CBP / Catenin Signaling Pathway Inhibitors and Their Uses

Abstract

Compounds of formula (Ia), (Ib), and (IIa), and pharmaceutically acceptable salts thereof, are provided. Pharmaceutical compositions comprising the compounds are also provided for treating conditions, diseases, or disorders (e.g., fibrosis, cancer, neurological conditions, metabolic disorders (e.g., diabetes, etc.), and skin conditions mediated by abnormal CBP / β-catenin signaling (e.g., dermatitis, psoriasis, scarring, alopecia, etc.). Methods for enhancing vaccine efficacy using the compounds and compositions are also provided. Methods for efficiently synthesizing clinical-grade drugs are also provided, including the use of intermediate 2-propynyl-compounds in the penultimate or final reaction step under GMP conditions to form clinical-grade isoxazole derivatives (e.g., by 3+2 cycloaddition).

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 62 / 988,827, filed March 12, 2020, the entire contents of which are incorporated herein by reference.

[0002] Aspects of the present invention relate generally to the modulation of the Wnt / β-catenin pathway in mammalian (human and non-human) cells and tissues, and more particularly to small molecule inhibitors of CREB-binding protein (CBP) / β-catenin signaling, and even more particularly to one or more of the following metabolic disorders, including, but not limited to, fibrosis, cancer, neurological disorders, diabetes, and fatty liver disease, e.g., alcoholic (ALD) and non-alcoholic fatty liver (ALD and NAFLD, respectively), and non-alcoholic steatohepatitis. The present invention relates to surprisingly active and bioavailable small molecule CBP / β-catenin inhibitors that have broad utility for modulating and treating conditions and disorders mediated by CBP / β-catenin signaling, including nonalcoholic steatohepatitis (NASH), skin conditions (e.g., dermatitis, psoriasis, alopecia, aging, etc.), wound healing, aging, and may further include one or more of pulmonary hypertension, congestive heart failure, chronic kidney disease, renal fibrosis, endometriosis, cardiac fibrosis, polycystic ovary syndrome (PCOS), and / or systemic fibrosis / scleroderma. Additional aspects relate to using the disclosed compounds and compositions to enhance vaccine efficacy. [Background technology]

[0003] The evolutionarily conserved Wnt / β-catenin signaling pathway plays a fundamental and essential role in embryonic development and adult homeostasis.In addition, considering the established and crucial role of dysregulated / hyperactive CBP / β-catenin signaling in fibrosis, cancer, neuropathy, skin disorders, and metabolic disorders (including diabetes and fatty liver disease) and aging, as well as in other Wnt / β-catenin-mediated conditions and disorders, there has been considerable interest in exploring therapeutic and cosmetic interventions by regulating (e.g., inhibiting) CBP / β-catenin signaling and / or enhancing p300 / β-catenin signaling, preferably using small molecule inhibitors of CBP / β-catenin interaction.However, to date, developing specific small molecule inhibitors of CBP / β-catenin interaction that are sufficiently active and bioavailable (preferably, orally available in most cases for therapeutic use) remains a challenge, which has substantially hindered the realization of their therapeutic and functional cosmetic potential. Particular aspects of the present invention provide such compounds, compositions comprising the compounds, pharmaceutical compositions, and methods for synthesizing and using the compounds and compositions therapeutically and cosmetically, as further described below. Summary of the Invention

[0004] Aspects of the present invention can be described in the following paragraphs. 1. Formula (Ia): [ka] {In the formula, R a is hydrogen or -CH3; R b is a monocyclic aryl group having 5 to 7 ring members, which may have 1 to 2 heteroatoms selected from nitrogen, oxygen, or sulfur, and which may have one or more substituents selected from the group consisting of halide, cyano, and lower alkyl; R is a phenyl group; a substituted phenyl group having one or more substituents (wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazonyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; benzyl groups; substituted benzyl groups having one or more substituents, wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; or a bicyclic aryl group, or a substituted bicyclic aryl having 8 to 11 ring members and optionally having 1 to 3 heteroatoms selected from nitrogen, oxygen, or sulfur (substituted bicyclic aryl rings are selected from amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-3 optionally bearing one or more substituents independently selected from one or more of alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; R 2 is hydrogen or -CH3; Y is selected from hydrogen, deuterium, or a halogen; W is hydrogen, phosphate or phosphoric acid salt, alkyl acid or fatty acid Ester of , or X, where X is [ka] wherein Z is OR4 (wherein R4 is hydrogen or C1-C6 alkyl), or Z is an amino acid or amino acid ester; n is 1 or 2; L is -CH2-, -CF2-, or -C(CH3)2-; Q is [ka] wherein A, B, and D are independently selected from O, S, N, or —CH; and Z1, Z2 are independently selected from hydrogen, deuterium, halogen, C1-C4 alkyl, —OH, —OC1-C6 alkyl; and [ka] wherein R1, R2, and R3 are independently selected from hydrogen, C1-C6 alkyl, or C1-C6 alkyl containing one or more -OH groups; Z3 is selected from hydrogen, halogen, or aryl, heteroaryl, cycloalkyl, heterocycloalkyl directly bonded or bonded to -NH-, bonded to -OC1-C3 alkyl, or bonded to -NHC1-C3 alkyl, or nitrogen-bonded cycloalkyl, heterocycloalkyl, or -NHC1-C4 alkyl, -N(C1-C4 alkyl), each of which is hydrogen, deuterium, halogen, C1-C4 alkyl, C1-C3 haloaryl; and pharmaceutically acceptable salts thereof; and pharmaceutically acceptable salts thereof.

[0005] 2. Alkyl or fatty acids Ester of but, [ka] (wherein m is 1 to 14), [ka] 2. The compound of claim 1, selected from:

[0006] 3. The compound of paragraph 1 or 2, wherein R is a bicyclic aryl selected from naphthyl, quinolinyl, isoquinolinyl, quinoxaline, phthalazine, quinazoline, cinnoline, or naphthyridine, or substituted variants thereof.

[0007] 4. The compound has the formula (Ib): [ka] (wherein X1 and X2 are independently selected from N, or -CH).

[0008] 5. L is -CH2-; Q is, [ka] wherein A, B, and D are independently selected from O, S, N, or —CH. 6. A is -CH, B is N, D is O (Q is Z3-isoxazole-), W is hydrogen, phosphate or phosphoric acid salt, alkyl acid or fatty acid Ester of , or X, X is [ka] wherein Z is OR4 (wherein R4 is hydrogen or C1-C6 alkyl), or Z is an amino acid or amino acid ester; n is 1 or 2.

[0009] 7. The compound of clause 6, wherein Z3 is selected from aryl or heteroaryl, each of which is substituted by 0 to 4 substituents independently selected from hydrogen, deuterium, halogen, C-C alkyl, C-C haloalkyl, -OH, -O-C alkyl, -O-C alkyl-C(O)NH-OH, -NH, -C(O)NH-C-C alkyl-heteroaryl, -NHC(O)C-C alkyl-C(O)NH-OH, heteroaryl, cycloalkyl, heterocycloalkyl, or cycloalkyl or heterocycloalkyl attached at nitrogen, -NHC-C alkyl, or -N(C-C alkyl). 8. The compound of paragraph 7, wherein Z3 is selected from aryl or heteroaryl substituted with 0-4 substituents independently selected from hydrogen, deuterium, halogen, C1-C4 alkyl, C1-C3 haloalkyl, —OH, —O—C1-C6 alkyl, or heterocycloalkyl attached to nitrogen.

[0010] 9. The compound is [ka] TIFF0007750851000011.tif229150 The compound described in item 8, which is TIFF0007750851000012.tif117170.

[0011] 10. A composition or pharmaceutical composition comprising the compound according to any one of items 1 to 9 and a pharmaceutically acceptable carrier. 11. A method of treating a disease or disorder, comprising administering to a patient or warm-blooded mammal having a disease or disorder mediated by aberrant CREB binding protein (CBP) / β-catenin signaling an amount of a compound described in any one of paragraphs 1-10 sufficient to inhibit CBP / catenin-mediated signaling and / or enhance p300 / catenin-mediated signaling.

[0012] 12. The method of claim 11, wherein the amount of compound administered comprises a therapeutically effective amount. 13. In the compound, W is X and X is: [ka] 13. The method of claim 11 or 12, wherein Z is OR4 (wherein R4 is hydrogen or C1-C6 alkyl), or Z is an amino acid or amino acid ester, and n is 1 or 2.

[0013] 14. In the compound, X is [ka] 14. The method according to any one of items 11 to 13, wherein Z is OR4 (wherein R4 is hydrogen or C1-C6 alkyl), or Z is an amino acid or ester, and n is 1 or 2.

[0014] 15. The method of any one of paragraphs 11 to 14, wherein the disease or disorder comprises one or more of fibrosis, cancer, a neurological condition, a metabolic disorder, and a skin condition. 16. The method of paragraph 15, wherein the metabolic disorder comprises one or more of diabetes and / or fatty liver disease. 17. The method of claim 16, wherein the fatty liver disease comprises one or more of alcoholic fatty liver disease (ALD), non-alcoholic fatty liver disease (NAFLD), and / or non-alcoholic steatohepatitis (NASH).

[0015] 18. The method of claim 15, wherein the fibrosis is pulmonary, hepatic, renal, cardiac, endometrial, cutaneous or systemic fibrosis. 19. The method of claim 18, wherein the fibrosis comprises fibrosis in tissue of a SARS-CoV-2 (COVID-19) patient. 20. The method of paragraph 15, wherein treating cancer comprises administering a CBP / β-catenin antagonist in combination with or as adjunctive therapy with one or more of the following: cytotoxic and / or directed chemotherapy, and / or radiation therapy, and / or immunotherapy, including checkpoint inhibition (e.g., with anti-PD1, anti-PD-L1, or anti-CTLA4), chimeric antigen receptor (CAR-T) and / or CAR-NK cell-based therapy. 21. The method of paragraph 15, wherein the neurological condition comprises one or more of Huntington's disease (HD), Parkinson's disease (PD), Alzheimer's disease (AD), multiple sclerosis (MS), and / or amyotrophic lateral sclerosis (ALS), muscular dystrophy (MD), and / or spinal muscular atrophy (SMA).

[0016] 22. The method of clause 15, wherein the skin condition comprises one or more of atopic dermatitis, psoriasis, acne, fibrosis, wounds, scars, burns, sun or UV damage, diabetic ulcerations, chronic ulcerations, and / or alopecia. 23.W is an alkyl or fatty acid Ester of and administering comprises topical administration. 24. A cosmetic method for treating a skin condition, comprising administering to a patient or warm-blooded mammal having the skin condition a cosmetically effective amount of a compound according to any one of paragraphs 1-10, wherein W is an alkyl or fatty acid. Ester of and administering comprises topical administration. 25. The method of paragraph 24, wherein the skin condition comprises one or more aging skin conditions selected from wrinkles, hyperpigmentation, redness, rosacea, dryness, cracking, loss of vibrance, loss of elasticity, thinning, loss of color, scarring, acne, sun damage, hair loss, loss of hair color, reduced cuticle growth, and reduced nail growth. 26. A method for the efficient synthesis of clinical grade drugs, comprising the use of an intermediate 2-propynyl-compound in the penultimate or final reaction step under GMP conditions to form a clinical grade isoxazole derivative by 3+2 cycloaddition. 27. The step of preparing clinical grade isoxazole derivatives comprises, in the penultimate or final reaction step, reacting a compound of formula (IIa): [ka] {In the formula, R a is hydrogen or -CH3; R b is a monocyclic aryl group having 5 to 7 ring members, which may have 1 to 2 heteroatoms selected from nitrogen, oxygen, or sulfur, and which may have one or more substituents selected from the group consisting of halide, cyano, and lower alkyl; R is a phenyl group; a substituted phenyl group having one or more substituents (wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-4 Alkyl, C 1-3alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; benzyl groups; substituted benzyl groups having one or more substituents, wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; or a bicyclic aryl group having 8 to 11 ring members and optionally having 1 to 3 heteroatoms selected from nitrogen, oxygen, or sulfur; R 2 is hydrogen or -CH3; Y is selected from hydrogen, deuterium, or a halogen; W is hydrogen, alkyl acid or fatty acid Ester of , or X, X is [ka] 27. The method of claim 26, comprising the step of preparing a compound using an intermediate 2-propynyl-compound of formula {wherein Z is OR4 (wherein R4 is hydrogen or C1-C6 alkyl), or Z is an amino acid or amino acid ester, and n is 1 or 2} by the method of any one of claims 6 to 9.

[0017] 28. A method according to paragraph 26 or 27, wherein the last or penultimate step carried out under GMP conditions is preceded by one or more reaction steps under non-GMP conditions as part of an overall reaction scheme for preparing clinical grade isoxazole derivatives.

[0018] 29.Formula (IIa): [ka] {In the formula, R a is methyl or hydrogen; R b is a monocyclic aryl group having 5 to 7 ring members, which may have 1 to 2 heteroatoms selected from nitrogen, oxygen, or sulfur, and which may have one or more substituents selected from the group consisting of halide, cyano, and lower alkyl; R is a phenyl group; a substituted phenyl group having one or more substituents (wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; benzyl groups; substituted benzyl groups having one or more substituents, wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; or a bicyclic aryl group having 8 to 11 ring members and optionally having 1 to 3 heteroatoms selected from nitrogen, oxygen, or sulfur; R 2 is hydrogen or -CH3; Y is selected from hydrogen, deuterium, or a halogen; W is X, and X is [ka] wherein Z is OR4 (wherein R4 is hydrogen or C1-C6 alkyl), or Z is an amino acid or amino acid ester; n is 1 or 2}.

[0019] 30. A method for enhancing vaccine efficacy, comprising administering to a subject, before vaccination, and / or during vaccination, and / or after vaccination, an amount of a compound described in any one of claims 1 to 10 sufficient to inhibit CBP / β-catenin-mediated signaling and / or enhance p300 / catenin-mediated signaling.

[0020] 31. The method of claim 31, wherein the amount of compound administered comprises a therapeutically effective amount. 32. The method of claim 30 or 31, wherein enhancing vaccine efficacy comprises one or more of: an increase in the level of vaccine antigen-specific antibodies; an increase in the percentage of protection generated; an increase in the number and / or persistence of differentiated memory T-cells; and / or an increase in the duration of protection. 33. The method of any one of claims 30 to 32, wherein inhibiting CBP / β-catenin-mediated signaling and / or enhancing p300 / catenin-mediated signaling comprises one or more of: metabolic maintenance of post-mitotic cellular asymmetry in activated T cells in the subject; enhancing antigen-specific immunity by increasing the number and / or persistence of differentiated memory T cells; and / or enhancing presentation of antigens to T cells by antigen-presenting cells to enhance cooperation between the innate and adaptive immune systems.

[0021] 34. The method of any one of claims 30 to 33, wherein vaccination comprises administration of an antiviral vaccine. 35. The method of any one of claims 30 to 34, wherein the vaccination comprises administration of an antiviral vaccine selected from influenza, SARS, SARS-CoV-2, HPV-A, HPV-B, and / or shingles. 36. The method of any one of claims 30 to 35, wherein the subject is a human aged 55 to 75 years, 55 to 85 years, ≥ 50 years, ≥ 60 years, or ≥ 65 years.

[0022] 37. The method of any one of claims 30 to 36, wherein the administration comprises administration as an initiator before vaccination; and / or simultaneous administration with vaccination; and / or administration after or simultaneous with the first vaccination. [Brief explanation of the drawings]

[0023] [Figure 1]

[0023] Figure 1 shows the results of a SuperTOPFLASH cell-based luciferase assay (Wnt-driven luciferase activity in stably transfected cell lines, Hek293 and STF1.1) comparing the CBP / β-catenin inhibitory activity of three exemplary isoxazole compounds of the present invention (encoded [3+2]-101, [3+2]-117, and [3+2]-229; each compared at concentrations between 16 and 1000 nM) with the art-recognized specific CBP / β-catenin inhibitor ICG-001 used as a positive control (at 1.25, 2.5, 5, and 10 μM), according to a non-limiting embodiment of the present invention. As is readily apparent, the three isoxazole compounds of the present invention are generally approximately 10x or more potent in this assay than the control compound ICG-001. [Figure 2]

[0023] Figure 1 shows the results of a survivin promoter-driven luciferase activity assay (assay of human survivin 1Kb promoter-driven luciferase activity in a stably transfected Hek293 cell line; 1Kb Hu-survivin / luc-Hek293) comparing the CBP / β-catenin inhibitory activity of three exemplary isoxazole compounds of the present invention (encoded [3+2]-101, [3+2]-117, and [3+2]-229; each compared at concentrations between 0.1 and 10 μM) with the art-recognized specific CBP / β-catenin inhibitor ICG-001 used as a positive control (at 1.25, 2.5, 5, and 10 μM). As is readily apparent, the three isoxazole compounds of the present invention have IC values ​​that are generally at least approximately 10x more potent in this assay than the control compound ICG-001. [Figure 3A-3B]According to yet a further non-limiting embodiment of the present invention, the results of a SYBR-Green qPCR assay for survivin / BIRC5 (CBP-specific) and EphB2 (p300-specific) gene expression using GAPDH as a control gene are shown. Figure 3A compares the compound [3+2]-101 of the present invention (at 1 μM) with the art-recognized positive control ICG-001 (at 10 μM) in inhibiting CBP / β-catenin-specific gene (H. Ma et al., Oncogene 2005, 24, 3619-31) survivin / Birc5 gene expression (reflected as an increase in ΔΔCt). As is readily apparent, the CBP / β-catenin expression inhibitory activity of the isoxazoles of the present invention is approximately 10x greater than that of the control. Figure 3B compares the compound of the present invention, [3+2]-101 (at 1 μM), with the art-recognized positive control, ICG-001 (at 10 μM), in stimulating EphB2 (p300 / β-catenin-specific gene (Kumar S, et. al. Cancer Res. 2009, 69, 3736-45) gene expression (reflected as a decrease in ΔΔCt). As is readily apparent, the EphB2 gene expression stimulating activity of the isoxazole of the present invention is at least 10× greater than that of the control. This reflects a decrease in CBP / β-catenin-based transcription accompanied by an increase in p300 / β-catenin-based transcription, mediated by the specific CBP / β-catenin inhibitory activity of [3+2]-101 and ICG-001. [Figure 4A-4B]According to a further non-limiting embodiment of the present invention, to further confirm that ICG-001 and [3+2]-117 selectively interfere with β-catenin binding to CBP (FIG. 4A) while enhancing β-catenin binding to p300 (FIG. 4B), a co-immunoprecipitation assay (Emami K. et al. PNAS USA, 2004 Aug 24;101(34):12682-7) was performed in SW480 colorectal cancer cells as described above. Immunoprecipitation of β-catenin by CBP was inhibited by ICG-001 at 10 μM (FIG. 3A) and by [3+2]-117 at 500 nM. [3+2]-117 was approximately 20-fold more active than ICG-001 (compare lane 1 (DMSO control) with lanes 2 and 3). Despite the fact that CBP and p300 are highly homologous, the minimal interaction between β-catenin and p300 observed in SW480 cells was not blocked by ICG-001. Indeed, treatment with 10 μM ICG-001 slightly increased the amount of β-catenin coimmunoprecipitated with p300, consistent with a switch from CBP / β-catenin-mediated transcription to p300 / β-catenin-mediated transcription associated with the initiation of differentiation (Figure 4B, compare p300 IP, lanes 1 and 2). [3+2]-117 at 500 nM dramatically increased the interaction of β-catenin with p300 (Figure 4B, compare p300 IP, lanes 1 and 3). Quantification of the immunoblot data based on pixelation is shown below the immunoblots. [Figure 5] FIG. 1 shows, according to a further non-limiting embodiment of the present invention, a significant (Bonferroni's multiple comparison test; mean + / - SD) increase in body weight was observed in mice treated with [3+2]-120A and mice treated with [3+2]-120B in a bleomycin-induced pulmonary fibrosis model study in mice (treatment period: days 7-20). [Figure 6]According to a further non-limiting embodiment of the present invention, in the study of FIG. 5, all mice survived (so that the survival curves for the different treatment groups are superimposed; log-rank test), whereas conventionally approximately 30% of mice die before day 21, even when treated in this bleomycin-induced pulmonary fibrosis model. [Figures 7A-7D] According to further non-limiting embodiments of the present invention, Figures 5 and 6 show that there was a significant (Bonferroni's multiple comparison test; mean + / - SD) reduction in lung mass and Ashcroft score in mice treated with [3+2]-120A and [3+2]-120B based on histological analysis in this study. Figures 7A-7D show: body weight on the day of sacrifice (7A); left lung mass (7B); post-caval lobe mass (7C); and Ashcroft score (7D). [Figures 8A-8E] According to a further non-limiting embodiment of the present invention, representative histological data (photomicrographs of Masson's trichrome-stained lung sections; initial magnification ×100) for the study of Figures 5-7 are shown, demonstrating that the [3+2] series compounds are effective in reducing extracellular collagen deposition in a bleomycin-induced pulmonary fibrosis model. Figures 8A-8E are as follows: historical bleomycin-only control (8A, "ID:103"); [3+2]-120A low dose (8B, "ID:102"); [3+2]-120A high dose (8C, "ID:201"); [3+2]-120B low dose (8D, "ID:302"); and [3+2]-120B high dose (8E, "ID:401"). [Figure 9] FIG. 1 shows, according to a further non-limiting embodiment of the present invention, that in the NG / Nga mouse model of atopic dermatitis, there was no significant difference in mean body weight between the vehicle group and the treatment (representative compounds of the present invention [3+2]-121A (low dose) and [3+2]-121B) (high dose compound [3+2]-121A)) groups on any day during the treatment period, demonstrating the safety of the treatment. [Figure 10]FIG. 10 shows, according to a further non-limiting embodiment of the present invention, that there was no significant difference in the amount of food consumed per mouse ("food intake") between the vehicle control group and the treatment groups in the study of FIG. [Figures 11A-11B] According to a further non-limiting embodiment of the present invention, the [3+2]-121A group (low dose) showed a significant reduction in total dermatitis severity score, primarily for the back, on days 10 and 14 compared to the vehicle control group in the study of Figures 9 and 10. Figures 11A and 11B show: back region (11A); and total (dorsal region plus atrial appendage region) (11B). [Figure 12] According to a further non-limiting embodiment of the present invention, in the test of Figures 9 to 11, the [3+2]-121A group and the [3+2]-121B group showed a significant decrease in TEWL on day 14, demonstrating an improvement in barrier function. [Figures 13A-13C]

[0023] According to further non-limiting embodiments of the present invention, representative tissue sections taken in the study of Figures 9-12 confirm a significant reduction in inflammation and inflammatory cell influx in the [3+2]-121A and [3+2]-121B groups, as shown in representative photomicrographs. Figures 13A-13C are as follows: vehicle control (13A); [3+2]-121A (13B); and [3+2]-121B (13C). DETAILED DESCRIPTION OF THE INVENTION

[0024] Formula (Ia): [ka] [In the formula, R a is hydrogen or -CH3; R b is a monocyclic aryl group having 5 to 7 ring members, which may have 1 to 2 heteroatoms selected from nitrogen, oxygen, or sulfur, and which may have one or more substituents selected from the group consisting of halide, cyano, and lower alkyl; R is a phenyl group; a substituted phenyl group having one or more substituents (wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; benzyl groups; substituted benzyl groups having one or more substituents, wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; or a bicyclic aryl group, or a substituted bicyclic aryl having 8 to 11 ring members and optionally having 1 to 3 heteroatoms selected from nitrogen, oxygen, or sulfur (substituted bicyclic aryl is an amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-3 optionally bearing one or more substituents independently selected from alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; R 2 is hydrogen or -CH3; Y is selected from hydrogen, deuterium, or a halogen; W is hydrogen, phosphate or phosphoric acid salt, alkyl acid or fatty acid Ester of , or X, X is [ka] wherein Z is OR4 (wherein R4 is hydrogen or C1-C6 alkyl), or Z is an amino acid or amino acid ester; n is 1 or 2; L is -CH2-, -CF2-, or -C(CH3)2-; Q is [ka] wherein A, B, and D are independently selected from O, S, N, or —CH; and Z1, Z2 are independently selected from hydrogen, deuterium, halogen, C1-C4 alkyl, —OH, —OC1-C6 alkyl; and [ka] wherein R1, R2, and R3 are independently selected from hydrogen, C1-C6 alkyl, or C1-C6 alkyl containing one or more -OH groups; Z3 is selected from hydrogen, halogen, or aryl, heteroaryl, cycloalkyl, heterocycloalkyl, directly bonded or bonded to -NH-, bonded to -OC1-C3 alkyl, or bonded to -NHC1-C3 alkyl, or nitrogen-bonded cycloalkyl, heterocycloalkyl, or -NHC1-C4 alkyl, -N(C1-C4 alkyl)2, each of which is hydrogen, deuterium, halogen, C1-C4 alkyl, C1-C3 haloalkyl, -OH, -OC1-C6 alkyl, -OC1-C6 alkyl-C(O)NH-O H, -NH, -C(O)NH-C1-C6 alkyl-heteroaryl, -NHC(O)C1-C6 alkyl-C(O)NH-OH, heteroaryl or cycloalkyl, heterocycloalkyl, or cycloalkyl or heterocycloalkyl attached to a nitrogen, -NHC1-C4 alkyl, or -N(C1-C4 alkyl)2; and any salts thereof, including stereoisomers, geometric isomers, tautomers, solvates (e.g., hydrates), metabolites, prodrugs, isotopically labeled derivatives, and pharmaceutically acceptable salts thereof, are provided.

[0025] In this compound, alkyl or fatty acids Ester of teeth, [ka] (wherein m is 1 to 14), [ka] It can be preferably selected from:

[0026] In the present compounds, R can be a bicyclic aryl selected from naphthyl, quinolinyl, isoquinolinyl, quinoxaline, phthalazine, quinazoline, cinnoline, naphthyridine, or substituted variants thereof.

[0027] In the present compound, the compound preferably has the formula (Ib): [ka] wherein X1 and X2 are independently selected from N, or -CH.

[0028] In a preferred embodiment of the compound, L is -CH2-; Q is [ka] wherein A, B, and D are independently selected from O, S, N, or -CH [preferably, A is -CH, B is N, D is O (Q is Z3-isoxazole-), and W is hydrogen, phosphate, alkyl acid, or fatty acid Ester of , or X, X is [ka] wherein Z is OR4 (wherein R4 is hydrogen or C1-C6 alkyl), or Z is an amino acid or amino acid ester; n is 1 or 2}.

[0029] In this compound, Z3 can be selected from aryl or heteroaryl, each of which is substituted by 0-4 substituents independently selected from hydrogen, deuterium, halogen, C1-C4 alkyl, C1-C3 haloalkyl, -OH, -O1-C6 alkyl, -O1-C6 alkyl-C(O)NH-OH, -NH, -C(O)NH-C1-C6 alkyl-heteroaryl, -NHC(O)C1-C6 alkyl-C(O)NH-OH, or heteroaryl, cycloalkyl, heterocycloalkyl, or cycloalkyl or heterocycloalkyl bonded to nitrogen, -NHC1-C4 alkyl, or -N(C1-C4 alkyl)2 (preferably, Z3 is selected from aryl or heteroaryl).

[0030] Particularly preferred compounds are [ka] TIFF0007750851000029.tif249166 TIFF0007750851000030.tif244170 and a composition or pharmaceutical composition comprising a compound disclosed herein and a pharmaceutically acceptable excipient or carrier.

[0031] Another aspect of the present invention provides compounds of Formulas (Ia), (Ib) and (IIa) that are potent modulators of the Wnt / β-catenin pathway. Provided are potent compounds that inhibit CREB-binding protein (CBP) / β-catenin-mediated signaling, compositions and pharmaceutical compositions comprising these compounds, and the use of these compounds for the treatment of any abnormal CBP / β-catenin-mediated signaling disease or disorder, including, but not limited to, fibrosis, cancer, neurological disorders, metabolic disorders (including diabetes and fatty liver disease, e.g., alcoholic (ALD) and non-alcoholic fatty liver disease (ALD and NAFLD, respectively), and non-alcoholic steatohepatitis (NASH)), skin conditions (e.g., dermatitis, psoriasis, alopecia, skin aging, etc.), and may further include one or more of pulmonary hypertension, congestive heart failure, chronic kidney disease, renal fibrosis, cardiac fibrosis, polycystic ovary syndrome (PCOS), endometriosis, and / or systemic fibrosis / scleroderma. Both therapeutic and cosmetic methods are provided.

[0032] The compounds of formula (Ia), (Ib), and (IIa) of the present invention may contain chiral centers and therefore may exist in different enantiomeric and diastereomeric forms. The present invention relates to all optical isomers and all stereoisomers of compounds having the structures defined above, as racemic mixtures and as individual enantiomers and diastereoisomers of such compounds, and mixtures thereof, and to all pharmaceutical compositions and methods of treatment, as defined below, containing or using them, respectively. In some embodiments, the compound is the (S)-enantiomer. In other embodiments, the compound is the (R)-enantiomer.

[0033] As the compounds of the present invention may have at least two asymmetric centers, they can occur in various stereoisomeric forms or configurations. Therefore, the compounds may exist in separated (+)- and (-)-optically active forms, as well as mixtures thereof. The present invention includes all such forms within its scope. Individual isomers can be obtained by known methods, such as optical resolution, optically selective reactions, or chromatographic separation in the preparation of the final product or its intermediate. The present invention also provides pharmaceutical compositions and formulations comprising one or more of the disclosed compounds in an amount sufficient to specifically inhibit CBP / catenin-mediated signal transduction in a warm-blooded mammalian subject suffering from a disease or disorder mediated by abnormal CREB-binding protein (CBP) / β-catenin signal transduction in the warm-blooded mammalian subject when administered to the subject. The amount of compound administered preferably comprises a therapeutically effective amount, and in such cases, the pharmaceutical compositions and formulations may comprise a therapeutically effective amount of a compound having a structure disclosed herein or a therapeutically acceptable salt thereof and a pharmaceutically acceptable carrier, diluent, or excipient therefor. All of these forms are encompassed within the scope of the present invention.

[0034] definition "Lower" means that the number of carbon atoms making up a given group is between 1 and 6, unless otherwise indicated. "Halogen" means fluorine, chlorine, bromine or iodine. "Halo" means fluoro, chloro, bromo or iodo. "Alkyl" means a straight or branched, saturated, aliphatic group having a chain of carbon atoms. "Alkenyl" means a straight or branched carbon chain containing at least one carbon-carbon double bond. "Alkynyl" means a straight or branched carbon chain containing at least one carbon-carbon triple bond. "Alkylene" means a straight or branched saturated aliphatic polyvalent carbon chain, unless otherwise indicated. "Oxy" means an -O- group. It is noted that the oxy radical may be further substituted with a variety of substituents to form different oxy groups including hydroxy, alkoxy, aryloxy, heteroaryloxy, and the like. "Phosphate" or "phosphate salt" means PO3H2 or PO3 -- and an appropriate counterion (e.g., 1-2Na + , 1-2K + , or Ca ++ , etc.).

[0035] "Thio" refers to the group -S-. It is noted that the thio group can be further substituted with a variety of substituents to form different thio groups including mercapto, alkylthio, arylthio, heteroarylthio, and the like. "Sulfinyl" means an -SO- group. It is noted that the sulfinyl radical may be further substituted with a variety of substituents to form different sulfinyl groups including alkylsulfinyl, arylsulfinyl, heteroarylsulfinyl, and the like. "Sulfonyl" means a -SO- group. It is noted that the sulfonyl radical may be further substituted with a variety of substituents to form different sulfonyl groups including alkylsulfonyl, arysulfonyl, heteroarylsulfonyl, and the like.

[0036] "Alkoxy" means an oxygen moiety having a further alkyl substituent. "Heteroatom" means an atom that is not a carbon or hydrogen atom. Particular examples of heteroatoms include, but are not limited to, nitrogen, oxygen, and sulfur. "Aryl" means a monocyclic or polycyclic group in which each ring is aromatic or, when fused with one or more rings, forms an aromatic ring. "Heteroaryl" means a monocyclic or polycyclic aromatic group in which at least one ring atom is a heteroatom and the remaining ring atoms are carbon. "Cycloalkyl" means a non-aromatic, saturated or partially unsaturated, monocyclic, fused bicyclic, or bridged polycyclic ring group.

[0037] "Heterocycloalkyl" means cycloalkyl, as defined in this Application, provided that one or more of the atoms forming the ring is a heteroatom independently selected from N, O, or S. As used herein, "fused ring" refers to a ring that is bonded to another ring to form a compound having a bicyclic structure when ring atoms common to both rings are directly bonded to each other. As used herein, a "bridged ring" refers to a ring that is joined to another ring to form a compound having a bicyclic structure, provided that two ring atoms common to both rings are not directly bonded to each other. "Protected derivatives" means derivatives of compounds in which one or more reactive sites are blocked with protecting groups. A comprehensive list of suitable protecting groups can be found in T.W. Greene, Protecting Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, Inc. 1999.

[0038] "Isomers" means all compounds that have identical molecular formulae but differ in the nature or sequence of bonding of their atoms or in the arrangement of their atoms in space. Isomers that differ in the arrangement of their atoms in space are called "stereoisomers." Stereoisomers that are not mirror images of one another are called "diastereomers," and stereoisomers that are non-superimposable mirror images are called "enantiomers" or sometimes "optical isomers." A carbon atom bonded to four non-identical substituents is called a "chiral center." A compound with one chiral center has two enantiomeric forms of opposite chirality. A mixture of two enantiomeric forms is called a "racemic mixture." A compound with more than two chiral centers has two enantiomeric forms.n-1 It has an enantiomeric pair (where n is the number of chiral centers). Compounds with two or more chiral centers can exist as individual diastereomers or as a mixture of diastereomers, called a "diastereomeric mixture." When one chiral center is present, a stereoisomer can be characterized by the absolute configuration of that chiral center. Absolute configuration refers to the arrangement in space of the substituents attached to the chiral center. Enantiomers are characterized by the absolute configuration of their chiral centers and are described by the R- and S-sequencing rules of Cahn, Ingold, and Prelog. Conventions for stereochemical nomenclature, methods for determining stereochemistry, and methods for separating stereoisomers are well known in the art (see, for example, "Advanced Organic Chemistry," 4th edition, March, Jerry, John Wiley & Sons, New York, 1992).

[0039] "Animal" includes humans, non-human mammals (e.g., mice, rats, dogs, cats, rabbits, cows, horses, sheep, goats, pigs, deer, etc.), and non-mammals (e.g., birds, etc.). "Disease" specifically includes any ill-health of an animal or part thereof, including ill-health that may be caused by or is prone to medical or veterinary therapy administered to that animal, i.e., a "side effect" of such therapy. "Pharmaceutically acceptable" generally means something that is safe, non-toxic, and not biologically undesirable and useful in preparing pharmaceutical compositions, and includes something that is acceptable for veterinary use as well as human pharmaceutical use.

[0040] "Pharmaceutically acceptable salt" or "salt" refers to a salt of a compound of the present invention that is pharmaceutically acceptable, as defined above, and has the desired pharmacological activity. Such salts include acid addition salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or acid addition salts formed with organic acids, such as acetic acid, propionic acid, hexanoic acid, heptanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, o-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxybenzoylbenzoic acid, ... Pharmaceutically acceptable salts include ethanesulfonic acid, benzenesulfonic acid, p-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, p-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]oct-2-ene-1-carboxylic acid, glucoheptonic acid, 4,4'-methylenebis(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, and the like. Pharmaceutically acceptable salts also include base addition salts, which can be formed when an acidic proton present can react with an inorganic or organic base. Acceptable inorganic bases include sodium hydroxide, sodium carbonate, potassium hydroxide, aluminum hydroxide, and calcium hydroxide. Acceptable organic bases include ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like.

[0041] By "therapeutically effective amount" or "therapeutically effective amount" is meant the amount that, when administered to an animal for treating a disease, is sufficient to effect such treatment for the disease. By "prophylactically effective amount" is meant the amount that, when administered to an animal for preventing a disease, is sufficient to effect such prevention for the disease. "Treatment" or "treating" refers to any administration of a compound of the invention, and includes (i) preventing disease in an animal that may be predisposed to the disease but has not yet experienced or exhibited the pathology or symptomology of the disease; (ii) inhibiting disease (i.e., halting further development of pathology and / or symptomology) in an animal experiencing or exhibiting the diseased pathology or symptomology; or (iii) ameliorating disease (i.e., reversing the pathology and / or symptomology) in an animal experiencing or exhibiting the diseased pathology or symptomology.

[0042] A "cosmetically effective amount" is an amount that, when administered (e.g., transdermally, topically), is sufficient to affect the cosmetic treatment of a cosmetic condition (e.g., wrinkles, hyperpigmentation, redness, rosacea, dryness, cracking, loss of color, loss of elasticity, thinning hair, loss of color, scarring, acne, sun damage, hair loss, loss of hair color, reduced cuticle growth, reduced nail growth).

[0043] Therapeutic Uses and Pharmaceutical Compositions of the Compounds of the Invention According to embodiments of the present invention, the exemplary diseases and conditions discussed below, regardless of cause or tissue manifestation, can be treated by modulating a common pathway (WNT-β-catenin signaling) shared by the pathologies. For example, treating a disease or disorder can include administering to a patient or warm-blooded mammal having a disease or disorder mediated by aberrant CREB-binding protein (CBP) / β-catenin signaling an amount of a compound of the present invention sufficient to inhibit CBP / catenin signaling and / or enhance p300 / catenin-mediated signaling. The WNT-β-catenin pathway plays a critical role in a wide range of diseases. Given the critical role of WNT signaling in virtually every organ system in normal homeostasis and repair after injury, it is not surprising that aberrant regulation of this signaling cascade is associated with a range of diseases (see, e.g., Kahn, M., NATURE REVIEWS | DRUG DISCOVERY VOLUME 13 | JULY 2014 | 513). Beyond its clear role in multiple malignancies, aberrant WNT signaling has been implicated as an important part of a variety of other diseases, including neurological disorders, inflammatory and fibrotic diseases, and disorders of endocrine function and bone metabolism in adults.

[0044] In particular aspects, aberrant WNT signaling is implicated in cancers (e.g., involvement of cancer stem cells in minimal residual disease in solid (e.g., colon, pancreas, lung, liver, bladder, prostate, melanoma, glioma, medulloblastoma, osteosarcoma, uterine, endometrial, and breast, etc.) and liquid tumors (e.g., CML, CLL, AML, ALL, etc.); multiple myeloma; autoimmune disorders including type 1 diabetes, rheumatoid arthritis, inflammatory bowel disease; focal disorders (e.g., psoriasis, vitiligo, and atopic dermatitis); fibrosis including cardiac, liver, lung, kidney, systemic, and peritoneal (endometriosis), and ocular fibrosis). fibrosis); osteoarthritis and osteoporosis; metabolic diseases including type II diabetes; hypertension; familial adenomatous polyposis; myelodysplastic syndromes (MDS); myeloproliferative neoplasms (MPN); CHIP (clonal hematopoiesis of undetermined potential); prefibrotic conditions (e.g., AFLD, NASH, NAFLD, cirrhosis); polycystic kidney disease; polycystic ovary syndrome (PCOS), endometriosis, metabolic diseases; type II diabetes; hypertension; pulmonary disorders (e.g., asthma and COPD); neurological diseases including, but not limited to, neurogenesis, autism, spectrum disorders, schizophrenia, and neurodegenerative diseases including ALS, Huntington's disease, Parkinson's disease, Alzheimer's disease, frontotemporal dementia, multiple sclerosis (MS); and more generally, diseases linked to aging. The involvement of WNT signaling in specific representative diseases is outlined in detail below.

[0045] Cancer. Aberrant regulation of WNT signaling has emerged as a recurring theme in cancer biology. Components of WNT signaling can be essentially characterized as positively or negatively acting components, which primarily act to suppress tumorigenesis; negatively acting components are found in mutated or loss-of-function forms in cancer, while positive components are activated. The discovery in 1991 that mutations in the tumor suppressor APC, through aberrant activation of WNT signaling, were associated with the majority of sporadic colorectal cancers, provided considerable impetus for therapeutically targeting this pathway. Germline defects in APC cause familial adenomatous polyposis, in which affected individuals develop hundreds of polyps in the colon at an early age and ultimately progress to colorectal cancer with 100% penetrance. Loss of function of both APC alleles is required for tumorigenesis and, in conjunction with the protein's ability to regulate β-catenin protein stability as well as chromosomal stability, currently highlights APC as one of the most frequently mutated genes in human cancer. Mutations affecting the WNT pathway are not limited to colon cancer. For example, loss-of-function mutations in AXIN have been found in hepatocellular carcinoma, and oncogenic β-catenin mutations, first described in colon cancer and melanoma, have subsequently been found to occur in a variety of solid tumors, including hepatocellular carcinoma, thyroid tumors, and ovarian endometrioid adenocarcinoma. Epigenetic silencing is also frequently observed to alter the expression levels of negative regulators of the WNT-β-catenin pathway. For example, methylation of genes encoding putative extracellular WNT antagonists, such as secreted Frizzled-related proteins (SFRPs), has been described in colon, breast, prostate, lung, and other cancers. Increased expression of WNT ligands or effector proteins, including Dishevelled (DVL), has also been described.In particular, aberrant WNT signaling has been linked to cancer stem cells involved in minimal residual disease and recurrence of solid (e.g., colon, pancreas, lung, liver, bladder, prostate, melanoma, glioma, medulloblastoma, osteosarcoma, uterine, endometrial, and breast) and liquid tumors (e.g., CML, CLL, AML, ALL, etc.) Specific small molecule CBP / β-catenin antagonists are effective in eliminating cancer stem cells, minimal residual disease, and disease recurrence (e.g., Kim, et al. Exp. Hematol. 2017 doi: 10.1016 / j.exphem.2017.04.010). Furthermore, premalignancies and syndromes such as clonal hematopoiesis of undetermined potential (CHIP), myelodysplastic syndromes (MDS), myelofibrosis (MF) and myeloproliferative neoplasms (MPN), driven by defective stem cells, and Barrett's esophagus can be prophylactically eliminated by specific small molecule CBP / β-catenin antagonists (Thomas and Kahn Cell Biol. Toxicol. 2016 doi: 10:1007 / s10565-016-9318-0).

[0046] Fibrosis. Fibrosis is characterized by the excessive accumulation of extracellular matrix components, which disrupts physiological tissue assembly and leads to dysfunction of the affected organ. It is generally suggested that fibrosis accounts for approximately 45% of deaths in industrialized countries, thereby highlighting the great medical need for effective antifibrotic therapies. Activated WNT-β-catenin signaling has been linked to fibrosis in multiple organ systems, including the lung, indicating that this developmental pathway can be reactivated in adult tissues after injury. Specific small molecule WNT modification in several mouse models of fibrosis (e.g., lung and kidney models) has proven highly effective.Specific inhibition of CBP-β-catenin interaction has been shown to not only ameliorate but also reverse late fibrotic injury in mouse models of lung, kidney, liver, heart, and systemic fibrosis, and endometriosis (Sci Rep. 2019 Dec 27;9(1):20056. doi:10.1038 / s41598-019-56302-4; Akcora et al., Biochim Biophys Acta Mol Basis Dis. 2018 Mar;1864(3):804-818. doi: 10.1016 / j.bbadis.2017.12.001; Xiao et al., Biochim Biophys Acta Mol Basis Dis. 2019 Jun 1;1865(6):1313-1322. doi: 10.1016 / j.bbadis.2019.01.027. Epub 2019 Jan 30;Zhao et al., Sci Rep. 2018 Jun 12;8(1):8996. doi:10.1038 / s41598-018-27064-2; Kimura et al., EBioMedicine 2017 Sep;23:79-87. doi: 10.1016 / j.ebiom.2017.08.016. Epub 2017 Aug 19. Safety, Tolerability, and Preliminary Efficacy of the Anti-Fibrotic Small Molecule PRI-724, a CBP / β-Catenin Inhibitor, in Patients with Hepatitis C Virus-related Cirrhosis: A Single-Center, Open-Label, Dose In this study, 10 and 40 mg / m 2 The CBP / β-catenin antagonist PRI-724, administered intravenously at a dose of 1 / day for 12 weeks (1 week on, 1 week off) to patients with HCV cirrhosis, appeared safe, provided dose-dependent plasma exposure of the drug, and resulted in improvement in liver histology and Child-Pugh classification in some patients.

[0047] Pulmonary fibrosis. Pulmonary fibrosis destroys the lungs' ability to transport oxygen and other gases into and out of the blood. The disease alters the lungs' delicate, elastic tissue, transforming them into thicker, tougher, fibrous tissue. This transformation or replacement of original tissue is similar to the permanent scarring that can occur in other damaged tissues. Scarring in the lungs reduces the lungs' ability to transfer gases (i.e., oxygen, carbon dioxide) into and out of the blood. Gradually, the alveolar sacs are replaced by fibrotic tissue. As scarring forms, the tissue thickens, causing an irreversible loss of the tissue's ability to transfer oxygen into the bloodstream. Symptoms include, among others, shortness of breath with exertion; a chronic, dry, hacking cough; fatigue and weakness; chest discomfort; loss of appetite; and rapid weight loss. Several causes of pulmonary fibrosis are known, including occupational and environmental exposures. Many jobs, especially those involving mining or those that expose workers to asbestos or metal dust, can cause pulmonary fibrosis. Workers performing these types of jobs inhale small particles (such as silica dust or asbestos fibers) that can damage the lungs, especially the small airways and alveolar sacs, resulting in scarring associated with fibrosis. Farmers are also at risk. Some organic substances, such as moldy hay, can trigger an allergic reaction in the lungs. This reaction, called farmer's lung, can lead to pulmonary fibrosis. Other gases found on farms are directly toxic to the lungs. Another cause is sarcoidosis, a disease characterized by the formation of granulomas (areas of inflammatory cells), which can attack any area of ​​the body but most frequently affect the lungs. Some medications, such as radiation, can have the undesirable side effect of causing pulmonary fibrosis, as can treatments for breast cancer. Connective tissue or collagen diseases, such as systemic sclerosis, are also associated with pulmonary fibrosis. Genetic or familial factors may be involved, but this cause is less common than the other causes listed above. In chronic obstructive pulmonary disease (COPD), connective tissue proliferation and fibrosis may characterize severe COPD. COPD may develop as a result of smoking or chronic asthma.

[0048] Idiopathic Pulmonary Fibrosis (IPF). When all known causes of interstitial lung disease have been ruled out, the condition is called "idiopathic" (of unknown cause) pulmonary fibrosis (IPF). More than 83,000 Americans live with IPF, with more than 31,000 new cases occurring each year. Lung scarring contributes to this debilitating condition. Lung sacs develop scarring, or fibrotic tissue, which gradually interferes with the body's ability to transfer oxygen into the bloodstream, preventing vital organs and tissues from receiving enough oxygen to function normally. There are several potential causes of IPF, including viral illnesses such as SARS-CoV-2 and allergies or environmental exposures (including tobacco smoke). There is also a familial form of the disease known as familial idiopathic pulmonary fibrosis. Patients with IPF suffer from symptoms similar to those of patients with pulmonary fibrosis when their lungs lose their ability to transfer oxygen into the bloodstream. These symptoms include, among others, shortness of breath during or after physical activity; a spasmodic, dry cough; gradual, involuntary weight loss; fatigue and weakness; chest discomfort; clubbing, or swelling of the tips of the fingers (or occasionally toes) due to tissue buildup. These symptoms can greatly reduce the quality of life of patients with IPF. Pulmonary rehabilitation and oxygen therapy can reduce the lifestyle-altering effects of IPF but do not provide a cure.

[0049] Diabetes and metabolic diseases. WNT signaling is crucial not only for stem cell maintenance, differentiation, and migration, but also for organogenesis. WNT signaling also plays a crucial role in various endocrine functions and has therefore been implicated in several endocrine disorders. WNT signaling is important in controlling insulin sensitivity, and its dysregulation has been linked to the development of diabetes. In particular, WNT10B increases insulin sensitivity in skeletal muscle cells. Overexpression of WNT5B induces adipogenesis. Decreased expression of β-catenin-independent WNT5B, which has been demonstrated in patients with type 2 diabetes, can increase susceptibility to type 2 diabetes. β-catenin / TCF7L2-dependent WNT signaling (canonical pathway) is involved in pancreatic development, islet function, and insulin production and secretion. Glucagon-like peptide (GLP-1) and the chemokine stromal cell-derived factor-1 (SDF1) modify canonical WNT signaling. Furthermore, polymorphisms in the transcription factor TCF7L2 (also known as TCF4) are associated with increased susceptibility to type 2 diabetes. Individuals with risk alleles of TCF7L2 exhibit impaired insulin secretion, and TCF7L2 in pancreatic β-cells appears to play a critical role in glucose metabolism through the control of pancreatic β-cell mass.

[0050] Experimental loss of TCF7L2 function in pancreatic islets impairs glucose-stimulated insulin secretion, suggesting that disruptions in the Wnt signaling pathway may contribute substantially to the susceptibility and pathogenesis of T2D. Interestingly, nicotine has been shown to enhance renal cell proliferation and fibronectin production under high glucose conditions, in part through activating the Wnt / β-catenin pathway. Although increased Wnt / CBP / β-catenin signaling can initially induce pancreatic β-cell proliferation, continuous Wnt-driven mitogenic signaling can ultimately lead to a loss of differentiation potential and functionality.

[0051] Recently, researchers treated intact human islets derived from cadavers with conditioned medium derived from L-cells constitutively producing WNT3A, R-spondin 3, and noggin, and then added inhibitors of RHO-associated protein kinase (ROCK) and RHOA to enhance cell survival. This resulted in an approximately 20-fold increase in β-cell proliferation compared to glucose alone. Importantly, treatment with this conditioned medium did not impair glucose-stimulated insulin secretion or reduce cellular insulin content. Transcriptome-wide gene expression profiling and follow-up signaling studies demonstrated that conditioned medium treatment specifically promoted WNT signaling.

[0052] Neurological Disorders. The importance of WNT signaling during embryonic development of the central nervous system is well established. The WNT pathway also regulates nervous system patterning and neuroplasticity. WNTs also play a role in axon guidance and in influencing synaptogenesis. Therefore, it is not surprising that abnormalities in WNT signaling have been observed in neurological disorders in adulthood. For example, a Scottish family with a high incidence of schizophrenia, depression, and bipolar disorder was found to carry a balanced chromosomal translocation involving the DISC1 gene (disrupted in schizophrenia 1). Subsequently, the protein product of DISC1 was found to play an important role in neurogenesis and neural progenitor cell proliferation. DISC1 directly interacts with and inhibits GSK3β activity, thus enhancing β-catenin-mediated transcription.

[0053] Neuroanatomical observations and functional magnetic resonance imaging (MRI) have shown that a major pathological feature in individuals with autism may be premature overgrowth of the cerebral cortex, hippocampus, amygdala, and cerebellum. Interestingly, transgenic mice expressing a constitutively active form of β-catenin in neuronal progenitor cells developed macroscopically enlarged cerebral cortex, hippocampus, and amygdala. Importantly, copy number variations of microdeletions and microduplications of genes involved in the canonical WNT signaling pathway (e.g., Frizzled 9 (FZD9), B-cell lymphoma 9 (BCL9), or cadherin 8 (CDH8)) are found in patients with autism spectrum disorder. Related studies investigating WNT2, DISC1, MET, dedicator of cytokinesis protein 4 (DOCK4) or Abelson helper integration site 1 (AHI1; also known as jouberin) provide further evidence that the canonical WNT pathway may be affected in autism.

[0054] The WNT signaling cascade has also been implicated in Alzheimer's disease. Presenilin proteins, which are associated with early-onset Alzheimer's disease, are negative regulators of canonical WNT signaling. Variant alleles of the WNT receptor LRP6 (low-density lipoprotein receptor-related protein 6) have been associated with Alzheimer's disease in population-based linkage studies. This suggests that multiple mechanisms leading to abnormal WNT-mediated regulation of adult neurogenesis may be associated with Alzheimer's disease. Because the underlying cause(s) of Alzheimer's disease have not been clearly elucidated, the mechanisms by which abnormal WNT regulation may play a role in Alzheimer's disease are also unknown. WNT signaling is involved in cerebral angiogenesis and blood-brain barrier formation, synaptogenesis, amyloid-β-induced neuroinflammation and neurotoxicity, and neurodegeneration. Abnormal regulation of any or all of these processes may contribute to the initiation and progression of disease. Skin. The WNT signaling cascade has been implicated in skin development and maintenance (see, e.g., STEM CELLS 2018;36:22-35). Secreted Wnt proteins can stimulate multiple intracellular signaling pathways and act as growth factors to regulate a variety of processes, including cell proliferation, differentiation, migration, and polarity. Among Wnt-stimulated pathways, Wnt / β-catenin signaling is known as a key regulatory pathway governing fate choices during development and tissue morphogenesis. Wnt signaling is one of the major cues directing skin development and maintenance. While Wnt signaling is primarily linked to HF (hair follicle) induction during skin development, it has also recently been shown to regulate epidermal stratification. In primary human keratinocytes, Wnt5a acts as an autocrine stimulus to promote extracellular calcium-induced keratinocyte differentiation by coupling with the Wnt / β-catenin pathway. Throughout life, the skin epidermis is periodically renewed. Capable of self-renewal and differentiation, skin epidermal SCs provide an unlimited source of cells to maintain tissue homeostasis, as well as to regenerate HFs and repair the epidermis after injury. Wnt signaling is crucial in all of these processes, and Wnt-dependent signaling plays a crucial role in the maintenance, activation, and fate determination of SC populations.

[0055] Vaccine (e.g., SARS-CoV-2 (COVID-19), influenza, etc.) enhancement Age. With age, the immune system loses some of its momentum (reflected by immunosenescence, fewer naive T cells and B cells), which may contribute to a higher vulnerability to COVID-19 and infection more generally in older subject groups (e.g., humans ≥ 60 or ≥ 65 years old). Furthermore, vaccines may perform poorly in this elderly subject population, which often experiences inflammaging (Willyard, C., Nature Vol. 586, 2020; Akbar & Gilroy, Science 369 (6501), 256-257, 2020, DOI: 10.1126 / science.abb0762; A. Parmigiani et al., PLOS ONE 8, e79816 (2013)), characterized by chronic inflammation (impaired clearance of dead and dying cells from sites of immune activity and high baseline serum levels of C-reactive protein (CRP) and cytokines, e.g., interleukin-6 (IL-6), and IL-8, factors that can inhibit antigen-specific (e.g., anti-viral) immunity, e.g., influenza virus).

[0056] Current influenza vaccination strategies that prioritize older adults (55–75 years of age) have proven less effective than expected in reducing severe morbidity and mortality in this population, suggesting that additional strategies may be needed to enhance vaccination efficacy, which inherently relies on immunological memory (Anderson et al., Ann Intern Med. 2020;172:445–452. doi:10.7326 / M19-3075).

[0057] Similarly, SARS-CoV-2 primarily induces mild to moderate symptoms in younger individuals but causes a severe respiratory disease (coronavirus disease 2019, COVID-19) that induces devastating morbidity and mortality in older individuals. A key feature of severe disease is hyperproliferative inflammation in the patient's airways (Merad & Martin, Nat. Rev. Immunol. 20, 355, 2020). Memory T cells. A hallmark of the aging immune system is the inability to induce long-lived memory (Kim, Chulwoo, et al., Cell Reports 25, 2148-2162, November 20, 2018). Furthermore, promoting the rate of asymmetric cell division (ACD) can improve the long-term survival and function of T cells, opening new perspectives for vaccination (Borsa, et al., Sci. Immunol. 4, eaav1730 (2019)). It has been demonstrated that asymmetric cell division is responsible for the dichotomy between memory T cells and effector cells generated from a common precursor activated by antigen recognition in the context of antigen-presenting cells (Morrot, Alexandre, Ann Transl Med 2017;5(5):121; citing Verbist KC, Guy CS, Milasta S, et al. Metabolic maintenance of cell asymmetry following division in activated T lymphocytes (Nature 2016;532:389-93)). Furthermore, memory T cells appear to use asymmetric cell division to generate cellular heterogeneity when faced with pathogen rechallenge (Ciocca, Maria, L. et al., The Journal of Immunology, 2012, 188: 4145-4148).

[0058] Currently proposed approaches to vaccine enhancement may involve the use of vaccine adjuvants, higher doses of viral antigens, or the identification of drugs that can improve vaccine responses (e.g., rejuvenate the immune system) in older populations. For example, mTOR inhibitors (e.g., RTB101, rapamycin, metformin) have been proposed (Mannick, JB et al. Sci. Transl. Med. 10, 449, eaaq1564, 2020). Anti-inflammatory drugs (e.g., losmapimod, dexamethasone) and senolytics (e.g., fisetin) have also been proposed to boost immunity.

[0059] However, there is a further urgent need for more effective compositions and methods for enhancing vaccine (e.g., antiviral vaccines for influenza, SARS, SARS-CoV-2, HPV, HEP-A, HEP-B, shingles, etc.) responses, for example, by, for example; maintaining metabolic asymmetry in post-mitotic activated T cells in a subject; and / or enhancing antigen-specific immunity by increasing the number and / or persistence of differentiated memory T cells; and / or by enhancing presentation of antigens to T cells by antigen-presenting cells to enhance cooperation between the innate and adaptive immune systems, particularly in elderly subjects (e.g., 55-75 years old; >60 years old; >65 years old) (Ljungberg, Johanna K. et al, Front. of Immunol. 10;2521, 2019). CBP / catenin inhibitor. Treatment of ART-suppressed SIVmac251-infected RM with PRI-724 (a specific CBP / β-catenin inhibitor) has been shown to reduce memory stem cell (SCM) and central memory (CM) T-cell proliferation and modify the SCM and CM CD4+ T-cell transcriptomes toward more differentiated memory T-cell profiles, demonstrating that the stemness pathway of long-lived memory CD4+ T-cells can be pharmacologically modified in vivo, thus establishing a novel strategy for targeting HIV persistence (Mavigner, M. et al., J. Virol. doi:10.1128 / JVI.01094-19).

[0060] In accordance with particular aspects of the present invention, there is a need for safe and effective methods of treatment (eg, prophylactic and / or therapeutic) to target underlying aging mechanisms at approximately the time of "vaccination."

[0061] According to detailed embodiments of the present invention, the disclosed CBP / catenin inhibitors have substantial utility for enhancing vaccination (e.g., antiviral vaccines, e.g., for influenza, SARS, SARS-CoV-2, HPV-A, HPV-B, shingles, etc.), particularly in elderly subjects (e.g., 55-75 years; >60 years; >65 years), e.g., by maintaining metabolic asymmetry in post-mitotic activated T cells in a subject; and / or for enhancing antigen-specific immunity by increasing the number and / or persistence of differentiated memory T cells; and / or for enhancing presentation of antigens to T cells by antigen-presenting cells to enhance cooperation between the innate and adaptive immune systems, particularly in elderly subjects (e.g., 55-75 years; >60 years; >65 years). According to further embodiments, the compounds can be administered prophylactically and / or therapeutically, including as a starter before vaccination and / or co-administered with vaccination and / or co-administered after a primary vaccination or (e.g., with a vaccine booster). According to a further aspect, the compounds can be used to enhance vaccination in a mammalian (eg, human) subject with any vaccine.

[0062] SARS-CoV-2 (COVID-19) tissue (lung, liver, kidney, heart, etc.) destruction (e.g., pulmonary fibrosis, ARDS) As discussed above, inflammaging has many implications for COVID-19 patients (e.g., as discussed previously by Akbar & Gilroy). The accumulation of senescent cells in the airways of elderly patients may be involved in initiating an inflammatory cascade that can inhibit T cell responses to existing virus-infected cells. Massive inflammation alone does not explain the extensive tissue destruction observed in the lungs of COVID-19 patients with severe disease, suggesting that age-related changes in T cells may play a role in immunopathology. T lymphocytes that are highly differentiated and exhibit senescence-like characteristics accumulate in elderly individuals. Although these senescent T cells lose the ability to proliferate after activation and express multiple markers of senescence, including DNA damage-associated proteins [e.g., phosphorylated histone H2AX (gH2AX)] and cyclin-dependent kinase inhibitors (e.g., p16INK4A), they are still highly efficient cytotoxic cells, able to kill different cell types that express NKRs and NKR ligands, including senescent nonlymphoid cells. Another consequence of inflammation is the induction of NKR ligand expression by cells in the lung, which should make them more susceptible to killing by infiltrating NKR-expressing T cells (Id).

[0063] Pulmonary fibrosis. Almost all severe COVID-19-related outcomes are characterized by pneumonia, many with ground-glass opacities, and many (approximately 40%) develop acute respiratory distress syndrome (ARDS). There is concern that some organs, including the lungs, may suffer from long-term postinfection dysfunction, inflammation, and lack of resolution (e.g., pulmonary fibrosis, a recognized sequela of ARDS). Mechanical ventilation is the most important supportive therapy for patients with ARDS, including those with COVID-19, but it can induce or exacerbate lung injury, termed ventilator-induced lung injury (VILI) (Slutsky and Ranieri, NEJM, 2013). Although the virus is eradicated in patients recovering from COVID-19, removing the cause of lung injury does not inherently prevent the development of progressive, fibrotic, irreversible interstitial lung disease. Furthermore, even relatively small residual, non-progressive fibrosis can result in significant morbidity and mortality in the elderly population of patients with COVID-19, many of whom have pre-existing pulmonary conditions (Bem, Reinout A.; https: / / doi.org / 10.1016 / S2213-2600(20)30222-8). The description of the most lethal group of patients for whom SARS-CoV-2 infection is most severe is also highly representative of patients with idiopathic pulmonary fibrosis (IPF). Anti-fibrotic therapies available or in development may be of value in preventing severe COVID-19 in patients with IPF, have the potential to treat severe COVID-19 in patients without IPF, and may have a role in preventing fibrosis after SARS-CoV-2 infection (George et al., Lancet August 2020).

[0064] Acute lung injury and ARDS are the main causes of mortality in COVID-19. Conventional therapy may be possible with drugs such as pirfenidone and nintedanib, which are currently only available in oral form and therefore cannot be used in intubated and mechanically ventilated patients, thereby limiting their use in individuals with severe COVID-19 in intensive care units (ICUs). Furthermore, pirfenidone can be used to treat patients with an estimated glomerular filtration rate of 1.73 m / s. 2 It should be avoided if the flow rate is less than 30 mL / min per minute. Furthermore, both pirfenidone and nintedanib may be associated with hepatotoxicity, and liver dysfunction is common in patients infected with SARS-CoV-2. Further uncertainty concerns the rapidity with which antifibrotic agents act, and known agents may be of little value in ventilated patients, after the window of opportunity for effective treatment has already passed. In accordance with particular aspects of the present invention, there is a need for safe and effective methods of treating (e.g., prophylactically and / or therapeutically) SARS-CoV-2 (COVID-19) lung tissue destruction (e.g., pulmonary fibrosis, ARDS). As summarized above under "Therapeutic Uses and Pharmaceutical Compositions of the Invention" (pages 29-34), the utility of CBP / β-catenin inhibitors for treating aspects of fibrosis is recognized.

[0065] Thus, according to detailed aspects of the invention, the disclosed CBP / catenin inhibitors have substantial utility in treating SARS-CoV-2 (COVID-19) tissue (e.g., lung, liver, etc.) destruction (e.g., pulmonary fibrosis, ARDS), including both during the acute phase of disease, and in preventing long-term complications, particularly in elderly individuals (e.g., 55-75; ≥60 years; ≥65 years). According to further aspects, the compounds may be administered prophylactically and / or therapeutically, in either case preferably beginning before or within the first 1-3 weeks, preferably before or within the first week of ARDS onset.

[0066] Dosage: The pharmaceutical composition of the present invention is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., oral (e.g., in capsule or tablet form), intravenous, intradermal, subcutaneous, inhalation, transdermal (topical), transmucosal, and rectal administration. Solutions or suspensions (e.g., injections) used for parenteral (particularly intravenous), intradermal, or subcutaneous application may contain the following components: a sterile diluent, such as water for injection, saline, fixed oils, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; an antibacterial agent, such as benzyl alcohol or methylparaben; an antioxidant, such as ascorbic acid or sodium bisulfite; a chelating agent, such as ethylenediaminetetraacetic acid; a buffer, such as acetic acid, citric acid, or phosphate, and an agent for adjusting tonicity, such as sodium chloride or dextrose. In addition, the pH may be adjusted with an acid or base, such as hydrochloric acid or sodium hydroxide. Parenteral preparations can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

[0067] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, NJ), or phosphate-buffered saline (PBS). In all cases, the composition must be sterile and fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof. Proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. Prevention of microbial action can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, etc. In many cases, it is preferable to include an isotonic agent in the composition, for example, sugars, polyalcohols such as mannitol, sorbitol, sodium chloride, etc. Prolonged absorption of injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, aluminum monostearate and gelatin.

[0068] Sterile injectable preparations can be prepared by incorporating the active compound, for example, the compound having general formula (Ia), in the required amount in a suitable solvent containing one or a combination of the above-listed ingredients, and then optionally sterilizing and filtering.Generally, dispersion is prepared by incorporating the active compound into a sterile vehicle that contains dispersion medium and other necessary ingredients obtained from the above-listed ingredients.In the case of sterile powder for preparing sterile injectable preparations, the preferred method of preparation is vacuum drying and freeze-drying, which produces the powder of active ingredient plus any additional desired ingredients obtained from its solution that has been previously sterile-filtered.

[0069] Oral compositions generally include an inert diluent or an edible carrier. They can be sealed in gelatin capsules or compressed into tablets. For oral therapeutic administration, the active compound can be incorporated with an excipient and used in the form of tablets, lozenges, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, where the compound in the fluid carrier is applied orally, used topically, expectorated, or swallowed. Pharmaceutically compatible binders and / or adjuvant materials can be included as part of the composition. Tablets, pills, capsules, lozenges, etc. can contain any of the following ingredients or compounds of similar nature: That is, binders such as microcrystalline cellulose, gum tragacanth, or gelatin; excipients such as starch or lactose; disintegrants such as alginic acid, Primogel, or corn starch; lubricants such as magnesium stearate or Sterotes; glidants such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; or flavoring agents such as peppermint, methyl salicylate, or orange flavoring.

[0070] For administration by inhalation, the compounds are delivered in the form of an aerosol spray from pressured container or dispenser which contains a suitable propellant, eg, a gas such as carbon dioxide, or a nebulizer. Systemic administration can also be via transmucosal or transdermal means. For transmucosal or transdermal administration, a penetrant suitable for penetrating the barrier is used in the formulation. Such penetrants are generally known in the art, and include, for example, detergents, bile salts, and fusidic acid derivatives for transmucosal administration. Transmucosal administration can be achieved by using nasal sprays or suppositories. For transdermal administration, the active compound is formulated into ointments, salves, gels, or creams as generally known in the art.

[0071] The compounds can also be prepared in the form of suppositories (eg, with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery. In a specific embodiment, the active compound is prepared with a carrier that protects the compound from rapid elimination from the body, such as a controlled-release formulation, including implants and microencapsulated delivery systems.Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparing such formulations are clear to those skilled in the art.These materials can also be commercially obtained from Alza Corporation and Nova Pharmaceuticals, Inc.Liposomal suspensions can also be used as pharmaceutically acceptable carriers. It may be advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. As used herein, dosage unit form refers to a physically discrete unit suitable as a unitary dose for the subject to be treated; each unit contains a predetermined amount of active compound calculated to produce a desired therapeutic effect in association with the necessary pharmaceutical carrier. The specification for the dosage unit forms of the present invention is dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, as well as the limitations inherent in the art of formulating such active compounds for the treatment of individuals. For example, in some embodiments, pharmaceutical compositions of the present invention are suitable for oral administration in a unit dosage form, e.g., a tablet or capsule containing about 1 mg to about 10 g of a compound of the present invention. In some other embodiments, pharmaceutical compositions of the present invention are suitable for intravenous, subcutaneous, or intramuscular injection. Patients can receive, for example, about 1 μg / kg to about 1 g / kg of a compound of the present invention intravenously, subcutaneously, or intramuscularly. Intravenous, subcutaneous, and intramuscular administration can be by bolus injection or by continuous infusion over a period of time. Alternatively, the patient may receive a daily oral dose approximately equal to the daily parenteral dose, with the composition being administered 1 to 4 times per day.

[0072] The compound can be administered intravenously to mammals, including humans (e.g., by continuous drip infusion or rapid intravenous administration). In such cases, the dosage can be appropriately selected depending on various factors, such as the patient's weight and / or age, and / or the severity of symptoms and the route of administration. For example, the dosage of a compound for intravenous administration is generally about 1 / 2 of a human body surface area m 2 The daily dose is in the range of 1 to 10,000 mg per m of human body surface area. 2 The range is 1 to 5000 mg per day per m of human body surface area, and more preferably 2 The recommended daily intake is 10 to 5000 mg.

[0073] These therapeutic agents can be administered according to how often they are administered at specific times per day (e.g., one or more times per 24 hours), including the time between doses (e.g., every 6 hours), the time when the doses are administered (e.g., at 8:00 AM and 4:00 PM daily), and the amount of therapeutic agent (e.g., number of capsules).

[0074] Examples with data: Illustrative preparations of exemplary compounds of the present invention are shown in the following representative examples and schemes. The specific non-limiting data-supported examples of compounds are intended to illustrate certain exemplary embodiments of the present invention and are not intended to limit the scope of the specification or claims in any way. The compounds of the present invention may be prepared by the synthetic sequences shown in the following non-limiting examples and schemes. Those skilled in the art will appreciate that other routes of synthesis can be used as well. In particular, other routes of synthesis may actually be applied to some aspects of the present invention. Those skilled in the art will be referred to general textbooks, such as March's Advanced Organic Chemistry (Michael B. Smith & Jerry March, Wiley-Interscience, 2000), The Practice of Medicinal Chemistry (Camile G. Wermuth, Academia Press, 2003), and Protective Groups in Organic Synthesis (Theosora W. Greene & Peter GM Wuts; John Wiley & Sons Inc, 1999), all of which are incorporated herein by reference for their respective teachings.

[0075] Example 1 Reagents, synthesis methods, and biological property assays used Unless otherwise indicated, all reagents, starting materials, and solvents were obtained from commercial suppliers and used without further purification. Concentration or evaporation refers to evaporation under reduced pressure using a Buchi rotary evaporator and / or subsequent evaporation to dryness under high vacuum. Reaction products were purified by silica gel chromatography using the indicated solvent system or by HPLC purification using a C18 reverse-phase semi-preparative HPLC column with solvent A (0.1% TFA in water) and solvent B (0.1% TFA in CH3CN) as eluents. All final products are at least 95% pure as determined by analytical HPLC analysis with UV detection at 210 nm and / or 254 nm. Reported yields are isolated yields.

[0076] Analytical HPLC analysis was performed on an Agilent 1100 HPLC using a Phenomenex Luna C18(2) column (3 μm, 150 × 4.6 mm id) at a flow rate of 0.6 mL / min, with an initial operating pressure ranging from 120 to 140 bar, using binary solvent systems A and B: gradient elution from 10% to 90% B in 20 min, followed by 90% to 95% B in 5 min (gradient elution 1), or gradient elution from 70% to 95% B in 25 min, followed by 95% to 100% B in 3 min (gradient elution 2) (A: Milli-Q water with 0.1% TFA; B: CH3CN with 0.1% TFA). NMR spectra were recorded on a Bruker AV-300 or AV-301 300 MHz NMR instrument using DMSO-d6 or CDCl3 with TMS as the internal standard. Mass spectral data were obtained on a Bruker Esquire liquid chromatography-ion trap mass spectrometer.

[0077] The following abbreviations are used in the synthetic examples: aq (aqueous), h (hours), min (minutes), sat'd (saturated), THF (tetrahydrofuran), rt (room temperature), Et3N (triethylamine), NaCl (sodium chloride), MgSO4 (magnesium sulfate), CDCl3 (deuterated chloroform), H2O (water), HCl (hydrochloric acid), MeOH (methanol), NaOH (sodium hydroxide), TFA (trifluoroacetic acid), Na2CO3 (sodium carbonate), CHCl2 (methylene chloride), EtOAC (ethyl acetate), DMF (dimethylformamide), EtOH (ethanol), DMSO (dimethyl sulfoxide), DMSO-d6 (dimethyl sulfoxide-d6), NaHCO3 (sodium bicarbonate), HPLC (high performance liquid chromatography), ESI-MS or MS(ESI) (electrospray ionization mass spectrometry). Standard abbreviations used herein include NMR (nuclear magnetic resonance), DIEA (diisopropylethylamine), brine (saturated aqueous NaCl), NHCl (ammonium chloride), BocO (di-tert-butyl carbonate), HATU (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate), NaI (sodium iodide), KI (potassium iodide), Pd(OAc) (palladium(II) acetate), BINAP (2,2′-bis(diphenylphosphino)-1,1′-binaphthyl), DIEA or DIPEA (N,N-diisopropylethylamine), DCC (dicyclohexylcarbodiimide), NCS (N-chlorosuccinimide), PPTS (pyridinium p-toluenesulfonate), and other similar abbreviations.

[0078] The biological properties of exemplary compounds of the present invention were determined in at least the following assays. SuperTOPFLASH cell-based luciferase assay. Hek-293, STF1.1 cells are maintained in DMEM, 10% FBS, and Pen-Strep supplemented with 200 μg / mL G418. The day before the assay, cells are split into white, opaque 96-well plates at 10,000 cells per well in 50 μL of complete medium without G418 (for screening of Wnt signaling inhibitors, G418 can be omitted during the screening process). After allowing cells to stabilize and attach overnight, 40 μL of complete medium (without G418) containing 2.5x the final concentration of compound or DMSO control is added to the cells and allowed to incubate at 37°C and 5% CO2 for 1 hour before adding 10 μL of a 100 mM LiCl solution prepared in complete medium (without G418). After 24 hours, 100 μL of BrightGlo (Promega, Cat. No. G7573) is added to each well and the plate is shaken for 5 minutes before reading on a Perkin-Elmer EnVision plate reader.For example, the day before the assay: split cells into white opaque 96-well plates at 10,000 cells per well in 50 μL of complete growth medium; incubate plates overnight at 37° C., 5% CO2, allowing cells to adhere; the next day, prepare inhibitors to be tested at 2.5× the desired final concentration in complete growth medium (all conditions are performed in duplicate), and add 40 μL of medium containing the 2.5× concentration of compound to each well (including 2 wells for stimulation control, 2 wells for DMSO control, and wells for the positive control ICG-001 (e.g., 2, 5, and 10 micromolar)); after adding all inhibitors and controls, incubate plates at 37° C., 5% CO2 for 1 hour (prepare fresh 100 mM LiCl in complete growth medium while plates are incubating); after 1 hour, remove plates from the incubator and add 100 mM LiCl. 10 μL of medium containing LiCl is added to each well (except for the two unstimulated control wells, to which just 50 microliters of complete medium is added); the plate is incubated at 37°C and 5% CO2 for 24 hours; after 24 hours, 100 microliters of BrightGlo (Promega, catalog number: G7573) is added to each well, the plate is shaken for 5 minutes to ensure complete dissolution, and then the plate is read on a Perkin-Elmer EnVision 96-well plate reader.

[0079] Assay of human survivin 1Kb promoter-driven luciferase activity in the stably transfected cell line, 1Kb Hu-survivin\luc-Hek293. Briefly, 1Kb Hu-survivin\luc-Hek293 cells are maintained in DMEM, 10% FBS, and Pen-Strep supplemented with 1 μg / mL puromycin. The day before the assay, cells are split into white, opaque 96-well plates at 10,000 cells per well in 50 μL of complete medium without puromycin (for screening of CBP / β-catenin interaction inhibitors, puromycin can be omitted during the screening process). Cells are allowed to stabilize and attach overnight, after which 50 μL of complete medium (without G418) containing 2x the final concentration of compound or DMSO control is added to the cells. After 24 hours, 100 μL of BrightGlo (Promega, Cat. No. G7573) is added to each well and the plate is shaken for 5 minutes before reading on a Perkin-Elmer EnVision plate reader. For example, the day before the assay, cells are split into white opaque 96-well plates at 10,000 cells per well in 50 μL of complete growth medium; the plates are incubated overnight at 37°C and 5% CO2, allowing the cells to adhere; the next day, inhibitors to be tested are prepared in complete growth medium at 2x the desired final concentration (all conditions are performed in duplicate), and 50 μL of medium containing 2x the compound concentration is added to each well (2 wells for stimulation control, 2 wells for DMSO control, and wells for the positive control ICG-001 (e.g., containing 2, 5, 10, and 20 μM)). The plates are then incubated at 37°C and 5% CO2 for 24 hours; after 24 hours, 100 μL of BrightGlo (Promega, catalog number: G7573) is added to each well, the plates are shaken for 5 minutes to ensure complete dissolution, and the plates are read on a Perkin-Elmer EnVision 96-well plate reader.

[0080] Example 2 Synthesis of (S)-2-amino-3-(4-tert-butoxyphenyl)-N-(2,2-diethoxyethyl)-N-(quinolin-5-ylmethyl)propanamide (1) Compound 1, as a pale yellow oil, was prepared according to the procedure disclosed in US Pat. No. 7,671,054, starting from quinoline-5-carboxaldehyde, as shown in Scheme 1. MS (ESI): m / z 494.2 (M+H). + . Scheme 1: [ka]

[0081] Example 3 Synthesis of (S)-2-amino-3-(4-tert-butoxyphenyl)-N-(2,2-diethoxyethyl)-N-(naphthalen-1-ylmethyl)propanamide (2) Compound 2 as a colorless oil was prepared according to the procedure disclosed in US Pat. No. 7,671,054, starting from 1-naphthaldehyde, as shown in Scheme 2. MS (ESI): m / z 493.3 (M+H) + .

[0082] Scheme 2: [ka]

[0083] Example 4 Synthesis of tert-butyl 2-(2-(benzylcarbamoyl)hydrazinyl)acetate (3) Compound 3 was prepared according to the procedure described in steps 1-3 of Scheme 3 below.

[0084] Scheme 3: [ka] Step 1: tert-Butyl 2-(benzylcarbamoyl)hydrazinecarboxylate To a solution of tert-butylhydrazinecarboxylate (2.5 g, 19 mmol) in dry CHCl (50 mL) at 0° C., DIEA was added, followed by dropwise addition of benzyl isocyanate (2.35 mL, 19 mmol). The reaction mixture was stirred overnight at room temperature under argon and then poured into CHCl (150 mL) and 10% KHSO (100 mL). The organic layer was washed with saturated NaHCO (100 mL), saturated NaCl (100 mL), dried (MgSO), and evaporated to dryness. The title compound (5.4 g) as a white solid was used in the next step without further purification. 1 H-NMR (300 MHz, DMSO-d6) δ 8.54 (br s, 1H), 7.70 (s, 1H), 7.26 (m, 5H), 6.81 (br s, 1H), 4.22 (d, J = 6 Hz, 2H), 1.40 (s, 9H).

[0085] Step 2: N-benzylhydrazinecarboxamide tert-Butyl 2-(benzylcarbamoyl)hydrazinecarboxylate (5.4 g) was treated with CHCl (50 mL) and TFA (50 mL) at room temperature under argon for 3 h. Evaporation to dryness and coevaporation with CHCl twice to dryness gave the title compound (TFA salt; 5.6 g) as a pale yellow solid, which was used in the next step without further purification. 1 H-NMR (300 MHz, DMSO-d6) δ 7.24 (m, 5H), 7.03 (br s, 1H), 6.82 (br s, 1H), 4.24 (m, 4H). Step 3: tert-Butyl 2-(2-(benzylcarbamoyl)hydrazinyl)acetate To a suspension of N-benzylhydrazinecarboxamide (5.6 g) in dry CHCN (38 mL) was added NaHCO (4.8 g, 57 mmol), tert-butyl 2-bromoacetate (4.2 mL, 28.5 mmol), and NaI (0.28 g). The reaction mixture was stirred under argon at 60 °C for 24 h and poured into EtOAc (100 mL). The insoluble material was filtered. The filtrate was washed with water (100 mL). The aqueous layer was further extracted with EtOAc. The combined organic extracts were dried (NaSO) and evaporated to dryness. Purification by silica gel chromatography using EtOAc / CHCl (25% and 50%) afforded the title compound (2.3 g) as a white solid. 1 H-NMR (300 MHz, CDCl3) δ 7.30 (m, 5H), 6.38 (br s, 1H), 6.29 (s, 1H), 4.43(d, J = 6 Hz, 2H), 4.11 (t, J = 6 Hz, 1H), 3.42 (d, J = 9 Hz, 2H), 1.47 (s, 9H).

[0086] Example 5 Synthesis of (6S,9aS)-N-benzyl-6-(4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-ynyl)-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide (4) Compound 4 was prepared according to the procedure described in steps 1-4 of Scheme 4 below.

[0087] Scheme 4: [ka]

[0088] Step 1: tert-Butyl 2-(2-(benzylcarbamoyl)-1-(prop-2-ynyl)hydrazinyl)acetate A mixture of tert-butyl 2-(2-(benzylcarbamoyl)hydrazinyl)acetate (1.0 g, 3.58 mmol), propargyl bromide (0.7 mL, 9.2 M in toluene), NaHCO (0.6 g, 7.14 mmol), and KI (60 mg) in dry CHCN (30 mL) was stirred at 75 °C in a sealed tube for 3 days. The reaction mixture was cooled to room temperature and filtered. The filtrate was evaporated and purified by silica gel chromatography using 50% EtOAc / hexane to yield the title compound (0.63 g) as a colorless oil. MS (ESI): m / z 318.0 (M+H). + ;Analytical HPLC: 17.0min.

[0089] Steps 2-3: (S)-N-benzyl-2-(2-(3-(4-tert-butoxyphenyl)-1-((2,2-diethoxyethyl)(quinolin-5-ylmethyl)amino)-1-oxopropan-2-ylamino)-2-oxoethyl)-2-(prop-2-ynyl)hydrazinecarboxamide tert-Butyl 2-(2-(benzylcarbamoyl)-1-(prop-2-ynyl)hydrazinyl)acetate (0.31 g, 0.98 mmol) was treated with CHCl (10 mL) and TFA (10 mL) at room temperature under argon for 2 hours. Evaporation to dryness and coevaporation with CHCl twice to dryness gave 2-(2-(benzylcarbamoyl)-1-(prop-2-ynyl)hydrazinyl)acetic acid (TFA salt) as a yellow oil, which was used in the next step without further purification. MS (ESI): m / z 261.9 (M+H). + ;Analytical HPLC: 12.1min.

[0090] To a solution of (S)-2-amino-3-(4-tert-butoxyphenyl)-N-(2,2-diethoxyethyl)-N-(quinolin-5-ylmethyl)propanamide (0.48 g, 0.98 mmol) in DMF (7 mL) was added DIEA (0.51 mL, 2.93 mmol) and the above-prepared 2-(2-(benzylcarbamoyl)-1-(prop-2-ynyl)hydrazinyl)acetic acid (TFA salt). After cooling to 0 °C, HATU (0.37 g, 0.98 mmol) was added. The reaction mixture was stirred under argon at 0 °C for 0.5 h and at room temperature overnight, then poured into ethyl acetate (50 mL) and water (50 mL). The aqueous layer was extracted with ethyl acetate (25 mL × 2), and the combined organic extracts were washed with brine (25 mL), dried (Na SO ), and evaporated. Purification by silica gel chromatography using 5% MeOH / CH2Cl2 gave the desired product (0.79 g) as a pale yellow foam. MS (ESI): m / z 737.4 (M+H). + ;Analytical HPLC: 16.1min.

[0091] Step 4: (6S,9aS)-N-benzyl-6-(4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-ynyl)-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide A solution of (S)—N-benzyl-2-(2-(3-(4-tert-butoxyphenyl)-1-((2,2-diethoxyethyl)(quinolin-5-ylmethyl)amino)-1-oxopropan-2-ylamino)-2-oxoethyl)-2-(prop-2-ynyl)hydrazinecarboxamide (0.79 g, 1.07 mmol) in formic acid (25 mL) was stirred at room temperature for 20 hours. The solvent was removed and the resulting residue was taken up in EtOAc (25 mL) and saturated NaHCO3 (25 mL). The aqueous layer was further extracted with EtOAc (25 mL x 1). The combined organic extracts were dried (MgSO4) and evaporated. The crude material was purified by silica gel chromatography using EtOAc to give the desired product (0.36 g) as a pale yellow foam. MS (ESI): m / z 589.2 (M+H)+ ;Analytical HPLC: 12.5min.

[0092] Example 6 Synthesis of (6S,9aS)-N-benzyl-6-(4-hydroxybenzyl)-4,7-dioxo-2-((3-phenylisoxazol-5-yl)methyl)-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide (5) Compound 5 was prepared according to the procedure described in steps 1-6 of Scheme 5 below.

[0093] Scheme 5: [ka]

[0094] Steps 1-2: 5-(bromomethyl)-3-phenylisoxazole To a solution of benzaldehyde (2.0 g, 18.8 mmol) in MeOH (18 mL) were added hydroxylamine hydrochloride (1.6 g, 22.6 mmol) and K2CO3 (3.1 g, 22.6 mmol). The reaction mixture was stirred overnight at room temperature, diluted with water (50 mL), and extracted with EtOAc (50 mL × 1, 25 mL × 2). The organic extract was dried (Na2SO4) and evaporated to dryness. The obtained benzaldehyde oxime (2.4 g) as a colorless liquid was used in the next step without further purification. To a mixture of benzaldehyde oxime (2.4 g, 19.4 mmol) in water (60 mL) at room temperature was added KCl (1.5 g, 20.1 mmol) and propargyl bromide (5.6 mL, 9.2 M in toluene). The resulting mixture was cooled to 0 °C, and oxone (9.2 g, 15 mmol) was added portionwise. The reaction mixture was stirred at 0 °C for 1 h and at room temperature overnight and extracted with EtOAc (50 mL × 2). The organic extract was washed with saturated NaCl (25 mL), dried (MgSO4), and evaporated to dryness. The title compound (4.2 g) was obtained as a pale yellow solid and used in the next step without further purification. 1H-NMR (300 MHz, CDCl3) δ 7.80 (m, 2H), 7.47 (m, 3H), 6.63 (s, 1H), 4.52 (s, 2H).

[0095] Step 3: tert-butyl 2-(2-(benzylcarbamoyl)-1-((3-phenylisoxazol-5-yl)methyl)hydrazinyl)acetate A mixture of 5-(bromomethyl)-3-phenylisoxazole (56 mg, 0.24 mmol), tert-butyl 2-(2-(benzylcarbamoyl)hydrazinyl)acetate (60 mg, 0.22 mmol), NaHCO (36 mg, 0.43 mmol), and KI (3.7 mg) in dry CH CN (1.5 mL) was stirred at 65° C. for 20 h. The reaction mixture was cooled to room temperature and filtered. The filtrate was evaporated, and the resulting residue was purified by silica gel chromatography using EtOAc / hexane (ratio: 1:1 to 2:1) to yield the title compound (61 mg) as a white solid. MS (ESI): m / z 437.2 (M+H). + .

[0096] Step 4: 2-(2-(benzylcarbamoyl)-1-((3-phenylisoxazol-5-yl)methyl)hydrazinyl)acetic acid tert-Butyl 2-(2-(benzylcarbamoyl)-1-((3-phenylisoxazol-5-yl)methyl)hydrazinyl)acetate (61 mg, 0.14 mmol) was treated in CHCl (1.5 mL) and TFA (1.5 mL) at room temperature under argon for 3 h. Evaporation to dryness and co-evaporation with CHCl twice to dryness gave the title compound (TFA salt) as an off-white solid, which was used in the next step without further purification. 1H-NMR (300 MHz, DMSO-d6) δ 7.82 (m, 2H), 7.53 (m, 3H), 7.37 (s, 1H), 7.13 (s, 5H), 7.05 (s, 1H), 6.94 (t, J = 6 Hz, 1H), 4.20 (s, 2H), 4.17 (d, J = 6 Hz, 2H), 3.67 (s, 2H).

[0097] Step 5: (S)-N-benzyl-2-(2-(3-(4-tert-butoxyphenyl)-1-((2,2-diethoxyethyl)(quinolin-5-ylmethyl)amino)-1-oxopropan-2-ylamino)-2-oxoethyl)-2-((3-phenylisoxazol-5-yl)methyl)hydrazinecarboxamide To a solution of (S)-2-amino-3-(4-tert-butoxyphenyl)-N-(2,2-diethoxyethyl)-N-(quinolin-5-ylmethyl)propanamide (69 mg, 0.14 mmol) in DMF (1 mL) was added DIEA (73 μL, 0.42 mmol) and 2-(2-(benzylcarbamoyl)-1-((3-phenylisoxazol-5-yl)methyl)hydrazinyl)acetic acid (TFA salt; prepared in Step 4). After cooling to 0 °C, HATU (53 mg, 0.14 mmol) was added. The reaction mixture was stirred under argon at 0 °C for 0.5 h and at room temperature overnight, then poured into ethyl acetate (10 mL) and water (10 mL). The aqueous layer was extracted with ethyl acetate (10 mL × 2), and the combined organic extracts were washed with brine (10 mL), dried (NaSO), and evaporated. Purification by silica gel chromatography using MeOH / CH2Cl2 (5% and 10%) gave the desired product (104 mg) as a white foam. MS (ESI): m / z 856.7 (M+H). + ;Analytical HPLC: 18.0min.

[0098] Step 6: (6S,9aS)-N-benzyl-6-(4-hydroxybenzyl)-4,7-dioxo-2-((3-phenylisoxazol-5-yl)methyl)-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide A solution of (S)-N-benzyl-2-(2-(3-(4-tert-butoxyphenyl)-1-((2,2-diethoxyethyl)(quinolin-5-ylmethyl)amino)-1-oxopropan-2-ylamino)-2-oxoethyl)-2-((3-phenylisoxazol-5-yl)methyl)hydrazinecarboxamide (100 mg) in formic acid (5 mL) was stirred at room temperature for 20 hours. Evaporation to dryness, purification by silica gel chromatography (CHCl / CHOH / NHOH: 270:9:1 and 180:9:1) and lyophilization afforded the desired product (67 mg) as a white solid. MS (ESI): m / z 708.3 (M+H). + Analytical HPLC: 14.9 min (>99% pure).

[0099] Example 7 Synthesis of (6S,9aS)-N-benzyl-6-(4-hydroxybenzyl)-2-((3-(6-morpholinopyridin-2-yl)isoxazol-5-yl)methyl)-4,7-dioxo-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide (6) Compound 6 was prepared according to the procedure described in steps 1-9 of Scheme 6 below.

[0100] Scheme 6: [ka]

[0101] Step 1: 2-Bromo-6-(1,3-dioxolan-2-yl)pyridine A reaction mixture of 6-bromopicolinaldehyde (0.4 g, 2.15 mmol), ethylene glycol (0.62 mL, 11.1 mmol), and p-toluenesulfonic acid monohydrate (0.46 g, 2.67 mmol) in 1,2-dichloroethane (14.4 mL) and DMSO (0.32 mL) was stirred at 95 °C overnight under argon, cooled to room temperature, and evaporated to dryness. The resulting residue was diluted with EtOAc (25 mL), washed with 0.5 N NaOH (20 mL), saturated NaCl (15 mL), dried (NaSO), and evaporated. The crude material was purified by silica gel chromatography (25% EtOAc / hexanes) to give the desired product (0.32 g) as a colorless oil. 1 H-NMR (300 MHz, CDCl3) δ 7.61 (t, J = 4 Hz, 1H), 7.51 (m, 2H), 5.83 (s, 1H), 4.21 - 4.07 (m, 4H).

[0102] Step 2: 4-(6-(1,3-dioxolan-2-yl)pyridin-2-yl)morpholine A mixture of 2-bromo-6-(1,3-dioxolan-2-yl)pyridine (0.32 g, 1.38 mmol), morpholine (0.14 mL, 1.65 mmol), Pd(OAc) (40 mg), BINAP (104 mg), and CsCO (0.62 g) in toluene (14 mL) was purged with argon for 2 minutes and then stirred in a sealed tube at 100° C. for 22 hours. The reaction mixture was filtered through Celite and evaporated. Purification by silica gel chromatography using 25% and 50% EtOAc / hexanes afforded the title compound (0.26 g) as a brown solid. 1 H-NMR (300 MHz, CDCl3) δ 7.53 (t, J = 4 Hz, 1H), 6.88 (d, J = 4 Hz, 1H), 6..61 (d, J = 6 Hz, 1H), 5.72 (s, 1H), 4.19 - 4.05 (m, 4H), 3.81 (t, J = 4 Hz, 4H), 3.52 (t, H = 4 Hz, 4H).

[0103] Step 3: 6-Morpholinopicolinaldehyde To a suspension of 4-(6-(1,3-dioxolan-2-yl)pyridin-2-yl)morpholine (0.26 g, 1.10 mmol) in acetone (7 mL) and water (7 mL) was added pyridinium tosylate (83 mg). The reaction mixture was refluxed overnight and concentrated in vacuo. EtOAc (30 mL) was added, and the mixture was washed with saturated NaHCO (15 mL), saturated NaCl (15 mL), and dried (Na SO ). Evaporation and purification by silica gel chromatography (25% EtOAc / hexane) gave the desired product (0.19 g) as a pale yellow solid. 1 H-NMR (300 MHz, CDCl3) δ 10.1 (s, 1H), 7.71 (t, J = 9, and 4 Hz, 1H), 7.37 (d, J = 9 Hz, 1H), 6..93 (d, J = 6 Hz, 1H), 3.89 (t, J = 4.5 Hz, 4H), 3.68 (t, H = 4.5 Hz, 4H).

[0104] Step 4: 6-Morpholinopicolinaldehyde oxime To a solution of 6-morpholinopicolinaldehyde (0.19 g, 0.99 mmol) in MeOH (18 mL) was added hydroxylamine hydrochloride (81 mg, 1.17 mmol) and K2CO3 (0.16 g, 1.16 mmol). The reaction mixture was stirred overnight at room temperature, diluted with water (15 mL), and extracted with EtOAc (10 mL x 3). The organic extract was dried (Na2SO4) and evaporated to dryness. Purification by silica gel chromatography (30% EtOAc / hexane) afforded the desired product (0.18 g) as a white solid. 1H-NMR (300 MHz, DMSO-d6) δ 11.5 (s, 1H), 7.90 (s, 1H), 7.58 (d, J = 9, and 4 Hz, 1H), 7.08 (d, J = 6 Hz, 1H), 6.81 (d, J = 6 Hz, 1H), 3.70 (t, J = 4.5 Hz, 4H), 3.46 (t, H = 4.5 Hz, 4H).

[0105] Step 5: 4-(6-(5-(bromomethyl)isoxazol-3-yl)pyridin-2-yl)morpholine To a suspension of 6-morpholinopicolinaldehyde oxime (62 mg, 0.30 mmol) in 50% t-BuOH / water (2 mL) was added chloramine-T trihydrate (88.5 mg, 0.31 mmol), copper(II) sulfate pentahydrate (3 mg), and copper turnings (1 mg), followed by propargyl bromide (35.9 μL, 9.2 M in toluene). The reaction mixture was stirred overnight at room temperature and poured into EtOAc (10 mL) and water (10 mL). The aqueous layer was extracted with EtOAc (10 mL × 2). The combined organic extracts were washed with saturated NaCl (10 mL) and dried (NaSO). Evaporation and purification by silica gel chromatography (25% EtOAc / hexane) afforded the title compound (34 mg) as a white solid. MS (ESI): m / z 324.0, 326.0 (M+H). + ;Analytical HPLC: 17.8min. Steps 6-9: (6S,9aS)-N-benzyl-6-(4-hydroxybenzyl)-2-((3-(6-morpholinopyridin-2-yl)isoxazol-5-yl)methyl)-4,7-dioxo-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide. The title compound 6 was obtained according to the procedure described in Steps 3-6 of Example 6. MS (ESI): m / z 794.4 (M+H). + Analytical HPLC: 14.1 min (>99% pure).

[0106] Example 8 Synthesis of (6S,9aS)-N-benzyl-2-((3-(5-fluoropyridin-2-yl)isoxazol-5-yl)methyl)-6-(4-hydroxybenzyl)-4,7-dioxo-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide methanesulfonate (7) Compound 7 was prepared according to the procedure described in steps 1-5 of Scheme 7 below.

[0107] Scheme 7: [ka]

[0108] Step 1: 5-Fluoropicolinaldehyde oxime To a solution of 5-fluoropicolinaldehyde (0.5 g, 4.0 mmol) in MeOH (4 mL) was added hydroxylamine hydrochloride (0.33 g, 4.8 mmol) and K2CO3 (0.67 g, 4.8 mmol). The reaction mixture was stirred overnight at room temperature, diluted with water (10 mL), and extracted with EtOAc (10 mL x 3). The organic extract was dried (Na2SO4) and evaporated to dryness. The title compound (0.52 g) was used as a white solid in the next step without further purification. 1 H-NMR (300 MHz, DMSO-d6) δ 11.7 (s, 1H), 8.59 (d, J = 3 Hz, 1H), 8.09 (s, 1H), 7.87 (m, 1H), 7.77 (m, 1H).

[0109] Step 2: 5-Fluoro-N-hydroxypicolinimidoyl chloride To a solution of 5-fluoropicolinaldehyde oxime (0.4 g, 2.85 mmol) in DMF (3 mL) was added N-chlorosuccinimide (0.42 g, 3.14 mmol). The reaction mixture was stirred under argon at 50 °C overnight, cooled to room temperature, and poured into EtO (15 mL) and water (15 mL). The aqueous layer was extracted with EtO (15 mL × 2). The organic extract was washed with water (10 mL), saturated NHCl (10 mL), and saturated NaCl (10 mL), dried (NaSO), and evaporated to dryness. The title compound (0.49 g) as a white solid was used in the next step without further purification. 1 H-NMR (300 MHz, DMSO-d6) δ 12.7 (s, 1H), 8.68 (d, J = 2.8 Hz, 1H), 7.96 (m, 1H), 7.84 (m, 1H).

[0110] Step 3: 4-(((6S,9aS)-1-(benzylcarbamoyl)-2-((3-(5-fluoropyridin-2-yl)isoxazol-5-yl)methyl)-4,7-dioxo-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazin-6-yl)methyl)phenylacetate. To a solution of 4-(((6S,9aS)-1-(benzylcarbamoyl)-4,7-dioxo-2-(prop-2-ynyl)-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazin-6-yl)methyl)phenyl acetate (0.60 g, 0.95 mmol; prepared according to the procedure disclosed in Example 11, using acetyl chloride instead) in dry CHCl (27 mL) at 0° C., 5-fluoro-N-hydroxypicolinimidoyl chloride (0.16 g, 0.90 mmol) was added, followed by dropwise addition of EtN (0.25 mL, 1.80 mmol) in CHCl (1 mL). The reaction mixture was stirred under argon at 0° C. for 1 hour and at room temperature for 24 hours. Evaporation to dryness and purification by silica gel chromatography using CHOH / CHCl gave the partially purified title compound (0.54 g) as an oily residue. MS (ESI): m / z 769.3 (M+H). + ;Analytical HPLC: 14.9min.

[0111] Step 4: (6S,9aS)-N-benzyl-2-((3-(5-fluoropyridin-2-yl)isoxazol-5-yl)methyl)-6-(4-hydroxybenzyl)-4,7-dioxo-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide To a solution of 4-(((6S,9aS)-1-(benzylcarbamoyl)-2-((3-(5-fluoropyridin-2-yl)isoxazol-5-yl)methyl)-4,7-dioxo-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazin-6-yl)methyl)phenyl acetate (0.54 g; prepared as described in Step 3 above) in CHOH (16 mL) was added dropwise sat'd NaHCO (5.3 mL). The reaction mixture was stirred at room temperature for 3 h, evaporated in vacuo, and taken up in EtOAc (15 mL) and water (15 mL). The aqueous layer was extracted with EtOAc (15 mL × 2). The organic extract was washed with sat'd NaCl (15 mL), dried (NaSO), and evaporated to dryness. Purification by silica gel chromatography using EtOAc / CH2Cl2 gave the title compound (0.26 g) as a white solid. MS (ESI): m / z 727.3 (M+H) + Analytical HPLC: 13.8 min (>96% pure).

[0112] Step 5: (6S,9aS)-N-benzyl-2-((3-(5-fluoropyridin-2-yl)isoxazol-5-yl)methyl)-6-(4-hydroxybenzyl)-4,7-dioxo-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide methanesulfonate To a solution of (6S,9aS)-N-benzyl-2-((3-(5-fluoropyridin-2-yl)isoxazol-5-yl)methyl)-6-(4-hydroxybenzyl)-4,7-dioxo-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide (176 mg, 0.24 mmol) in CHCl (6 mL) was slowly added a solution of methanesulfonic acid (23.3 mg, 0.24 mmol) in acetone and CHCl. ​​After stirring for several minutes at room temperature, hexane (approximately 10 mL) was slowly added. The white precipitate that formed was collected by filtration and washed with hexane and EtOAc. Air drying and lyophilization from Milli-Q water afforded the desired product (173 mg) as a pale yellow solid. MS (ESI): m / z 727.2 (M+H). + Analytical HPLC: 13.8 min (>97% pure).

[0113] Example 9 Synthesis of (6S,9aS)-N-benzyl-6-(4-hydroxybenzyl)-4,7-dioxo-2-((3-(pyrazin-2-yl)isoxazol-5-yl)methyl)-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide (8) Compound 8 was prepared according to the procedure described in steps 1-2 of Scheme 8 below. Scheme 8: [ka]

[0114] Step 1: N-Hydroxypyrazine-2-carbimidoyl chloride The title compound as a pale yellow solid was obtained following the procedure described in Step 2 of Example 8. 1 H-NMR (300 MHz, DMSO-d6) δ 9.10 (s, 1H), 8.77 - 8.73 (m, 3H). Step 2: (6S,9aS)-N-benzyl-6-(4-hydroxybenzyl)-4,7-dioxo-2-((3-(pyrazin-2-yl)isoxazol-5-yl)methyl)-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide To a solution of 4-(((6S,9aS)-1-(benzylcarbamoyl)-4,7-dioxo-2-(prop-2-ynyl)-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazin-6-yl)methyl)phenyl acetate (0.30 g, 0.51 mmol) in dry CHCl (15 mL) at 0 °C, N-hydroxypyrazine-2-carbimidoyl chloride (76.1 mg, 0.48 mmol) was added, followed by dropwise addition of EtN (134 μL, 0.97 mmol). The reaction mixture was stirred at 0 °C for 1 h and at room temperature for 20 h under argon. Evaporation to dryness, purification by silica gel chromatography using CHOH / EtOAc, and lyophilization in CHCN and Milli-Q water afforded the title compound (0.12 g) as a white solid. MS(ESI): m / z 710.3(M+H) + Analytical HPLC: 12.6 min (>95% pure).

[0115] Example 10 Synthesis of (6S,9aS)-N-benzyl-6-(4-hydroxybenzyl)-4,7-dioxo-2-((3-(pyrrolidin-2-yl)isoxazol-5-yl)methyl)-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide (9) Compound 9 was prepared according to the procedure described in steps 1-4 of Scheme 9 below.

[0116] Scheme 9: [ka]

[0117] Steps 1-2: tert-Butyl 2-(chloro(hydroxyimino)methyl)pyrrolidine-1-carboxylate The title compound was obtained according to the procedure described in Steps 1-2 of Example 8. Analytical HPLC: 15.4 min (>97% pure). Step 3: tert-Butyl 2-(5-(((6S,9aS)-1-(benzylcarbamoyl)-6-(4-hydroxybenzyl)-4,7-dioxo-8-(quinolin-5-ylmethyl)dihydro-1H-pyrazino[2,1-c][1,2,4]triazine-2(6H,7H,8H,9H,9aH)-yl)methyl)isoxazol-3-yl)pyrrolidine-1-carboxylate To a solution of (6S,9aS)-N-benzyl-6-(4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-ynyl)-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide (70 mg, 0.12 mmol) in dry CHCl (5 mL) was added tert-butyl 2-(chloro(hydroxyimino)methyl)pyrrolidine-1-carboxylate (118 mg, 0.48 mmol), followed by dropwise addition of EtN (0.13 mL, 0.93 mmol). The reaction mixture was stirred under argon at 40 °C for 2 days. Evaporation to dryness and purification by silica gel chromatography with EtOAc afforded the title compound (26 mg) as a white residue. MS (ESI): m / z 801.5 (M+H). + ;Analytical HPLC: 14.7min (94% pure).

[0118] Step 4: (6S,9aS)-N-benzyl-6-(4-hydroxybenzyl)-4,7-dioxo-2-((3-(pyrrolidin-2-yl)isoxazol-5-yl)methyl)-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide tert-Butyl 2-(5-(((6S,9aS)-1-(benzylcarbamoyl)-6-(4-hydroxybenzyl)-4,7-dioxo-8-(quinolin-5-ylmethyl)dihydro-1H-pyrazino[2,1-c][1,2,4]triazine-2(6H,7H,8H,9H,9aH)-yl)methyl)isoxazol-3-yl)pyrrolidine-1-carboxylate (26 mg, 0.032 mmol) was stirred with CHCl (1 mL) and TFA (1 mL) at room temperature under argon for 2 hours. Evaporation to dryness and coevaporation with CHCl twice, purification by silica gel chromatography (CHCl / CHOH / NHOH: 180:9:1 and 90:9:1), followed by lyophilization using CHCN and Milli-Q water gave the desired product (14 mg) as a white powder. MS(ESI): m / z 701.3(M+H) + Analytical HPLC: 10.6 min (>99% pure).

[0119] Example 11 Synthesis of 4-(((6S,9aS)-1-(benzylcarbamoyl)-2-((3-(4-fluorophenyl)isoxazol-5-yl)methyl)-4,7-dioxo-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazin-6-yl)methyl)phenyl dodecanoate (10) Compound 10 was prepared according to Scheme 10 using the procedures disclosed below. Scheme 10: [ka]

[0120] To a solution of (6S,9aS)-N-benzyl-2-((3-(4-fluorophenyl)isoxazol-5-yl)methyl)-6-(4-hydroxybenzyl)-4,7-dioxo-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide (1.2 g, 1.67 mmol) in dry CHCl (47 mL) at 0° C. was added EtN (0.47 mL, 3.34 mmol), followed by the slow addition of lauroyl chloride (0.58 mL, 2.51 mmol). The reaction mixture was stirred under argon at 0° C. for 1 hour and at room temperature overnight and evaporated to dryness. The resulting residue was taken up in EtOAc (100 mL) and saturated NaHCO (50 mL), and the organic layer was washed with saturated NaCl (50 mL) and dried (MgSO). Evaporation, purification by silica gel chromatography using EtOAc / CH Cl and lyophilization gave the title product (1.1 g) as a white foam. MS (ESI): m / z 908.7 (M+H). + .

[0121] Example 12 Synthesis of ethyl 2-(2-((4-(((6S,9aS)-1-(benzylcarbamoyl)-4,7-dioxo-2-((3-(pyridin-2-yl)isoxazol-5-yl)methyl)-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazin-6-yl)methyl)phenoxy)carbonylamino)acetamido)acetate (11) Compound 11 was prepared according to Scheme 11 using the procedures disclosed below.

[0122] Scheme 11: [ka]

[0123] Step 1: Ethyl 2-(2-(tert-butoxycarbonylamino)acetamido)acetate To a suspension of 2-(2-(tert-butoxycarbonylamino)acetamido)acetic acid (0.5 g, 2.15 mmol) in CHCl (8 mL) was added EtOH (0.14 mL, 2.37 mmol), DCC (0.49 g, 2.37 mmol), and DMAP (29 mg, 0.24 mmol). The reaction mixture was stirred overnight at room temperature under argon and filtered. The filtrate was diluted with CHCl (25 mL), washed with water (25 mL) and 10% KHSO (25 mL), dried (MgSO), and evaporated. The crude material was purified by silica gel chromatography using EtOAc / hexanes to yield the title product (0.53 g) as a white oily residue. 1 H-NMR (300 MHz, DMSO-d6) δ 8.17 (t, J = 6 Hz, 1H), 7.0 (t, J = 6 Hz, 1H), 4.09 (q, J = 6 Hz, 2H), 3.82 (d, J = 6 Hz, 2H), 3.56 (d, J = 6 Hz, 2H), 1.38 (s, 9H), 1.16 (t, J = 6 Hz, 3H).

[0124] Step 2: 2-(2-ethoxy-2-oxoethylamino)-2-oxoethanaminium chloride Ethyl 2-(2-(tert-butoxycarbonylamino)acetamido)acetate (0.53 g, 2.04 mmol) was treated with 6 N HCl (5 mL) in EtOH (5 mL) at room temperature overnight. All volatiles were removed by evaporation to give the desired product (0.36 g) as a white solid. 1 H-NMR (300 MHz, DMSO-d6) δ 8.84 (br s, 3H), 8.71 (br s, 1H), 4.11 (q, J = 6 Hz, 2H), 3.94 (d, J = 6 Hz, 2H), 3.6 (d, J = 3 Hz, 2H), 1.20 (t, J = 7.5 Hz, 3H).

[0125] Step 3: Ethyl 2-(2-((4-(((6S,9aS)-1-(benzylcarbamoyl)-4,7-dioxo-2-((3-(pyridin-2-yl)isoxazol-5-yl)methyl)-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazin-6-yl)methyl)phenoxy)carbonylamino)acetamido)acetate To a solution of (6S,9aS)-N-benzyl-6-(4-hydroxybenzyl)-4,7-dioxo-2-((3-(pyridin-2-yl)isoxazol-5-yl)methyl)-8-(quinolin-5-ylmethyl)octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide (0.13 g, 0.18 mmol) in CHCl under argon at −15° C., diphosgene (31 μL, 0.26 mmol) and DIEA (44.6 μL, 0.26 mmol) were added. The reaction mixture was stirred at room temperature under argon for 2 hours. To this reaction mixture at -5 °C was added 2-(2-ethoxy-2-oxoethylamino)-2-oxoethanaminium chloride (0.22, 1.13 mmol), DIEA (0.29 mL, 1.67 mmol), and CHCl (2.3 mL). The reaction mixture was continued stirring at room temperature under argon for another 2 h before EtO (5 mL) was added. Insoluble material was removed by filtration. Evaporation to dryness and purification by silica gel chromatography using EtOAc / CHOH afforded the title product (116 mg) as a white solid. MS (ESI): m / z 895.3 (M+H). + .

[0126] Example 13 Compounds 12-34 shown in Table 1 were prepared based on procedures similar to those used in Examples 6-12, using compounds 1-4 exemplified in Examples 2-5 and the starting materials shown in Table 2 accordingly.

[0127] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5]

[0128] Example 14 The starting materials used for the preparation of these examples and exemplary compounds of the invention shown in Table 1 are listed in Table 2.

[0129] [Table 2] TIFF0007750851000048.tif156146

[0130] Example 15 Three representative isoxazole compounds ([3+2]-101, [3+2]-117, and [3+2]-229) generally demonstrated approximately 10x or greater potency than the positive control compound ICG-001 in the art-recognized SuperTOPFLASH cell-based luciferase assay. 1 shows, according to a non-limiting embodiment of the present invention, the results of a SuperTOPFLASH cell-based luciferase assay (Wnt-driven luciferase activity in stably transfected cell lines, Hek293, STF1.1) comparing the CBP / β-catenin inhibitory activity of three exemplary isoxazole compounds of the present invention (encoded [3+2]-101, [3+2]-117, and [3+2]-229; each compared at concentrations between 16 and 1000 nM) with the art-recognized CBP / β-catenin inhibitor ICG-001 used as a positive control (at 1.25, 2.5, 5, and 10 μM). As is readily apparent, the three representative isoxazole compounds of the present invention are at least approximately 10× or more potent in this assay than the control compound ICG-001. The potency of most of the disclosed CBP / β-catenin inhibitor compounds was determined to generally be at least approximately 3× or greater than the potency of the control compound ICG-001 in this assay.

[0131] Example 16 Three representative isoxazole compounds ([3+2]-101, [3+2]-117, and [3+2]-229) generally demonstrated approximately 10x or greater potency than the positive control compound ICG-001 in the art-recognized survivin promoter-driven luciferase activity assay.

[0132] In Figure 2, according to additional non-limiting embodiments of the present invention, the results of a survivin promoter-driven luciferase activity assay (assay of human survivin 1Kb-promoter-driven luciferase activity in a stably transfected Hek293 cell line; 1Kb Hu-survivin / luc-Hek293) are shown, comparing the CBP / β-catenin inhibitory activity of three exemplary isoxazole compounds of the present invention (encoded [3+2]-101, [3+2]-117, and [3+2]-229; each compared at concentrations between 0.1 and 10 μM) with the art-recognized CBP / β-catenin inhibitor ICG-001 used as a positive control (at 1.25, 2.5, 5, and 10 μM). As is readily apparent, these three isoxazole compounds of the present invention exhibited high IC 50 The IC values ​​for most of the disclosed CBP / β-catenin inhibitor compounds are at least approximately 10× or more potent in this assay than those of the control compound ICG-001. 50 Values ​​were generally determined to be at least approximately 3x or greater than the potency of the control compound ICG-001 in this assay.

[0133] Example 17 The representative isoxazole compound [3+2]-101 was generally shown to be at least approximately 10x or more potent for reducing CBP / β-catenin-based transcription than the potency of the positive control compound ICG-001 in an art-accepted SYBR-Green qPCR assay for survivin / BIRC5.

[0134] According to further non-limiting embodiments of the present invention, Figures 3A and 3B show the results of SYBR-Green qPCR assays for survivin / BIRC5 (CBP-specific) and EphB2 (p300-specific) gene expression using GAPDH as a control gene. Figure 3A shows the results of SYBR-Green qPCR assays for CBP / β-catenin-specific gene (H. Ma et al. Oncogene 2005, 24, 3619-31) and survivin / Birc5 gene expression (ΔΔCt Figure 3B compares the compound of the present invention [3+2]-101 (at 1 μM) with the art-recognized positive control ICG-001 (at 10 μM) in inhibiting CBP / β-catenin expression (reflected as an increase in ΔΔC). As is readily apparent, the CBP / β-catenin expression inhibitory activity of the isoxazoles of the present invention exceeds approximately 10× that of the control. Figure 3B shows the effect of EphB2 (p300 / β-catenin-specific gene (Kumar S, et. al. Cancer Res. 2009, 69, 3736-45) gene expression (ΔΔC t The compound of the present invention, [3+2]-101 (at 1 μM), is compared to the art-recognized positive control, ICG-001 (at 10 μM), in stimulating EphB2 gene expression (reflected as a decrease in EphB2 activity). As is readily apparent, the EphB2 gene expression stimulating activity of representative isoxazoles of the present invention is at least 10× greater than that of the control. This reflects a decrease in CBP / β-catenin-based transcription accompanied by an increase in p300 / β-catenin-based transcription, mediated by the specific CBP / β-catenin inhibitory activity of [3+2]-101 and ICG-001.

[0135] Example 18 Co-immunoprecipitation assays were performed in SW480 cells to further confirm that the positive control compound ICG-001 and the representative inventive compound [3+2]-117 selectively interfered with β-catenin binding to CBP, while enhancing the binding of β-catenin to p300. According to a further non-limiting embodiment of the present invention, Figures 4A and 4B show co-immunoprecipitation assays performed in SW480 cancer cells as previously described (Emami K. et al. PNAS USA, 2004 Aug 24;101(34):12682-7) to further confirm that the positive controls ICG-001 and [3+2]-117 selectively interfere with β-catenin binding to CBP (Figure 4A) while enhancing β-catenin binding to p300 (Figure 4B). Immunoprecipitation of β-catenin by CBP was inhibited by ICG-001 at 10 μM (Figure 3A) and by [3+2]-117 at 500 nM. [3+2]-117 was approximately 20-fold more active than ICG-001 (compare lane 1 (DMSO control) with lanes 2 and 3). Despite the fact that CBP and p300 are highly homologous, the minimal interaction between β-catenin and p300 observed in SW480 cells was not blocked by ICG-001. Indeed, treatment with 10 μM ICG-001 slightly increased the amount of β-catenin coimmunoprecipitated with p300, consistent with the switch from CBP / β-catenin transcription to p300 / β-catenin transcription associated with the onset of differentiation (Figure 4B, compare p300 IP, lanes 1 and 2). [3+2]-117 at 500 nM dramatically increased the interaction of β-catenin with p300 (Figure 4B, compare p300 IP, lanes 1 and 3). Quantification of the immunoblot data based on pixelation is shown below the immunoblots.

[0136] Example 19 Using a bleomycin-induced mouse model of idiopathic pulmonary fibrosis (IPF), we demonstrated that the efficacy of representative compounds of the present invention, [3+2]-120A and [3+2]-120B, compared favorably with historical control data for pirfenidone and nintedanib.

[0137] Bleomycin-induced model of IPF. As previously shown by first-generation (ICG-001) and second-generation (PRI-724) CBP / β-catenin antagonists, the representative compound of the present invention, [3+2]-120A (compound 20), demonstrated efficacy in a bleomycin-induced model of IPF in mice (SMC Laboratories, Tokyo, Japan). Specifically, oral (po) administration of [3+2]-120A by gavage at 100 mg / kg / day (A-low) and 400 mg / kg / day (A-high) was initiated on day 7 after bleomycin administration and continued for 2 weeks. Historical control data and Ashcroft scoring for overall survival were compared with those for both FDA-approved treatments for IPF, namely, pirfenidone (400 mg / kg / day po) and nintedanib (100 mg / kg / day po). A representative compound of the present invention, [3+2]-120B (Compound 11), at 400 mg / kg / day (high) also compared favorably with historical control data.

[0138] Results. Figure 5 shows, according to a further non-limiting embodiment of the present invention, that there was a significant increase in weight gain in mice treated with [3+2]-120A (both dose groups) and mice treated with [3+2]-20B (high dose group), and all mice that survived (Figure 6), whereas traditionally, approximately 30% of mice die before day 21 when treated in this model. Furthermore, significant decreases in lung mass and Ashcroft score (Table 3) were demonstrated in mice treated with [3+2]-120A (both dose groups) and mice treated with [3+2]-120B (high dose group) (Figures 8A-8E) based on histological analysis (Figures 7A-7D). Figures 8A-8E show the following: historical bleomycin-saline treated control (8A, "ID: 103"); [3+2]-120A low dose (8B, "ID: 102"); [3+2]-120A high dose (8C, "ID: 201"); [3+2]-120B low dose (8D, "ID: 302"); and [3+2]-120B high dose (8E, "ID: 401"). In all treatment groups, there is a significant reduction in extracellular collagen deposition compared to the bleomycin-saline treated historical control, as judged by Masson's trichrome blue staining. Based on these results, the [3+2] series compounds are effective in the bleomycin-induced model of idiopathic pulmonary fibrosis when administered orally (per os, po).

[0139] [Table 3]

[0140] Example 20 The STAM™ mouse model of NASH-HCC was used to demonstrate the efficacy of a representative compound of the present invention, [3+2]-120A.

[0141] A representative compound of the present invention, [3+2]-120A (compound 20), has demonstrated efficacy in mice (SMC Laboratories, Tokyo, Japan) in the proprietary STAM™ mouse model of NASH-HCC. Specifically, in ongoing studies, oral (po) gavage administration of [3+2]-120A at 100 mg / kg / day (A-low) and 400 mg / kg / day (A-high) has been favorably compared with vehicle control data for overall survival time after 4 weeks of treatment. In the vehicle control treatment group, 5 out of 8 mice survived, while 7 out of 8 and 8 out of 8 mice survived with [3+2]-120A at 100 mg / kg / day (A-low) and 400 mg / kg / day (A-high), respectively.

[0142] Example 21 The NG / Nga mouse model of atopic dermatitis was used to demonstrate the efficacy of a representative compound of the present invention, [3+2]-121A, in reducing TEWL scores and dermatitis scores compared to vehicle controls.

[0143] NG / Nga Mouse Model of Atopic Dermatitis: NC / Nga mice (female) were obtained from Charles River Laboratories Japan Inc. Mice were randomized into four groups of eight mice each on day 0 based on mouse weight and dermatitis score. Eight NC / Nga mice were topically administered a test compound-containing [3+2]-121A (compound 34) ointment or vehicle control (hydrophilic petrolatum) once daily (at a volume of 100 mg) from day 0 to day 13. Representative compound [3+2]-121A at low (100 μM) and high (500 μM) dose levels significantly reduced TEWL scores and dermatitis scores compared to the vehicle control.

[0144] Representative test substance [3+2]-121A was formulated in hydrophilic petrolatum at low (100 μM) and high (500 μM) doses (referred to as [3+2]-121A and [3+2]-121B in this study, respectively). Seven-week-old female NC / Nga mice were used. The first induction of atopic dermatitis was performed 3 weeks before compound administration, during which the hair on the back and behind the ears of the mice was shaved using clippers and a razor. Then, the hair of the mice was completely removed with a depilatory (Epilat™, Kracie Home Products, Ltd.), and 100 mg of Biostir AD® ointment was evenly applied with a plastic spoon to the shaved dorsal skin and the surface of each ear. A second induction of atopic dermatitis was then performed. The hair on the backs and behind the ears of the mice was shaved, and then 150 μl of 4% sodium lauryl sulfate solution was applied to the backs and behind the ears with a plastic spoon. This was then dried with a hair dryer (cold air) and then allowed to dry naturally for approximately 1-2 hours. 100 mg of Biostir AD® Ointment was evenly applied with a plastic spoon to the shaved dorsal skin and the surface of each ear. This procedure was repeated twice a week for 3 weeks. Twenty-four atopic dermatitis model mice were sorted into three groups of 8 mice each based on their weight and dermatitis severity score on the morning of Day 0. Vehicle control (hydrophilic petrolatum), [3+2]-121A, or [3+2]-121B was topically administered at a volume of 100 mg once daily from Day 0 to Day 13. Body weight and food intake were measured once a week on Days 0, 7, and 14. The severity of dermatitis was evaluated on days 0, 3, 7, 10, and 14. The occurrence of 1) erythema / bleeding, 2) scarring / dryness, 3) edema, and 4) peeling / erosion was scored as 0 (none), 1 (mild), 2 (moderate), and 3 (severe). The sum of the individual scores was used as the dermatitis score. TEWL was evaluated on days 0 and 14 after administration. Histological analysis was performed on skin from the dorsal lower neck and auricles that were fixed in 10% neutral formalin, embedded in paraffin, sectioned at 3-4 μm, and stained with hematoxylin-eosin using Caracci hematoxylin solution.For analysis of skin areas, bright-field images of HE-stained sections were randomly recorded at 2- and 4-fold magnification using a digital camera (DS-Fi3; Nikon). Statistical analysis was performed using Prism Software 6 (GraphPad Software, USA). Statistical analysis was performed using Dunnett's comparison test. Comparisons were made between the following groups: Group 1 (vehicle) vs. Group 2 ([3+2]-121A) and Group 3 ([3+2]-121B). A P value of <0.05 was considered statistically significant. Results were expressed as mean ± SD. There was no significant difference in mean body weight between the vehicle and treatment groups on any day during the treatment period, demonstrating the lack of toxicity of the treatment (Figure 9). There was no significant difference in food intake per mouse between the vehicle and treatment groups (Figure 10).

[0145] Dermatitis severity score: The [3+2]-121A group showed a significant reduction in dermatitis severity score on the back compared to the vehicle group on days 10 and 14 (FIG. 11A back 11B whole).

[0146] TEWL analysis (Transepidermal water loss (TEWL) is the amount of water that passively evaporates through the skin to the external environment due to the water vapor pressure gradient across the skin barrier and is used to characterize skin barrier function): [3+2]-121A and 121B groups showed a significant decrease in TEWL on day 14 (Figure 12), demonstrating an increase in barrier function associated with treatment. Histological analysis: As shown in representative photomicrographs, the [3+2]-121A and [3+2]-121B groups demonstrated a significant reduction in inflammation and inflammatory cell influx (FIGS. 13A-13C).

[0147] The present invention and the manner and process of making and using it are now described in such full, clear, concise and exact terms as to enable any person skilled in the art to which it pertains to make and use the same. Although specific embodiments of the present invention have been described herein for purposes of illustration, it will be understood that various modifications can be made without departing from the spirit and scope of the invention. Accordingly, the invention is not limited except as by the appended claims.

[0148] All of the above U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications and non-patent publications referenced herein are hereby incorporated by reference. Another aspect of the present invention may be as follows. [1] Formula (Ia): [C1] TIFF0007750851000050.tif6570 (Ia) {In the formula, R a is hydrogen or -CH 3 and; R b is a monocyclic aryl group having 5 to 7 ring members, which may have 1 to 2 heteroatoms selected from nitrogen, oxygen, or sulfur, and which may have one or more substituents selected from the group consisting of halide, cyano, and lower alkyl; R is a phenyl group; a substituted phenyl group having one or more substituents (wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; benzyl groups; substituted benzyl groups having one or more substituents, wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; or a bicyclic aryl group, or a substituted bicyclic aryl having 8 to 11 ring members and optionally having 1 to 3 heteroatoms selected from nitrogen, oxygen, or sulfur (substituted bicyclic aryl is an amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-3 optionally bearing one or more substituents independently selected from alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; R 2 is hydrogen or -CH 3 and; Y is selected from hydrogen, deuterium, or a halogen; W is hydrogen, phosphate or phosphoric acid salt, ester of alkyl or fatty acid, or X, where X is [Case 2] TIFF0007750851000051.tif33146 wherein Z is OR 4 (In the formula, R 4 is hydrogen or C 1 -C 6 alkyl), or Z is an amino acid or amino acid ester, and n is 1 or 2; L is -CH 2 -, -CF 2 - or -C(CH 3 ) 2 - and; Q is [C3] TIFF0007750851000052.tif26150 wherein A, B, and D are independently selected from O, S, N, or —CH; 1 、Z 2 are hydrogen, deuterium, halogens, C 1 -C 4 Alkyl, -OH, -OC 1 -C 6 alkyl, and [C4] TIFF0007750851000053.tif43146 [In the formula, R 1 、R 2 and R 3 is hydrogen, C 1 -C 6 Alkyl or C containing one or more -OH 1 -C 6 alkyl; Z 3 is hydrogen, halogen, or -OC directly bonded or bonded to -NH- 1 -C 3 bonded to alkyl or -NHC 1 -C 3 Aryl, heteroaryl, cycloalkyl, heterocycloalkyl bound to an alkyl, or cycloalkyl, heterocycloalkyl bound to a nitrogen, or -NHC 1 -C 4 Alkyl, -N(C 1 -C 4 alkyl) 2 each of which is selected from hydrogen, deuterium, a halogen, C 1 -C 4 Alkyl, C 1 -C 3 Haloalkyl, -OH, -OC 1 -C 6 Alkyl, -OC 1 -C 6 Alkyl-C(O)NH-OH, -NH 2 , -C(O)NH-C 1 -C 6 Alkyl-heteroaryl, -NHC(O)C 1 -C 6 alkyl-C(O)NH—OH, heteroaryl, cycloalkyl, heterocycloalkyl, or cycloalkyl or heterocycloalkyl attached to nitrogen, —NHC 1 -C 4 Alkyl, or -N(C 1 -C 4 alkyl) 2 and pharmaceutically acceptable salts thereof, wherein R is a 5- or 6-membered nitrogen-containing heteroaryl substituted by 0-2 substituents selected from: [2] The ester of an alkyl acid or a fatty acid is [5] TIFF0007750851000054.tif2376 (wherein m is 1 to 14), [6] TIFF0007750851000055.tif39150 The compound according to [1] above, selected from: [3] The compound according to [1] or [2] above, wherein R is a bicyclic aryl selected from naphthyl, quinolinyl, isoquinolinyl, quinoxaline, phthalazine, quinazoline, cinnoline, naphthyridine, or substituted variants thereof. [4] The compound is represented by formula (Ib): [7] TIFF0007750851000056.tif6868 (Ib) (In the formula, X 1 and X 2 The compound according to any one of the above [1] to [3], wherein: [5] L is -CH 2 - and; Q is, [8] TIFF0007750851000057.tif2032 (wherein A, B, and D are independently selected from O, S, N, or —CH). [6] A is -CH, B is N, and D is O(Z 3 -isoxazole-), W is hydrogen, phosphate or phosphoric acid salt, ester of alkyl or fatty acid, or X, and X is [9] TIFF0007750851000058.tif33146 wherein Z is OR 4 (In the formula, R 4 is hydrogen or C 1 -C 6 or Z is an amino acid or amino acid ester, and n is 1 or 2. 〔7〕Z 3 is selected from aryl or heteroaryl, each of which is hydrogen, deuterium, halogen, C 1 -C 4 Alkyl, C 1 -C 3 Haloalkyl, -OH, -OC 1 -C 6 Alkyl, -OC 1 -C 6 Alkyl-C(O)NH-OH, -NH 2 , -C(O)NH-C 1 -C 6 Alkyl-heteroaryl, -NHC(O)C 1 -C 6 alkyl-C(O)NH—OH, heteroaryl, cycloalkyl, heterocycloalkyl, or cycloalkyl or heterocycloalkyl attached to nitrogen, —NHC 1 -C 4 Alkyl, or -N(C 1 -C 4 alkyl) 2 The compound according to [6] above, substituted with 0 to 4 substituents independently selected from: 〔8〕Z 3 However, hydrogen, deuterium, halogens, C 1 -C 4 Alkyl, C 1 -C 3 Haloalkyl, -OH, -OC 1 -C 6 The compound according to [7] above, wherein the aryl or heteroaryl is substituted by 0 to 4 substituents independently selected from alkyl, or heterocycloalkyl bonded to nitrogen. 〔9〕 The compound is [C10] TIFF0007750851000059.tif229147 TIFF0007750851000060.tif232111 TIFF0007750851000061.tif143123 The compound according to [8] above,

[10] A composition or pharmaceutical composition comprising the compound according to any one of [1] to [9] above and a pharmaceutically acceptable carrier.

[11] A method for treating a disease or disorder, comprising administering to a patient or warm-blooded mammal having a disease or disorder mediated by CREB binding protein (CBP) / β-catenin signaling an amount of a compound described in any one of [1] to

[10] above sufficient to inhibit CBP / catenin signaling and / or enhance signaling mediated by p300 / catenin.

[12] The method according to

[11] above, wherein the amount of the compound administered comprises a therapeutically effective amount.

[13] In the compound, W is X, and X is [C11] TIFF0007750851000062.tif15150 wherein Z is OR 4 (In the formula, R 4 is hydrogen or C 1 -C 6 The method according to

[11] or

[12] , wherein Z is an amino acid or an amino acid ester, and n is 1 or 2.

[14] In the compound, X is [C12] TIFF0007750851000063.tif30142 wherein Z is OR 4 and R 4 is hydrogen or C 1 -C 6 The method according to any one of

[11] to

[13] above, wherein Z is alkyl, or Z is an amino acid or amino acid ester, and n is 1 or 2.

[15] The method according to any one of

[11] to

[14] above, wherein the disease or disorder comprises one or more of fibrosis, cancer, a neurological condition, a metabolic disorder, a skin condition, and aging.

[16] The method according to

[15] , wherein the metabolic disorder includes one or more of diabetes and / or fatty liver disease.

[17] The method according to

[16] , wherein the fatty liver disease includes one or more of alcoholic fatty liver disease (ALD), non-alcoholic fatty liver disease (NAFLD), and / or non-alcoholic steatohepatitis (NASH).

[18] The method according to

[15] above, wherein the fibrosis is pulmonary, hepatic, renal, cardiac, endometrial, cutaneous or systemic fibrosis.

[19] The method according to

[18] , wherein the fibrosis includes fibrosis in tissues of a SARS-CoV-2 (COVID-19) patient.

[20] The method of

[15] , wherein treating cancer comprises administering a CBP / β-catenin antagonist in combination with or as adjunctive therapy to one or more of cytotoxic and / or directed chemotherapy, and / or radiation therapy, and / or immunotherapy, including checkpoint inhibition, chimeric antigen receptor (CAR-T) and / or CAR-NK.

[21] The method of

[15] , wherein the neurological condition comprises one or more of Huntington's disease (HD), Parkinson's disease (PD), Alzheimer's disease (AD), multiple sclerosis (MS), and / or amyotrophic lateral sclerosis (ALS), muscular dystrophy (MD), and / or spinal muscular atrophy (SMA).

[22] The method of

[15] , wherein the skin condition includes one or more of atopic dermatitis, psoriasis, acne, fibrosis, wounds, scars, burns, sun or UV damage, diabetic ulceration, chronic ulceration, and / or alopecia.

[23] The method of

[22] above, wherein W is an ester of an alkyl acid or a fatty acid, and administration includes topical or transdermal administration.

[24] A cosmetic method for treating a skin condition, comprising the step of topically administering a cosmetically effective amount of the compound according to any one of [1] to

[10] to a patient or a warm-blooded mammal having the skin condition, wherein W is an ester of an alkyl acid or a fatty acid, and preferably the ester of the alkyl acid or the fatty acid is [C13] TIFF0007750851000064.tif2272 (wherein m is 1 to 14), [C14] TIFF0007750851000065.tif38137 A beauty method to be selected from.

[25] The method of

[24] , wherein the skin condition comprises one or more aging skin conditions selected from wrinkles, hyperpigmentation, redness, rosacea, dryness, cracking, loss of color, loss of elasticity, thinning hair, loss of color, scarring, acne, sun damage, hair loss, loss of hair color, reduced cuticle growth, and reduced nail growth.

[26] A method for the efficient synthesis of clinical grade drugs, comprising the use of an intermediate 2-propynyl-compound in the penultimate or final reaction step under GMP conditions to form a clinical grade isoxazole derivative by 3+2 cycloaddition.

[27] The step of preparing a clinical grade isoxazole derivative comprises, in the penultimate or final reaction step, reacting a final pharmaceutically acceptable salt, phosphate or phosphoric acid salt, of formula (IIa): [C15] TIFF0007750851000066.tif6070 (IIa) [In the formula, R a is hydrogen or -CH 3 and; R b is a monocyclic aryl group having 5 to 7 ring members, which may have 1 to 2 heteroatoms selected from nitrogen, oxygen, or sulfur, and which may have one or more substituents selected from the group consisting of halide, cyano, and lower alkyl; R is a phenyl group; a substituted phenyl group having one or more substituents (wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; benzyl groups; substituted benzyl groups having one or more substituents, wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; or a bicyclic aryl group having 8 to 11 ring members and optionally having 1 to 3 heteroatoms selected from nitrogen, oxygen, or sulfur; R 2 is hydrogen or -CH 3 and; Y is selected from hydrogen, deuterium, or a halogen; W is hydrogen, an ester of an alkyl or fatty acid, or X, where X is [C16] TIFF0007750851000067.tif32149 (wherein Z is OR 4 and R 4 is hydrogen or C 1 -C 6

[26] The method according to

[26] , comprising a step of preparing a compound by the method according to any one of [6] to [9] using the step before any optional formation of an intermediate 2-propynyl-compound of the formula (I) wherein Z is alkyl, or Z is an amino acid or amino acid ester, and n is 1 or 2).

[28] The method according to

[26] or

[27] , wherein the final or penultimate step carried out under GMP conditions is preceded by one or more reaction steps carried out under non-GMP conditions as part of an overall reaction scheme for preparing a clinical-grade isoxazole derivative.

[29] Formula (IIa): [C17] TIFF0007750851000068.tif5564 (IIa) {In the formula, R a is methyl or hydrogen; R b is a monocyclic aryl group having 5 to 7 ring members, which may have 1 to 2 heteroatoms selected from nitrogen, oxygen, or sulfur, and which may have one or more substituents selected from the group consisting of halide, cyano, and lower alkyl; R is a phenyl group; a substituted phenyl group having one or more substituents (wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; benzyl groups; substituted benzyl groups having one or more substituents, wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; or a bicyclic aryl group having 8 to 11 ring members and optionally having 1 to 3 heteroatoms selected from nitrogen, oxygen, or sulfur; R 2 is hydrogen or -CH 3 and; Y is selected from hydrogen, deuterium, or a halogen; W is X, where X is an alkyl or fatty acid; [C18] TIFF0007750851000069.tif33144 wherein Z is OR 4 (In the formula, R 4 is hydrogen or C 1 -C 6 or Z is an amino acid or amino acid ester, and n is 1 or 2.

[30] A method for enhancing vaccine efficacy, comprising administering to a subject, before vaccination, during vaccination, and / or after vaccination, an amount of a compound described in any one of [1] to

[10] sufficient to inhibit CBP / β-catenin-mediated signaling and / or enhance p300 / catenin-mediated signaling.

[31] The method according to

[31] above, wherein the amount of the compound administered comprises a therapeutically effective amount.

[32] The method of

[30] or

[31] , wherein enhancing vaccine efficacy includes one or more of: increasing the level of vaccine antigen-specific antibodies; increasing the percentage of protection generated; increasing the number and / or persistence of differentiated memory T-cells; and / or increasing the duration of protection.

[33] The method according to any one of

[30] to

[32] above, wherein inhibiting CBP / β-catenin-mediated signal transduction and / or enhancing p300 / catenin-mediated signal transduction comprises one or more of: maintaining metabolic asymmetry of activated T cells after their division in the subject; enhancing antigen-specific immunity by increasing the number and / or persistence of differentiated memory T cells; and / or enhancing presentation of antigens to T cells by antigen-presenting cells to enhance cooperation between the innate immune system and the adaptive immune system.

[34] The method according to any one of

[30] to

[33] above, wherein the vaccination comprises administration of an antiviral vaccine.

[35] The method according to any one of

[30] to

[34] , wherein the vaccination comprises administering an antiviral vaccine selected from influenza, SARS, SARS-CoV-2, HPV-A, HPV-B, and / or shingles.

[36] The method according to any one of

[30] to

[35] above, wherein the subject is a human aged 55 to 75 years, 55 to 85 years, ≥ 50 years, ≥ 60 years, or ≥ 65 years.

[37] The method according to any one of

[30] to

[36] , wherein the administration includes administration as an initiator before vaccination; and / or simultaneous administration with vaccination; and / or administration after the first vaccination or simultaneous administration.

Claims

1. Formula (Ia): 【Chemical 1】 (Ia) {During the ceremony, R a is hydrogen or -CH 3 and R b is a monocyclic aryl group having 5 to 7 ring members, which optionally has 1 to 2 heteroatoms selected from nitrogen, oxygen, or sulfur, and which optionally has one or more substituents selected from the group consisting of halide, cyano, and lower alkyl; R is a phenyl group; a substituted phenyl group having one or more substituents (wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; benzyl groups; substituted benzyl groups having one or more substituents, wherein the one or more substituents are selected from amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-3 or a bicyclic aryl group or a substituted bicyclic aryl having 8 to 11 ring members and optionally having 1 to 3 heteroatoms selected from nitrogen, oxygen, or sulfur (substituted bicyclic aryl is selected from amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-3 optionally bearing one or more substituents independently selected from alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; R 2 is hydrogen or -CH 3 and Y is selected from hydrogen, deuterium, or halogen; W is hydrogen, phosphate or phosphoric acid; 【change】 wherein m is 1 to 14. 【change】 or X, X is 【Chemistry 2】 wherein Z is OR 4 (In the formula, R 4 is hydrogen or C 1 -C 6 alkyl), and n is 1 or 2; L is -CH 2 -, -CF 2 -, or -C(CH 3 ) 2 - and; Q is a 5- or 6-membered nitrogen-containing heteroaryl substituted by 0-2 substituents; 【Chemistry 3】 wherein A, B, and D are independently selected from O, S, N, or —CH; 1 and Z 2 is hydrogen, deuterium, halogen, C 1 -C 4 Alkyl, —OH, —OC 1 -C 6 Alkyl, 【Chemistry 4】 (In the formula, R 1 , R 2 and R 3 is hydrogen, C 1 -C 6 alkyl, or C containing one or more —OH 1 -C 6 alkyl), and Z 3 is the following (i) hydrogen, (ii) halogens, (iii) a directly bonded aryl, heteroaryl, cycloalkyl, or heterocycloalkyl; (iv) an aryl, heteroaryl, cycloalkyl, or heterocycloalkyl bonded to —NH—; (v)-OC 1 -C 3 an aryl, heteroaryl, cycloalkyl, or heterocycloalkyl bonded to an alkyl; (vi)-NHC 1 -C 3 an aryl, heteroaryl, cycloalkyl, or heterocycloalkyl bonded to an alkyl; (vii) a cycloalkyl or heterocycloalkyl attached to a nitrogen atom; (viii)-NHC 1 -C 4 alkyl, or (ix)-N(C 1 -C 4 alkyl) 2 and each of (iii) to (ix) is selected from hydrogen, deuterium, halogen, C 1 -C 4 Alkyl, C 1 -C 3 Haloalkyl, —OH, —OC 1 -C 6 Alkyl, —OC 1 -C 6 Alkyl-C(O)NH-OH, -NH 2 , -C(O)NH-C 1 -C 6 Alkyl-heteroaryl, —NHC(O)C 1 -C 6 substituted by 0 to 4 substituents independently selected from alkyl-C(O)NH—OH, heteroaryl, cycloalkyl, or heterocycloalkyl; or a pharmaceutically acceptable salt thereof.

2. W is 2. The compound of claim 1, wherein m is 1 to 14.

3. 3. The compound of claim 1 or 2, wherein R is a bicyclic aryl selected from naphthyl, quinolinyl, isoquinolinyl, quinoxaline, phthalazine, quinazoline, cinnoline, naphthyridine, or substituted variants thereof.

4. The compound has the formula (Ib): 【Chemistry 7】 (Ib) (In the formula, X 1 and X 2 are independently selected from N, or —CH; R 2 4. The compound according to any one of claims 1 to 3, wherein Y, W, L and Q are as defined in claim 1.

5. L is -CH 2 - and; Q is, 【Chemistry 8】 wherein A, B, and D are independently selected from O, S, N, or —CH; 3 is as defined in claim 1).

6. A is —CH, B is N, and D is O(Z 3 -isoxazole-), W is hydrogen, phosphate or phosphoric acid; 【change】 wherein m is 1 to 14. 【change】 or X, where X is 【Chemistry 9】 wherein Z is OR 4 (In the formula, R 4 is hydrogen or C 1 -C 6 6. The compound of claim 5, wherein n is selected from the group consisting of:

7. Z 3 is selected from aryl or heteroaryl, and each of the aryl and heteroaryl is selected from hydrogen, deuterium, halogen, C 1 -C 4 Alkyl, C 1 -C 3 Haloalkyl, —OH, —OC 1 -C 6 Alkyl, —OC 1 -C 6 Alkyl-C(O)NH-OH, -NH 2 , -C(O)NH-C 1 -C 6 Alkyl-heteroaryl, —NHC(O)C 1 -C 6 7. The compound of claim 6, substituted with 0 to 4 substituents independently selected from alkyl-C(O)NH-OH, heteroaryl, cycloalkyl, or heterocycloalkyl.

8. Z 3 However, hydrogen, deuterium, halogens, C 1 -C 4 Alkyl, C 1 -C 3 Haloalkyl, —OH, —OC 1 -C 6 8. The compound of claim 7, wherein the aryl or heteroaryl is substituted with 0 to 4 substituents independently selected from alkyl, or heterocycloalkyl attached to the nitrogen.

9. The compound is 【Chemistry 10】 【change】 【change】 6. The compound of claim 5, wherein:

10. A composition or pharmaceutical composition comprising a compound according to any one of claims 1 to 9 and a pharmaceutically acceptable carrier.

11. A pharmaceutical composition for treating a disease or disorder mediated by CREB binding protein (CBP) / β-catenin signaling, comprising a compound according to any one of claims 1 to 9 or a pharmaceutical composition according to claim 10 in an amount sufficient to inhibit CBP / catenin signaling and / or enhance signaling mediated by p300 / catenin.

12. 12. The pharmaceutical composition of claim 11, wherein the amount of the compound comprises a therapeutically effective amount.

13. In the compound, W is X, and X is 【Chemistry 11】 wherein Z is OR 4 (In the formula, R 4 is hydrogen or C 1 -C 6 alkyl), 13. The pharmaceutical composition according to claim 11 or 12, wherein n is 1 or 2.

14. In the compound, X is 【Chemistry 12】 wherein Z is OR 4 and R 4 is hydrogen or C 1 -C 6 The pharmaceutical composition according to any one of claims 11 to 13, wherein n is alkyl and n is 1 or 2.

15. The pharmaceutical composition of any one of claims 11 to 14, wherein the disease or disorder comprises one or more of fibrosis, cancer, a neurological condition, a metabolic disorder, a skin condition and aging.

16. 16. The pharmaceutical composition of claim 15, wherein the metabolic disorder comprises one or more of diabetes and / or fatty liver disease.

17. 17. The pharmaceutical composition of claim 16, wherein the fatty liver disease comprises one or more of alcoholic fatty liver (ALD), non-alcoholic fatty liver (NAFLD), and / or non-alcoholic steatohepatitis (NASH).

18. 16. The pharmaceutical composition of claim 15, wherein the fibrosis is pulmonary, hepatic, renal, cardiac, endometrial, cutaneous or systemic fibrosis.

19. 19. The pharmaceutical composition of claim 18, wherein the fibrosis comprises fibrosis in tissue of a SARS-CoV-2 (COVID-19) patient.

20. 16. The pharmaceutical composition of claim 15, wherein treating cancer comprises administering a CBP / β-catenin antagonist in combination with or as adjunctive therapy with one or more of cytotoxic and / or directed chemotherapy, and / or radiotherapy, and / or immunotherapy including checkpoint inhibition, chimeric antigen receptor (CAR-T) and / or CAR-NK.

21. 16. The pharmaceutical composition of claim 15, wherein the neurological condition comprises one or more of Huntington's disease (HD), Parkinson's disease (PD), Alzheimer's disease (AD), multiple sclerosis (MS), and / or amyotrophic lateral sclerosis (ALS), muscular dystrophy (MD), and / or spinal muscular atrophy (SMA).

22. 16. The pharmaceutical composition of claim 15, wherein the skin condition comprises one or more of atopic dermatitis, psoriasis, acne, fibrosis, wounds, scars, burns, sun or U.V. damage, diabetic ulcerations, chronic ulcerations, and / or alopecia.

23. W is, wherein m is 1 to 14. and administration comprises topical or transdermal administration.

24. A cosmetic composition for treating a skin condition, comprising a cosmetically effective amount of a compound according to any one of claims 1 to 9 or a pharmaceutical composition according to claim 10, wherein W is wherein m is 1 to 14. A cosmetic composition.

25. 25. The cosmetic composition of claim 24, wherein the skin condition comprises one or more aging skin conditions selected from wrinkles, hyperpigmentation, redness, rosacea, dryness, cracking, loss of complexion, loss of elasticity, thinning hair, loss of complexion, scarring, acne, sun damage, hair loss, loss of hair color, reduced cuticle growth, and reduced nail growth.

26. 1. A method for the efficient synthesis of drugs, comprising the use of an intermediate 2-propynyl-compound of formula (IIa) in the penultimate or final reaction step under GMP conditions to form an isoxazole derivative by 3+2 cycloaddition, wherein the step of preparing the isoxazole derivative comprises the use of an intermediate 2-propynyl-compound of formula (IIa) in the penultimate or final reaction step, prior to any formation of a final pharmaceutically acceptable salt, phosphate or phosphoric acid salt: 【Chemistry 13】 (IIa) [In the formula, R a is hydrogen or -CH 3 and R b is a monocyclic aryl group having 5 to 7 ring members, which optionally has 1 to 2 heteroatoms selected from nitrogen, oxygen, or sulfur, and which optionally has one or more substituents selected from the group consisting of halide, cyano, and lower alkyl; R is a phenyl group; a substituted phenyl group having one or more substituents (wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; benzyl groups; substituted benzyl groups having one or more substituents, wherein the one or more substituents are selected from amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-3 or a bicyclic aryl group having 8 to 11 ring members and optionally having 1 to 3 heteroatoms selected from nitrogen, oxygen, or sulfur; R 2 is hydrogen or -CH 3 and Y is selected from hydrogen, deuterium, or halogen; W is 【change】 wherein m is 1 to 14. 【change】 or X, X is 【Chemistry 14】 (Wherein Z is OR 4 and R 4 is hydrogen or C 1 -C 6 is alkyl, and and n is 1 or 2), to prepare a compound according to any one of claims 6 to 9.

27. 27. The method of claim 26, wherein the last or penultimate step carried out under GMP conditions is preceded by one or more reaction steps under non-GMP conditions as part of an overall reaction scheme for preparing an isoxazole derivative.

28. Formula (IIa): 【Chemistry 15】 (IIa) {During the ceremony, R a is methyl or hydrogen; R b is a monocyclic aryl group having 5 to 7 ring members, which optionally has 1 to 2 heteroatoms selected from nitrogen, oxygen, or sulfur, and which optionally has one or more substituents selected from the group consisting of halide, cyano, and lower alkyl; R is a phenyl group; a substituted phenyl group having one or more substituents (wherein the one or more substituents are amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-4 Alkyl, C 1-3 alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl groups; benzyl groups; substituted benzyl groups having one or more substituents, wherein the one or more substituents are selected from amino, amidino, guanidino, hydrazino, amidazolyl, C 1-4 Alkylamino, C 1-4 Dialkylamino, halogen, perfluoro C 1-4 Alkyl, C 1-3 or a bicyclic aryl group having 8 to 11 ring members and optionally having 1 to 3 heteroatoms selected from nitrogen, oxygen, or sulfur; R 2 is hydrogen or -CH 3 and Y is selected from hydrogen, deuterium, or halogen; W is 【change】 wherein m is 1 to 14. 【change】 or X, X is 【Chemistry 16】 wherein Z is OR 4 (In the formula, R 4 is hydrogen or C 1 -C 6 alkyl), and n is 1 or 2}.

29. A pharmaceutical composition for enhancing vaccine efficacy, comprising a compound according to any one of claims 1 to 9 or a pharmaceutical composition according to claim 10 in an amount sufficient to inhibit CBP / β-catenin-mediated signaling and / or enhance p300 / catenin-mediated signaling, wherein the enhancement of vaccine efficacy is achieved by administering said amount of compound to a subject before, during, and / or after vaccination.

30. 30. The pharmaceutical composition of claim 29, wherein the amount of the compound comprises a therapeutically effective amount.

31. 31. The pharmaceutical composition of claim 29 or 30, wherein enhancing vaccine efficacy comprises one or more of: increasing the level of vaccine antigen-specific antibodies; increasing the percentage of protection generated; increasing the number and / or persistence of differentiated memory T-cells; and / or increasing the duration of protection.

32. 32. The pharmaceutical composition of any one of claims 29 to 31, wherein inhibiting CBP / β-catenin mediated signaling and / or enhancing p300 / catenin mediated signaling comprises one or more of: maintaining metabolic cellular asymmetry after division of activated T cells in a subject; enhancing antigen-specific immunity by increasing the number and / or persistence of differentiated memory T cells; and / or enhancing presentation of antigens to T cells by antigen presenting cells to enhance cooperation between the innate and adaptive immune systems.

33. The pharmaceutical composition according to any one of claims 29 to 32, wherein the vaccination comprises the administration of an antiviral vaccine.

34. 34. The pharmaceutical composition of any one of claims 29 to 33, wherein the vaccination comprises the administration of an antiviral vaccine selected from influenza, SARS, SARS-CoV-2, HPV-A, HPV-B, and / or shingles.

35. 35. The pharmaceutical composition of any one of claims 29 to 34, wherein the subject is a human aged 55-75 years, 55-85 years, ≥ 50 years, ≥ 60 years, or ≥ 65 years.

36. The pharmaceutical composition of any one of claims 29 to 35, wherein administration comprises administration as a starter before vaccination; and / or simultaneous administration with vaccination; and / or administration after or simultaneous administration with the first vaccination.

37. 11. A method for treating a disease or disorder, comprising administering to a non-human animal having a disease or disorder mediated by CREB binding protein (CBP) / β-catenin signaling an amount of a compound according to any one of claims 1 to 9 or a pharmaceutical composition according to claim 10 sufficient to inhibit CBP / catenin signaling and / or enhance signaling mediated by p300 / catenin.

38. 11. A method for enhancing vaccine efficacy, comprising administering to a non-human subject, before and / or during and / or after vaccination, an amount of a compound according to any one of claims 1 to 9 or a pharmaceutical composition according to claim 10 sufficient to inhibit CBP / β-catenin mediated signaling and / or enhance p300 / catenin mediated signaling.

39. 【Catalog 17】 A compound represented by the formula:

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