Compounds for targeting ULK3 and uses of the same

Abstract

The present disclosure relates to compounds and to their pharmaceutically acceptable salts, pharmaceutical compositions, methods of use, and methods for their preparation. The compounds disclosed herein are useful for modulating ULK3 activity and may be used in the treatment of disorders in which ULK3 activity is implicated, such as skin cancer.

Description

MGL-06625COMPOUNDS FOR TARGETING ULK3 AND USES OF THE SAMECROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 649,764, filed May 20, 2024, and U.S. Provisional Application No. 63 / 696,317, filed September 18, 2024, the entire contents of which are incorporated herein by reference.STATEMENT CONCERNING GOVERNMENT SUPPORT

[0002] This invention was made with government support under Agreement No. 2020A016550, Fund No. 239253 awarded by the National Institutes of Health-National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIH-NIAMS). The government has certain rights in the invention.BACKGROUND

[0003] The present disclosure relates to small molecules and their use as modulators of Unc-51 like kinase 3 (ULK3) for the treatment of a disease or disorder. ULK3 is a nuclear kinase with elevated expression levels in squamous cell carcinomas (SCCs) and is a determinant of the selfrenewal and oncogenic potential of human primary keratinocytes (HKCs) and SCC cells. Mechanistically, ULK3 directly binds and regulates the activity of two histone arginine methyltransferases, PRMT1 and PRMT5 (PRMT1 / 5), with ULK3 loss compromising PRMT1 / 5 chromatin association to specific genes and overall methylation of histone H4, a shared target of these enzymes. Downmodulating ULK3 by RNA interference or locked antisense nucleic acids (LNAs) blunts the proliferation and tumorigenic potential of SCC cells and promotes differentiation in two orthotopic models of skin cancer. Despite the highly important therapeutic implications provided by these insights, therapeutic developments toward modulating ULK3 expression during oncogenesis remain poorly understood.

[0004] The disclosure arises from a need to provide compounds for the modulation of ULK3. In particular, compounds with improved physicochemical, pharmacological and pharmaceutical properties are desirable.MGL-06625SUMMARY

[0005] In some embodiments, the present disclosure relates to a method of treating a disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of a compound selected from the group consisting of:p ,MGL-06625 or a pharmaceutically acceptable salt thereof; and wherein the disease or disorder is a cancer, a pre-cancerous condition, skin fibrosis, or skin aging.

[0006] In some aspects, the present disclosure provides a pharmaceutical composition comprising a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients, diluents, or carriers, or a combination thereof.

[0007] In some aspects, the present disclosure provides a method of modulating ULK3 activity (e.g., in vitro or in vivo) with a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0008] In some aspects, the present disclosure provides a method of treating or preventing a disease or disorder disclosed herein in a subject in need thereof, comprising administering to the subject a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0009] In some aspects, the present disclosure provides a method of treating a disease or disorder disclosed herein in a subject in need thereof, comprising administering to the subject a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0010] In some aspects, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof for use in modulating ULK3 activity (e.g., in vitro or in vivo).

[0011] In some aspects, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof for use in treating or preventing a disease or disorder disclosed herein.

[0012] In some aspects, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof for use in treating a disease or disorder disclosed herein.

[0013] In some aspects, the present disclosure provides use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for modulating ULK3 activity (e.g., in vitro or in vivo).

[0014] In some aspects, the present disclosure provides use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating or preventing a disease or disorder disclosed herein.MGL-06625

[0015] In some aspects, the present disclosure provides use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a disease or disorder disclosed herein.

[0016] In some embodiments, the disease or disorder is a squamous cell carcinoma, a skin melanoma, or a pre-cancerous condition.

[0017] In some embodiments, the disease or disorder is skin fibrosis.

[0018] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. In the specification, the singular forms also include the plural unless the context clearly dictates otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described below. All publications, patent applications, patents and other references mentioned herein are incorporated by reference. The references cited herein are not admitted to be prior art to the claimed invention. In the case of conflict, the present specification, including definitions, will control. In addition, the materials, methods and examples are illustrative only and are not intended to be limiting. In the case of conflict between the chemical structures and names of the compounds disclosed herein, the chemical structures will control.

[0019] Other features and advantages of the disclosure will be apparent from the following detailed description and claims.BRIEF DESCRIPTION OF DRAWINGS

[0020] FIGs. 1A-1C show the growth curves for the SCC13 (FIG. 1A), Cal27 (FIG. IB), and SCCO28 (FIG. 1C) cell lines for Compounds A, B, C, D, F, and G and the SU6668 positive control.

[0021] FIG. 2 shows the Incucyte time- lapse cell proliferation analysis of skin (SCC13) and oral (Cal27) SCC cell lines treated with Compounds A, B, C, D, F, and G.

[0022] FIGs. 3A-3D show (FIG. 3A) cell growth of the control (shCT) and ULK3-silenced SCC 13 cells, (FIG. 3B) WB on the same cells, and (FIGs. 3C-3D) the corresponding cell growth profiles of shULK3 SCC 13 cells with Compounds A, B, C, D, F, and G (blue) compared to control- silenced (black) SCC13 cells, DMSO-treated SCC13 shULK3 (red), and SCC13 shULK3 cells treated with SU6668 (purple).MGL-06625

[0023] FIGs. 4A-4B show (FIG. 4A) quantification of organoids formed by SCC13 cells and (FIG. 4B) representative images of the cells after treatment with Compound A, B, D, F, and G.

[0024] FIG. 5 shows the quantification of the sphere number (%) formed by Cal27 SCC cells after treatment with Compounds A, B, C, D, F, and G.

[0025] FIG. 6 shows growth curves of A375 melanoma cell line treated with vehicle or Compounds A, B, C, D, F, and G.

[0026] FIG. 7 shows the RT-qPCR analysis of the indicated genes in two cancer-associated fibroblast strains treated with vehicle or Compounds A, B, C, D, F, and G.

[0027] FIG. 8 shows plots demonstrating dose-dependent ULK3 autophosphorylation assays with the Compound B and Compound F (8 dilutions, 0.01-120pM range) versus DMSO. ULK3 activity (%) measured with ADP-GLO system (Promega), Mean+SD, n(assays)=3. Non-linear regression (curve fit) was calculated using PrismlO.

[0028] FIGs. 9A-9B show plots demonstrating data obtained in in vitro specificity assays of the ULK3 inhibitors Compound B and Compound F vs ULK1 kinase. FIG. 9 A) Dose-dependent ULK1 and ULK3 autophosphorylation assays with Compound B, Compound F (0.1-200pM range), or DMSO. Activity measured with ADP-GLO system (Promega). FIG. 9B) Dosedependent ULK1 kinase assays in the presence of MBP (Ipg) with Compound B, Compound F (0-60pM range), or DMSO. Mean n(assays)=2. Activity measured with ADP-GLO system (Promega).

[0029] FIGs. 10A-10B show data obtained in in vivo specificity assays of the ULK3 inhibitors Compounds B and F. FIG. 10A) SCC13 cells were treated for 2h with Compounds B or F (lOpM) or the ULK1 / 2 kinase inhibitor SBL0206965 (lOpM), or DMSO followed by immuno blot analysis using antibodies specific for ULK1 / 2 target sites on ATG1338 and ATG1439, total ATG13, ATG14, ULK3 and ACTIN. FIG. 10B) SCC 13 cells were treated for 2 hrs with Compounds B, F, or the broad specificity SU6668 compound followed by analysis with a commercially available antibody array to assess phosphorylation levels of various proliferation and stress kinases.

[0030] FIG. 11 is a schematic depiction of in silico prediction of Compounds B and F binding to the ULK3 protein.

[0031] FIG. 12 shows graphs demonstrating inhibition of SCC cell proliferation by ULK3 inhibitors. Live cell proliferation assays over the indicated times (hours) of SCC 13 cells treatedMGL-06625ULK3 inhibitors (40-20-10-5 pM) vs DMSO. Confluency normalized to Day 0 (triplicate, 5 images / well, 4-hourly, 4 days).

[0032] FIGs. 13A-13B show graphs demonstrating no growth inhibitory effects of ULK3 inhibitors Compounds A, B, C, D, F, and G on SCC13 cells with ULK3 silencing. Live cell proliferation assays of SCC13 cells with silenced ULK3 by ( proliferation analysis: (shULK3 #1 — FIG. 13A, and shULK3#2 — FIG. 13B) with the indicated ULK3 inhibitors (blue lines) versus the non-specific SU6668 compound (red line) and DMSO control (black line), (triplicate, 5 images / well, 4h intervals, 4 days).

[0033] FIGs. 14A-14B show graphs demonstrating absence of toxicity by the ULK3 inhibitors Compounds A, B, C, D, F, and G in SCCs and primary HDFs. FIG. 14A) SCC13 cell toxicity assessed with Caspase 3 / 7 dye (Sartorius) over 72h with compounds (lOpM), DMSO (negative control), and Bleomycin (lOpM, positive control). Live cell imaging recorded green object count / mm2every 3h (4 images / time point), mean, n(wells / compound)=3. FIG. 14B) A similar experiment in primary HDFs with Etoposide (25ng / ml) as a positive control. Green object count / mm2was recorded every 3h (4 images / time point), mean, n=( wells / compound)=2.

[0034] FIGs. 15A-15B show plots demonstrating FIG. 15 A) ULK3 expression in TCGA profiles of primary melanoma versus acquired nevi and melanoma profiles clustered after the ulceration status of melanoma, z-scores values, mean ± SEM, two-tailed unpaired, t-test; and FIG. 15B) Kaplan-Meier curves of melanoma patients’ long-term survival (% vs months) from the TCGA dataset, divided by high (red line) versus low (blue line) levels of ULK3 expression logrank of survival, Hazard Ratio, P- value are indicated. n(low ULk3)=229 n(high ULK3)=229.

[0035] FIGs. 16A-16C show graphs and images demonstrating FIG. 16A) efficient ULK3 silencing in melanoma. RT-PRC analysis of ULK3 expression normalized to 36B4 and western blot analysis using anti ULK3 and GAPH antibodies, in three melanoma cell lines (SKMEL28, M14 and A375) infected with two ULK3 silencing lentiviruses, or a control virus. FIG. 16B) ULK3 silencing blunts colony formation. The indicated melanoma cell lines infected with two ULK3 silencing lentiviruses, or a control virus, were plated at limited cell density for one week. Crystal violet stained colonies were quantified using ImageJ. n(dishes)=3, mean ± SEM, P<0.0001, one-way ANOVA. FIG. 16C) ULK3 silencing inhibits sphere-forming capabilities. The indicated melanoma cell lines infected with two ULK3 silencing lentiviruses, or a control virus were cultured in triplicate dishes under diluted conditions in Matrigel suspension for one week,MGL-06625 followed by quantification of spheres. Shown are the number of sphere number, n(dishes)=3, mean ± SEM, P<0.0001, one-way ANOVA.

[0036] FIG. 17 shows plots demonstrating live cell proliferation assays over the indicated times (hours) of WM989 melanoma cells treated ULK3 inhibitors Compounds A, B, C, D, F, and G (20- 10-5 pM) vs DMSO. Confluency normalized to Day 0 (triplicate, 5 images / well, 4-hourly, 4 days).

[0037] FIGs. 18A-18B show an image and a plot demonstrating treatment with Compound B or F decrease WM989 multicellular spheroid outgrowth. FIG. 18 A) Quantification of WM989 spheroid volumes (3,000 cells, 2 weeks) in 5% Matrigel. Compounds (lOpM), SU6668, or DMSO were added at 48h and 1 week. FIG. 18B) The ratio at time 0 (To) versus 14 days is plotted. Mean+SEM, n(sphere / concentration))=5 / dilution, one-way ANOVA, **P<0.001. A representative image for each condition at the end of the experiment is shown below. (To=time zero)

[0038] FIG. 19 shows plots demonstrating down-modulation of CAF gene expression by UEK3 inhibitors Compounds A, B, C, D, F, and G. RT-qPCR analysis of the indicated CAF effector genes in two cancer-associated fibroblast strains (CAF#8 and CAF# 19) treated with all UEK3 kinase inhibitors, SU6668 (lOpM) or DMSO for 72 h. Gene expression was normalized to 36B4 and expression is relative to DMSO-treated cells.

[0039] FIG. 20 is a table summarizing CAF-related gene signatures suppressed by treatment of three different CAF strains with Compounds B or F. Gene set enrichment analysis (GSEA) of global transcriptomic profiles of CAF #8, CAF#26, CAF#27 (characterized in ref. 14) after treatment (24h) with Compounds B or F (10 pM), or DMSO.

[0040] FIG. 21 is a plot showing decreased SCC cell expansion of co-cultured SCC / CAFs treated with UEK3 inhibitors Compounds A, B, C, D, F, and G. Red stained CAF#8 cells (3000 / well) were co-cultured with 500 SCC13 EGFP cells with six UEK3 kinase inhibitors (lOpM), or DMSO and imaged for red / green fluorescence every 6 hours (4 images / well) using Incucyte. Green fluorescent clusters (>4000 pixels), quantified over time (Al-Sartorius) and analyzed in PrismlO. Mean+SE, n(images / time point)=12, n(cultures)=3, one-way ANOVA, **P<0.01.

[0041] FIGs. 22A-22B are a schematic representation and a plot showing decreased in vivo SCC tumor growth by UEK3 inhibitors. FIG. 22A) SCC lesions from EGFP-expressing SCC 13 and CAFs co-injected in NOD / SCID IER2g- / - mouse ears (2 sites / ear). After 5 days, ears were treated with 15 pM compounds in EtOH, 2x / week for 2 weeks. Contralateral ears: EtOH controls (CTRE).MGL-06625Images show ear lesions in 2 animals (fluorescent + phase-contrast) and FIG. 22B) final tumor volume quantification ((L*lA2)*0.52). Mean ± SEM. n=4 mice / condition; *P<0.05, ***P<0.001, two-tailed unpaired t-test.

[0042] FIGs. 23A-23B Are an image and a plot demonstrating that ULK3 is up-regulated by UVA in human skin and in HDFs. FIG. 23A) Representative images (left) of double IF analysis with anti-UEK3 and keratin 10 (KRT10) antibodies of three independent human skin samples 24 h after UVA irradiation (18J / cm2) versus mock control, together with quantification of double positive UEK3 and KRT10 cells (right). > 100 cells per sample were counted. Mean ± SEM, n(samples)=3, two-tailed unpaired t-test, p<0.05; FIG. 23B) Western blot analysis with anti-UEK3 and ACTIN antibodies of HDFs at the indicated times after exposure to either 1 J / cm2 or 2J / cm2 UVA, or mock treated (Ctr).

[0043] FIG. 24 is a plot showing suppression of the transcriptional response of HDFs to UVA by treatment with Compound B or Compound F. HDFs were exposed to UVA ( 10J ) in PBS, followed by the addition of a fresh medium and addition of the Compound B or Compound F compounds (15pM), or DMSO vehicle. Cells were collected 24 hours later and analysed by global transcriptomics (Clariom D cDNA hybridization). Shown are the total number of genes up- or down-modulated by UVA exposure (> 2 fold change) in control versus Compound B or Compound F treated cells.

[0044] FIG. 25 shows a plot demonstrating GSEA analysis of transcriptomic profiles derived from the GTEX data base of 1,305 human skin samples derived from sun-exposed (n=701) versus non- sun-exposed (n=604) areas. Transcriptomic profiles of HDFs treated Compound B for 24 hours versus DMSO controls were used to build a signature of > 200 Compound B repressed genes. Shown are enrichment plots of the signature of Compound B repressed genes with corresponding normalized enrichment scores (NES) and nominal p values.

[0045] FIG. 26 shows a plot demonstrating GSEA analysis of transcriptomic profiles derived from the GTEX data base of 1,305 human skin samples derived from sun-exposed (n=701) versus non- sun-exposed (n=604) areas. Transcriptomic profiles of HDFs treated Compound F for 24 hours versus DMSO controls were used to build a signature of > 200 Compound F repressed genes. Shown are enrichment plots of the signature of Compound F repressed genes with corresponding normalized enrichment scores (NES) and nominal p values.MGL-06625

[0046] FIG. 27 shows a plot demonstrating GSEA analysis of transcriptomic profiles derived from the GTEX data base of 1,305 human skin samples derived from individuals of older (>50 years, n=894) versus younger age (<50 years, n=411), pooling together samples from sun- exposed and non-exposed areas and from males and females. Transcriptomic profiles of HDFs treated with Compound B for 24 hours versus DMSO controls were used to build a signature of > 200 Compound B repressed genes. Shown are enrichment plots of the signature of Compound B repressed genes with corresponding normalized enrichment scores (NES) and nominal p values.

[0047] FIG. 28 shows a plot demonstrating GSEA analysis of transcriptomic profiles derived from the GTEX data base of 1,305 human skin samples derived from individuals of older (>50 years, n=894) versus younger age (<50 years, n=411), pooling together samples from sun- exposed and non-exposed areas and from males and females. Transcriptomic profiles of HDFs treated with Compound F for 24 hours versus DMSO controls were used to build a signature of > 200 Compound F repressed genes. Shown are enrichment plots of the signature of Compound F repressed genes with corresponding normalized enrichment scores (NES) and nominal p values.DETAILED DESCRIPTION

[0048] The present disclosure relates to compounds which may modulate ULK3 expression or activity and are accordingly useful in methods of treatment of the human or animal body. The present disclosure also relates to pharmaceutical compositions comprising the compounds and to their use in the treatment of disorders in which ULK3 is implicated, such as skin cancer.

[0049] Skin photoaging and field cancerization result from cumulative alterations in the epidermal and dermal compartments, primarily affecting keratinocytes and dermal fibroblasts. These changes contribute to increased skin fragility, reduced regenerative capacity, and a heightened risk of tumorigenesis. Given the critical role of epigenetic regulation in these processes, targeting key epigenetic regulators represents a promising therapeutic approach for both prevention and treatment.

[0050] In some embodiments, the present disclosure relates to inhibition of the nuclear kinase ULK3, which has been identified as a pivotal regulator in both keratinocytes and dermal fibroblasts. ULK3 plays a role in their tumorigenic conversion and contributes to the pathogenesis of skin cancer and photoaging.MGL-06625

[0051] In some embodiments, the present disclosure relates to the treatment of cancer-associated fibroblasts (CAFs) with compounds of Table 1A.Table 1A.MGL-06625

[0052] The presently disclosed data show that the compounds of Table 1A suppresses the pro- tumorigenic activity of cancer-associated fibroblasts (CAFs), reversing their transcriptional profile to that of normal human dermal fibroblasts (HDFs). Moreover, these compounds block the transcriptional response of HDFs to UVA exposure, a key driver of skin photoaging and cancer progression. The relevance of the findings for skin aging is further supported by an integrated analysis comparing the gene signature affected by the two ULK3 inhibitors with the transcriptomic profiles of 1,305 skin samples, stratified by UV exposure (exposed vs. non-exposed) or age (irrespective of UV exposure).These findings position ULK3 inhibition as a viable strategy for addressing both skin cancer prevention and rejuvenation.Definitions

[0053] Unless otherwise stated, the following terms used in the specification and claims have the following meanings set out below.

[0054] Without wishing to be limited by this statement, it is understood that, while various options for variables are described herein, the disclosure intends to encompass operable embodiments having combinations of the options. The disclosure may be interpreted as excluding the non- operable embodiments caused by certain combinations of the options. For example, while various options for variables are described herein, the disclosure may be interpreted as excluding structures for non-operable compound caused by certain combinations of variables.

[0055] As used herein, the term “about” refers to a recited amount, value, or duration ± 10 % or less of said amount, value, or duration. In some embodiments, “about” refers to a recited amount, value, or duration ± 10 %, ± 8 %, ± 6 %, ± 5 %, ± 4 %, ± 2 %, ± 1 %, or ± 0.5 %. In other embodiments, “about” refers to a recited amount, value, or duration ± 10 %, ± 8 %, ± 6 %, ± 5 %, ± 4 %, or ± 2 %. In other embodiments, “about” refers to a recited amount, value, or duration ± 5 %. In some embodiments, “about” refers to a listed amount, value, or duration ± 2 % or ± 1 %. For example, in some embodiments, when the term “about” is used when reciting a temperature or temperature range, these terms refer to the recited temperature or temperature range ± 5 °C, ± 2 °C, or ± 1 °C. In other embodiments, the term “about” refers to the recited temperature or temperature range ± 2 °C.

[0056] As used herein, the expressions “one or more of A, B, or C,” “one or more A, B, or C,” “one or more of A, B, and C,” “one or more A, B, and C,” “selected from the group consisting ofMGL-06625A, B, and C”, “selected from A, B, and C”, and the like are used interchangeably and all refer to a selection from a group consisting of A, B, and / or C, i.e., one or more As, one or more Bs, one or more Cs, or any combination thereof, unless indicated otherwise.

[0057] It is to be understood that, throughout the description, where compositions are described as having, including, or comprising specific components, it is contemplated that compositions also consist essentially of, or consist of, the recited components. Similarly, where methods or processes are described as having, including, or comprising specific process steps, the processes also consist essentially of, or consist of, the recited processing steps. Further, it should be understood that the order of steps order for performing certain actions is immaterial so long as the invention remains operable. Moreover, two or more steps or actions can be conducted simultaneously.

[0058] It is to be understood that, unless otherwise stated, any description of a method of treatment or prevention includes use of the compounds to provide such treatment or prevention as is described herein. It is to be further understood, unless otherwise stated, any description of a method of treatment or prevention includes use of the compounds to prepare a medicament to treat or prevent such condition. The treatment or prevention includes treatment or prevention of human or non-human animals including rodents and other disease models.

[0059] It is to be understood that, unless otherwise stated, any description of a method of treatment includes use of the compounds to provide such treatment as is described herein. It is to be further understood, unless otherwise stated, any description of a method of treatment includes use of the compounds to prepare a medicament to treat such condition. The treatment includes treatment of human or non-human animals including rodents and other disease models.

[0060] As used herein, the term “subject” includes human and non-human animals, as well as cell lines, cell cultures, tissues, and organs. In some embodiments, the subject is a mammal. The mammal can be e.g. a human or appropriate non-human mammal, such as primate, mouse, rat, dog, cat, cow, horse, goat, camel, sheep or a pig. The subject can also be a bird or fowl. In some embodiments, the subject is a human.

[0061] As used herein, the term “subject in need thereof’ refers to a subject having a disease or having an increased risk of developing the disease. A subject in need thereof can be one who has been previously diagnosed or identified as having a disease or disorder disclosed herein. A subject in need thereof can also be one who is suffering from a disease or disorder disclosed herein. Alternatively, a subject in need thereof can be one who has an increased risk of developing suchMGL-06625 disease or disorder relative to the population at large (i.e., a subject who is predisposed to developing such disorder relative to the population at large). A subject in need thereof can have a refractory or resistant disease or disorder disclosed herein (i.e., a disease or disorder disclosed herein that does not respond or has not yet responded to treatment). The subject may be resistant at start of treatment or may become resistant during treatment. In some embodiments, the subject in need thereof received and failed all known effective therapies for a disease or disorder disclosed herein. In some embodiments, the subject in need thereof received at least one prior therapy.

[0062] As used herein, the term “treating” or “treat” describes the management and care of a patient for the purpose of combating a disease, condition, or disorder and includes the administration of a compound of the present disclosure, or a pharmaceutically acceptable salt, polymorph or solvate thereof, to alleviate the symptoms or complications of a disease, condition or disorder, or to eliminate the disease, condition or disorder. The term “treat” can also include treatment of a cell in vitro or an animal model. It is to be appreciated that references to “treating” or “treatment” include the alleviation of established symptoms of a condition. “Treating” or “treatment” of a state, disorder or condition therefore includes: (1) preventing or delaying the appearance of clinical symptoms of the state, disorder or condition developing in a human that may be afflicted with the state, disorder or condition but does not yet experience or display clinical or subclinical symptoms of the state, disorder or condition, (2) inhibiting the state, disorder or condition, i.e., arresting, reducing or delaying the progression of the disease or a relapse thereof (in case of maintenance treatment) or at least one clinical or subclinical symptom thereof, or (3) relieving or attenuating the disease, i.e., causing regression of the state, disorder or condition or at least one of its clinical or subclinical symptoms.

[0063] It is to be understood that a compound of the present disclosure, or a pharmaceutically acceptable salt, polymorph or solvate thereof, can or may also be used to prevent a relevant disease, condition or disorder, or used to identify suitable candidates for such purposes.

[0064] As used herein, the term “preventing,” “prevent,” or “protecting against” describes reducing or eliminating the onset of the symptoms or complications of such disease, condition or disorder.

[0065] It is to be understood that one skilled in the art may refer to general reference texts for detailed descriptions of known techniques discussed herein or equivalent techniques. These texts include Ausubel et al., Current Protocols in Molecular Biology, John Wiley and Sons, Inc. (2005);MGL-06625Sambrook et al., Molecular Cloning, A Laboratory Manual (3rdedition), Cold Spring Harbor Press, Cold Spring Harbor, New York (2000); Coligan et al., Current Protocols in Immunology, John Wiley & Sons, N.Y.; Enna et al., Current Protocols in Pharmacology, John Wiley & Sons, N.Y.; Fingl et al., The Pharmacological Basis of Therapeutics (1975), Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, 18thedition (1990). These texts can, of course, also be referred to in making or using an aspect of the disclosure.

[0066] It is to be understood that the present disclosure also provides pharmaceutical compositions comprising any compound described herein in combination with one or more pharmaceutically acceptable excipient, diluent, adjuvant, carrier, or a combination thereof.

[0067] As used herein, the term “pharmaceutical composition” is a formulation containing the compounds of the present disclosure in a form suitable for administration to a subject. In one embodiment, the pharmaceutical composition is in bulk or in unit dosage form. The unit dosage form is any of a variety of forms, including, for example, a capsule, an IV bag, a tablet, a single pump on an aerosol inhaler or a vial. The quantity of active ingredient (e.g., a formulation of the disclosed compound or salt, hydrate, solvate or isomer thereof) in a unit dose of composition is an effective amount and is varied according to the particular treatment involved. One skilled in the art will appreciate that it is sometimes necessary to make routine variations to the dosage depending on the age and condition of the patient. The dosage will also depend on the route of administration. A variety of routes are contemplated, including oral, pulmonary, rectal, parenteral, transdermal, subcutaneous, intravenous, intramuscular, intraperitoneal, inhalational, buccal, sublingual, intrapleural, intrathecal, intranasal, and the like. Dosage forms for the topical or transdermal administration of a compound of this disclosure include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. In one embodiment, the active compound is mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants that are required.

[0068] As used herein, the term “pharmaceutically acceptable” refers to those compounds, anions, cations, materials, compositions, carriers, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.MGL-06625

[0069] As used herein, the term “pharmaceutically acceptable excipient” means an excipient that is useful in preparing a pharmaceutical composition that is generally safe, non-toxic and neither biologically nor otherwise undesirable, and includes excipient that is acceptable for veterinary use as well as human pharmaceutical use. A “pharmaceutically acceptable excipient” as used in the specification and claims includes both one and more than one such excipient.

[0070] It is to be understood that a pharmaceutical composition of the disclosure is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., ingestion), inhalation, transdermal (topical), and transmucosal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates, and agents for the adjustment of tonicity such as sodium chloride or dextrose. The pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

[0071] It is to be understood that a compound or pharmaceutical composition of the disclosure can be administered to a subject in many of the well-known methods currently used for chemotherapeutic treatment. For example, a compound of the disclosure may be injected into the blood stream or body cavities or taken orally or applied through the skin with patches. The dose chosen should be sufficient to constitute effective treatment but not so high as to cause unacceptable side effects. The state of the disease condition (e.g., a disease or disorder disclosed herein) and the health of the patient should preferably be closely monitored during and for a reasonable period after treatment.

[0072] As used herein, the term “therapeutically effective amount”, refers to an amount of a pharmaceutical agent to treat or ameliorate an identified disease or condition, or to exhibit a detectable therapeutic or inhibitory effect. The effect can be detected by any assay method known in the art. The precise effective amount for a subject will depend upon the subject’s body weight, size, and health; the nature and extent of the condition; and the therapeutic or combination ofMGL-06625 therapeutics selected for administration. The dosage may vary within this range depending upon the dosage form employed, sensitivity of the patient, and the route of administration.

[0073] Dosage and administration are adjusted to provide sufficient levels of the active agent(s) or to maintain the desired effect. Factors which may be taken into account include the severity of the disease state, general health of the subject, age, weight, and gender of the subject, diet, time and frequency of administration, drug combination(s), reaction sensitivities, and tolerance / response to therapy. Long-acting pharmaceutical compositions may be administered every 3 to 4 days, every week, or once every two weeks depending on half-life and clearance rate of the particular formulation.

[0074] The pharmaceutical compositions containing active compounds of the present disclosure may be manufactured in a manner that is generally known, e.g. by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilizing processes. Pharmaceutical compositions may be formulated in a conventional manner using one or more pharmaceutically acceptable carriers comprising excipients and / or auxiliaries that facilitate processing of the active compounds into preparations that can be used pharmaceutically. Of course, the appropriate formulation is dependent upon the route of administration chosen.

[0075] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringeability 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 (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), cyclodextrins and suitable mixtures thereof. The 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 dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to includeMGL-06625 isotonic agents, for example, sugars, polyalcohols such as mannitol and sorbitol, and sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

[0076] Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, methods of preparation are vacuum drying and freeze-drying that yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

[0077] Oral compositions generally include an inert diluent or an edible pharmaceutically acceptable carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, capsules or sachets. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed. Pharmaceutically compatible binding agents, and / or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, orange flavoring.

[0078] 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, e.g. a gas such as carbon dioxide, or a nebulizer.

[0079] Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, forMGL-06625 example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays, powders or suppositories. For transdermal administration, the active compounds are formulated into ointments, salves, gels, or creams as generally known in the art.

[0080] The active compounds can be prepared with pharmaceutically acceptable carriers that will protect the compound against 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, poly anhydrides, polyglycolic acid, collagen, poly orthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. The materials can also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted to infected cells with monoclonal antibodies to viral antigens) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No. 4,522,811.

[0081] It is especially advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the disclosure are dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved.

[0082] It is to be understood that the pharmaceutical compositions can be included in a container, pack, or dispenser together with instructions for administration.

[0083] It is to be understood that, for the compounds of the present disclosure being capable of further forming salts, all of these forms are also contemplated within the scope of the claimed disclosure.

[0084] As used herein, the term “pharmaceutically acceptable salts” refer to derivatives of the compounds of the present disclosure wherein the parent compound is modified by making acid or base salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral organic acid salts of basic residues such as amines, alkali organic salts of acidic residues such as carboxylic acids, and the like. The pharmaceutically acceptable salts include theMGL-06625 conventional non-toxic salts or the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic organic acids. For example, such conventional non-toxic salts include, but are not limited to, those derived from inorganic and organic acids selected from 2-acetoxybenzoic, 2-hydroxyethane sulfonic, acetic, ascorbic, benzene sulfonic, benzoic, bicarbonic, carbonic, citric, edetic, ethane disulfonic, 1,2-ethane sulfonic, fumaric, glucoheptonic, gluconic, glutamic, glycolic, glycollyarsanilic, hexylresorcinic, hydrabamic, hydrobromic, hydrochloric, hydroiodic, hydroxymaleic, hydroxynaphthoic, isethionic, lactic, lactobionic, lauryl sulfonic, maleic, malic, mandelic, methane sulfonic, napsylic, nitric, oxalic, pamoic, pantothenic, phenylacetic, phosphoric, polygalacturonic, propionic, salicylic, stearic, subacetic, succinic, sulfamic, sulfanilic, sulfuric, tannic, tartaric, toluene sulfonic, and the commonly occurring amine acids, e.g., glycine, alanine, phenylalanine, arginine, etc.

[0085] In some embodiments, the pharmaceutically acceptable salt is a sodium salt, a potassium salt, a calcium salt, a magnesium salt, a diethylamine salt, a choline salt, a meglumine salt, a benzathine salt, a tromethamine salt, an ammonia salt, an arginine salt, or a lysine salt.

[0086] Other examples of pharmaceutically acceptable salts include hexanoic acid, cyclopentane propionic acid, pyruvic acid, malonic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, 4- chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo-[2.2.2]-oct-2-ene-l -carboxylic acid, 3 -phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, muconic acid, and the like. The present disclosure also encompasses salts formed when an acidic proton present in the parent compound either is replaced by a metal ion, e.g., an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinates with an organic base such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like. In the salt form, it is understood that the ratio of the compound to the cation or anion of the salt can be 1:1, or any ratio other than 1:1, e.g., 3: 1, 2: 1, 1:2, or 1:3.

[0087] The compounds, or pharmaceutically acceptable salts thereof, are administered orally, nasally, transdermally, pulmonary, inhalationally, buccally, sublingually, intraperitoneally, subcutaneously, intramuscularly, intravenously, rectally, intrapleurally, intrathecally and parenterally. In one embodiment, the compound is administered orally.

[0088] The dosage regimen utilizing the compounds is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severityMGL-06625 of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound or salt thereof employed.

[0089] Techniques for formulation and administration of the disclosed compounds of the disclosure can be found in Remington: the Science and Practice of Pharmacy, 19thedition, Mack Publishing Co., Easton, PA (1995). In an embodiment, the compounds described herein, and the pharmaceutically acceptable salts thereof, are used in pharmaceutical preparations in combination with a pharmaceutically acceptable carrier or diluent. Suitable pharmaceutically acceptable carriers include inert solid fillers or diluents and sterile aqueous organic solutions. The compounds will be present in such pharmaceutical compositions in amounts sufficient to provide the desired dosage amount in the range described herein.

[0090] All percentages and ratios used herein, unless otherwise indicated, are by weight. Other features and advantages of the present disclosure are apparent from the different examples. The provided examples illustrate different components and methodology useful in practicing the present disclosure. The examples do not limit the claimed disclosure. Based on the present disclosure the skilled artisan can identify and employ other components and methodology useful for practicing the present disclosure.

[0091] All publications and patent documents cited herein are incorporated herein by reference as if each such publication or document was specifically and individually indicated to be incorporated herein by reference. Citation of publications and patent documents is not intended as an admission that any is pertinent prior art, nor does it constitute any admission as to the contents or date of the same. The invention having now been described by way of written description, those of skill in the art will recognize that the invention can be practiced in a variety of embodiments and that the foregoing description and examples below are for purposes of illustration and not limitation of the claims that follow.

[0092] As use herein, the phrase “compound of the disclosure” refers to those compounds which are disclosed herein, both generically and specifically.Compounds of the Present Disclosure

[0093] In some embodiments, the compound is selected from the compounds described in Table 1, or a prodrug or pharmaceutically acceptable salt thereof.MGL-06625

[0094] In some embodiments, the compound is selected from the compounds described in Table 1, or a pharmaceutically acceptable salt thereof.

[0095] In some embodiments, the compound is selected from the compounds described in Table 1.

[0096] In some embodiments, the compound is a hydrochloride salt of a compound in Table 1.

[0097] In some embodiments, the compound is selected from the prodrugs of compounds described in Table 1, or a pharmaceutically acceptable salt thereof.Table 1.MGL-06625

[0098] In some embodiments, the compound is a pharmaceutically acceptable salt of any one of the compounds described in Table 1.

[0099] In some embodiments, the compound is Compound A or a pharmaceutically acceptable salt thereof.

[0100] In some embodiments, the compound is Compound A.

[0101] In some embodiments, the compound is Compound B or a pharmaceutically acceptable salt thereof.

[0102] In some embodiments, the compound is Compound B.

[0103] In some embodiments, the compound is Compound C or a pharmaceutically acceptable salt thereof.

[0104] In some embodiments, the compound is Compound C.

[0105] In some embodiments, the compound is Compound D or a pharmaceutically acceptable salt thereof.

[0106] In some embodiments, the compound is Compound D.

[0107] In some embodiments, the compound is Compound E or a pharmaceutically acceptable salt thereof.

[0108] In some embodiments, the compound is Compound E.

[0109] In some embodiments, the compound is Compound F or a pharmaceutically acceptable salt thereof.

[0110] In some embodiments, the compound is Compound F.

[0111] In some embodiments, the compound is Compound G or a pharmaceutically acceptable salt thereof.

[0112] In some embodiments, the pharmaceutically acceptable salt of Compound G is the hydrochloride salt.

[0113] In some embodiments, the compound is Compound G.MGL-06625

[0114] In some aspects, the present disclosure provides a compound being an isotopic derivative (e.g., isotopically labeled compound) of any one of the compounds of the compounds disclosed herein.

[0115] In some embodiments, the compound is an isotopic derivative of any one of the compounds described in Table 1, or a prodrug or pharmaceutically acceptable salt thereof.

[0116] In some embodiments, the compound is an isotopic derivative of any one of the compounds described in Table 1, or a pharmaceutically acceptable salt thereof.

[0117] In some embodiments, the compound is an isotopic derivative of any one of prodrugs of the compounds described in Table 1, or a pharmaceutically acceptable salt thereof.

[0118] In some embodiments, the compound is an isotopic derivative of any one of the compounds described in Table 1.

[0119] It is understood that the isotopic derivative can be prepared using any of a variety of art- recognized techniques.

[0120] In some embodiments, the isotopic derivative is a deuterium labeled compound.

[0121] In some embodiments, the isotopic derivative is a deuterium labeled compound of any one of the compounds disclosed herein.

[0122] The term “isotopic derivative”, as used herein, refers to a derivative of a compound in which one or more atoms are isotopically enriched or labelled. For example, an isotopic derivative of a compound described in Table 1 is isotopically enriched with regard to, or labelled with, one or more isotopes as compared to the corresponding compound described in Table 1. In some embodiments, the isotopic derivative is enriched with regard to, or labelled with, one or more atoms selected from2H,13C,14C,15N,18O,29Si,31P, and34S. In some embodiments, the isotopic derivative is a deuterium labeled compound (i.e. being enriched with2H with regard to one or more atoms thereof). In some embodiments, the compound is a18F labeled compound. In some embodiments, the compound is a123I labeled compound, a124I labeled compound, a125I labeled compound, a129I labeled compound, a131I labeled compound, a135I labeled compound, or any combination thereof. In some embodiments, the compound is a33S labeled compound, a34S labeled compound, a35S labeled compound, a36S labeled compound, or any combination thereof.

[0123] It is understood that the18F,123I,124I,125I,129I,131I,135I,32S,34S,35S, and / or36S labeled compound, can be prepared using any of a variety of art-recognized techniques.MGL-06625

[0124] A compound of the invention or a pharmaceutically acceptable salt or solvate thereof that contains one or more of the aforementioned18F,123I,124I,125I,129I,131I,135I,32S,34S,35S, and36S atom(s) is within the scope of the invention. Further, substitution with isotope (e.g,,18F,123I,124I,125I,129I,1311,1351,3S,34S,35S, and / or36S) may afford certain therapeutic advantages resulting from greater metabolic stability, e.g., increased in vivo half-life or reduced dosage requirements.

[0125] For the avoidance of doubt it is to be understood that, where in this specification a group is qualified by “described herein”, the said group encompasses the first occurring and broadest definition as well as each and all of the particular definitions for that group.

[0126] A suitable pharmaceutically acceptable salt of a compound of the disclosure is, for example, an acid-addition salt of a compound of the disclosure which is sufficiently basic, for example, an acid-addition salt with, for example, an inorganic organic acid, for example hydrochloric, hydrobromic, sulfuric, phosphoric, trifluoroacetic, formic, citric methane sulfonate or maleic acid. In addition, a suitable pharmaceutically acceptable salt of a compound of the disclosure which is sufficiently acidic is an alkali metal salt, for example a sodium or potassium salt, an alkaline earth metal salt, for example a calcium or magnesium salt, an ammonium salt or a salt with an organic base which affords a pharmaceutically acceptable cation, for example a salt with methylamine, dimethylamine, diethylamine, trimethylamine, piperidine, morpholine or tris-(2-hydroxyethyl)amine.

[0127] It will be understood that the compounds of any one of the compounds disclosed herein and any pharmaceutically acceptable salts thereof, comprise stereoisomers, mixtures of stereoisomers, polymorphs of all isomeric forms of said compounds.

[0128] It will be understood that while compounds disclosed herein may be presented in one particular configuration. Such particular configuration is not to be construed as limiting the disclosure to one or another isomer, tautomer, regioisomer or stereoisomer, nor does it exclude mixtures of isomers, tautomers, regioisomers or stereoisomers. In some embodiments, the presentation of a compound herein in a particular configuration intends to encompass, and to refer to, each of the available isomers, tautomers, regioisomers, and stereoisomers of the compound, or any mixture thereof; while the presentation further intends to refer to the specific configuration of the compound.

[0129] It will be understood that while compounds disclosed herein may be presented without specified configuration (e.g., without specified stereochemistry). Such presentation intends toMGL-06625 encompass all available isomers, tautomers, regioisomers, and stereoisomers of the compound. In some embodiments, the presentation of a compound herein without specified configuration intends to refer to each of the available isomers, tautomers, regioisomers, and stereoisomers of the compound, or any mixture thereof.

[0130] As used herein, the term “isomerism” means compounds that have identical molecular formulae but differ in the 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 termed “stereoisomers.” Stereoisomers that are not mirror images of one another are termed “diastereoisomers,” and stereoisomers that are non-superimposable mirror images of each other are termed “enantiomers” or sometimes optical isomers. A mixture containing equal amounts of individual enantiomeric forms of opposite chirality is termed a “racemic mixture.”

[0131] As used herein, the term “chiral center” refers to a carbon atom bonded to four nonidentical substituents.

[0132] As used herein, the term “chiral isomer” means a compound with at least one chiral center. Compounds with more than one chiral center may exist either as an individual diastereomer or as a mixture of diastereomers, termed “diastereomeric mixture.” When one chiral center is present, a stereoisomer may be characterized by the absolute configuration (R or S) of that chiral center. Absolute configuration refers to the arrangement in space of the substituents attached to the chiral center. The substituents attached to the chiral center under consideration are ranked in accordance with the Sequence Rule of Cahn, Ingold and Prelog. (Cahn et al., Angew. Chem. Inter. Edit. 1966, 5, 385; errata 511; Cahn et al., Angew. Chem. 1966, 78, 413; Cahn and Ingold, J. Chem. Soc. 1951 (London), 612; Cahn et al., Experientia 1956, 12, 81; Cahn, J. Chem. Educ. 1964, 41, 116).

[0133] As used herein, the term “geometric isomer” means the diastereomers that owe their existence to hindered rotation about double bonds or a cycloalkyl linker (e.g., 1,3 -cyclobutyl). These configurations are differentiated in their names by the prefixes cis and trans, or Z and E, which indicate that the groups are on the same or opposite side of the double bond in the molecule according to the Cahn-Ingold-Prelog rules.

[0134] It is to be understood that the compounds of the present disclosure may be depicted as different chiral isomers or geometric isomers. It is also to be understood that when compounds have chiral isomeric or geometric isomeric forms, all isomeric forms are intended to be includedMGL-06625 in the scope of the present disclosure, and the naming of the compounds does not exclude any isomeric forms, it being understood that not all isomers may have the same level of activity.

[0135] It is to be understood that the structures and other compounds discussed in this disclosure include all atropic isomers thereof. It is also to be understood that not all atropic isomers may have the same level of activity.

[0136] As used herein, the term “atropic isomers” are a type of stereoisomer in which the atoms of two isomers are arranged differently in space. Atropic isomers owe their existence to a restricted rotation caused by hindrance of rotation of large groups about a central bond. Such atropic isomers typically exist as a mixture, however as a result of recent advances in chromatography techniques, it has been possible to separate mixtures of two atropic isomers in select cases.

[0137] As used herein, the term “tautomer” is one of two or more structural isomers that exist in equilibrium and is readily converted from one isomeric form to another. This conversion results in the formal migration of a hydrogen atom accompanied by a switch of adjacent conjugated double bonds. Tautomers exist as a mixture of a tautomeric set in solution. In solutions where tautomerization is possible, a chemical equilibrium of the tautomers will be reached. The exact ratio of the tautomers depends on several factors, including temperature, solvent and pH. The concept of tautomers that are interconvertible by tautomerisations is called tautomerism. Of the various types of tautomerism that are possible, two are commonly observed. In keto-enol tautomerism a simultaneous shift of electrons and a hydrogen atom occurs. Ring-chain tautomerism arises as a result of the aldehyde group (-CHO) in a sugar chain molecule reacting with one of the hydroxy groups (-OH) in the same molecule to give it a cyclic (ring-shaped) form as exhibited by glucose.

[0138] It is to be understood that the compounds of the present disclosure may be depicted as different tautomers. It should also be understood that when compounds have tautomeric forms, all tautomeric forms are intended to be included in the scope of the present disclosure, and the naming of the compounds does not exclude any tautomer form. It will be understood that certain tautomers may have a higher level of activity than others.

[0139] Compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed “isomers”. Isomers that differ in the arrangement of their atoms in space are termed “stereoisomers”. Stereoisomers that are not mirror images of one another are termed “diastereomers” and those that areMGL-06625 non-superimposable mirror images of each other are termed “enantiomers”. When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers is possible. An enantiomer can be characterised by the absolute configuration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and designated as dextrorotatory or levorotatory (i.e., as (+) or (-)-isomers respectively). A chiral compound can exist as either individual enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a “racemic mixture”.

[0140] The compounds of this disclosure may possess one or more asymmetric centers; such compounds can therefore be produced as individual (R)- or (S)-stereoisomers or as mixtures thereof. Unless indicated otherwise, the description or naming of a particular compound in the specification and claims is intended to include both individual enantiomers and mixtures, racemic or otherwise, thereof. The methods for the determination of stereochemistry and the separation of stereoisomers are well-known in the art (see discussion in Chapter 4 of “Advanced Organic Chemistry”, 4th edition J. March, John Wiley and Sons, New York, 2001), for example by synthesis from optically active starting materials or by resolution of a racemic form. Some of the compounds of the disclosure may have geometric isomeric centers (E- and Z- isomers). It is to be understood that the present disclosure encompasses all optical, diastereoisomers and geometric isomers and mixtures thereof that possess ULK3 modulatory activity.

[0141] The present disclosure also encompasses compounds of the disclosure as defined herein which comprise one or more isotopic substitutions.

[0142] It is to be understood that the compounds described herein include the compounds themselves, as well as their salts, and their solvates, if applicable. A salt, for example, can be formed between an anion and a positively charged group (e.g., amino) on a substituted compound disclosed herein. Suitable anions include chloride, bromide, iodide, sulfate, bisulfate, sulfamate, nitrate, phosphate, citrate, methanesulfonate, trifluoroacetate, glutamate, glucuronate, glutarate, malate, maleate, succinate, fumarate, tartrate, tosylate, salicylate, lactate, naphthalenesulfonate, and acetate (e.g., trifluoroacetate).

[0143] As used herein, the term “pharmaceutically acceptable anion” refers to an anion suitable for forming a pharmaceutically acceptable salt. Likewise, a salt can also be formed between a cation and a negatively charged group (e.g., carboxylate) on a substituted compound disclosedMGL-06625 herein. Suitable cations include sodium ion, potassium ion, magnesium ion, calcium ion, and an ammonium cation such as tetramethylammonium ion or diethylamine ion. The substituted compounds disclosed herein also include those salts containing quaternary nitrogen atoms.

[0144] It is to be understood that the compounds of the present disclosure, for example, the salts of the compounds, can exist in either hydrated or unhydrated (the anhydrous) form or as solvates with other solvent molecules. Nonlimiting examples of hydrates include monohydrates, dihydrates, etc. Nonlimiting examples of solvates include ethanol solvates, acetone solvates, etc.

[0145] As used herein, the term “solvate” means solvent addition forms that contain either stoichiometric or non- stoichiometric amounts of solvent. Some compounds have a tendency to trap a fixed molar ratio of solvent molecules in the crystalline solid state, thus forming a solvate. If the solvent is water the solvate formed is a hydrate; and if the solvent is alcohol, the solvate formed is an alcoholate. Hydrates are formed by the combination of one or more molecules of water with one molecule of the substance in which the water retains its molecular state as H2O.

[0146] As used herein, the term “analog” refers to a chemical compound that is structurally similar to another but differs slightly in composition (as in the replacement of one atom by an atom of a different element or in the presence of a particular functional group, or the replacement of one functional group by another functional group). Thus, an analog is a compound that is similar or comparable in function and appearance, but not in structure origin to the reference compound.

[0147] As used herein, the term “derivative” refers to compounds that have a common core structure and are substituted with various groups as described herein.

[0148] As used herein, the term “bioisostere” refers to a compound resulting from the exchange of an atom or of a group of atoms with another, broadly similar, atom or group of atoms. The objective of a bioisosteric replacement is to create a new compound with similar biological properties to the parent compound. The bioisosteric replacement may be physicochemically or topologically based. Examples of carboxylic acid bioisosteres include, but are not limited to, acyl sulfonamides, tetrazoles, sulfonates and phosphonates. See, e.g., Patani and LaVoie, Chem. Rev. 96, 3147-3176, 1996.

[0149] It is also to be understood that certain compounds disclosed herein may exist in solvated as well as unsolvated forms such as, for example, hydrated forms. A suitable pharmaceutically acceptable solvate is, for example, a hydrate such as hemi-hydrate, a mono-hydrate, a di-hydrateMGL-06625 or a tri-hydrate. It is to be understood that the disclosure encompasses all such solvated forms that possess ULK3 modulatory activity.

[0150] It is also to be understood that certain compounds disclosed herein may exhibit polymorphism, and that the disclosure encompasses all such forms, or mixtures thereof, which possess ULK3 modulatory activity. It is generally known that crystalline materials may be analysed using conventional techniques such as X-Ray Powder Diffraction analysis, Differential Scanning Calorimetry, Thermal Gravimetric Analysis, Diffuse Reflectance Infrared Fourier Transform (DRIFT) spectroscopy, Near Infrared (NIR) spectroscopy, solution and / or solid state nuclear magnetic resonance spectroscopy. The water content of such crystalline materials may be determined by Karl Fischer analysis.

[0151] Compounds disclosed herein may exist in a number of different tautomeric forms and references to Compounds A-G include all such forms. For the avoidance of doubt, where a compound can exist in one of several tautomeric forms, and only one is specifically described or shown, all others are nevertheless embraced. Examples of tautomeric forms include keto-, enol-, and enolate-forms, as in, for example, the following tautomeric pairs: keto / enol (illustrated below), imine / enamine, amide / imino alcohol, amidine / amidine, nitroso / oxime, thioketone / enethiol, and nitro / aci-nitro.keto enol enolate

[0152] Compounds disclosed herein containing an amine function may also form N-oxides. A reference herein to a compound that contains an amine function also includes the N-oxide. Where a compound contains several amine functions, one or more than one nitrogen atom may be oxidised to form an N-oxide. Particular examples of N-oxides are the N-oxides of a tertiary amine or a nitrogen atom of a nitrogen-containing heterocycle. N-oxides can be formed by treatment of the corresponding amine with an oxidizing agent such as hydrogen peroxide or a peracid (e.g. a peroxycarboxylic acid), see for example Advanced Organic Chemistry, by Jerry March, 4th Edition, Wiley Interscience, pages. More particularly, N-oxides can be made by the procedure of L. W. Deady (Syn. Comm. 1977, 7, 509-514) in which the amine compound is reacted with metachloroperoxybenzoic acid (mCPBA), for example, in an inert solvent such as dichloromethane.MGL-06625

[0153] The compounds disclosed herein may be administered in the form of a prodrug which is broken down in the human or animal body to release a compound of the disclosure. A prodrug may be used to alter the physical properties and / or the pharmacokinetic properties of a compound of the disclosure. A prodrug can be formed when the compound of the disclosure contains a suitable group or substituent to which a property-modifying group can be attached. Examples of prodrugs include derivatives containing in vivo cleavable alkyl or acyl substituents at the ester or amide group in any one of the compounds disclosed herein.

[0154] Accordingly, the present disclosure includes those compounds disclosed herein as defined hereinbefore when made available by organic synthesis and when made available within the human or animal body by way of cleavage of a prodrug thereof. Accordingly, the present disclosure includes those compounds disclosed herein that are produced by organic synthetic means and also such compounds that are produced in the human or animal body by way of metabolism of a precursor compound, that is a compound disclosed herein may be a synthetically-produced compound or a metabolically -produced compound.

[0155] A suitable pharmaceutically acceptable prodrug of a compound disclosed herein is one that is based on reasonable medical judgment as being suitable for administration to the human or animal body without undesirable pharmacological activities and without undue toxicity. Various forms of prodrug have been described, for example in the following documents: a) Methods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder, et al. (Academic Press, 1985); b) Design of Pro-drugs, edited by H. Bundgaard, (Elsevier, 1985); c) A Textbook of Drug Design and Development, edited by Krogsgaard-Larsen and H. Bundgaard, Chapter 5 “Design and Application of Pro-drugs”, by H. Bundgaard p. 113-191 (1991 ); d) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992); e) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77, 285 (1988); f) N. Kakeya, et al., Chem. Pharm. Bulk, 32, 692 (1984); g) T. Higuchi and V. Stella, “Pro-Drugs as Novel Delivery Systems”, A.C.S. Symposium Series, Volume 14; and h) E. Roche (editor), “Bioreversible Carriers in Drug Design”, Pergamon Press, 1987.

[0156] A suitable pharmaceutically acceptable prodrug of a compound disclosed herein that possesses a hydroxy group is, for example, an in vivo cleavable ester or ether thereof. An in vivo cleavable ester or ether of a compound disclosed herein containing a hydroxy group is, for example, a pharmaceutically acceptable ester or ether which is cleaved in the human or animal body to produce the parent hydroxy compound. Suitable pharmaceutically acceptable esterMGL-06625 forming groups for a hydroxy group include inorganic esters such as phosphate esters (including phosphoramidic cyclic esters). Further suitable pharmaceutically acceptable ester forming groups for a hydroxy group include C1-C10 alkanoyl groups such as acetyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl groups, C1-C10 alkoxycarbonyl groups such as ethoxycarbonyl, N,N-(Ci-Ce alkyl)2carbamoyl, 2-dialkylaminoacetyl and 2-carboxyacetyl groups. Examples of ring substituents on the phenylacetyl and benzoyl groups include aminomethyl, N- alkylaminomethyl, N,N-dialkylaminomethyl, morpholinomethyl, piperazin- 1-ylmethyl and 4-(Ci- C4 alkyl)piperazin- 1 -ylmethyl. Suitable pharmaceutically acceptable ether forming groups for a hydroxy group include oc-acyloxyalkyl groups such as acetoxymethyl and pivaloyloxymethyl groups.

[0157] A suitable pharmaceutically acceptable prodrug of a compound disclosed herein that possesses a carboxy group is, for example, an in vivo cleavable amide thereof, for example an amide formed with an amine such as ammonia, a Ci-4alkylamine such as methylamine, a (C1-C4 alkyl)2amine such as dimethylamine, N-ethyl-N-methylamine or diethylamine, a C1-C4 alkoxy-C2- C4 alkylamine such as 2-methoxy ethylamine, a phenyl-Ci-C4 alkylamine such as benzylamine and amino acids such as glycine or an ester thereof.

[0158] A suitable pharmaceutically acceptable prodrug of a compound disclosed herein that possesses an amino group is, for example, an in vivo cleavable amide derivative thereof. Suitable pharmaceutically acceptable amides from an amino group include, for example an amide formed with C1-C10 alkanoyl groups such as an acetyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl groups. Examples of ring substituents on the phenylacetyl and benzoyl groups include aminomethyl, N-alkylaminomethyl, N,N-dialkylaminomethyl, morpholinomethyl, piperazin- 1 -ylmethyl and 4-(CI-C4 alkyl)piperazin- 1-ylmethyl.

[0159] The in vivo effects of a compound disclosed herein may be exerted in part by one or more metabolites that are formed within the human or animal body after administration of a compound disclosed herein. As stated hereinbefore, the in vivo effects of a compound disclosed herein may also be exerted by way of metabolism of a precursor compound (a prodrug).

[0160] Suitably, the present disclosure excludes any individual compounds not possessing the biological activity defined herein.Biological AssaysMGL-06625

[0161] Compounds designed, selected and / or optimized by methods described above, once produced, can be characterized using a variety of assays known to those skilled in the art to determine whether the compounds have biological activity. For example, the molecules can be characterized by conventional assays, including but not limited to those assays described below, to determine whether they have a predicted activity, binding activity and / or binding specificity.

[0162] Furthermore, high-throughput screening can be used to accelerate analysis using such assays. As a result, it can be possible to rapidly screen the molecules described herein for activity, using techniques known in the art. General methodologies for performing high-throughput screening are described, for example, in Devlin (1998) High Throughput Screening, Marcel Dekker; and U.S. Patent No. 5,763,263. High-throughput assays can use one or more different assay techniques including, but not limited to, those described below.

[0163] Various in vitro or in vivo biological assays are may be suitable for detecting the effect of the compounds of the present disclosure. These in vitro or in vivo biological assays can include, but are not limited to, enzymatic activity assays, electrophoretic mobility shift assays, reporter gene assays, in vitro cell viability assays, and the assays described herein.

[0164] In some embodiments, the biological assay is described in the Examples herein.

[0165] In some embodiments, the biological assay is an in vitro SCC cell confluency assay.

[0166] In some embodiments, the SCC cell confluency assays are conducted on skin SCC cell lines. In some embodiments, SCC cell confluency assays are conducted on oral SCC cell lines.

[0167] In some embodiments, SCC cell confluency assays are conducted on SCC 13 cell lines. In some embodiments, the SCC cell confluency assays are conducted on Cal27 cell lines. In some embodiments, SCC cell confluency assays are conducted on SCCO28 cell lines.

[0168] In some embodiments, IC50 is determined from the assay conducted on the SCC cell lines.

[0169] In some embodiments, the SCC cell confluency assays are conducted with SCC cell lines comprising silenced ULK3.

[0170] In some embodiments, the SCC cell confluency assays are conducted with a SCC13 cell line comprising silenced ULK3.

[0171] In some embodiments, the biological assay measures the gene expression of cancer associated fibroblasts.

[0172] In some embodiments, the biological assay monitors ULK3 inhibition.MGL-06625

[0173] In some embodiments, the biological assays described herein are conducted with a compound selected from Table 1.Pharmaceutical Compositions

[0174] In some aspects, the present disclosure provides a pharmaceutical composition comprising a compound of the present disclosure as an active ingredient. In some embodiments, the present disclosure provides a pharmaceutical composition comprising at least one compound of each of the compounds described herein, or a pharmaceutically acceptable salt or solvate thereof, and one or more pharmaceutically acceptable carrier, diluent, adjuvant, excipient, or a combination thereof. In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound described in Table 1. In some embodiments, the present disclosure provides a pharmaceutical composition comprising at least one compound selected from Table 1.

[0175] As used herein, the term “composition” is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts.

[0176] The compounds of present disclosure can be formulated for oral administration in forms such as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powders, granules, elixirs, tinctures, suspensions, syrups and emulsions. The compounds of present disclosure on can also be formulated for intravenous (bolus or in- fusion), intraperitoneal, topical, subcutaneous, intramuscular or transdermal (e.g., patch) administration, all using forms well known to those of ordinary skill in the pharmaceutical arts.

[0177] The formulation of the present disclosure may be in the form of an aqueous solution comprising an aqueous vehicle. The aqueous vehicle component may comprise water and at least one pharmaceutically acceptable excipient. Suitable acceptable excipients include those selected from the group consisting of a solubility enhancing agent, chelating agent, preservative, tonicity agent, viscosity / suspending agent, buffer, and pH modifying agent, and a mixture thereof.

[0178] Any suitable solubility enhancing agent can be used. Examples of a solubility enhancing agent include cyclodextrin, such as those selected from the group consisting of hydroxypropyl-P- cyclodextrin, methyl-P-cyclodextrin, randomly methylated- P-cyclodextrin, ethylated-P- cyclodextrin, triacetyl-P-cyclodextrin, peracetylated-P-cyclodextrin, carboxymethyl-P- cyclodextrin, hydroxyethyl- P-cyclodextrin, 2-hydroxy-3-(trimethylammonio)propyl-P-MGL-06625 cyclodextrin, glucosyl-P-cyclodextrin, sulfated P-cyclodextrin (S-P-CD), maltosyl-P-cyclodextrin, P-cyclodextrin sulfobutyl ether, branched-P-cyclodextrin, hydroxypropyl-y-cyclodextrin, randomly methylated-y-cyclodextrin, and trimethyl-y-cyclodextrin, and mixtures thereof.

[0179] Any suitable chelating agent can be used. Examples of a suitable chelating agent include those selected from the group consisting of ethylenediaminetetraacetic acid and metal salts thereof, disodium edetate, trisodium edetate, and tetrasodium edetate, and mixtures thereof.

[0180] Any suitable preservative can be used. Examples of a preservative include those selected from the group consisting of quaternary ammonium salts such as benzalkonium halides (preferably benzalkonium chloride), chlorhexidine gluconate, benzethonium chloride, cetyl pyridinium chloride, benzyl bromide, phenylmercury nitrate, phenylmercury acetate, phenylmercury neodecanoate, merthiolate, methylparaben, propylparaben, sorbic acid, potassium sorbate, sodium benzoate, sodium propionate, ethyl p-hydroxybenzoate, propylaminopropyl biguanide, and butyl- p-hydroxybenzoate, and sorbic acid, and mixtures thereof.

[0181] The aqueous vehicle may also include a tonicity agent to adjust the tonicity (osmotic pressure). The tonicity agent can be selected from the group consisting of a glycol (such as propylene glycol, diethylene glycol, triethylene glycol), glycerol, dextrose, glycerin, mannitol, potassium chloride, and sodium chloride, and a mixture thereof.

[0182] The aqueous vehicle may also contain a viscosity / suspending agent. Suitable viscosity / suspending agents include those selected from the group consisting of cellulose derivatives, such as methyl cellulose, ethyl cellulose, hydroxyethylcellulose, polyethylene glycols (such as polyethylene glycol 300, polyethylene glycol 400), carboxymethyl cellulose, hydroxypropylmethyl cellulose, and cross-linked acrylic acid polymers (carbomers), such as polymers of acrylic acid cross-linked with polyalkenyl ethers or divinyl glycol (Carbopols - such as Carbopol 934, Carbopol 934P, Carbopol 971, Carbopol 974 and Carbopol 974P), and a mixture thereof.

[0183] In order to adjust the formulation to an acceptable pH (typically a pH range of about 5.0 to about 9.0, more preferably about 5.5 to about 8.5, particularly about 6.0 to about 8.5, about 7.0 to about 8.5, about 7.2 to about 7.7, about 7.1 to about 7.9, or about 7.5 to about 8.0), the formulation may contain a pH modifying agent. The pH modifying agent is typically a mineral acid or metal hydroxide base, selected from the group of potassium hydroxide, sodium hydroxide, and hydrochloric acid, and mixtures thereof, and preferably sodium hydroxide and / or hydrochloricMGL-06625 acid. These acidic and / or basic pH modifying agents are added to adjust the formulation to the target acceptable pH range. Hence it may not be necessary to use both acid and base - depending on the formulation, the addition of one of the acid or base may be sufficient to bring the mixture to the desired pH range.

[0184] The aqueous vehicle may also contain a buffering agent to stabilize the pH. When used, the buffer is selected from the group consisting of a phosphate buffer (such as sodium dihydrogen phosphate and disodium hydrogen phosphate), a borate buffer (such as boric acid, or salts thereof including disodium tetraborate), a citrate buffer (such as citric acid, or salts thereof including sodium citrate), and s-aminocaproic acid, and mixtures thereof.

[0185] The formulation may further comprise a wetting agent. Suitable classes of wetting agents include those selected from the group consisting of polyoxypropylene-polyoxyethylene block copolymers (poloxamers), polyethoxylated ethers of castor oils, polyoxyethylenated sorbitan esters (polysorbates), polymers of oxyethylated octyl phenol (Tyloxapol), polyoxyl 40 stearate, fatty acid glycol esters, fatty acid glyceryl esters, sucrose fatty esters, and polyoxyethylene fatty esters, and mixtures thereof.

[0186] Oral compositions generally include an inert diluent or an edible pharmaceutically acceptable carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed. Pharmaceutically compatible binding agents, and / or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, orange flavoring.

[0187] According to a further aspect of the disclosure there is provided a pharmaceutical composition which comprises a compound of the disclosure as defined hereinbefore, or aMGL-06625 pharmaceutically acceptable salt, hydrate or solvate thereof, in association with a pharmaceutically acceptable diluent or carrier.

[0188] The compositions of the disclosure may be in a form suitable for oral use (for example as tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), for topical use (for example as creams, ointments, gels, or aqueous or oily solutions or suspensions), for administration by inhalation (for example as a finely divided powder or a liquid aerosol), for administration by insufflation (for example as a finely divided powder) or for parenteral administration (for example as a sterile aqueous or oily solution for intravenous, subcutaneous, intramuscular, intraperitoneal or intramuscular dosing or as a suppository for rectal dosing).

[0189] The compositions of the disclosure may be obtained by conventional procedures using conventional pharmaceutical excipients, well known in the art. Thus, compositions intended for oral use may contain, for example, one or more coloring, sweetening, flavoring and / or preservative agents.

[0190] An effective amount of a compound of the present disclosure for use in therapy is an amount sufficient to treat or prevent a ULK3 related condition referred to herein, slow its progression and / or reduce the symptoms associated with the condition.

[0191] An effective amount of a compound of the present disclosure for use in therapy is an amount sufficient to treat a ULK3 related condition referred to herein, slow its progression and / or reduce the symptoms associated with the condition.

[0192] The size of the dose for therapeutic or prophylactic purposes of a compound described in Table 1 will naturally vary according to the nature and severity of the conditions, the age and sex of the animal or patient and the route of administration, according to well-known principles of medicine.Methods of Use

[0193] In some embodiments, the present disclosure relates to a method of treating a disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of a compound selected from the group consisting of:MGL-06625p ,p , or a pharmaceutically acceptable salt thereof; and wherein the disease or disorder is a cancer, a pre-cancerous condition, skin fibrosis, or skin aging.MGL-06625In some embodiments, the compound is(Compound F). In, ,p

[0194] In some embodiments, the disease or disorder is cancer.

[0195] In some embodiments, the cancer is metastatic cancer.MGL-06625

[0196] In some embodiments, the method further comprises administering to the subject chemotherapy, immunotherapy, radiation therapy, biological therapy, surgical intervention, or any combination thereof.

[0197] In some embodiments, the subject has been previously treated with chemotherapy, immunotherapy, radiation therapy, biological therapy, surgical intervention, or any combination thereof.

[0198] In some embodiments, the disease or disorder is a squamous cell carcinoma, a skin melanoma, or a pre-cancerous condition.

[0199] In some embodiments, the disease or disorder is a squamous cell carcinoma.

[0200] In some embodiments, the squamous cell carcinoma is a skin cancer, a lung cancer, a head and neck cancer, a lung cancer, an esophageal cancer, or a cervical cancer.

[0201] In some embodiments, the squamous cell carcinoma is adenoid / pseudoglandular squamous cell carcinoma, intraepidermal squamous cell carcinoma, large cell keratinizing squamous cell carcinoma, large cell non-keratinizing squamous cell carcinoma, lymphoepithelial carcinoma, papillary squamous cell carcinoma, papillary thyroid carcinoma, small cell keratinizing squamous cell carcinoma, spindle cell squamous cell carcinoma, or verrucous squamous-cell carcinoma.

[0202] In some embodiments, the disease or disorder is a skin melanoma.

[0203] In some embodiments, the skin melanoma is superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma, amelanotic melanoma, acral lentiginous melanoma, mucosal melanoma, melanoma of the eye, or desmoplastic melanoma.

[0204] In some embodiments, the disease or disorder is a pre-cancerous condition.

[0205] In some embodiments, the pre-cancerous condition is actinic keratosis, squamous cell carcinoma in situ, or keratoacanthoma.

[0206] In some embodiments, the disease or disorder is skin fibrosis.

[0207] In some embodiments, the compound modulates ULK3; optionally, wherein modulation is inhibition.

[0208] In some embodiments, the compound inhibits ULK3.

[0209] In some embodiments, the compound is administered topically.

[0210] In some embodiments, the compound is administered orally.

[0211] In some embodiments, the compound is administered parenterally.MGL-06625

[0212] In some aspects, the present disclosure provides a method of modulating ULK3 expression with a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0213] In some aspects, the present disclosure provides a method of modulating ULK3 expression (e.g., in vitro or in vivo) with an effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0214] In some aspects, the present disclosure provides a method of modulating ULK3 expression (e.g., in vitro or in vivo) with a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0215] In some aspects, the present disclosure provides a method of modulating ULK3 expression (e.g., in vitro or in vivo) with an effective amount of a pharmaceutical composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0216] In some aspects, the present disclosure provides a method of modulating ULK3 expression (e.g., in vitro or in vivo) with a pharmaceutical composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0217] In some aspects, the present disclosure provides a method of inhibiting ULK3 expression with a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0218] In some aspects, the present disclosure provides a method of inhibiting ULK3 expression (e.g., in vitro or in vivo) with an effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0219] In some aspects, the present disclosure provides a method of inhibiting ULK3 expression (e.g., in vitro or in vivo) with a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0220] In some aspects, the present disclosure provides a method of inhibiting ULK3 expression (e.g., in vitro or in vivo) with an effective amount of a pharmaceutical composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0221] In some aspects, the present disclosure provides a method of inhibiting ULK3 expression (e.g., in vitro or in vivo) with a pharmaceutical composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0222] In some aspects, the present disclosure provides a method of treating or preventing a disease or disorder disclosed herein in a subject in need thereof, comprising administering to theMGL-06625 subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0223] In some aspects, the present disclosure provides a method of treating a disease or disorder disclosed herein in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0224] In some aspects, the present disclosure provides a method of treating or preventing a disease or disorder disclosed herein in a subject in need thereof, comprising administering to the subject a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0225] In some aspects, the present disclosure provides a method of treating a disease or disorder disclosed herein in a subject in need thereof, comprising administering to the subject a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0226] In some embodiments, the disease or disorder is associated with an implicated ULK3 expression. In some embodiments, the disease or disorder is a disease or disorder in which ULK3 expression is implicated.

[0227] In some embodiments, a modulation in expression is a modulation in activity.

[0228] In some embodiments, the disease or disorder is a squamous cell carcinoma, a skin melanoma, or a pre-cancerous condition.

[0229] In some embodiments, the disease or disorder is a squamous cell carcinoma.

[0230] In some embodiments, the squamous cell carcinoma is a skin cancer, a lung cancer, a head and neck cancer, a lung cancer, an esophageal cancer, or a cervical cancer.

[0231] In some embodiments, the squamous cell carcinoma is a skin cancer.

[0232] In some embodiments, the squamous cell carcinoma is adenoid / pseudoglandular squamous cell carcinoma, intraepidermal squamous cell carcinoma, large cell keratinizing squamous cell carcinoma, large cell non-keratinizing squamous cell carcinoma, lymphoepithelial carcinoma, papillary squamous cell carcinoma, papillary thyroid carcinoma, small cell keratinizing squamous cell carcinoma, spindle cell squamous cell carcinoma, or verrucous squamous-cell carcinoma.

[0233] In some embodiments, the disease or disorder is a skin melanoma.MGL-06625

[0234] In some embodiments, the skin melanoma is superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma, amelanotic melanoma, acral lentiginous melanoma, mucosal melanoma, melanoma of the eye, or desmoplastic melanoma.

[0235] In some embodiments, the disease or disorder is a pre-cancerous condition.

[0236] In some embodiments, the pre-cancerous condition is actinic keratosis, squamous cell carcinoma in situ, or keratoacanthoma.

[0237] In some embodiments, the disease or disorder is skin fibrosis.

[0238] In some aspects, the present disclosure provides a method of treating a squamous cell carcinoma, a skin melanoma, or a pre-cancerous condition.

[0239] In some embodiments, the squamous cell carcinoma, skin melanoma, or pre-cancerous condition is from an environmental effect.

[0240] In some embodiments, the environmental effect is UV exposure.

[0241] In some embodiments, the environmental effect is an immunosuppressive therapy.

[0242] In some embodiments, the immunosuppressive therapy is a calcineurin inhibitor.

[0243] In some embodiments, the environmental effect is an infection.

[0244] In some embodiments, the infection is human papillomavirus. In some embodiments, the infection is Mycobacterium tuberculosis. In some embodiments, the infection is Epstein-Barr virus.

[0245] In some embodiments, the environmental effect is chemical exposure.

[0246] In some embodiments, the chemical is cigarette smoke. In some embodiments, the chemical is radon gas. In some embodiments, the chemical is asbestos. In some embodiments, the chemical is air pollution. In some embodiments, the chemical is an organic solvent. In some embodiments, the chemical is alcohol. In some embodiments, the chemical is a metal.

[0247] In some embodiments, the metal is arsenic. In some embodiments, the metal is cadmium. In some embodiments, the metal is chromium.

[0248] In some embodiments, the environmental effect is a nutritional deficiency.

[0249] In some embodiments, the nutritional deficiency is from a deficiency in vitamins or minerals.

[0250] In some embodiments, the environmental effect is contraceptive use.

[0251] In some embodiments, the contraceptive use is long-term oral contraceptive use.

[0252] In some embodiments, the environmental effect is childbirth.MGL-06625

[0253] In some embodiments, the environmental effect is a high number of childbirths.

[0254] In some embodiments, the disease or disorder is aging.

[0255] In some embodiments, the disease or disorder is natural aging.

[0256] In some embodiments, natural aging is associated with a decrease in collagen abundance and increase in fibril fragmentation.

[0257] In some embodiments, natural aging is associated with a decrease in collagen abundance.

[0258] In some embodiments, natural aging is associated with an increase in fibril fragmentation.

[0259] In some embodiments, increased levels of metalloproteases (MMPs), and lower production of ECM, contribute to overall ECM decline.

[0260] In some embodiments, increased levels of MMPs degrade extracellular matrix (ECM) components.

[0261] In some embodiments, ECM modulation contributes to aging. In some embodiments, modulation is a decrease in production.

[0262] In some embodiments, modulating transforming growth factor (TGF-P) modulates ECM production and breakdown by MMPs. In some embodiments, modulating ULK3 modulates ECM production and breakdown by MMPs.

[0263] In some embodiments, modulating TGF-P modulates ECM production and breakdown by MMPs.

[0264] In some embodiments, the disease or disorder is extrinsic aging.

[0265] In some embodiments, extrinsic aging is photoaging.

[0266] In some embodiments, photoaging is a skin process accelerated by exposure to ultraviolet (UVA) radiation from sunlight.

[0267] In some embodiments, the disease or disorder is a symptom of photoaging.

[0268] In some embodiments, a symptom of photoaging is increased wrinkles, dryness, pigmentation irregularities, and / or a reduction in skin elasticity.

[0269] In some embodiments, ULK3 is up-regulated in the human dermal fibroblasts by UVA irradiation.

[0270] In some embodiments, the compounds of the present disclosure decrease DNA damage associated with genomic instability in activated fibroblasts.

[0271] In some embodiments, aging is attributable to sun exposure.MGL-06625

[0272] In some embodiments, aging is attributable to environmental factors (e.g., pollution and lifestyles).

[0273] In some embodiments, an environmental factor is smoking or exposure to smoking.

[0274] In some embodiments, aging is attributable to genetic and / or epigenetic mechanisms.

[0275] In some embodiments, aging is skin aging.

[0276] In some embodiments, accumulation of genetic damage over time, the diminished efficiency of DNA repair mechanisms, and the loss of telomere integrity all lead to cellular senescence and contribute to aging.

[0277] In some embodiments, the compounds of the present disclosure inhibit DNA damage.

[0278] In some embodiments, the compounds of the present disclosure inhibit genomic instability.

[0279] In some embodiments, the DNA damage is caused by skin exposure to UV, sunlight, smoking, or an external genotoxic agent.

[0280] In some embodiments, the genomic instability is caused by skin exposure to UV, sunlight, smoking, or an external genotoxic agent.

[0281] In some embodiments, the disease or disorder is multiple myeloma.

[0282] In some aspects, the present disclosure provides a method of treating or preventing a squamous cell carcinoma in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0283] In some aspects, the present disclosure provides a method of treating a squamous cell carcinoma in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0284] In some aspects, the present disclosure provides a method of treating or preventing a squamous cell carcinoma in a subject in need thereof, comprising administering to the subject a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0285] In some aspects, the present disclosure provides a method of treating a squamous cell carcinoma in a subject in need thereof, comprising administering to the subject a compound of theMGL-06625 present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0286] In some aspects, the present disclosure provides a method of treating or preventing a skin melanoma in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0287] In some aspects, the present disclosure provides a method of treating a skin melanoma in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0288] In some aspects, the present disclosure provides a method of treating or preventing a skin melanoma in a subject in need thereof, comprising administering to the subject a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0289] In some aspects, the present disclosure provides a method of treating a skin melanoma in a subject in need thereof, comprising administering to the subject a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0290] In some aspects, the present disclosure provides a method of treating or preventing a pre- cancerous condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0291] In some aspects, the present disclosure provides a method of treating a pre-cancerous condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0292] In some aspects, the present disclosure provides a method of treating or preventing a pre- cancerous condition in a subject in need thereof, comprising administering to the subject a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.MGL-06625

[0293] In some aspects, the present disclosure provides a method of treating a pre-cancerous condition in a subject in need thereof, comprising administering to the subject a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0294] In some aspects, the present disclosure provides a method of treating or preventing skin fibrosis condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0295] In some aspects, the present disclosure provides a method of treating skin fibrosis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0296] In some aspects, the present disclosure provides a method of treating or preventing skin fibrosis in a subject in need thereof, comprising administering to the subject a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0297] In some aspects, the present disclosure provides a method of treating skin fibrosis in a subject in need thereof, comprising administering to the subject a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

[0298] In some aspects, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure for use in modulating ULK3 expression.

[0299] In some aspects, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof for use in modulating ULK3 expression (e.g., in vitro or in vivo).

[0300] In some aspects, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof for use in modulating ULK3 expression (e.g., in vitro or in vivo).MGL-06625

[0301] In some aspects, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of a disease in a patient, wherein the disease is characterized by an increase in ULK3 expression.

[0302] In some aspects, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure for use in treating or preventing a disease or disorder.

[0303] In some aspects, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof for use in treating or preventing a disease or disorder disclosed herein.

[0304] In some aspects, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof for use in treating a disease or disorder disclosed herein.

[0305] In some aspects, the present disclosure provides use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for modulating ULK3 expression (e.g., in vitro or in vivo).

[0306] In some aspects, the present disclosure provides use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating or preventing a disease or disorder disclosed herein.

[0307] In some aspects, the present disclosure provides use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a disease or disorder disclosed herein.

[0308] In some aspects, the present disclosure provides use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating or preventing a squamous cell carcinoma in a subject in need thereof.

[0309] In some aspects, the present disclosure provides use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a squamous cell carcinoma in a subject in need thereof.

[0310] In some aspects, the present disclosure provides use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating or preventing a skin melanoma in a subject in need thereof.MGL-06625

[0311] In some aspects, the present disclosure provides use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a skin melanoma in a subject in need thereof.

[0312] In some aspects, the present disclosure provides use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating or preventing a pre-cancerous condition in a subject in need thereof.

[0313] In some aspects, the present disclosure provides use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a pre-cancerous condition in a subject in need thereof.

[0314] In some aspects, the present disclosure provides use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating or preventing skin fibrosis in a subject in need thereof.

[0315] In some aspects, the present disclosure provides use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating skin fibrosis in a subject in need thereof.

[0316] In some embodiments, the present disclosure provides compounds that function as modulators of ULK3 activity.

[0317] In some embodiments, modulation results in an inhibition of ULK3 expression.

[0318] Effectiveness of compounds of the disclosure can be determined by industry-accepted assays / disease models according to standard practices of elucidating the same as described in the art and are found in the current general knowledge.

[0319] The present disclosure also provides a method of treating a disease or disorder in which ULK3 activity is implicated in a subject in need of such treatment, said method comprising administering to said subject a compound, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as defined herein.Routes of Administration

[0320] The compounds of the disclosure or pharmaceutical compositions comprising these compounds may be administered to a subject by any convenient route of administration, whether systemically / peripherally or topically (i.e., at the site of desired action).MGL-06625

[0321] Routes of administration include, but are not limited to, oral (e.g. by ingestion); buccal; sublingual; transdermal (including, e.g., by a patch, plaster, etc.); transmucosal (including, e.g., by a patch, plaster, etc.); intranasal (e.g., by nasal spray or powder); ocular (e.g., by eye drops); pulmonary (e.g., by inhalation or insufflation therapy using, e.g., via an aerosol, e.g., through the mouth or nose); rectal (e.g., by suppository or enema); vaginal (e.g., by pessary); parenteral, for example, by injection, including subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal, subcuticular, intraarticular, subarachnoid, and intrasternal; by implant of a depot or reservoir, for example, subcutaneously or intramuscularly.EXAMPLESExample 1. Compounds in the Present Disclosure.

[0322] Compounds of the disclosure can be purchased from a vendor (e.g., Hit21ead.com) and are as shown in Table A.Table A.MGL-06625Example 2. Treatment of SCC Cells in Vitro with Compounds A, B, C, D, F, and G.SCC Cell Confluency

[0323] Cells were seeded in a 96-well microtiter plate and the next day treated with 1-10 pM of compound, DMSO as negative control, or a reported ULK3 inhibitor SU6668 as positive control.

[0324] The microtiter was placed in an incubator with an Incucyte time-lapse microscope, and for each well, five images were taken every four hours for four days. The confluency over time was evaluated by using the Al Confluence algorithm included in the Incucyte. Growth curves of the indicated cell lines treated with DMSO (vehicle) or the indicated concentrations of the six ULK3 inhibitory compounds were generated using an Incucyte (Sartorius). Confluency was normalized to Day 0. SCC 13 corresponds to skin SCC; Cal27 and SCCO28 correspond to oral SCC. Cells were seeded at 500 cells / well in triplicate wells. 5 images / well were acquired every 4 h over 4 days.MGL-06625

[0325] The normalized SCC cell confluency growth curves for the SCC13, Cal27, and SCCO28 SCC cell lines attenuated upon treatment with a compound of the present disclosure (i.e. Compound A, B, C, D, F, and G). For the SCC13 and Cal27 cell lines, the cell confluency decreased with increasing compound concentration. The normalized SCCO28 cell confluency showed variability as a function of compound concentration, the total cell confluency decreased from the DMSO negative control for Compounds A, B, C, D, F, and G. FIGs. 1A-1C shows the growth curves for the SCC13 (FIG. 1A), Cal27 (FIG. IB), and SCCO28 (FIG. 1C) cell lines for Compounds A, B, C, D, F, and G and the SU6668 positive control. SCC13 and Cal27 cell confluency experiments measured at 3 days with a static concentration of 10 pM of compound showed a maximum confluency of 60% the confluency of the DMSO negative control, demonstrating the effect of the compounds to attenuate cell confluency. FIG. 2 shows the Incucyte time-lapse cell proliferation analysis of skin (SCC13) and oral (Cal27) SCC cell lines treated with 10 pM of Compounds A, B, C, D, F, and G (Mean+SE, n(experiments)=3).

[0326] Table B shows the IC50 (pM) of the six ULK3 inhibitory compounds and SU6668 for the SCC13, Cal27, and SCCO28 cell lines. IC50 values of: >1 pM and <3 pM are labelled as “+++++”, >3 pM and <5 pM are labelled as “++++”, >5 pM and <7 pM are labelled as “+++”, >7 and <9 are labelled as “++”, and >10 pM are labelled as “+”.Table B.SCC 13 with Silenced ULK3

[0327] Cells were treated with DMSO or lOpM of Compounds A, B, C, D, F, and G or SU6668 at 500 cells / well in triplicate wells. Five images / well were acquired every four hours over four days.

[0328] SCC cell confluency assays were performed on ULK3-silenced SCC13 cell lines. Two ULK3-silenced SCC13 cell lines (shULK3 #1 and shULK3 #2) showed a normalized cellMGL-06625 confluency of 75% and 50%, respectively, compared to a shCT control, demonstrating the influence of ULK3 on SCC cells. FIG. 3A shows cell growth of the control (shCT) and ULK3- silenced SCC 13 cells and FIG. 3B shows WB on the same cells confirming the loss of epigenetic marker H4R3. Treatment of the ULK3-silenced cell lines with Compounds A, B, C, D, F, and G resulted in a negligible change in the cell confluency, suggesting that the compounds attenuated SCC 13 cell confluency through ULK3 inhibition. FIG. 3C and FIG. 3D show the corresponding cell growth profiles of shULK3 SCC13 cells plus Compounds A, B, C, D, F, and G (blue) compared to control-silenced (black) SCC 13 cells, DMSO-treated SCC 13 shULK3 (red), and SCC 13 shULK3 cells treated with SU6668 (purple).Example 3. Compounds A, B, C, D, F, and G Activity.SCC 13 Organoids Growth

[0329] SCC13 cells were treated with two concentrations of Compounds A, B, D, F, and G (10 and 20 pM), or DMSO (vehicle), and monitored after 24 h to determine organoid formation in 3% Matrigel. (Mean+ / -SEM, One way ANOVA, n(organoids / compound dilution)=3) *P<0.01. FIG. 4A shows (a) quantification of organoids formed by 6.000 SCC 13 cells grown in suspension for two weeks and FIG. 4B shows representative images of the cells.Cal27 Spheroid Formation

[0330] Cal27 SCC cells were treated with three concentrations of Compounds A, B, C, D, F, and G (5, 10 and 20 pM), or DMSO (vehicle), once per week. (Mean+ / -SEM, One way ANOVA, n(well / compound dilution)=3), *P<0.01. FIG. 5 shows the quantification of the sphere number (%) formed by Cal27 SCC cells grown in Matrigel for two weeks after treatment with Compounds A, B, C, D, F, and G.A375 Growth

[0331] A375 melanoma cell lines were seeded at 500 cells / well in triplicate wells and treated with Compounds A, B, C, D, F, and G or DMSO (vehicle). 5 images / well were acquired every four hours over four days. Confluency was normalized to Day 0 (SU (SU6668): positive control). FIG. 6 shows growth curves using an Incucyte of a A375 melanoma cell line treated with DMSO (vehicle) or Compound A (5, 10, 20, or 40 pM), B (3.8, 7.5, 15, or 30 pM), C (5, 10, 20, or 40 pM), D (3.8, 7.5, 15, or 30 pM), F (5, 10, 20, or 40 pM), and G (5, 10, 20, or 40 pM).Cancer Associated Fibroblasts Effector Genes ExpressionMGL-06625

[0332] Fibroblast strains were treated with Compound A, B, C, D, F, and G or DMSO (vehicle) and monitored. FIG. 7 shows the RT-qPCR analysis of CAF1 and CAF21 genes in two cancer- associated fibroblast strains treated with DMSO (vehicle) or Compounds A, B, C, D, F, and G (lOpM). Gene expression was normalized to the 36B4 gene, and the fold induction to DMSO (heatmap: white no change value=l, red valued, blue: value>l; SU (SU6668): positive control (10pM)).Example 4. ULK3 kinase inhibitors: kinase activity assays.Compounds A, B, C, D, F, and G were studied in dose-dependent in vitro ULK3 autophosphorylation kinase assays with decreasing concentrations of compounds (120-0.01 pM). The assays identified in particular Compounds B and F as having the lowest IC50 values (0.8 and 8 pM. respectively (FIG. 8).For the dose-dependent inhibition of ULK3 autophosphorylation by each compound, 8 dilutions (0.01-120pM range) or DMSO were incubated in kinase buffer (Cell Signaling) with 150 ng of recombinant ULK3 (Abeam) and 100pm ATP for Ih at 30 °C. ULK3 activity (%) was measured with the ADP-GLO system (Promega) as indicated by the provider. Mean, SD, nonlinear regression (curve fit) and IC50S were calculated using Prism 10.FIG. 8 shows ULK3 autophosphorylation assays with the Compound B and Compound F (8 dilutions, 0.01-120pM range) versus DMSO. ULK3 activity (%) measured with ADP-GLO system (Promega), Mean+SD, n(assays)=3. Non-linear regression (curve fit) was calculated using PrismlO. Table 2 shows the IC50 (pM) for all similarly tested ULK3 inhibitors.Table 2.IC50 values of: <3 pM are labelled as “+++++”, >3 pM and <5 pM are labelled as “++++”, >5 pM and <7 pM are labelled as “+++”, >7 and < 10 are labelled as “++”, and >10 pM are labelled as “+”.Example 5. ULK3 kinase inhibitors: kinase specificity assays.MGL-06625The specificity of Compounds B and F was further assessed by in vitro kinase assays with purified ULK1 kinase. Neither compound inhibited ULK1 autophosphorylation at concentrations (1-10 pM) that suppressed ULK3 activity by more than 75% (FIG. 9A) and only slightly reduced ULKl-mediated phosphorylation of the MBP substrate (FIG. 9B).To compare the effects of Compounds B and F on ULK3 vs ULK1 activity, Compounds B and F (0.1-200pM range), or DMSO, were incubated in kinase buffer (Cell Signaling) with 150 ng of recombinant ULK3 (Abeam) or 100 ng of ULK1 (Abeam) and 100pm ATP for Ih at 30 °C. ULK3 and ULK1 autophosphorylation activity (%) was measured with the ADP-GLO system (Promega) as indicated by the provider. For the effects on the kinase activity towards an exogenous substrate, recombinant ULK3 (150ng) (Abeam) and ULK1 (lOOng) (Promega) were incubated with Compounds B and F (0-60pM range), or DMSO, in the presence of Ipg myelin basic protein (MBP, Promega), and 100 pM ATP. Phosphorylation activity (%) was measured with the ADP- GLO system (Promega), as indicated by the provider.FIGs. 9A-9B show in vitro specificity assays of the ULK3 inhibitors Compounds B and F vs ULK1 kinase. FIG. 9 A shows Dose-dependent ULK1 and ULK3 autophosphorylation assays with Compounds B and F (0.1-200pM range), or DMSO. Activity measured with ADP-GLO system (Promega). FIG. 9B shows dose-dependent ULK1 kinase assays in the presence of MBP (Ipg) with Compounds B and F (0-60pM range), or DMSO. Mean n(assays)=2. Activity measured with ADP-GLO system (Promega).Additional in vivo specificity assays were based on treatment of cultured SCC13 cells with Compounds B, F or the ULKl / 2-specific inhibitor SBL0206965 (SBI).SBI-0206965Immunoblot analysis showed that, unlike SBI, neither Compound B nor Compounds F reduced basal phosphorylation of ULK 1 / 2- specific sites on ATG14 or ATG13 proteins (FIG. 10A). More global analysis using a commercially available panel showed that treatment of SCC13 cells with the Compounds B or F affected phosphorylation levels of various cell signaling kinasesMGL-06625 to a much lesser extent than SU6668 (FIG. 10B). The data in FIG. 10A SCC13 cells were treated for 2h with Compounds B and F (lOpM) or the ULK1 / 2 kinase inhibitor SBI-0206965 (lOpM), or DMSO followed by immuno blot analysis using antibodies specific for ULK1 / 2 target sites on ATG13 and ATG14, total ATG13, ATG14, ULK3 and ACTIN. FIG. 10B SCC13 cells were treated with Compounds B and F or the broad specificity SU6668 compound followed by analysis with a commercially available antibody array to assess phosphorylation levels of various proliferation and stress kinases. SCC13 cells were treated with Compound B, Compound F, or SU8886 (lOpM) for 2hr before lysis. An equal amount of total proteins, as assessed by Bradford quantification (BioRad), were overlayed overnight on the antibody arrays (Abeam) to assess the phosphorylation levels of 17 kinases and kinase substrates. Arrays were incubated with secondary antibodies, washed, and developed using ECL substrates as indicated by the provider (Abeam). Images of the membranes were quantified using ImageJ.Example 6. In silico prediction of Compounds B and F binding to the ULK3 protein.To gain deeper insights into the binding of Compounds B and F to ULK3, which could facilitate compound optimization, the Dynamic Bind software (alphafold2) was utilized. The Cn3D NIH web tool was employed to analyze the best docking results, predicting interactions of the two compounds with conserved amino acids within the ATP-binding site (FIG. 11). Analysis found both shared and compound-specific interacting amino acids between Compounds B and F. FIG. 11 shows in silico prediction of Compounds B and F binding to the ULK3 protein. Best suggested interactions of Compound B (left) and Compound F (right) within the ULK3 kinase domain by using computer-assisted docking with the Dynamic Bind tool, using the information of the ULK3 (Q6PHR2_F1) from protein data bank (UniProt). Interactions with conserved amino acids within the ATP-binding site using the Cn3D NIH web tool are indicated below, with black arrows showing Compounds B and F common interactions while the red arrows show the compound-specific interactions. Color map of the scheme: Magenta: halogen bonds; Grey lines: contacts; Green lines: H-bonds; Red lines: n-cations.Example 7. Inhibition of SCC cell proliferation by Compounds A, B, C, D, F, and G|Compounds A, B, C, D, F, and G were tested for their impact on multiple skin and Head / Neck squamous cell lines (SCC13, Cal27 and SCC028 cells). Proliferative activity of cells exposed at different concentrations of these compounds was assessed by an advanced imaging andMGL-06625 analysis platform designed for real-time, continuous monitoring of live cells for several days (Incucyte).For the measurement of the effects on cell proliferation using the IncuCyte Live-Cell Imaging System (Sartorius) (cell proliferation assays), 500-1000 cells of each cell line (Cal27, SCC13 and SCCO28) were seeded in 96-wells plates. The next day, (40-20-10-5 pM) of each compound, SU6668 or DMSO were added in triplicate wells per condition, and the cells were allowed to proliferate for the indicated times (up to 7 days). At the time intervals (3 / 4 hours), the Live-Cell Imaging System captured 5 images per well. Artificial Intelligence (Al)-driven cell confluence analysis software quantified the cell confluency over time, normalized to the time 0 image for each well, and the Mean, SE, and IC50 were calculated using Graphpad Prism.As shown in FIG. 12 and Table 3, all compounds inhibited effectively proliferation of the tested cell lines with an IC50 that varied, among cell lines, between 1.2 and 10 pM. The data in FIG. 12 shows live cell proliferation assays over the indicated times (hours) of SCC13 cells treated ULK3 inhibitors (40-20-10-5 pM) vs DMSO. Confluency normalized to Day 0 (triplicate, 5 images / well, 4-hourly, 4 days).Table 3.IC50 values of: >1 pM and <3 pM are labelled as “+++++”, >3 pM and <5 pM are labelled as “++++”, >5 pM and <7 pM are labelled as “+++”, >7 and <9 are labelled as “++”, and >10 pM are labelled as “+”.Example 8. Compounds A, B, C, D, F, and G show no growth inhibitory effects on SCC13 cells with ULK3 silencing.Silencing of the ULK3 gene slows but does not completely abrogate the growth of SCC cells. In contrast to treatment with the broad specificity SU6668 inhibitors, none of Compounds A, B, C, D, F, and G suppressed the proliferation of ULK3-silenced SCC 13 cells, indicating that the growth inhibitory effects of the compounds are specifically ULK3 -dependent and not due to aMGL-06625 specific toxicity (FIGs. 13A-13B). In fact, none of these compounds exhibited toxicity effects on SCC 13 cells or primary human dermal fibroblasts (HDFs), as detected by live cell imaging with a fluorescent active caspase 3 / 7 dye for 72h (FIGs. 14A-14B).FIGs. 13A-13B show that Compounds A, B, C, D, F, and G have no growth inhibitory effects on SCC 13 cells with ULK3 silencing.For the measurement of the effects on cell proliferation using the IncuCyte Live-Cell Imaging System (Sartorius) (cell proliferation assays), 500-1000 cells of each cell line (Cal27, SCC13 and SCCO28) were seeded in 96-wells plates. The next day, (40-20-10-5 pM) of each compound, SU6668 or DMSO, were added in triplicate wells per condition, and the cells were allowed to proliferate for the indicated times (up to 7 days). At the time intervals (3 / 4 hours), the Live-Cell Imaging System captured 5 images per well. Artificial Intelligence (Al)-driven cell confluence analysis software quantified the cell confluency over time, normalized to the time 0 image for each well, and the Mean, SE, and IC50 were calculated using Graphpad Prism.FIGs. 13A-13B show data obtained in live cell proliferation assays of SCC 13 cells with silenced ULK3 by ( proliferation analysis: (shULK3#l and #2) with the indicated ULK3 inhibitors (blue lines) versus the non-specific SU6668 compound (red line) and DMSO control (black line), (triplicate, 5 images / well, 4h intervals, 4 days).FIGs. 14A-14B show absence of toxicity by Compounds A, B, C, D, F, and G in SCCs and primary HDFs. FIG. 14A SCC13 cell toxicity assessed with Caspase 3 / 7 dye (Sartorius) over 72h with compounds (lOpM), DMSO (negative control), and Bleomycin (lOpM, positive control). Live cell imaging recorded green object count / mm2every 3h (4 images / time point), mean, n(wells / compound)=3. FIG. 14B A similar experiment in primary HDFs with Etoposide (25ng / ml) as a positive control. Green object count / mm2was recorded every 3h (4 images / time point), mean, n=(wells / compound)=2.The data in Examples 7 and 8 show that Compounds A, B, C, D, F, and G suppress proliferation and stem cell potential of squamous cancer cells without aspecific toxic effects.Example 9. Compounds A, B, C, D, F, and G suppress proliferation and stem cell potential of melanoma cells.Aging is a major risk factor for both the development and the severity of melanoma, with older adults facing higher rates of recurrence, metastasis, and death, partly due to age-related changes in skin structure and immune function. While field cancerization is well established inMGL-06625 actinic keratosis (AK) and squamous cell carcinoma (SCC), it also contributes to melanoma development through UV-induced DNA damage, photoaging, chronic inflammation, and immunosuppression. Additionally, cancer field effects have been proposed for acral melanomas (which arise on the palms, soles, and beneath the nails), where multiple gene amplifications in melanocytes have been observed in clinically normal-appearing skin. Analysis of the large Cancer Genome Atlas (TCGA; https: / / www.cancer.gov / ccg / research / genome-sequencing / tcga) shows that the ULK3 gene is amplified in 8% of metastatic melanoma cases and has elevated expression in primary melanoma compared to nevi. Furthermore, its expression is associated with more aggressive behavior and worst outcome. (FIGs. 15A-15B).Silencing of the ULK3 gene in multiple melanoma lines (A375, SKMEL28, and M14) strongly suppressed their proliferation and self-renewal potential, as assessed by clonogenicity and sphere forming assays (FIGs. 16A-16C). FIG. 16A shows efficient ULK3 silencing in melanoma. RT-PRC analysis of ULK3 expression normalized to 36B4 and western blot analysis using anti ULK3 and GAPH antibodies, in three melanoma cell lines (SKMEL28, M14 and A375) infected with two ULK3 silencing lentiviruses, or a control virus. FIG. 16B shows ULK3 silencing blunts colony formation. The indicated melanoma cell lines infected with two ULK3 silencing lentiviruses, or a control virus, were plated at limited cell density for one week. Crystal violet stained colonies were quantified using ImageJ. n(dishes)=3, mean ± SEM, P<0.0001, one-way ANOVA. FIG. 16C shows ULK3 silencing inhibits sphere-forming capabilities. The indicated melanoma cell lines infected with two ULK3 silencing lentiviruses, or a control virus were cultured in triplicate dishes under diluted conditions in Matrigel suspension for one week, followed by quantification of spheres. Shown are the number of sphere number, n(dishes)=3, mean ± SEM, P<0.0001, one-way ANOVA.Similar growth inhibitory effects were observed by live cell imaging assays of multiple melanoma cells treated with decreasing concentrations of the Compounds A, B, C, D, F, and G, with IC50S that depend on the cell line (FIG. 17 and Table 4).For the measurement of the effects on melanoma cell proliferation using the IncuCyte Live- Cell Imaging System (Sartorius) (cell proliferation assays), 1000 cells of each cell line (SKLMEL28, WM989, and A375) were seeded in 96- well plates. The next day, (40-20-10-5 pM) of each compound, SU6668 or DMSO were added in triplicate wells per condition, and the cells were allowed to proliferate for 4 / 5 days. At 4-hour time intervals, the Live-Cell Imaging SystemMGL-06625 captured 5 images per well. Al-driven cell confluence analysis software quantified the cell confluency over time, normalized to the time 0 image for each well, and the Mean, SE, and IC50 were calculated using Graphpad Prism.FIG. 17 shows data obtained in live cell proliferation assays over the indicated times (hours) of WM989 melanoma cells treated ULK3 inhibitors (20-10-5 pM) vs DMSO. Confluency normalized to Day 0 (triplicate, 5 images / well, 4-hourly, 4 days)Table 4.IC50 values of: <5 pM are labelled as “++++”, >5 pM and <10 pM are labelled as “+++”, >10 pM and <15 pM are labelled as “++”, and >15 pM are labelled as “+”.Outgrowth and matrix invasion of multicellular tumor spheroids formed by WM898 melanoma cells by a hanging drop method 13 was also significantly reduced by treatment with the Compounds B and F inhibitors even if to a lesser extent than by treatment with the less specific SU6668 compound (FI Gs. 18A-18B).Multicellular melanoma spheroids were made as follows: 3000 WM989 cells were seeded in ultralow- adherence prime surface 96- well tissue culture plates (SBio, Japan) in 10% DMEM containing 5% Matrigel and centrifuged at 2000 rpm at 4 °C before culturing in a CO2 incubator for 2 weeks. On 5 spheroids for each condition (one per well), Compound B, Compound F, SU6668 (lOpM), or DMSO were added at 48h and 1 week. Images were taken at 48h and after 2 weeks, and the volume (calculated as a sphere) ratio was plotted at time 0 (To) versus 2 weeks, and statistical analysis performed using Graphpad Prism.The data in FIGs. 18A-18B show that treatment with Compounds B and F decreases WM989 multicellular spheroid outgrowth. FIG. 18A: Quantification of WM989 spheroid volumes (3,000 cells, 2 weeks) in 5% Matrigel. Compounds (lOpM), SU6668, or DMSO were added at 48h and 1 week. FIG. 18B: The ratio at time 0 (To) versus 14 days is plotted. Mean+SEM,MGL-06625 n(sphere / concentration))=5 / dilution, one-way ANOVA, **P<0.001. A representative image for each condition at the end of the experiment is shown. (To=time zero)Example 10. Compounds A, B, C, D, F, and G revert the transcriptional program of cancer associated fibroblast (CAF) activation.ULK3 levels are higher in tumor promoting cancer associated fibroblasts (CAFs) derived from head and neck, prostate, and breast cancers and in skin, head and neck, and cervical SCCs. Paralleling the effects of silencing the ULK3 gene, treatment of multiple CAF strains with Compounds A, B, C, D, F, and G was found to inhibit ULK3 kinase activity down-modulated expression of key CAF effector genes (ACTA2, IL6, TNC and ANKRD1) (FIG.19) and, at the global transcriptomic level, suppressed gene signatures related to CAF activation, similarly to what observed upon ULK3 gene silencing (FIG. 20). The data in FIG. 19 show down-modulation of CAF gene expression by Compounds A, B, C, D, F, and G. RT-qPCR analysis of the indicated CAF effector genes in two cancer-associated fibroblast strains (CAF#8 and CAF#19) treated with all ULK3 kinase inhibitors, SU6668 (lOpM) or DMSO for 72 h. Gene expression was normalized to 36B4 and expression is relative to DMSO-treated cells.Total RNA was extracted by using Directzol RNA Miniprep assay Kit (Zymo Research #R2071) 24h from the addition of 10 pM Compound B, Compound F, or DMSO to CAFs (CAF #8, CAF#26, CAF#27). Library probe preparation was done with I pg of total RNA. Clariom D arrays (Applied Biosystem) were used on the GeneChip™ 3000 instrument system, and the results were processed with Transcriptome Analysis Console software (Thermofisher) to analyze and visualize global expression patterns of genes and pathways.Gene set enrichment analysis (GSEA) for Clariom D array expression profiles was performed by using GSAA-SeqSP software (gene set association analysis for array expression data with sample permutation) from the Gene Set Association Analysis (GSAA) platform (version GSAA_2.0). Curated gene sets were obtained from the Molecular Signatures Database (MSigDB v5.2).The data in FIG. 20 show CAF-related gene signatures suppressed by treatment of three different CAF strains with Compounds B and F. Gene set enrichment analysis (GSEA) of global transcriptomic profiles of CAF #8, CAF#26, CAF#27 after treatment (24h) with Compounds B or F (10 pM), or DMSO.MGL-06625Example 11. Compounds A, B, C, D, F, and G suppress expansion of cancer / stromal cells in in vitro and in vivo models of field cancerization.Field cancerization results from the interaction between cancer cells and CAFs. ULK3 inhibitors might be used to simultaneously target SCC and stromal cell. An in vitro model of field cancerization based on the co-culturing of Red Fluorescent-labeled CAFs with green fluorescent- labeled SCC cells allows live imaging and quantification over the time of cancer cell expansion2. In such a system Compounds A, B, C, D, F, and G significantly diminished the expansion of SCC 13 cells vs DMSO (FIG. 21). The data in FIG 21 show decreased SCC cell expansion of cocultured SCC / CAFs treated with Compounds A, B, C, D, F, and G. Red stained CAF#8 cells (3000 / well) were co-cultured with 500 SCC 13 EGFP cells with six ULK3 kinase inhibitors (lOpM), or DMSO and imaged for red / green fluorescence every 6 hours (4 images / well) using Incucyte. Green fluorescent clusters (>4000 pixels), quantified over time (Al-Sartorius) and analyzed in Prism 10. Mean+SE, n(images / time point)=12, n(cultures)=3, one-way ANOVA, **P<0.01.The effects of targeting in vivo ULK3 activity in an orthotopic NOD / SCID mouse model of cancer / stromal cell expansion based on ear injection of fluorescently labeled cells was further tested. Expansion of co-injected SCC 13 cells and CAFs was significantly decreased in this model by the topical treatment with Compound Bor the non-specific SU6668 compound (FIGs. 22A- 22B). The data in FIGs. 22A-22B show decreased in vivo SCC tumor growth by ULK3 inhibitors. FIG. 22A: SCC lesions from EGFP-expressing SCC 13 and CAFs co-injected in NOD / SCID ILR2g- / - mouse ears (2 sites / ear). After 5 days, ears were treated with 15 pM compounds in EtOH, 2x / week for 2 weeks. Contralateral ears: EtOH controls (CTRL). Images show ear lesions in 2 animals (fluorescent + phase-contrast) and FIG. 22B: final tumor volume quantification ((L*lA2)*0.52). Mean ± SEM. n=4 mice / condition; *P<0.05, ***P<0.001, two-tailed unpaired t- test.Example 12. Compounds B and F suppress the transcriptional response of human dermal fibroblasts to UVA exposure, as well as the expression of genes upregulated in photo-exposed skin and in the skin of aged versus young individuals.Changes in the dermal fibroblasts play a central role in various aspects of skin aging, including (i) extracellular matrix alterations, with loss of elasticity and wrinkling, (ii) chronic inflammation, (iii) altered communication and differentiation of overlying epidermal cells withMGL-06625 scaling and cancer. UVA radiation significantly contributes to skin aging and cancer development by directly targeting dermal fibroblasts and disrupting their function, with the induction of senescence and DNA methylation changes and an impact on distinct fibroblast lineages and immune cells.A possible role of ULK3 in the skin UVA response is indicated by the induction of this gene in both intact human skin explants and cultured dermal fibroblasts in response to UVA treatment (FIGs. 23A-23B). FIG. 23 shows that ULK3 is up-regulated by UVA in human skin and in HDFs. Representative images (FIG. 23 A) of double IF analysis with anti- ULK3 and keratin 10 (KRT10) antibodies of three independent human skin samples 24 h after UVA irradiation (18J / cm2) versus mock control, together with quantification of double positive ULK3 and KRT10 cells (FIG. 23B). > 100 cells per sample were counted. Mean ± SEM, n(samples)=3, two-tailed unpaired t-test, p<0.05. B) Western blot analysis with anti-ULK3 and ACTIN antibodies of HDFs at the indicated times after exposure to either lJ / cm2 or 2J / cm2 UVA, or mock treated (Ctr).As shown in FIG 24, the global transcriptomic response of human dermal fibroblasts to UVA was suppressed to large extent by treatment with Compounds B and F administered immediately after the UVA irradiation. In detail, 1210 transcripts were up-regulated upon UVA irradiation (>2FC), of which only 10% (257 and 254 transcripts for Compounds B and F, respectively) were still up-regulated (>2FC) in the HDFs treated with the compounds. The data in FIG. 24 show uppression of the transcriptional response of HDFs to UVA by treatment with Compounds B and F. HDFs were exposed to UVA (10J ) in PBS, followed by the addition of a fresh medium and addition of the Compound B or F (15pM), or DMSO vehicle. Cells were collected 24 hours later and analyzed by global transcriptomics (Clariom D cDNA hybridization). Shown are the total number of genes up- or down-modulated by UVA exposure (> 2 fold change) in control versus Compounds B or F treated cells.Example 23. Compounds B and F suppress gene signature that are upregulated in photoexposed skin samples and in skin samples derived from people >50 years old.The Genotype-Tissue Expression (GTEx) dataset is a comprehensive resource established by the National Institutes of Health (NIH), designed to study the relationship between genetic variation and gene expression across multiple human tissues. The probed gene expression patterns (TPM, Transcripts per Million) were derived from bulk RNA sequencing of 1,305 human skin samples, encompassing both sun-exposed and non- sun-exposed areas, utilizing data obtained fromMGL-06625 the GTEx Portal on 05 / 09 / 2024. The sun-exposed samples were collected from the lower leg (n=701), while the non-sun-exposed samples were obtained from the suprapubic area (n=604). The cohort included 427 female and 878 male samples, with an additional age-based classification into young (<50 years, n=411) and older (>50 years, n=894) groups. To assess differential gene expression patterns, Gene Set Enrichment Analysis (GSEA), a widely used statistical approach for identifying coordinated changes in gene transcription between biological conditions was employed. GSEA was applied to assess the expression pattern of genes suppressed by Compounds B and F in samples stratified according to sun exposure and age group.As shown in FIGs. 25 and 26, Compounds B and F suppressed gene signature is significantly enriched in sun-exposed skin samples, indicating that ULK3 inhibitors effectively repress genes upregulated by photo-exposure. GSEA analysis of transcriptomic profiles derived from the GTEX data base of 1,305 human skin samples derived from sun-exposed (n=701) versus non-sun-exposed (n=604) areas. Transcriptomic profiles of HDFs treated with either Compound B (FIG. 25) or Compound F (FIG. 26) for 24 hours versus DMSO controls were used to build a signature of > 200 Compound B or F repressed genes. FIGs. 25 and 26 show the enrichment plots of the signature of Compounds B and F repressed genes with corresponding normalized enrichment scores (NES) and nominal p values.A similar GSEA conducted on transcriptomic profiles of skin samples from older versus younger individuals revealed significant enrichment of the Compound B- and Compound F- repressed genes in the older group (FIGs. 27 and 28). GSEA analysis of transcriptomic profiles derived from the GTEX data base of 1,305 human skin samples derived from individuals of older (>50 years, n=894) versus younger age (<50 years, n=411), pooling together samples from sun- exposed and non-exposed areas and from males and females. Transcriptomic profiles of HDFs treated with either Compound B (FIG. 27) or Compound F (FIG. 28) for 24 hours versus DMSO controls were used to build a signature of > 200 Compound B or F repressed genes. FIGs. 27 and 28 show enrichment plots of the signature of Compounds B and F repressed genes with corresponding normalized enrichment scores (NES) and nominal p values.MGL-06625EQUIVALENTS

[0333] The details of one or more embodiments of the disclosure are set forth in the accompanying description above. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. Other features, objects, and advantages of the disclosure will be apparent from the description and from the claims. In the specification and the appended claims, the singular forms include plural referents unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents and publications cited in this specification are incorporated by reference.

[0334] The foregoing description has been presented only for the purposes of illustration and is not intended to limit the disclosure to the precise form disclosed, but by the claims appended hereto.

Claims

MGL-06625CLAIMS:

1. A method of treating a disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of a compound selected from the group consisting of:p ,MGL-06625(Compound G), or a pharmaceutically acceptable salt thereof; and wherein the disease or disorder is a cancer, a pre-cancerous condition, skin fibrosis, or skin aging.

2. The method of claim 1, wherein the compound is:or a pharmaceutically acceptable salt thereof.

3. The method of claim 1, wherein the compound is:or a pharmaceutically acceptable salt thereof.

4. The method of claim 1, wherein the compound is:or a pharmaceutically acceptable salt thereof.MGL-066255. The method of claim 1, wherein the compound is:or a pharmaceutically acceptable salt thereof.

6. The method of claim 1, wherein the compound is:or a pharmaceutically acceptable salt thereof.

7. The method of claim 1, wherein the compound is:or a pharmaceutically acceptable salt thereof.

8. The method of claim 1, wherein the compound is:or a pharmaceutically acceptable salt thereof.

9. The method of any one of claims 1-8, wherein the disease or disorder is cancer.

10. The method of claim 9, wherein the cancer is metastatic cancer.MGL-0662511. The method of any one of claims 1-10, wherein the method further comprises administering to the subject chemotherapy, immunotherapy, radiation therapy, biological therapy, surgical intervention, or any combination thereof.

12. The method of any one of claims 1-11, wherein the subject has been previously treated with chemotherapy, immunotherapy, radiation therapy, biological therapy, surgical intervention, or any combination thereof.

13. The method of any one of claims 1-8, wherein the disease or disorder is a squamous cell carcinoma, a skin melanoma, or a pre-cancerous condition.

14. The method of any one of claims 1-13, wherein the disease or disorder is a squamous cell carcinoma.

15. The method of claim 14, wherein the squamous cell carcinoma is a skin cancer, a lung cancer, a head and neck cancer, a lung cancer, an esophageal cancer, or a cervical cancer.

16. The method of claim 14, wherein the squamous cell carcinoma is adenoid / pseudoglandular squamous cell carcinoma, intraepidermal squamous cell carcinoma, large cell keratinizing squamous cell carcinoma, large cell non-keratinizing squamous cell carcinoma, lymphoepithelial carcinoma, papillary squamous cell carcinoma, papillary thyroid carcinoma, small cell keratinizing squamous cell carcinoma, spindle cell squamous cell carcinoma, or verrucous squamous-cell carcinoma.

17. The method of any one of claims 1-13, wherein the disease or disorder is a skin melanoma.

18. The method of claim 17, wherein the skin melanoma is superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma, amelanotic melanoma, acral lentiginous melanoma, mucosal melanoma, melanoma of the eye, or desmoplastic melanoma.MGL-0662519. The method of any one of claims 1-8, wherein the disease or disorder is a pre-cancerous condition.

20. The method of any one of claim 19, wherein the pre-cancerous condition is actinic keratosis, squamous cell carcinoma in situ, or keratoacanthoma.

21. The method of any one of claims 1-8, wherein the disease or disorder is skin fibrosis.

22. The method of any one of claims 1-8, wherein the disease or disorder is skin aging.

23. The method of any one of claims 1-22, wherein the compound modulates ULK3; optionally, wherein modulation is inhibition.

24. The method of any one of claims 1-23, wherein the compound is administered topically.

25. The method of any one of claims 1-23, wherein the compound is administered orally.

26. The method of any one of claims 1-23, wherein the compound is administered parenterally.

27. A pharmaceutical composition comprising:(Compound A), or a pharmaceutically acceptable salt thereof, and one or more of an excipient, a diluent, an adjuvant, or a carrier.MGL-0662528. A pharmaceutical composition comprising:(Compound B), or a pharmaceutically acceptable salt thereof, and one or more of an excipient, a diluent, an adjuvant, or a carrier.

29. A pharmaceutical composition comprising:(Compound C), or a pharmaceutically acceptable salt thereof, and one or more of an excipient, a diluent, an adjuvant, or a carrier.

30. A pharmaceutical composition comprising:(Compound D), or a pharmaceutically acceptable salt thereof, and one or more of an excipient, a diluent, an adjuvant, or a carrier.MGL-0662531. A pharmaceutical composition comprising:(Compound E), or a pharmaceutically acceptable salt thereof, and one or more of an excipient, a diluent, an adjuvant, or a carrier.

32. A pharmaceutical composition comprising:(Compound F), or a pharmaceutically acceptable salt thereof, and one or more of an excipient, a diluent, an adjuvant, or a carrier.

33. A pharmaceutical composition comprising:(Compound G), or a pharmaceutically acceptable salt thereof, and one or more of an excipient, a diluent, an adjuvant, or a carrier.MGL-0662534. A method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound, wherein the compound is:or a pharmaceutically acceptable salt of any of the foregoing.

35. A composition comprising a compound selected from Compound A, Compound B, Compound C, Compound D, Compound E, Compound F, and Compound G, or a pharmaceutically acceptable salt thereof, for use in treating a disease or disorder in a subject in need thereof.

36. A pharmaceutical composition comprising a compound selected from Compound A, Compound B, Compound C, Compound D, Compound E, Compound F, and Compound G, or a pharmaceutically acceptable salt thereof, for use in treating a disease or disorder in a subject in need thereof.MGL-0662537. Compound A, Compound B, Compound C, Compound D, Compound E, Compound F, or Compound G, or a pharmaceutically acceptable salt thereof, for use in treating a disease or disorder in a subject in need thereof.

38. Use of a composition comprising a compound selected from Compound A, Compound B, Compound C, Compound D, Compound E, Compound F, and Compound G, or a pharmaceutically acceptable salt thereof, for the treatment of a disease or disorder in a subject in need thereof.

39. Use of a pharmaceutical composition comprising a compound selected from Compound A, Compound B, Compound C, Compound D, Compound E, Compound F, and Compound G, or a pharmaceutically acceptable salt thereof, for the treatment of a disease or disorder in a subject in need thereof.

40. Use of Compound A, Compound B, Compound C, Compound D, Compound E, Compound F, or Compound G, or a pharmaceutically acceptable salt thereof, for the treatment of a disease or disorder in a subject in need thereof.

41. Use of a composition comprising a compound selected from Compound A, Compound B, Compound C, Compound D, Compound E, Compound F, and Compound G, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disease or disorder in a subject in need thereof.

42. Use of a pharmaceutical composition comprising a compound selected from Compound A, Compound B, Compound C, Compound D, Compound E, Compound F, and Compound G, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disease or disorder in a subject in need thereof.

43. Use of Compound A, Compound B, Compound C, Compound D, Compound E, Compound F, or Compound G, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disease or disorder in a subject in need thereof.MGL-0662544. The method, composition, compound, or use of any one of claims 34-43, wherein the disease or disorder is a squamous cell carcinoma, a skin melanoma, or a pre-cancerous condition.

45. The method, composition, compound, or use of claim 44, wherein the disease or disorder is a squamous cell carcinoma.

46. The method, composition, compound, or use of any one of claims 44-45, wherein the squamous cell carcinoma is a skin cancer, a lung cancer, a head and neck cancer, a lung cancer, an esophageal cancer, or a cervical cancer.

47. The method, composition, compound, or use of any one of claims 44-46, wherein the squamous cell carcinoma is adenoid / pseudoglandular squamous cell carcinoma, intraepidermal squamous cell carcinoma, large cell keratinizing squamous cell carcinoma, large cell nonkeratinizing squamous cell carcinoma, lymphoepithelial carcinoma, papillary squamous cell carcinoma, papillary thyroid carcinoma, small cell keratinizing squamous cell carcinoma, spindle cell squamous cell carcinoma, or verrucous squamous-cell carcinoma.

48. The method, composition, compound, or use of claim 44, wherein the disease or disorder is a skin melanoma.

49. The method, composition, compound, or use of claim 44 or claim 48, wherein the skin melanoma is superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma, amelanotic melanoma, acral lentiginous melanoma, mucosal melanoma, melanoma of the eye, or desmoplastic melanoma.

50. The method, composition, compound, or use of claim 44, wherein the disease or disorder is a pre-cancerous condition.

51. The method, composition, compound, or use of claim 44 or claim 50, wherein the pre- cancerous condition is actinic keratosis, squamous cell carcinoma in situ, or keratoacanthoma.MGL-0662552. The method, composition, compound, or use of any one of claims 34-43, wherein the disease or disorder is skin fibrosis.

53. The method, composition, compound, or use of any one of claims 34-43, wherein the disease or disorder is skin aging.

54. The method, composition, compound, or use of any one of claims 34-43, wherein the compound modulates ULK3 ; optionally, wherein modulation is inhibition.

55. The method, composition, compound, or use of any one of claims 34-44, wherein the compound is:(Compound A), or a pharmaceutically acceptable salt thereof.

56. The method, composition, compound, or use of any one of claims 34-44, wherein the compound is:(Compound B), or a pharmaceutically acceptable salt thereof.

57. The method, composition, compound, or use of any one of claims 34-44, wherein the compound is:MGL-06625(Compound C), or a pharmaceutically acceptable salt thereof.

58. The method, composition, compound, or use of any one of claims 34-44, wherein the compound is:or a pharmaceutically acceptable salt thereof.

59. The method, composition, compound, or use of any one of claims 34-44, wherein the compound is:(Compound E), or a pharmaceutically acceptable salt thereof.

60. The method, composition, compound, or use of any one of claims 34-44, wherein the compound is:(Compound F), or a pharmaceutically acceptable salt thereof.MGL-0662561. The method, composition, compound, or use of any one of claims 34-44, wherein the compound is:(Compound G), or a pharmaceutically acceptable salt thereof.

62. The method, composition, compound, or use of any one of claims 34-44, wherein the pharmaceutically acceptable salt is the hydrochloride salt.

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