfenidone, nintedanib. N-acetylcysteine. phosphodiesterase inhibitors (e.g., sildenafil, vardenafil, tadalafiL and avanafil), lysophosphatidic acid receptor antagonists (e.g., promethazine, FTY720, AM152, BMS-986020, VPC 12249, AM966, and AM095), autotaxin inhibitors (e.g., taribavirin, BI-2545, GLPG1690, BBT 877, SAR100842, and BMS-986,020). DI receptor agonists (e.g.. minaprine, dopamine, levodopa, apomorphine, fenoldopam, pergolide, bromocriptine, and cabergoline), dasatinib, rho kinase inhibitors (e g., hydroxyfasudil, ripasudil, netarsudil, and belumosudil), IL-13 neutralizing antibodies (e.g., lebrikizumab, tralokinumab, and dupilumab), CTGF neutralizing antibodies (e.g., pamrevlumab), and any combinations thereof. Examples of agents that can be administered to a mammal to treat PSC disease include, without limitation, ursodeoxycholic acid (UDCA), corticosteroids (e.g., glucocorticoids such as prednisolone), bile acid sequestrants, antibiotics, antihistamines, and any combinations thereof.

[0164] In cases where one or more flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) are used in combination with additional agents used to treat one or more fibrotic and / or aberrant wound healing / scarring conditions, the one or more additional agents can be administered at the same time (e.g., in a single composition containing both one or more flavonoid compounds having the structure of Formula (I) and the one or more additional agents) or independently. For example, one or more flavonoid compounds having the structure of Formula (I) described herein can be administered first, and the one or more additional agents administered second, or vice versa.

[0165] In some cases, the methods for treating a mammal (e.g., a mammal such as a human having one or more fibrotic and / or aberrant wound healing / scarring conditions such as IPF, NASH, and PSC) as described herein (e.g., by administering one or more flavonoid compounds having the structure of Formula (I)) also can include performing one or more (e.g., one, two, three, four, five or more) additional therapies used to treat one or more fibrotic and / or aberrant wound healing / scarring conditions (e.g, IPF, NASH, and PSC) on the mammal. Examples of therapies used to treat the IPF disease include, without limitation, oxygen therapy, pulmonary rehabilitation, and / or lung transplantation. Examples of therapies used to treat the PSC disease include, without limitation, endoscopic therapy (e.g.. balloon dilatation and stent placement), percutaneous therapy, non-transplant surgery, liver transplantation, and / or fecal microbiome transplant. 07039-2361WO1 / 2025-008

[0166] In cases where one or more flavonoid compounds provided herein (e.g., one or more flavonoid compounds having the structure of Formula (I)) are used in combination with one or more additional therapies used to treat one or more fibrotic and / or aberrant wound healing / scarring conditions (e.g., IPF, NASH, and PSC), the one or more additional therapies can be performed at the same time or independently of the administration of one or more flavonoid compounds having the structure of Formula (I) described herein. For example, one or more flavonoid compounds having the structure of Formula (I) described herein can be administered before, during, or after the one or more additional therapies are performed.

[0167] The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.

[0168] EXAMPLES

[0169] The compounds herein may be prepared in a manner analogous to that disclosed in International PCT Patent Application Publication No. WO 2023 / 244619. incorporated by reference herein in its entirety. The compounds may also be prepared according to the following schemes.

[0170] Synthetic Schemes

[0171] Table 1. Synthesis and SAR of flavonol senolytics and Bcl-2 Inhibitors of the representative compounds of Formula (I) shown below:

[0172]

[0173] Representative compounds of Formula (I) may be prepared according to the scheme below:

[0174] Synthetic scheme and SAR of flavonol senolytics and Bcl-2 Inhibitors of representative compounds of Formula (I): 07039-2361WO1 / 2025-008

[0175]

[0176] In the above scheme, R3-R4, R8, and X are as shown below. CH2X corresponds to R2in the main embodiment of Formula (I).

[0177] All EC50 values in the Table 1 below are in micromolar (pM): Table 1:

[0178] Compound R3R4X R8Senescent Proliferating Fibroblast Fibroblast Fibroblast activation IC50 apoptosis apoptosis (pM) EC50 (pM) EC50 (pM)

[0179] F-19 H H CH3H 2.4 24.5 2.4

[0180] F-20 H H Phenyl H 1.5 27.6 2.0

[0181] F-2N H H 2-pyridyl H 21.8 29.8 inactive F-3N H H 3 -pyridyl H 11.2 14.9 13.8

[0182] F-4N H H 4-pyridyl H 0.8 48.9 1.8

[0183] 6-MeOH-F-19 H OCH3CH3 H 2.7 15.1 4.0

[0184] 6-MeOH-F-20 H OCH3 Phenyl H 0.8 5.1 2.0

[0185] F-19(Me) H H C2H5 H 3.4 19.5 4.0

[0186] F-19(3-MeOH) H H CH3 OCH3 5.9 10.5 6.4

[0187] F-CP H H cyclopropyl H 0.9 10.9 1.1

[0188] ABT-737 0.7 9.7 Not tested ABT- 199 3.2 76.5 Not tested ABT-263 1.9 14.9 Not tested

[0189]

[0190] 07039-2361WO1 / 2025-008

[0191] Examples:

[0192] Flavonol Synthesis

[0193] Equimolar amounts (2-8 mmol) of the hydroxyacetophenone were combined with the aldehyde in 10-50 mL of EtOH. 5-12 mL of 50% NaOH(aq) was added to the reaction and allowed to proceed for 6-48 hours at room temperature while monitoring by thin layer chromatography (TLC). After aldehyde depletion the resulting calchone was precipitated by 10% HCl(aq). Calchone was isolated, dried, weighed, and dissolved (1-4 mmol) in 8-30 mL of MeOH containing IM KOH. 5-15 mL of H2O2 was added to the reaction and allowed to proceed for 1-6 hours at room temperature while monitoring by TLC. Final product was precipitated by 10% HCl(aq) and purified by recrystallization. Structure and purity (>95%) were confirmed by proton nuclear magnetic resonance.

[0194] Cell Culture

[0195] Primary human lung fibroblasts (NHLF) were purchased from ScienCell and cultured in DMEM / F12 (Gibco) containing 10% fetal bovine serum (Invitrogen) and Antibiotic-Antimycotic (Gibco).

[0196] Mice

[0197] Both mTmG (strain 007576) and Colla2-CreER (strain 029567) were purchased from Jackson Labs to track fibroblast specific. All animal experiments and procedures were approved by Mayo Clinic’s Institutional Animal Care Committee (IACUC) and performed in accordance with the National institute of Health guidelines.

[0198] Senescence Induction in vitro

[0199] Senescence was induced experimentally by passaging the cells until replication exhaustion, typically passage 18-22, treating cells with 20 pM etoposide or 10 pM bleomycin for 48 hours, followed by 10 days in normal culture media as described in Gao, A. Y. et al. Pim-1 kinase is a positive feedback regulator of the senescent lung fibroblast inflammatory secretome. Am J Physiol-Lung C 323, L685-L697,

[0200] doi:10.1152 / ajplung.00023.2022 (2022), and in Schafer, M. J. et al. Cellular senescence mediates fibrotic pulmonary disease. Nature Communications 8, doi: ARTN 14532, 07039-2361WO1 / 2025-008

[0201] 10.1038 / ncomms14532 (2017). Radiation induced senescence was triggered by exposing cells to 20 Gy of X-ray irradiation and culturing for 10- and 12-days postirradiation with media refreshed every 48-72 hours as described in Victorelli, S. et al.

[0202] Apoptotic stress causes mtDNA release during senescence and drives the SASP. Nature 622, 627-636, doi:10.1038 / s41586-023-06621-4 (2023).

[0203] Cleaved Caspase-3 Staining and Analysis and Caspase 3 / 7 Activity Assay

[0204] Experiments were performed as described elsewhere (Schafer Nature Aging, 2024). Briefly, proliferating or senescent cells were added to 96-well plates and allowed to attach. After 6 hours media was changed to EMEM containing 0% FBS and the indicated concentration of compound was added. After 72 hours the cells were fixed with 4% formalin, permeabilized with 0.25% triton X-100, blocked with 1% BSA, and immunostained with an antibody for cleaved caspase-3 (Cell Signaling, 9661), an Alexa Fluor-488 secondary antirabbit (Thermo Fisher, A-11008) was incubated along with DAPI. Images were collected using a Cytation5 automated microscope (Biotek), using a 4X objective and cleaves caspase-3 intensity was quantified using onboard software (Gen5). Alternatively, caspase 3 / 7 activity was measured using a Caspase-Gio 3 / 7 assay from Promega following the manufacturer’s suggested protocol after the 48-hour incubation with compounds. Luminescence was measured using a Promega GloMax plate reader.

[0205] RNA isolation and qPCR analysis

[0206] Cells or lung tissue homogenates were treated as indicated in each experiment before RNA isolation using the RNeasy Plus Mini Kit (Qiagen, Germantown, MD) as per the manufacturer’s protocol. Isolated RNA was used to synthesize cDNA using SuperScript VILO (Invitrogen Life Technologies, Carlsbad, CA). Fast Start Essential DNA Green Master (Roche, Indianapolis, IN) was used to perform quantitative PCR analysis using a LightCycler 96 (Roche) machine. AACt calculation was used relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) to determine the fold change in gene expression.

[0207] SA-β-Gal staining 07039-2361WO1 / 2025-008

[0208] Growth media was removed from the cells before the plate was rinsed with PBS. 1 ml of the Fixative solution from the Senescence Beta-Galactosidase Staining Kit (Cell Signaling, Danvers, MA) was added to each well and incubated for 15 minutes. The plates were rinsed twice with PBS before 1 mL of Beta-Galactosidase Staining Solution (Cell Signaling, 9860) was added to each well. The plate was then incubated at room temperature overnight. The cells were then observed under a microscope and SA-β-Gal staining was detected at 20x objective to identify blue precipitants using an inverted bright field microscope.

[0209] Bleomycin Lung Fibrosis Model

[0210] Tamoxifen (75 mg / kg) was injected i.p into 10-week-old mixed sex Colla2-Cre; mTmG mice for five consecutive days. Three different bleomycin models were used in these studies- one-time in young mice, one-time in aged mice, and repeated (3X) in young mice. Mice were given bleomycin intratracheally 0.6 or 1.0 U / kg (novaplus) or saline control while under isoflurane anesthesia. Injections were administered to the Colla2-cre;mTmG at 2 months of age, FVB at 15-18 months of age (aged), and FVB at 2 months of age received three administrations, every two weeks. The Colla2-Cre mice were treated from day 10-14 with F-19 (10 mg / kg) via I. P injections and harvested at day 14 for flow analysis. In both the aged and chronic models’ mice received F-4N (10 mg / kg) i.p daily from days 14-28 post final bleomycin administration. Mice were sacrificed and samples were collected at day 28 for the aged and chronic studies.

[0211] Fluorescent activated cell sorting

[0212] Mouse primary lung cell suspensions were generated by a combination of mechanical digestion via razor blade and then enzymatic digestion in DMEM (Gibco) with Liberase TM (10 pg / mL, Sigma) and DNase I (10 U / mL, Sigma) for 35 minutes at 37°C and passed through a 40 pm strainer (Fisher Scientific). Digestion was inactivated by DMEM with 10% FBS and pelleted at 400g for 5 minutes. Samples underwent 90 seconds of red blood cell lysis (biolegend) which was inactivated using autoMACS rinsing solution (Miltenyi) and passed through a 30 pm mesh (Sysmex America). Cells were spun down stained with CD326: APC, CD45: PerCP-Cy5.5, CD31 (biolegend) and Dapi (Thermo) for 30 minutes on ice. The cells were sorted using a BD Aria II cell sorter. 07039-2361WO1 / 2025-008

[0213] Precision-Cut Lung Slice Model

[0214] Mice were anesthetized with an i.p. injection of ketamine / xylazine (150 mg / kg / 10 mg / kg). Mouse was perfused with PBS with 2 mM EDTA, a tracheotomy was performed, and the lungs were insufflated with 10% porcine gelatine (Sigma) in Hanks’ balanced salt solution (HBSS). Trachea was ligated and lungs removed from the thoracic cavity and placed in cold HBSS (Gibco). Once gelatin solidified, the left lobe was removed and sectioned into 300 pm slices. Slices were cultured for 3 days in 50% normal fibroblast culture media described above and 50% alveolar epithelial media from Sciencell containing the manufacturer’s suggested growth supplements. This media combination was effective at supporting markers of activated fibroblasts and maintaining alveolar epithelial markers ex vivo.

[0215] Results

[0216] B-ring, para-ethoxy substitution drives potency and selectivity of synthetic flavonols.

[0217] In order to develop and explore the structure-activity relationship (SAR) of flavonol senolytics human lung fibroblasts were cultured to replicative senescence (18-20 passages), confirmed by measuring proliferation, expression of CDKN1A and CDKN2A and SA-β-gal staining (Fig 1A-C) as described in Schafer, M. J. et al. Cellular senescence mediates fibrotic pulmonary disease. Nat Commun 8, 14532, doi:10.1038 / ncomms14532 (2017). A high-throughput assay was developed to quantify expression of cleaved caspase-3 (CC3), a hallmark of apoptosis, by high-content immunocytochemistry (Schafer, Nature Aging) and compared dose-response effects of natural flavonols quercetin (Q) and fisetin (F) (Fig. ID).

[0218] Consistent with published findings, Q and F displayed senolytic effects only above 30 pM. One of the limitations with naturally derived flavonols is the narrow chemical diversity presented on their A and B ring substitutions; with only the number and location of hydroxyl groups being the sole driver of differentiation. A custom diversity library of commercially available flavonols encompassing natural products, natural product metabolites, and synthetic flavonols, and a 2D structural overlay to visualize consistent and divergent moieties across the molecules, were generated. Tanimoto similarity indexes for each compound were then calculated relative to quercetin to visualize the unique diversity provided by synthetic 07039-2361WO1 / 2025-008

[0219] flavonols (Fig. IE). The senolytic potential of the flavonol diversity library was first triaged by measuring viable cell counts in senescent lung fibroblasts after 72 hours in culture with 10, 30, and 100 pM of each compound (Fig. 6). Any compounds that had a clear impact on reducing senescent cells at 10 pM were selected for follow-up dose-response experiments in senescent and proliferating lung fibroblasts. F-15, F-26, F-17, and F-19 were found to be the most potent and selective senolytics (Fig. IF) and intriguingly they all contained a paraethoxy substitution on their B-ring. F-19 was the most potent and selective senolytic flavonol that was identified, with an EC50 of 2.4 pM to induce CC3 expression in senescent lung fibroblasts, compared to 25 pM in proliferating lung fibroblasts. F-19 is 10-fold more potent than the natural products Q or F and has a much larger window of selectivity in this in vitro model.

[0220] F-19 promotes senescence clearance and supports alveolar regeneration ex vivo and in vivo.

[0221] Ex vivo lung slice cultures or precision-cut lung slices (PCLS) are useful models to study complex multicellular biology in environments more consistent with native tissue (Cedilak, M. et al. Precision-cut lung slices from bleomycin treated animals as a model for testing potential therapies for idiopathic pulmonary fibrosis. Pulm Pharmacol Ther 55, 75-83, doi:10.1016 / j.pupt.2019.02.005 (2019)). PCLS were generated from aged, + / - bleomycin induced fibrotic mouse lungs and treated with F-19 or a clinically approved antifibrotic, Nintedanib, ex vivo for 72 hours prior to collecting RNA from the lung slice for qPCR analysis. F-19 treatment promotes clearance of senescence associated markers Cdkn2a, Cdknla, and 116, where Nintedanib was ineffective, both compounds reduced expression of Col lai the gene critical for production of Type I Collagen, the most abundance ECM protein in the lung (Fig. 2A). A finding consistent with previously reported effects of senolytic targeting in fibrotic lungs ex vivo, F-19 increased expression of Sftpc and Hopx, markers of mature alveolar epithelium, suggesting the possibility that removal of senescent cells from fibrotic lung could both stop interstitial fibrosis and stimulate or facilitate alveolar regeneration (Lehmann, M. et al. Senolytic drugs target alveolar epithelial cell function and attenuate experimental lung fibrosis. European Respiratory Journal 50, doi: Artn 1602367, 07039-2361WO1 / 2025-008

[0222] 10.1183 / 13993003.02367-2016 (2017)) (Fig. 2A). Pathologically unique, mesenchymal and epithelial cells expressing markers of senescence have been identified in experimental models of lung fibrosis and IPF patient lungs(Xu, Y. et al. Single-cell RNA sequencing identifies diverse roles of epithelial cells in idiopathic pulmonary fibrosis. Jci Insight 1, doi: ARTN e90558, 10.1172 / jci.insight.90558 (2016); Alexander, M. J., Budinger, G. R. S. & Reyfman, P. A. Breathing fresh air into respiratory research with single-cell RNA sequencing. Eur Respir Rev 29, doi: Artn 200060, 10.1183 / 16000617.0060-2020 (2020); Pardo, A. & Selman, M. Lung Fibroblasts, Aging, and Idiopathic Pulmonary Fibrosis. Annals of the American Thoracic Society 13, S417-S421, doi:10.1513 / AnnalsATS.201605-341AW (2016); Xie, T. et al. Single-Cell Deconvolution of Fibroblast Heterogeneity in Mouse Pulmonary Fibrosis. CellRep 22, 3625-3640, doi:10.1016 / j.celrep.2018.03.010 (2018); Adams, T. S. et al. Single-cell RNA-seq reveals ectopic and aberrant lung-resident cell populations in idiopathic pulmonary fibrosis. Sci Adv 6, eaba1983,

[0223] doi: 10.1126 / sciadv.aba1983 (2020). Tsukui, T. et al. Collagen-producing lung cell atlas identifies multiple subsets with distinct localization and relevance to fibrosis. Nature Communications 11, doi: ARTN 1920, 10.1038 / s41467-020-15647-5 (2020)). In order to investigate the specific effect of F-19 on fibroblasts and epithelial cells, a mesenchymal lineage tracing mouse was generated by breeding Colla2-CreERT(2) mice (Jackson Laboratory) with Rosa26-mTmG mice. To permanently lineage-label the Colla2+ population in these mice, each mouse was administered tamoxifen (75mg / kg) for 5 days, starting 15 days prior to lung injury (Swonger, J. M., Liu, J. S., Ivey, M. J. & Tallquist, M. D. Genetic tools for identifying and manipulating fibroblasts in the mouse. Differentiation 92, 66-83, doi: 10.1016 / j.diff.2016.05.009 (2016)). (Fig. 2B). On day 10 after bleomycin injury, treatment was begun with 10 mg / kg F-19 i.p. daily injections for 4 days. Lungs were then harvested, and flow sorted to collect fibroblasts and epithelial cells. RNA from each population of cells was collected and analyzed by qPCR. Expression of senescence associated genes Cdkn2a (pl 6), and Cdknla (p21) were both repressed in fibroblasts and epithelials cells collected from mice treated with F-19 (Fig. 2C). Next, transcript analysis was expanded to include ECM associated genes enriched in fibrotic lung fibroblasts (Tsukui, T. et al. Collagen-producing lung cell atlas identifies multiple subsets with distinct localization and relevance to fibrosis. Nature Communications 11, doi: ARTN 1920 07039-2361WO1 / 2025-008

[0224] 10.1038 / s41467-020-15647-5 (2020); Jin, C. J. et al. Single-cell RNA sequencing reveals special basal cells and fibroblasts in idiopathic pulmonary fibrosis. Sci Rep-Uk 14, doi: ARTN 15778): Collal, Tnc, and Cthrcl, senescence associated soluble factors: 116, Ccl2, and Serpinel, and markers of the aberrant basal oid / intermediate epithelial population unique to fibrotic lungs (Xu, Y. et al. Single-cell RNA sequencing identifies diverse roles of epithelial cells in idiopathic pulmonary fibrosis. Jci Insight 1, doi: ARTN e90558

[0225] 10.1172 / jci. insight.90558 (2016); and Boesch, M. et al. Transcriptomic profiling reveals disease-specific characteristics of epithelial cells in idiopathic pulmonary fibrosis. Respir Res 21, 165, doi:10.1186 / sl2931-020-01414-z (2020)): Cldn4, Krtl9, and Snf. In all cases, F-19 treatment had a dramatic improvement in normalizing expression of these pathological markers of lung fibrosis (Fig.2D-E) Individual gene plots are also provided in Figure 7. Consistent with ex vivo findings, F-19 treatment stimulates expression or alveolar epithelial ATII markers: Sftpc, Sftpb, Sftpal, and ATII / ATI markers: Abcal, Abca3, and ATI markers: Hopx andXger. F-19 did not alter expression of TNF in either cellular compartment which is potentially advantageous as TNF-a has previously been shown to be beneficial in lung fibrosis resolution (Redente, E. F. et al. Tumor Necrosis Factor-a Accelerates the Resolution of Established Pulmonary Fibrosis in Mice by Targeting Profibrotic Lung Macrophages. Am J Resp Cell Mol 50, 825-837, doi:10.1165 / rcmb.2013-03860C (2014)), further supporting the potential of optimized senolytic flavonols and highlighting the nature of this molecule is not simply an “antiinflammatory”.

[0226] Flavonol F-4N potently and selectively induces apoptosis in senescent lung fibroblasts In an effort to improve solubility, potency, and selectivity, a series of additional paraethoxy substituted analogs of F-19 were designed and synthesized and their senolytic potential was measured using the CC3 assay (Table 1). To benchmark the synthetic flavonols against clinically developed and approved therapies also known to selectively induce apoptosis in senescent cells, the potency and selectivity of Bcl-2 family inhibitors ABT-263 (Navitoclax), ABT-199 (Venetoclax), and ABT-737 were also tested in the CC3 assay comparing senescent and proliferating lung fibroblasts (Table 1). Concerning the medicinal chemistry of the flavonols, the placement of the pyridine nitrogen in position 4 (F-4N) had a vastly superior potency (EC₅₀=900 nM) and produced an excellent senolytic window (~60- 07039-2361WO1 / 2025-008

[0227] fold selective) (Table 1, Fig.3A-B). Caspase 3 / 7 activity was confirmed using a luminescent substrate (Fig.3C). It was validated that F-4N led to selective apoptosis in senescent lung fibroblasts without an observable impact on proliferating lung fibroblasts (Fig.3D). It was also confirmed that F-4N could induce apoptosis in multiple inducible models of senescence including radiation and DNA damaging chemotherapeutics (Fig. 8).. Experiments with PCLS generated from 18-month-old mice + / -bleomycin injury were performed to compare F-4N to Bcl-2 inhibitors. It was found that either class of compounds supported clearance of senescence associated markers but only the flavonols stimulated expression of markers indicative of alveolar regeneration, suggesting not all senolytics are equal and beneficial or detrimental effects of these molecules can be scaffold or mechanistically unique (Fig. 9).

[0228] F-4N resolves lung fibrosis and improves mortality in vivo.

[0229] In order to assess the efficacy of F-4N in the most robust models of rodent lung fibrosis, a study was first performed in aged mice, delivering bleomycin one-time to produce chronic, nonresolving fibrosis (Caporarello, N. et al. Vascular dysfunction in aged mice contributes to persistent lung fibrosis. Aging Cell 19, el3196, doi: 10.1111 / acel.13196 (2020); Hecker, L. et al. Reversal of persistent fibrosis in aging by targeting Nox4-Nrf2 redox imbalance. Sci Transl Med 6, 231ra247, doi:10.1126 / scitranslmed.3008182 (2014); Redente, E. F. et al. Age and sex dimorphisms contribute to the severity of bleomycin-induced lung injury and fibrosis. Am J Physiol Lung Cell Mol Physiol 301, L510-518,

[0230] doi: 10.1152 / ajplung.00122.2011 (2011)). On day 14 after bleomycin injury, treatment was begun with 10 mg / kg F-4N or vehicle by i.p. daily injections for 14 days. At the completion of the study, a dramatic improvement in mouse survival was observed in the group treated with F-4N (Fig. 4A). F-4N also elicited marked improvements in overall lung architecture and collagen staining, and a nearly complete resolution of lung collagen content (Fig. 4B). In vivo efficacy measurements of whole lung transcript levels of pathological markers associated with ECM production and senescence associated genes were also consistent with the ex vivo studies, ECM and senescence genes were repressed by F-4N, and markers associated with alveolar regeneration are increased (Fig. 4C). Plasma levels of IL-6 were also measured - IL-6 is a senescence associated biomarker - and a nearly complete resolution was observed (Fig. 4D). An additional model of chronic lung fibrosis in mice is repeated 07039-2361WO1 / 2025-008

[0231] delivery of bleomycin, every two weeks for 6 weeks in young mice. An analogous dosing / treatment study was performed with F-4N using this “repeated bleomycin” model and observed consistent resolution features in the flavonol treated group with improvements in body mass, lung architecture, collagen content, and whole lung gene expression changes (Fig. 5). In particular, expression of mature alveolar epithelial markers Hopx, Ager, Sftpal, and Sftpc, was measured, and again F-4N was observed to enhance expression while repressing expression of markers of the aberrant basaloid / intermediate epithelial population unique to fibrotic lungs (Xu, Y. et al. Single-cell RNA sequencing identifies diverse roles of epithelial cells in idiopathic pulmonary fibrosis. Jci Insight 1, doi: ARTN e90558,

[0232] 10.1172 / jci. insight.90558 (2016); Boesch, M. et al. Transcriptomic profiling reveals diseasespecific characteristics of epithelial cells in idiopathic pulmonary fibrosis. Respir Res 21, 165, doi: 10.1186 / S12931-020-01414-z (2020))33’41: Snf, Cldn4, Krtl9, and Mmp7 (a putative biomarker that is predictive of lung fibrosis progression and mortality (Bauer, Y. et al. MMP-7 is a predictive biomarker of disease progression in patients with idiopathic pulmonary fibrosis. ERJ Open Res 3, doi:10.1183 / 23120541.00074-2016 (2017); Fainberg, H. P. et al. Cluster analysis of blood biomarkers to identify molecular patterns in pulmonary fibrosis: assessment of a multicentre, prospective, observational cohort with independent validation. Lancet Respir Med 12, 681-692, doi:10.1016 / S2213-2600(24)00147-4 (2024); Khan, F. A., Stewart, I., Saini, G., Robinson, K. A. & Jenkins, R. G. A systematic review of blood biomarkers with individual participant data meta-analysis of matrix metalloproteinase-7 in idiopathic pulmonary fibrosis. Eur Respir J 59, doi:10.1183 / 13993003.01612-2021 (2022)) (Fig. 5C)).

[0233] Additional Assays:

[0234] Fibroblast Activation

[0235] Human adult lung fibroblasts (passage 3) are plated into 96-well plates and treated with the indicated compound (6-point dose-response) +2 ng / mL TGFP to stimulated fibroblast activation. Cells are incubated for 96 hours, fixed using 4% PF A, permeabilized using 0.25% triton x-100, and stained using a primary antibody recognizing alpha-smooth muscle actin and a secondary fluorescent conjugated antibody, and DAPI. Cells are then 07039-2361WO1 / 2025-008

[0236] imaged using a 4X objective on a Cytation5 microscope and alpha-smooth muscle actin intensity is quantified using automated software (Biotek Gen5).

[0237] PSC

[0238] This Example describes the use of one or more flavonoid compounds having the structure of Formula (I) in treating PSC.

[0239] Methods

[0240] Hydroxyproline Assay

[0241] Hydroxyproline content is measured using a hydroxyproline assay kit (Biovision) according to the manufacturer's instructions with slight modification. The liver tissues are weighed, homogenized in sterile water (10 mg of tissue per 100 pL H2O) and hydrolyzed in 12 MHC1 in a pressure-tight, Teflon capped vial at 120°C for 3 hours followed by filtration through a 45 pm Spin-X Centrifuge Tube filter (Coming). Ten pL of the hydrolyzed samples are dried in a Speed- Vac for 2 hours, followed by incubation with 100 pL of Chloramine T reagent for 5 minutes at room temperature and 100 pL of 4-(dimethylamino)benzaldehyde (DMAB) for 90 minutes at 60°C. The absorbance of oxidized hydroxyproline is determined at 560 nm. Hydroxyproline concentrations are calculated from a standard curve generated using known concentrations of trans-4-hydroxyl-L-proline. The total amount of protein isolated from the weighed tissues is determined by using a protein assay kit (BioRad, absorbance at 595 nm).

[0242] Picrosirius Staining

[0243] Paraffin-embedded liver sections are de-paraffmized and re-hydrated by heating at 60°C for 10-30 minutes, then passaging in the following solutions:

[0244] • Xylene 1 - 2 x 10 minutes

[0245] • 100% EtOH - 1 x 2 minutes; 95% EtOH - 1 minute; 70% EtOH - 1 minute; 50% EtOH- 1 minute

[0246] • PBS - 3 x 3 minutes

[0247] The slides are then stained with a solution containing 0.1% picrosirius red (Direct Red 80) and 0.1% fast green (counterstain) in saturated aqueous solution of picric acid, for 07039-2361WO1 / 2025-008

[0248] 60 minutes at room temperature, followed by one wash in distilled water, de-hydration in 3 changes of 100% EtOH, clear in xylene and mount in resinous medium.

[0249] Liver Function Tests

[0250] Serum alanine aminotransferase (ALT), alkaline phosphatase (ALP) and total bile acids are measured using a commercially available veterinary chemistry analyzer (VetScan 2, Ab axis).

[0251] Reverse Transcription-Quantitative Polymerase Chain Reaction

[0252] RNA isolation using RNeasy Plus Mini Kit (Qiagen) according to the manufacturer's instructions. Isolated RNA (250 ng) is used to synthesize cDNA using SuperScript VILO (Invitrogen). Quantitative PCR is performed using FastStart Essential DNA Green Master (Roche) and analyzed using a LightCycler 96 (Roche). Data are expressed as a fold change by AACt relative to glyceraldehyde-3 -phosphate dehydrogenase (GAPDH).

[0253] Flavonol structure optimization

[0254] Materials and Methods

[0255] For senescent fibroblast survival experiments, human adult lung fibroblasts are replicatively passaged until senescence (confirmed by RNA and senescence associated beta galactosidase staining). Cells are then plated into 96-well plates and treated with the indicated compound and incubated for 96 hours. Cells are then fixed using 4% PF A, permeabilized using 0.25% triton x-100, and nuclear stained using DAPI. Cells are then imaged using a 4X objective on a Cytation5 microscope and cell counts are quantified using automated software (Biotek Gen5). Data shown are plotted as % survival, normalized to the vehicle treated well. Mean + / - SEM, N=3 independent experiments. For the TGFp stimulated collagen deposition experiments (square datapoints), human adult lung fibroblasts (passage 3) are plated into 96-well plates and treated with the indicated compound +2 ng / mL TGFp to stimulated collagen expression. Cells are incubated for 96 hours, fixed using 4% PF A, permeabilized using 0.25% triton x-100, and stained using a primary antibody recognizing type I collagen and a secondary infrared conjugated antibody. Wells are then imaged at IX 07039-2361WO1 / 2025-008

[0256] using a Odyssey Lx (LI-CORE) infrared imager and collagen intensity is quantified using automated software.

[0257] Treating IPF

[0258] A human identified as having IPF is administered or self-administers a composition including one or more flavonoid compounds having the structure of Formula (I).

[0259] Treating PSC

[0260] A human identified as having PSC is administered or self-administers a composition including one or more flavonoid compounds having the structure of Formula (I).

[0261] Non-alcoholic steatohepatitis (NASH)

[0262] Mice in a mouse model of NASH are treated with 10 mg / kg test compound daily i.p. for 2 weeks. Livers from treated mice are analyzed by H& E and Sirius Red staining. Sirius Red staining are quantified. Liver weight and colon weight changes are also assessed.

[0263] Ocular fibrosis

[0264] Conjunctival fibroblasts are cultured for 4 days in 2% FBS and with or without test compound and evaluated for proliferation. Treated cells are fixed, stained with DAPI, and counted using automated software on a Cytation 5.

[0265] Conjunctival fibroblasts are cultured for 3 days in 2% FBS with or without 2 ng / mL TGFp and with or without test compound and evaluated for the presence of fibroblasts.

[0266] Treated cells are fixed, stained with DAPT, stained with an aSMA antibody, and quantified using automated software on a Cytation 5.

[0267] Conjunctival fibroblasts cultured for 6 days in 2% FBS with or without 2 ng / mL TGFp and with or without test compound and evaluated for collagen deposition. Treated cells are fixed, stained with DAPI, stained with an antibody for collagen I, and quantified using automated software on a Cytation 5 (Figure 23C).

[0268] OTHER EMBODIMENTS

[0269] It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not 07039-2361WO1 / 2025-008

[0270] limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

07039-2361WO1 / 2025-008WHAT IS CLAIMED IS:

1. A compound of Formula (I)or a pharmaceutically acceptable salt thereof,whereinR1is selected from the group consisting of H, alkyl, substituted alkyl, cyclo-alkyl, substituted cyclo-alkyl, and OH;R2is selected from the group consisting of H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, and optionally substituted heterocyclyl;A, B, C are each independently N or CR3;each R3is independently H or halogen;D is N or CR4;R4is H, halogen, OR5, NR6R7, optionally substituted alkyl, or optionally substituted cycloalkyl;R5, R6, R7are each independently hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl;E and F are each independently N or CR8;each R8is independently H, halogen, OR9, NR10R11, optionally substituted alkyl, or optionally substituted cycloalkyl;R9, R10, R11are each independently hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl;G is O, NR12, S, SO, SO2;R12is hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl and optionally substituted heterocyclyl; and07039-2361WO1 / 2025-008each ring bond in the ring containing A, B, C and D is a single bond or a double bond, provided that no more than three ring bonds are double bonds, and provided that if all ring bonds are single bonds, then at least one of A, B, C or D is N.

2. The compound of claim 1, wherein R1is OH.

3. The compound of claim 1 or 2, wherein R2is optionally substituted alkyl.

4. The compound of claim 3, wherein R2is substituted alkyl.

5. The compound of claim 4, wherein R2is alkyl substituted with Ci-Ce alkoxy.

6. The compound of claim 5, wherein R2is ethoxy.

7. The compound of claim 4, wherein R2is substituted with carbocyclyl.

8. The compound of claim 7, wherein R2is alkyl substituted with phenyl.

9. The compound of claim 4, wherein R2is alkyl substituted with heterocyclyl.

10. The compound of claim 9, wherein R2is alkyl substituted with heteroaryl.

11. The compound of claim 10, wherein R2is alkyl substituted with pyridinyl.

12. The compound of claim 11, wherein R2is alkyl substituted with 2-pyridinyl.

13. The compound of claim 11, wherein R2is alkyl substituted with 3-pyridinyl.

14. The compound of claim 11, wherein R2is alkyl substituted with 4-pyridinyl.

15. The compound of any one of claims 1-14, wherein A is N.

16. The compound of any one of claims 1-14, wherein B is N.

17. The compound of any one of claims 1-14, wherein C is N.

18. The compound of any one of claims 1-14, wherein D is N.

19. The compound of any one of claims 1-14, wherein one of A, B, C or D is N, and the remaining three are not N.

20. The compound of any one of claims 1-19, wherein E is CR8.

21. The compound of any one of claims 1-19, wherein F is CR8.

22. The compound of any one of claims 1-21, wherein G is O.

23. The compound of claim 1, wherein the compound is selected from the group consisting of the compounds in Table 1 herein, or a pharmaceutically acceptable salt thereof.

24. The compound of claim 1, wherein the compound is selected from the group consisting of the following compounds, or a pharmaceutically acceptable salt thereof:07039-2361WO1 / 2025-00807039-2361WO1 / 2025-008(“F-CB”) and (“F-L132”).

25. A pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier..

26. A method of treating a fibrotic and / or aberrant wound healing / scarring condition, comprising administering a compound of claim 1, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising such compound or salt, to a subject in need thereof.

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