Liver receptor homolog-1 (LRH-1) agonist isosteres and uses

Abstract

This disclosure relates to a series of small molecule agonists that mimic liver receptor homolog-1 (LRH-1). The compounds display exceptional potency and efficacy. Also disclosed are pharmaceutical compounds and methods of treating or preventing various conditions including cancer, cardiovascular disease, diabetes, colitis, or ulcerative colitis. Further disclosed are methods of modulating activity of LRH-1 in cells.

Description

[0001] LIVER RECEPTOR HOMOLOG-1 (LRH-1) AGONIST ISOSTERES AND USES

[0002] CROSS-REFERENCES TO RELATED APPLICATION

[0003] The application claims the benefit of and the priority to U.S. Provisional Application No. 63 / 638,536, filed April 25, 2024, which is hereby incorporated by reference in its entirety for all purposes.

[0004] STATEMENT REGARDING FEDERALLY FUNDED RESEARCH

[0005] This invention was made with government support under Grant No. DK115213 awarded by the National Institutes of Health. The government has certain rights in the invention.

[0006] SEQUENCE LISTING

[0007] A Sequence Listing conforming to the rules of WIPO Standard ST.26 is hereby incorporated by reference. Said Sequence Listing has been filed as an electronic document via PatentCenter encoded as XML in UTF-8 text. The electronic document, created on April 16, 2025, is entitled “043150-1486567-093W01_ST26.xml”, and is 12,467 bytes in size.

[0008] BACKGROUND

[0009] Nuclear receptors (NRs) are ligand-regulated transcription factors that allow biological systems to sense and respond to lipophilic molecules by altering gene expression. Their functions are driven by a modular structure consisting of a ligand binding domain (LBD) and DNA-binding domain (DBD). Ligand binding to the LBD promotes NR association with coactivators, which drive target gene expression through recruitment of transcriptional machinery and remodeling of chromatin. Liver receptor homolog-1 (LRH-1; NR5A2) is a nuclear receptor that is primarily expressed in tissues of endodermal origin and has been thoroughly characterized in the liver, where it regulates lipid and glucose homeostasis. This receptor also plays a key role in cell renewal and local glucocorticoid biosynthesis in the gut, making it an attractive target for inflammatory bowel disease.

[0010] SUMMARY

[0011] Covered embodiments of the invention are defined by the claims, not by this summary. This summary is a high-level overview of various aspects of the invention and introduces some of the concepts that are described and illustrated in the present document and the accompanying figures. This summary is not intended to identify key or essential features of the claimed subject matter, nor is it intended to be used in isolation to determine the scope of the claimed subject matter. The subject matter should be understood by reference to appropriate portions of the entire specification, any or all figures and each claim.

[0012] Described herein are a series of small molecule agonists that mimic liver receptor homolog-1 (LRH-1). The small molecule agonists described herein are less metabolically labile while having improved potency and efficacy over known LRH-1 agonists. In some examples, this disclosure relates to methods of treating or preventing diabetes, cancer, or cardiovascular disease by administering to a subject in need thereof an effective amount of an LRH-1 agonist as described herein.

[0013] Described herein are compounds of the following formula:

[0014] Formula I or a prodrug, salt, or stereoisomer thereof, wherein n is 1 to 10; X is heteroaryl, amido, or sulfamate, wherein X is optionally substituted with one or more, the same or different, of hydrogen, alkyl, hydroxy, hydroxyalkyl, alkanoyl, carbocyclyl, benzyl, aryl, or heterocyclyl; R1is halogen, nitro, cyano, hydroxy, amino, sulfamoylamino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl, wherein R1is optionally substituted with one or more, the same or different, R10; R2is hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl, wherein R2is optionally substituted with one or more, the same or different, R10; R3is hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl, wherein R3is optionally substituted with one or more, the same or different, R10; R4is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl, wherein R4is optionally substituted with one or more, the same or different, R10; R5is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl, wherein R5is optionally substituted with one or more, the same or different, R10; R10is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl, wherein R10is optionally substituted with one or more, the same or different, R11; and R11is halogen, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy, isopropoxy, tert-butoxy, hydroxymethyl, hydroxyethyl, thiomethyl, thioethyl, aminomethyl, aminoethyl, acetyl, acetoxy, methylamino, ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino, acetylamino, N- methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N- methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulfmyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl, tert-butoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N- methyl-N-ethylsulfamoyl, benzoyl, benzyl, carbocyclyl, aryl, or heterocyclyl. In some examples, X is -C(O)NR6R7, -O-S(=O)2NR8R9, or a tetrazole, wherein R6, R7, R8, and R9are each independently hydrogen, alkyl, hydroxy, hydroxyalkyl, alkanoyl, carbocyclyl, benzyl, aryl, or heterocyclyl, or R6and R7or R8and R9, together with the nitrogen atom to which they are connected, form a heterocyclyl or heteroaryl, wherein R6, R7, R8, and / or R9is optionally substituted with one or more, the same or different, R10.

[0015] Optionally, the compound of Formula I is a compound of the following formula:

[0016] Formula I-A or a prodrug, salt, or stereoisomer thereof, wherein R6and R7are each independently hydrogen, alkyl, hydroxy, hydroxyalkyl, alkanoyl, carbocyclyl, benzyl, aryl, or heterocyclyl, or R6and R7together with the nitrogen atom to which they are connected form a heterocyclyl or heteroaryl, wherein R6and / or R7is optionally substituted with one or more, the same or different, R10. In some examples, the compound has the following formula:

[0017] Formula I- A3 Formula I-A4 or a prodrug, salt, or stereoisomer thereof.

[0018] In some examples, the compound has the following formula:

[0019] Formula I-B or a prodrug, salt, or stereoisomer thereof, wherein R8and R9are each independently hydrogen, alkyl, hydroxy, hydroxyalkyl, alkanoyl, carbocyclyl, benzyl, aryl, or heterocyclyl, or R8and R9together with the nitrogen atom to which they are connected form a heterocyclyl or heteroaryl, wherein R8and / or R9is optionally substituted with one or more, the same or different, R10.

[0020] Optionally, the compound has the following formula:

[0021] Formula I-Bl or a prodrug, salt, or stereoisomer thereof. In some examples, the compound has the following formula:

[0022] Formula I-C or a prodrug, salt, or stereoisomer thereof, wherein R8is hydrogen, alkyl, hydroxy, hydroxyalkyl, alkanoyl, carbocyclyl, benzyl, aryl, or heterocyclyl, wherein R8is optionally substituted with one or more, the same or different, R10.

[0023] In some examples, the compound has the following formula:

[0024] Formula I-Cl or a prodrug, salt, or stereoisomer thereof.

[0025] In some examples of the compounds as described herein, R4is 1-phenylvinyl, n is 2 to

[0026] 7, R1is hydroxy or sulfamoylamino, R2and R3are hydrogen, and / or R5is phenyl.

[0027] Also described herein are compounds according to the following structures: or a prodrug or salt thereof.

[0028] Further disclosed herein are pharmaceutical compositions containing a compound as described herein or a pharmaceutically acceptable salt and a pharmaceutically acceptable excipient. Also disclosed are methods of treating or preventing cancer. The methods of treating or preventing cancer as described herein comprise administering a therapeutically effective amount of a pharmaceutical composition as described herein to a subject in need thereof. Optionally, the cancer is selected from the group consisting of bladder cancer, breast cancer, colon cancer, rectal cancer, endometrial cancer, kidney cancer, leukemia, lung cancer, melanoma, non-Hodgkin lymphoma, pancreatic cancer, prostate cancer, and thyroid cancer.

[0029] Additionally disclosed are methods of treating or preventing diabetes. The methods of treating or preventing cancer as described herein comprise administering a therapeutically effective amount of a pharmaceutical composition as described herein to a subject in need thereof. Optionally, the diabetes is insulin-dependent diabetes mellitus, non-insulin-dependent diabetes mellitus, or gestational diabetes.

[0030] Further disclosed are methods of treating or preventing cardiovascular disease, inflammatory bowel diseases (IBD), colitis or ulcerative colitis, and / or diabetic nephropathy comprising administering a therapeutically effective amount of a pharmaceutical composition as described herein to a subject in need thereof.

[0031] Aso disclosed are methods of modulating steroidogenic factor-1 (SF-1) and / or liver receptor homolog-1 (LRH-1) activity in a cell, subject. The methods comprise contacting the cell with a therapeutically effective amount of a compound as described herein. The contacting can optionally be performed in vitro or in vivo.

[0032] The details of one or more examples are set forth in the drawings and the description below. Other features, objects, and advantages will be apparent from the description and drawings, and from the claims.

[0033] DESCRIPTION OF DRAWINGS

[0034] Figure 1 shows LRH-1 binding to activating ligands. Panel A. Structure of the LRH-1 LBD (white) bound to activating phospholipid DLPC (tan) and fragment of coactivator peptide TIF2 (green). Panel B. Binding orientation of DLPC (top; tan; PDB entry 4DOS), parent compound RJW100 (bottom; green; PDB entry 5L11), and phospholipid-mimicking small molecule 10CA (bottom; salmon; PDB entry 7JYD). Interacting side and main chains are shown as sticks (O = red, N = blue, P = orange, C = white). Hydrogen bond interactions are indicated with dotted lines and waters involved in mediating small molecule-LRH-1 interactions are represented as spheres. Panel C. Chemical structures of molecules shown in panel (B) with the PL-mimicking carboxylic acid of 1 OCA highlighted. Figure 2 Fluorescence polarization (FP) binding and FP competition assays are displayed independently. Top left curve represents the forward binding curve used to determine the binding affinity of 6N-FAM for the LRH-1 LBD. Data shown as means ± SEM from eight independent experiments. Kd = dissociation constant. 95% confidence intervals are shown in brackets. The remaining curves represent FP competition assays used to determine compound Ki (inhibition constant) values. FP competition data shown as means ± standard error of the mean (SEM) from two independent experiments, with 95% confidence intervals shown in brackets. Polarization values were normalized for each independent experiment in such a way that the highest value = 100 and the lowest value = 0.

[0035] Figure 3 shows moieties of the compounds as described herein.

[0036] Figure 4 shows that the Tet isostere maintains high affinity while improving compound potency and increasing LRH-1 target gene expression. Panel A. FP competition assay showing binding of compounds to the LRH-1 LBD (Ki = inhibition constant). Data shown as means from two independent experiments. Error bars represent 95% confidence intervals. Panel B. Comparison of ligand-driven thermal stability of the LRH-1 LBD. The inflection point corresponds to the temperature at which the protein unfolds. Data shown as means ± SD from two independent experiments. Panel C. Data from luciferase reporter assays. Relative efficacy (RE) was calculated by normalizing the fold change by that of 10CA. The quadrant representing improved potency and efficacy is shaded in green. Data shown as means from three biological replicates. Panel D. RT-qPCR analysis of HepG2 cells treated with agonists (10 pM: Tet, Sul; 30 pM: 10CA, Am, HA, Pip, Ser) for 24 hours. Data normalized to signal of DMSO control and shown as means ± SD from four biological replicates. Brown-Forsythe and Welch one-way ANOVA with Dunnett multiple comparisons rest, **p < 0.01, ***p < 0.001.

[0037] Figure 5 shows luciferase reporter assays displayed independently. Luciferase reporter assay (HeLa cells) showing effects of small molecules on LRH-1 activity. Relative luciferase activity corresponds to the calculated span of the curve + 1. Data were normalized relative to DMSO control and are shown as means ± SEM from three biological replicates. ECso = half maximal effective concentration. 95% confidence intervals are shown in brackets. ECso values reported here, along with relative luciferase activity normalized to that of 10CA, were used to construct Figure 4C.

[0038] Figure 6 contains the crystal structure of LRH-1 LBD complexed with Tet and fragment of coactivator TIF2, revealing interactions at both regions of binding pocket. A. Crystal structure of Tet (teal) bound to the LRH-1 LBD (PDB entry 8F8M). TIF2 coactivator peptide is shown in green. B. Ligand 2Fo-Fc map showing electron density for Tet contoured at Is. C. Overlay of Tet (teal) and 10CA (salmon; PDB entry 7JYD). D. Key deep pocket interactions made by Tet. E. Pocket mouth interactions made by Tet. Sidechains (along with backbones of G421, T423, and L424) of engaged residues are shown as sticks (O = red, N = blue, S = yellow, C = white). Water molecules are shown as spheres. Hydrogen bonds are represented as dotted lines. Note that K520 is slightly out of hydrogen bonding range for the tetrazole. The position of K520 in lOCA-bound LRH-1 (PDB entry 7JYD) is shown in salmon.

[0039] Figure 7 contains molecular dynamics simulations showing similarities and differences in compound binding and protein motions driven by 10CA and Tet. A. Percent of time that 10CA and Tet spent hydrogen bonding with mouth residues during MD simulations. Data shown as means ± SD from four 500 ns simulations. Data analyzed with unpaired two-tailed t- test, *p < 0.05, **p < 0.01, ***p < 0.001. B. Distances between residues (A) were determined for complexes and difference distance matrices were then calculated by subtracting distances from two complexes (between Ca atoms). Averaged after four 500 ns simulations. Matrices reflect structural differences between indicated complexes. C. Analysis showing communities of residues with correlated motion. Nodes represent Ca atoms, along with their associated backbone and side chain atoms. Edges represent correlated motion and are weighted by the degree of correlated motion. H6-H7 is indicated by a red oval.

[0040] Figure 8 shows that the addition of sulfamide improves compound affinity and potency while maintaining tail-mediated efficacy. A. Overlay of 6N-10CA (purple; PDB entry 7TT8) and 10CA (salmon; PDB entry 7JYD). Select residues that engage small molecules deep within the pocket and at the pocket mouth are indicated. Sidechains are shown as sticks (O = red, N = blue, S = yellow, C = white). Water molecules shown as spheres. Hydrogen bonds are represented as dotted lines. B. Chemical structure of Tet and hybrid molecule 6N-Tet, which incorporates the sulfamide moiety of LRH-1 small molecule agonist 6N. C. FP competition assay showing binding of compounds to the LRH-1 LBD. Ki = inhibition constant. Data shown as means ± 95% confidence intervals determined from two independent experiments (see Figure 2 for binding curves). D. Luciferase reporter assay comparing compound-driven activation of LRH-1. ECso = half maximal effective concentration. Efficacies of Tet and 6N- Tet were 1.9 and 2.1, respectively. Data shown as means from three (Tet) or five (6N-Tet) biological replicates. E. Comparison of ligand-driven thermal stability of the LRH-1 LBD. The inflection point corresponds to the temperature at which the protein unfolds. Data shown as means ± SD from two independent experiments. F. RT-qPCR analysis of HepG2 cells treated with agonists (6N, Tet, and 6N-Tet - 10 pM; 10CA - 30 pM) for 24 hrs. Data normalized to signal of DMSO control and shown as means ± SD from two (DMSO and 6N-Tet), three (6N), or four (10CA and Tet) biological replicates. Brown-Forsythe and Welch one-way ANOVA with Dunnett T3 multiple comparisons rest, #p < 0.01, *p < 0.05.

[0041] Figure 9 contains graphs showing that the small molecules described herein show specificity for NR5A receptors. A. LRH-1 and SF-1 luciferase reporter assays, indicating ligand-induced activity after addition of 10 pM of indicated small molecules. Data represented as means + SD from four biological replicates. B. 6N-Tet was introduced to reporter cells at 2 pM and the activity of respective nuclear receptors was tested. Agonism was examined for all receptors, aside from RORy, where inverse agonism was tested. Data was normalized relative to receptor activity induced by an agonist (or antagonist for RORy) for each receptor added at a concentration corresponding to the ECioo (or ICioo for RORy).

[0042] DETAILED DESCRIPTION

[0043] Described herein are small molecules that mimic polar interactions made by phospholipid ligands. Specifically, LRH-1 (also referred to as NR5A2), is a phospholipidbinding nuclear receptor that regulates metabolism in the liver and gut. The compounds described herein, display exceptional potency and show selectivity towards LRH-1 and its close homolog steroidogenic factor-1 (SF-1, NR5A1) making them effective drugs in modulating NR5A activity.

[0044] Before the present disclosure is described in greater detail, it is to be understood that this disclosure is not limited to particular examples described, and as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular examples only, and is not intended to be limiting, since the scope of the present disclosure will be limited only by the appended claims.

[0045] I. Definitions

[0046] 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. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, the preferred methods and materials are now described.

[0047] All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present disclosure is not entitled to antedate such publication by virtue of prior disclosure. Further, the dates of publication provided could be different from the actual publication dates that may need to be independently confirmed.

[0048] All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference and are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications and patents are cited.

[0049] Examples of the present disclosure will employ, unless otherwise indicated, techniques of medicine, organic chemistry, medicinal chemistry, biochemistry, molecular biology, pharmacology, and the like, which are within the skill of the art. Such techniques are explained fully in the literature, such as the references cited herein.

[0050] It must be noted that, as used in the specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. In this specification and in the claims that follow, reference will be made to a number of terms that shall be defined to have the following meanings unless a contrary intention is apparent.

[0051] Certain of the compounds described herein may contain one or more asymmetric centers and may give rise to enantiomers, diastereomers, and other stereoisomeric forms that can be defined, in terms of absolute stereochemistry at each asymmetric atom as (R)- or (S)-. The present chemical entities, pharmaceutical compositions and methods are meant to include all such possible isomers, including racemic mixtures, tautomer forms, hydrated forms, optically substantially pure forms, and intermediate mixtures.

[0052] Unless otherwise stated, structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement or enrichment of a hydrogen by deuterium or tritium at one or more atoms in the molecule, or the replacement or enrichment of a carbon by13C or14C at one or more atoms in the molecule, are within the scope of this disclosure. In one example, provided herein are isotopically labeled compounds having one or more hydrogen atoms replaced by or enriched by deuterium. In one example, provided herein are isotopically labeled compounds having one or more hydrogen atoms replaced by or enriched by tritium. In one example, provided herein are isotopically labeled compounds having one or more carbon atoms replaced or enriched by13C. In one example, provided herein are isotopically labeled compounds having one or more carbon atoms replaced or enriched by14C. The disclosure also embraces isotopically labeled compounds that are identical to those recited herein, except that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such as, e.g.,2H,3H,13C,14C,15N,18O,17O,31P,32P,35S,18F, and36C1, respectively. Certain isotopically labeled compounds (e.g., those labeled with3H and / or14C) are useful in compound and / or substrate tissue distribution assays. Tritiated (i.e.,3H) and carbon-14 (i.e.,14C) isotopes can allow for ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium (i.e.,2H) can afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements). Isotopically labeled disclosed compounds can generally be prepared by substituting an isotopically labeled reagent for a non-isotopically labeled reagent. In some examples, provided herein are compounds that can also contain unnatural proportions of atomic isotopes at one or more of atoms that constitute such compounds. All isotopic variations of the compounds as disclosed herein, whether radioactive or not, are encompassed within the scope of the present disclosure.

[0053] As used herein, a “lipid” group refers to a hydrophobic group that is naturally or non- naturally occurring that is highly insoluble in water. As used herein a lipid group is considered highly insoluble in water when the point of connection on the lipid is replaced with a hydrogen and the resulting compound has a solubility of less than 3 x 10'3w / w (at 25 °C) in water, e.g., 9.5 x 10'4% w / w (at 25 °C) which is the percent solubility of hexane in water by weight. See Solvent Recovery Handbook, 2ndEd, Smallwood, 2002 by Blackwell Science, page 193. Examples of naturally occurring lipids include saturated or unsaturated hydrocarbon chains found in fatty acids, glycerolipids, cholesterol, steroids, polyketides, and derivatives. Non- naturally occurring lipids include derivatives of naturally occurring lipids, acrylic polymers, and alkylated compounds and derivatives thereof.

[0054] As used herein, “alkyl” means a noncyclic straight chain or branched, unsaturated or saturated hydrocarbon such as those containing from 1 to 22 carbon atoms, while the term “lower alkyl” or “Ci-4 alkyl” has the same meaning as alkyl but contains from 1 to 4 carbon atoms. The term “higher alkyl” has the same meaning as alkyl but contains from 8 to 22 carbon atoms. Representative saturated straight chain alkyls include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-septyl, n-octyl, n-nonyl, and the like; while saturated branched alkyls include isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, and the like. Unsaturated alkyls contain at least one double or triple bond between adjacent carbon atoms (referred to as an “alkenyl” or “alkynyl”, respectively). Representative straight chain and branched alkenyls include ethylenyl, propylenyl, 1-butenyl, 2-butenyl, isobutylenyl, 1 -pentenyl, 2-pentenyl, 3 - methyl- 1-butenyl, 2-methyl-2-butenyl, 2,3- dimethyl-2-butenyl, and the like; while representative straight chain and branched alkynyls include acetylenyl, propynyl, 1-butynyl, 2- butynyl, 1 -pentynyl, 2-pentynyl, 3- methyl- 1-butynyl, and the like.

[0055] Non-aromatic mono or polycyclic alkyls are referred to herein as “carbocycles” or “carbocyclyl” groups. Representative saturated carbocycles include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like; while unsaturated carbocycles include cyclopentenyl and cyclohexenyl, and the like. Optionally, the carbocyclyl groups described herein have 3 to 30 carbon atoms, i.e., C3-C30 carbocyclyl. Optionally, the carbocyclyl groups described herein have 3 to 20 carbon atoms, i.e., C3-C20 carbocyclyl. Optionally, the carbocyclyl groups described herein have 3 to 12 carbon atoms, i.e., C3-C12 carbocyclyl.

[0056] “Heterocarbocycles” or “heterocarbocyclyl” groups are carbocycles which contain from 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur which may be saturated or unsaturated (but not aromatic), monocyclic or polycyclic, and wherein the nitrogen and sulfur heteroatoms may be optionally oxidized, and the nitrogen heteroatom may be optionally quaternized. Heterocarbocycles include morpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyridinyl, tetrahydroprimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like.

[0057] “Aryl” means an aromatic carbocyclic monocyclic or polycyclic ring such as phenyl or naphthyl. Polycyclic ring systems may, but are not required to, contain one or more non- aromatic rings, as long as one of the rings is aromatic. “Arylalkyl” means an alkyl substituted with an aryl, e.g., benzyl, methyl substituted with phenyl. Optionally, the aryl groups described herein have 6 to 30 carbon atoms, i.e., C6-C30 aryl. Optionally, the aryl groups have 6 to 20 carbon atoms, i.e., C6-C20 aryl. Optionally, the aryl groups have 6 to 12 carbon atoms, i.e., Ce- C12 aryl.

[0058] As used herein, “benzoyl” refers a group having the formula ~C(=O)“C6H5.

[0059] As used herein, “benzyl” refers to a group having the formula R-CH2-C6H5.

[0060] As used herein, “heteroaryl” refers to an aromatic heterocarbocycle having 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur, and containing at least 1 carbon atom, including both mono- and polycyclic ring systems. Polycyclic ring systems may, but are not required to, contain one or more non-aromatic rings, as long as one of the rings is aromatic. Representative heteroaryls are furyl, benzofuranyl, thiophenyl, benzothiophenyl, pyrrolyl, indolyl, isoindolyl, azaindolyl, pyridyl, quinolinyl, isoquinolinyl, oxazolyl, isooxazolyl, benzoxazolyl, pyrazolyl, imidazolyl, benzimidazolyl, thiazolyl, benzothiazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, cinnolinyl, phthalazinyl, and quinazolinyl. It is contemplated that the use of the term “heteroaryl” includes N-alkylated derivatives such as a l-methylimidazol-5-yl substituent. Optionally, the heteroaryl groups described herein have 3 to 30 carbon atoms, i.e., C3-C30 heteroaryl. Optionally, the heteroaryl groups have 3 to 20 carbon atoms, i.e., C3-C20 heteroaryl. Optionally, the heteroaryl groups have 3 to 11 carbon atoms, i.e., C3-C11 heteroaryl.

[0061] As used herein, “heterocycle” or “heterocyclyl” refers to mono- and polycyclic ring systems having 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur, and containing at least 1 carbon atom. The mono- and polycyclic ring systems may be aromatic, non-aromatic or mixtures of aromatic and non-aromatic rings. Heterocycle includes heterocarbocycles, heteroaryls, and the like.

[0062] “Alkylthio” refers to an alkyl group as defined above attached through a sulfur bridge. An example of an alkylthio is methylthio, (i.e., -S-CH3).

[0063] “Alkoxy” refers to an alkyl group as defined above attached through an oxygen bridge. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, i-propoxy, n- butoxy, s-butoxy, t-butoxy, n- pentoxy, and s-pentoxy. Preferred alkoxy groups are methoxy, ethoxy, n-propoxy, i- propoxy, n-butoxy, s-butoxy, and t-butoxy.

[0064] “Amino” refers to -NRaRb, wherein Ra and Rb are independently selected from hydrogen, alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl, wherein each of the alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl can be optionally and independently substituted by one or more substitution groups described herein. When Ra and Rb are hydrogen, the amino group is a primary amino.

[0065] “Alkylamino” refers to an alkyl group as defined above attached through an amino bridge. An example of an alkylamino is methylamino, (i.e., -NH-CH3).

[0066] “Alkanoyl” refers to an alkyl as defined above attached through a carbonyl bridge (i.e., -(C=O)alkyl).

[0067] “Alkyl sulfonyl” refers to an alkyl as defined above attached through a sulfonyl bridge (i.e., -S(=O)2alkyl) such as mesyl and the like, "aryl sulfonyl" refers to an aryl attached through a sulfonyl bridge (i.e., - S(=O)2aryl). “Aminosulfonyl” or “sulfamoyl” refers to an amino attached through a sulfonyl bridge (i.e., -S(=O)2NH2).

[0068] “Amido” refers to a functional group of an amide attached with an acid radical present.

[0069] “Sulfamoylamino” refers to a sulfamoyl attached through an amino bridge (i.e., -NH- S(=O)2NH2).

[0070] “Alkyl sulfinyl” refers to an alkyl as defined above attached through a sulfinyl bridge (i.e., -S(=O)alkyl).

[0071] “Aminoalkyl” refers to an amino group attached through an alkyl bridge. An example of an aminoalkyl is aminomethyl, (i.e., NH2-CH2-).

[0072] As used herein, “acyl” refers -C(=O)Ra, wherein Ra is selected from alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl, wherein each of the alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl can be optionally substituted by one or more substitution groups described herein.

[0073] As used herein, “alkoxycarbonyl” refers to an alkoxy group attached to a carbonyl bridge (e.g., RO-C(=O)-).

[0074] “Hydroxyalkyl” refers to a hydroxy group attached through an alkyl bridge. An example of a hydroxyalkyl is hydroxyethyl, (i.e., HO-CH2CH2-).

[0075] “Hydroxy amino” refers to -NH(OH).

[0076] “Hal” refers to a halogen such as F, Br, Cl, or I.

[0077] The term “substituted” refers to a molecule wherein at least one hydrogen atom is replaced with a substituent. When substituted, one or more of the groups are “substituents.” The molecule may be multiply substituted. In the case of an oxo substituent (“=O”), two hydrogen atoms are replaced. Example substituents within this context may include halogen, hydroxy, alkyl, alkoxy, nitro, cyano, oxo, carbocyclyl, carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, -NRaRb, -NRaC(=O)Rb, - NRaC(=O)NRaNRb, -NRaC(=O)ORb, -NRaSChRb, -C(=O)Ra, -C(=O)ORa, -C(=O)NRaRb, - OC(=O)NRaRb, -ORa, -SRa, -SORa, - S(=O)2Ra, -OS(=O)2Ra and -S(=O)2ORa. Ra and Rb in this context may be the same or different and independently hydrogen, halogen, hydroxy, alkyl, alkoxy, alkyl, amino, alkylamino, dialkylamino, carbocyclyl, carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl.

[0078] The term “optionally substituted,” as used herein, means that substitution is optional and therefore it is possible for the designated atom to be unsubstituted. As used herein, “salts” refer to derivatives of the disclosed compounds where the parent compound is modified making acid or base salts thereof. Examples of salts include, but are not limited to, mineral salts such as sodium, potassium, or zinc carboxylic acid salts, or organic acid salts of basic residues such as amines, alkylamines, or dialkylamines; alkali or organic salts of acidic residues such as carboxylic acids; and the like.

[0079] As used herein, “pharmaceutically acceptable salt” refers to the modification of the original compound by making the acid or base salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines, as well as alkali or organic salts of acidic residues such as carboxylic acids or phosphorus acids. For original compounds containing a basic residue, the pharmaceutically acceptable salts can be prepared by treating the original compounds with an appropriate amount of a non-toxic inorganic or organic acid; alternatively, the pharmaceutically acceptable salts can be formed in situ during preparation of the original compounds; alternatively, the pharmaceutically acceptable salts can be prepared via ion-exchange with existing salts of the original compounds. Exemplary salts of the basic residue include salts with an inorganic acid selected from hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, and nitric acids or with an organic acid selected from acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, naphthalenesulfonic, methanesulfonic, ethane disulfonic, oxalic, and isethionic acids. For original compounds containing an acidic residue, the pharmaceutically acceptable salts can be prepared by treating the original compounds with an appropriate amount of a non-toxic base; alternatively, the pharmaceutically acceptable salts can be formed in situ during preparation of the original compounds; alternatively, the pharmaceutically acceptable salts can be prepared via ionexchange with existing salts of the original compounds. Exemplary salts of the acidic residue include salts with a base selected from ammonium hydroxide, sodium hydroxide, potassium hydroxide, lithium hydroxide, calcium hydroxide, magnesium hydroxide, ferrous hydroxide, zinc hydroxide, copper hydroxide, aluminum hydroxide, ferric hydroxide, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2- dimethylaminoethanol, 2-diethylaminoethanol, lysine, arginine, and histidine. Optionally, the pharmaceutically acceptable salts can be prepared by reacting the free acid or base form of the original compounds with a stoichiometric amount or more of the appropriate base or acid, respectively, in water, in an organic solvent, or in a mixture thereof. Lists of suitable pharmaceutically acceptable salts can be found in Remington’s Pharmaceutical Sciences, 20th Ed., Lippincott Williams & Wilkins, Baltimore, MD, 2000; and Handbook of Pharmaceutical Salts: Properties, Selection, and Use, Stahl and Wermuth, Eds., Wiley-VCH, Weinheim, 2002.

[0080] In some examples, the salts are conventional nontoxic pharmaceutically acceptable salts including the quaternary ammonium salts of the parent compound formed, and non-toxic inorganic or organic acids. Preferred salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, and the like.

[0081] In some examples, the pharmaceutical acceptable salts of the disclosed compounds and prodrugs thereof are salts with ammonium hydroxide, i.e., ammonium salts. In some examples, the pharmaceutical acceptable salts of the disclosed compounds and prodrugs thereof are lithium salts.

[0082] As used throughout, subject can be a vertebrate, more specifically a mammal (e.g., a human, horse, cat, dog, cow, pig, sheep, goat, mouse, rabbit, rat, and guinea pig), birds, reptiles, amphibians, fish, and any other animal. The term does not denote a particular age or sex. Thus, adult and newborn subjects, whether male or female, are intended to be covered. As used herein, patient or subject may be used interchangeably and the term patient or subject includes human and veterinary subjects.

[0083] As used herein, the term “derivative” refers to a structurally similar compound that retains sufficient functional attributes of the identified analogue. The derivative may be structurally similar because it is lacking one or more atoms, substituted with one or more substituents, a salt, in different hydration / oxidation states, e.g., substituting a single or double bond, substituting a hydroxy group for a ketone, or because one or more atoms within the molecule are switched, such as, but not limited to, replacing an oxygen atom with a sulfur or nitrogen atom, or replacing an amino group with a hydroxy group or vice versa. Replacing a carbon with nitrogen in an aromatic ring is a contemplated derivative. The derivative may be a prodrug. Derivatives may be prepared by any variety of synthetic methods or appropriate adaptations presented in the chemical literature or as in synthetic or organic chemistry textbooks, such as those provide in March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, Wiley, 6th Edition (2007) Michael B. Smith or Domino Reactions in Organic Synthesis, Wiley (2006) Lutz F. Tietze hereby incorporated by reference. The term “prodrug” refers to an agent that is converted into a biologically active form in vivo. Prodrugs are often useful because, in some situations, they may be easier to administer than the parent compound. They may, for instance, be bioavailable by oral administration whereas the parent compound is not. The prodrug may also have improved solubility in pharmaceutical compositions over the parent drug. A prodrug may be converted into the parent drug by various mechanisms, including enzymatic processes and metabolic hydrolysis. Typical prodrugs are pharmaceutically acceptable esters or enol ethers. Prodrugs include compounds wherein a hydroxy, amino or mercapto group is bonded to any group that, when the prodrug of the active compound is administered to a subject, cleaves to form a free hydroxy, free amino or free mercapto group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate, and benzoate derivatives of an alcohol or acetamide, formamide and benzamide derivatives of an amine functional group in the active compound and the like.

[0084] For example, if a disclosed compound or a pharmaceutically acceptable form of the compound contains a carboxylic acid functional group, a prodrug can comprise a pharmaceutically acceptable ester formed by the replacement of the hydrogen atom of the acid group with a group such as (Ci-C4)alkyl, (Ci-Csjalkyl, (C2-Ci2)alkanoyloxymethyl, 1- (alkanoyloxy)ethyl having from 4 to 9 carbon atoms, 1 -methyl- l-(alkanoyloxy)-ethyl having from 5 to 10 carbon atoms, alkoxy carbonyloxymethyl having from 3 to 6 carbon atoms, 1- (alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms, 1 -methyl- 1- (alkoxycarbonyloxy)ethyl having from 5 to 8 carbon atoms, N-(alkoxycarbonyl)aminomethyl having from 3 to 9 carbon atoms, l-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10 carbon atoms, 3-phthalidyl, 4-crotonolactonyl, gamma-butyrolacton-4-yl, di-N,N-(Ci- C2)alkylamino(C2-C3)alkyl (such as beta-dimethylaminoethyl), carbarn oyl-(Ci-C2)alkyl, N,N- di(Ci-C2)alkylcarbamoyl-(Ci-C2)alkyl and piperidino-, pyrrolidino- or morpholino(C2- Csjalkyl.

[0085] If a disclosed compound or a pharmaceutically acceptable form of the compound contains an alcohol functional group, a prodrug can be formed by the replacement of the hydrogen atom of the alcohol group with a group such as (Ci-Cejalkanoyloxymethyl, l-(( Ci- Cejalkanoyloxy) ethyl, l-methyl-l((Ci-C6)alkanoyloxy)ethyl (Ci-

[0086] Cejalkoxycarbonyloxymethyl, -N-(Ci-C6)alkoxycarbonylaminomethyl, succinoyl, (Ci- Cejalkanoyl, alpha-amino(Ci-C4)alkanoyl, arylacyl and alpha-aminoacyl, or alpha-aminoacyl - alpha-aminoacyl, where each alpha-aminoacyl group is independently selected from naturally occurring L-amino acids P(O)(OH)2, -P(O)(O(Ci-Ce)alkyl)2, and glycosyl (the radical resulting from the removal of a hydroxy group of the hemiacetal form of a carbohydrate). If a disclosed compound or a pharmaceutically acceptable form of the compound incorporates an amine functional group, a prodrug can be formed by the replacement of a hydrogen atom in the amine group with a group such as R-carbonyl, RO-carbonyl, NRR'- carbonyl where R and R' are each independently (Ci-Cio)alkyl, (C3-C7)cycloalkyl, benzyl, a natural alpha-aminoacyl, -C(OH)C(O)OYi wherein Y1is H, (Ci-Ce)alkyl or benzyl, - C(OY2)Y3wherein Y2 is (C1-C4) alkyl and Y3 is (Ci-Ce)alkyl, carboxy(Ci-Ce)alkyl, amino(Ci- C4)alkyl or mono-Nor di-N,N-(Ci-C6)alkylaminoalkyl, -C(Y4)Ys wherein Y4 is H or methyl and Ys is mono-N- or di-N,N-( Ci-C6)alkylamino, morpholino, piperidin-l-yl or pyrrolidin-1- yi.

[0087] As used herein, “pharmaceutically acceptable esters” include, but are not limited to, alkyl, alkenyl, alkynyl, aryl, arylalkyl, and cycloalkyl esters of acidic groups, including, but not limited to, carboxylic acids, phosphoric acids, phosphinic acids, sulfonic acids, sulfinic acids, and boronic acids.

[0088] As used herein, “pharmaceutically acceptable enol ethers” include, but are not limited to, derivatives of formula -C=C(OR) where R can be selected from alkyl, alkenyl, alkynyl, aryl, aralkyl, and cycloalkyl. Pharmaceutically acceptable enol esters include, but are not limited to, derivatives of formula -C=C(OC(O)R) where R can be selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, aralkyl, and cycloalkyl.

[0089] As used herein, the term “therapeutically effective amount” or effective amount refers to an amount of a therapeutic composition that, when administered to a subject, is effective to treat a disease or disorder such that the symptoms of the disease or disorder are ameliorated, or the likelihood of the disease or disorder developing or progressing is decreased. A therapeutically effective amount is not, however, a dosage so large as to cause adverse side effects, such as hyperviscosity syndromes, pulmonary edema, congestive heart failure, and the like. A suitable dose of a therapeutic composition as described herein, which dose is capable of treating a cancer, diabetes, inflammatory bowel disease, colitis, ulcerative colitis, or diabetic nephropathy in a subject, can depend on a variety of factors including the particular therapeutic composition used and whether it is used concomitantly with other therapeutic agents. For example, a different dose of a compound described herein may be required to treat a subject with cancer diabetes, inflammatory bowel disease, colitis, ulcerative colitis, or diabetic nephropathy as compared to the dose of a currently available treatment option required to treat the same subject. Other factors affecting the dose administered to the subject include, e.g., the type or extent of cancer, the type or extent of diabetes, the type or extent of inflammatory bowel disease, the type or extent of colitis, the type or extent of ulcerative colitis, and the type or extent of diabetic nephropathy. For example, a subject that has had a previous cancer (e.g., a subject with relapsed or recurrent cancer) may require administration of a different dosage of the compounds described herein than a subject who has not previously had cancer. Generally, a therapeutically effective amount may vary with the subject’s age, condition, and sex, as well as the extent of the disease in the subject and can be determined by one of skill in the art. Other factors can include, e.g., other medical disorders concurrently or previously affecting the subject, the age and general health of the subject, the genetic disposition of the subject, diet, time of administration, the route of administration, and the size (body weight, body surface, or organ size), the rate of excretion, drug combination, and any other additional therapeutics that are administered to the subject. It should also be understood that a specific dosage and treatment regimen for any particular subject also depends upon the judgment of the treating medical practitioner (e.g., doctor or nurse). A therapeutically effective amount is also one in which any toxic or detrimental effects of the composition are outweighed by the therapeutically beneficial effects. The dosage of the therapeutically effective amount may be adjusted by the individual physician in the event of any complication. In some instances, a therapeutically effective amount may vary from about 0.01 mg / kg to about 50 mg / kg, preferably from about 0.1 mg / kg to about 20 mg / kg, most preferably from about 0.2 mg / kg to about 2 mg / kg, in one or more dose administrations daily, for one or several days. In some embodiments, the doses can be about 1, about 0.5, about 0.1, about 0.05, or about 0.01 mg / kg, or any intervening dose between about 0.01 mg / kg and 1 mg / kg. In some instances, the compounds described herein are administered for 2 to 5 or more consecutive days. In some instances, the compounds described herein are administered to a subject at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times.

[0090] “Cancer” refers any of various cellular diseases with malignant neoplasms characterized by the proliferation of cells. It is not intended that the diseased cells must actually invade surrounding tissue and metastasize to new body sites. Cancer can involve any tissue of the body and have many different forms in each body area. Within the context of certain examples, whether "cancer is reduced" may be identified by a variety of diagnostic manners known to one skill in the art including, but not limited to, observation the reduction in size or number of tumor masses or if an increase of apoptosis of cancer cells observed, e.g., if more than a 5 % increase in apoptosis of cancer cells is observed for a sample compound compared to a control without the compound. It may also be identified by a change in relevant biomarker or gene expression profile, such as PSA for prostate cancer, HER2 for breast cancer, or others.

[0091] A “chemotherapy agent,” “chemotherapeutic,” “anti-cancer agent” or the like, refer to molecules that are recognized to aid in the treatment of a cancer. Contemplated examples include the following molecules or derivatives such as temozolomide, carmustine, bevacizumab, procarbazine, lomustine, vincristine, gefitinib, erlotinib, cisplatin, carboplatin, oxaliplatin, 5 -fluorouracil, gemcitabine, tegafur, raltitrexed, methotrexate, cytosine arabinoside, hydroxyurea, adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin, mithramycin, vinblastine, vindesine, vinorelbine, paclitaxel, taxol, docetaxel, etoposide, teniposide, amsacrine, topotecan, camptothecin, bortezomib, anagrelide, tamoxifen, toremifene, raloxifene, droloxifene, idoxifene, fulvestrant, bicalutamide, flutamide, nilutamide, cyproterone, goserelin, leuprorelin, buserelin, megestrol, anastrozole, letrozole, vorozole, exemestane, finasteride, marimastat, trastuzumab, cetuximab, dasatinib, imatinib, combretastatin, thalidomide, azacitidine, azathioprine, capecitabine, chlorambucil, cyclophosphamide, cytarabine, daunorubicin, doxifluridine, epothilone, irinotecan, mechlorethamine, mercaptopurine, mitoxantrone, pemetrexed, tioguanine, valrubicin and / or lenalidomide or combinations thereof such as cyclophosphamide, methotrexate, 5 -fluorouracil (CMF); doxorubicin, cyclophosphamide (AC); mustine, vincristine, procarbazine, prednisolone (MOPP); adriamycin, bleomycin, vinblastine, dacarbazine (ABVD); cyclophosphamide, doxorubicin, vincristine, prednisolone (CHOP); bleomycin, etoposide, cisplatin (BEP); epirubicin, cisplatin, 5 -fluorouracil (ECF); epirubicin, cisplatin, capecitabine (ECX); methotrexate, vincristine, doxorubicin, cisplatin (MV AC).

[0092] As used herein, the terms “prevent” and “preventing” include the prevention of the recurrence, spread or onset. It is not intended that the present disclosure be limited to complete prevention. In some examples, the onset is delayed, or the severity of the disease is reduced.

[0093] As used herein, “treat,” “treating,” “treatment,” and the like mean obtaining a desired pharmacologic and / or physiologic effect. “Treating” or “treatment” may refer to any indicia of success in the treatment or amelioration of cancer. “Treating” or “treatment” includes the administration of an agent to impede growth of a cancer, to do one or more of the following: cause a cancer to shrink by weight or volume, extend the expected survival time of the subject, or extend the expected time to progression of the tumor, or the like. The effect of treatment can be compared to an individual or pool of individuals not receiving the treatment, or to the same patient prior to treatment or at a different time during treatment.

[0094] As used herein, the term “combination with” when used to describe administration with an additional treatment means that the agent may be administered prior to, together with, or after the additional treatment, or a combination thereof.

[0095] Further explanations of the terms recited above are provided below in the description. II. Compounds

[0096] Disclosed herein are agonists of liver receptor homolog-1 (LRH-1). The substitutions in the molecules described herein are designed with substituents (e.g., having polar residues) that bind deep in the receptor pocket, improving binding and agonism. The compounds as described herein having these features exhibit increased stability and are prepared by reproducible synthetic methods, resulting in promising therapeutics for the treatment of the conditions described herein.

[0097] To the extent that chemical formulas described herein contain one or more unspecified chiral centers, the formulas are intended to encompass all stable stereoisomers, enantiomers, and diastereomers. Such compounds can exist as a single enantiomer, a mixture of diastereomers, a racemic mixture, or combinations thereof. It is also understood that the chemical formulas encompass all tautomeric forms.

[0098] A class of LRH-1 agonists described herein is represented by Formula I:

[0099] Formula I including prodrugs, salts, and stereoisomers thereof.

[0100] In Formula I, n is 1 to 10. For example, n can be from 2 to 9, from 3 to 8, from 4 to 7, from 2 to 7, or from 2 to 4.

[0101] Also in Formula I, X is heteroaryl, amido, or sulfamate. Optionally, X can be substituted with one or more, the same or different, of hydrogen, alkyl, hydroxy, hydroxyalkyl, alkanoyl, carbocyclyl, benzyl, aryl, or heterocyclyl. In some examples, X is -C(O)NR6R7, -O- S(=O)2NR8R9, or a tetrazole, wherein R6, R7, R8, and R9are each independently hydrogen, alkyl, hydroxy, hydroxyalkyl, alkanoyl, carbocyclyl, benzyl, aryl, or heterocyclyl, or R6and R7or R8and R9, together with the nitrogen atom to which they are connected, form a heterocyclyl or heteroaryl. Optionally, R6, R7, R8, and / or R9can be substituted with one or more, the same or different, R10.

[0102] Further in Formula I, R1is halogen, nitro, cyano, hydroxy, amino, sulfamoylamino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl. Optionally, R1can be substituted with one or more, the same or different, R10(as described below). In some examples, R1is hydroxy. In some examples, R1is sulfamoylamino.

[0103] Also in Formula I, R2is hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl. Optionally, R2can be substituted with one or more, the same or different, R10. In some examples, R2is hydrogen.

[0104] Additionally in Formula I, R3is hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl. Optionally, R3can be substituted with one or more, the same or different, R10. In some examples, R3is hydrogen.

[0105] Further in Formula I, R4is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl. Optionally, R4can be substituted with one or more, the same or different, R10. In some examples, R4is 1- phenylvinyl.

[0106] Also in Formula I, R5is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl. Optionally, R5can be substituted with one or more, the same or different, R10. In some examples, R5is phenyl.

[0107] In Formula I, R10is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl. Optionally, R10can be substituted with one or more, the same or different, R11.

[0108] Also in Formula I, R11is halogen, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy, isopropoxy, tert-butoxy, hydroxymethyl, hydroxy ethyl, thiomethyl, thioethyl, aminomethyl, aminoethyl, acetyl, acetoxy, methylamino, ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl, tert-butoxycarbonyl, N-methylsulfamoyl, N- ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl, benzoyl, benzyl, carbocyclyl, aryl, or heterocyclyl.

[0109] Optionally, the compounds of Formula I have the structure of Formula I-A as shown

[0110] Formula I-A including prodrugs, salts, and stereoisomers thereof.

[0111] In Formula I-A, n, R1, R2, R3, R4, and R5are as defined above for Formula I.

[0112] Also in Formula I-A, R6and R7are each independently hydrogen, alkyl, hydroxy, hydroxyalkyl, alkanoyl, carbocyclyl, benzyl, aryl, or heterocyclyl, or R6and R7together with the nitrogen atom to which they are connected form a heterocyclyl or heteroaryl. Optionally, R6and / or R7can be substituted with one or more, the same or different, R10.

[0113] In some examples, n is 1 to 10 carbons. In some examples, R6is hydrogen and R7is a hydrogen. In some examples, R6is a hydroxy group and R7is hydrogen. In some examples, R6and R7together with the nitrogen atom to which they are connected form a heterocyclyl or heteroaryl, such as a piperazine. In some examples, R6is a hydroxyalkyl group.

[0114] Optionally, the compounds of Formula I-A have the structure of Formula I-Al as shown below:

[0115] Formula I-Al including prodrugs, salts, and stereoisomers thereof.

[0116] In Formula I-Al, n, R1, R2, R3, R4, and R5are as defined above for Formula I.

[0117] Optionally, the compounds of Formula I-A have the structure of Formula I-A2 as shown below:

[0118] Formula I-A2 including prodrugs, salts, and stereoisomers thereof.

[0119] In Formula I-A2, n, R1, R2, R3, R4, and R5are as defined above for Formula I.

[0120] Optionally, the compounds of Formula I-A have the structure of Formula I- A3 as shown below:

[0121] Formula I- A3 including prodrugs, salts, and stereoisomers thereof.

[0122] In Formula I-A3, n, R1, R2, R3, R4, and R5are as defined above for Formula I.

[0123] Optionally, the compounds of Formula I-A have the structure of Formula I-A4 as shown below:

[0124] Formula I-A4 including prodrugs, salts, and stereoisomers thereof.

[0125] In Formula I-A4, n, R1, R2, R3, R4, and R5are as defined above for Formula I.

[0126] Optionally, the compounds of Formula I have the structure of Formula I-B as shown

[0127] Formula I-B including prodrugs, salts, and stereoisomers thereof. In Formula I-B, n, R1, R2, R3, R4, and R5are as defined above for Formula I.

[0128] Also in Formula I-B, R8and R9are each independently hydrogen, alkyl, hydroxy, hydroxyalkyl, alkanoyl, carbocyclyl, benzyl, aryl, or heterocyclyl, or R8and R9together with the nitrogen atom to which they are connected form a heterocyclyl or heteroaryl. Optionally, R8and / or R9can be substituted with one or more, the same or different, R10. In some examples, R8and R9are hydrogen.

[0129] Optionally, the compounds of Formula I-B have the structure of Formula I-Bl as shown below:

[0130] Formula I-Bl including prodrugs, salts, and stereoisomers thereof.

[0131] In Formula I-Bl, n, R1, R2, R3, R4, and R5are as defined above for Formula I.

[0132] Optionally, the compounds of Formula I have the structure of Formula I-C as shown below:

[0133] Formula I-C including prodrugs, salts, and stereoisomers thereof.

[0134] In Formula I-C, n, R1, R2, R3, R4, and R5are as defined above for Formula I.

[0135] Also in Formula I-C, R8is hydrogen, alkyl, hydroxy, hydroxyalkyl, alkanoyl, carbocyclyl, benzyl, aryl, or heterocyclyl. Optionally, R8is substituted with one or more, the same or different, R10. In some examples, R8is hydrogen.

[0136] Optionally, the compounds of Formula I-C have the structure of Formula I-Cl as shown below: Formula I-Cl including prodrugs, salts, and stereoisomers thereof.

[0137] In Formula I-Cl, n, R1, R2, R3, R4, and R5are as defined above for Formula I.

[0138] Compounds according to the present disclosure include enantiomers, a mixture of enantiomers, a diastereomer, a mixture of two or more diastereomers, a tautomer, a mixture of two or more tautomers, or an isotopic variant thereof, or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

[0139] Exemplary Compounds

[0140] In some examples, a compound according to the present disclosure can be selected from the compounds below, including an enantiomer, a mixture of enantiomers, a mixture of two or more diastereomers, a tautomer, a mixture of two or more tautomers, or an isotopic variant thereof, or a pharmaceutically acceptable salt, solvate, or hydrate thereof:

[0141] Exemplary compounds described herein, include, but are not limited to, the following compounds and pharmaceutically acceptable salts (such as ammonium and lithium salts) thereof:

[0142] III. Methods of Making the Compounds

[0143] The compounds described herein can be prepared in a variety of ways. The compounds can be synthesized using various synthetic methods. At least some of these methods are known in the art of synthetic organic chemistry. The compounds described herein can be prepared from readily available starting materials. Optimum reaction conditions can vary with the particular reactants or solvent used, but such conditions can be determined by one skilled in the art by routine optimization procedures. For example, synthetic schemes may be followed according to the methods described below in the examples section. Variations on Formula I and other compounds as described herein include the addition, subtraction, or movement of the various constituents as described for each compound. Similarly, when one or more chiral centers are present in a molecule, all possible chiral variants are included. Additionally, compound synthesis can involve the protection and deprotection of various chemical groups. The use of protection and deprotection, and the selection of appropriate protecting groups can be determined by one skilled in the art. The chemistry of protecting groups can be found, for example, in Wuts, Greene’s Protective Groups in Organic Synthesis, 5th. Ed., Wiley & Sons, 2014, which is incorporated herein by reference in its entirety.

[0144] Methods of making pharmaceutical formulations are generally known in the art. Exemplary methods can be found in the following references and references cited therein: Lieberman, et al., Pharmaceutical Dosage Forms: Tablets, Marcel Dekker, Inc., New York, 1989; Ansel, et al., Pharmaceutical Dosage Forms and Drug Delivery Systems, 6th Ed., Williams & Wilkins, Media, PA, 1995; Remington - The Science and Practice of Pharmacy, 20th Ed., Lippincott Williams & Wilkins, Baltimore, MD, 2000. Delayed release, extended release, and / or pulsatile release formulations may be prepared as described in the references described above. These references provide information on carriers, materials, equipment, and processes for preparing tablets, capsules, and granules, as well as controlled release forms of the tablets, capsules, and granules.

[0145] Reactions to produce the compounds described herein can be carried out in solvents, which can be selected by one of skill in the art of organic synthesis. Solvents can be substantially nonreactive with the starting materials (reactants), the intermediates, or products under the conditions at which the reactions are carried out, i.e., temperature and pressure. Reactions can be carried out in one solvent or a mixture of more than one solvent. Product or intermediate formation can be monitored according to any suitable method known in the art. For example, product formation can be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g.,JH or13C) infrared spectroscopy, spectrophotometry (e.g., UV-visible), or mass spectrometry, or by chromatography such as high-performance liquid chromatography (HPLC) or thin layer chromatography.

[0146] IV. Pharmaceutical Compositions

[0147] While it is possible that, for use in therapy, a therapeutically effective amount of a compound disclosed herein may be administered as the raw chemical, it is typically presented as the active ingredient of a pharmaceutical composition or formulation. Accordingly, the disclosure further provides a pharmaceutical composition comprising a compound disclosed herein. The pharmaceutical composition may further comprise one or more pharmaceutically acceptable carriers, diluents, and / or excipients. The carrier(s), diluent(s) and / or excipient(s) must be acceptable in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof. In accordance with another aspect of the disclosure there is also provided a process for the preparation of a pharmaceutical formulation including admixing a compound disclosed herein with one or more pharmaceutically acceptable carriers, diluents and / or excipients.

[0148] Pharmaceutical formulations may be presented in unit dose forms containing a predetermined amount of active ingredient per unit dose. Such a unit may contain, for example, 0.5 mg to 1 g, preferably 1 mg to 700 mg, more preferably 5 mg to 100 mg of a compound disclosed herein (as a free-base, solvate (including hydrate) or salt, in any form), depending on the condition being treated, the route of administration, and the age, weight and condition of the patient. Preferred unit dosage formulations are those containing a daily dose, weekly dose, or monthly dose, of an active ingredient. Furthermore, such pharmaceutical formulations may be prepared by any of the methods well known in the pharmacy art.

[0149] Pharmaceutical formulations may be adapted for administration by any appropriate route, for example by the oral (including capsules, tablets, liquid-filled capsules, disintegrating tablets, immediate, delayed and controlled release tablets, oral strips, solutions, syrups, buccal and sublingual), rectal, nasal, inhalation, topical (including transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal) route. Such formulations may be prepared by any method known in the art of pharmacy, for example by bringing into association the active ingredient with the carrier(s), excipient(s) or diluent. Generally, the carrier, excipient or diluent employed in the pharmaceutical formulation is "non-toxic," meaning that it / they is / are deemed safe for consumption in the amount delivered in the pharmaceutical composition, and "inert" meaning that it / they does / do not appreciably react with or result in an undesired effect on the therapeutic activity of the active ingredient.

[0150] Pharmaceutical formulations adapted for oral administration may be presented as discrete units such as liquid-filled or solid capsules; immediate, delayed, or controlled release tablets; powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; edible foams or whips; oil-in-water liquid emulsions, water-in-oil liquid emulsions or oral strips, such as impregnated gel strips.

[0151] For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral pharmaceutically acceptable carrier such as ethanol, glycerol, sterilized water, and the like. Powders are prepared by comminuting the compound to a suitable fine size and mixing with a similarly comminuted pharmaceutical carrier such as an edible carbohydrate, as, for example, starch or mannitol. Flavoring, preservative, dispersing and coloring agent can also be present.

[0152] Solid capsules are made by preparing a powder mixture, as described above, and filling formed gelatin sheaths. Glidants and lubricants such as colloidal silica, talc, magnesium stearate, calcium stearate or solid polyethylene glycol can be added to the powder mixture before the filling operation. A disintegrating or solubilizing agent such as agar-agar, calcium carbonate or sodium carbonate can also be added to improve the availability of the medicament when the capsule is ingested.

[0153] Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents, and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth, or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. Tablets are formulated, for example, by preparing a powder mixture, granulating, slugging, and / or adding a lubricant and disintegrant and pressing into tablets. A powder mixture may be prepared by mixing the compound, suitably comminuted, with a diluent or base as described above, and optionally, with a binder such as carboxymethylcellulose, an alginate, gelatin, or polyvinyl pyrrolidone, a solution retardant such as paraffin, a resorption accelerator such as a quaternary salt and / or an absorption agent such as bentonite, kaolin or dicalcium phosphate. The powder mixture can be granulated by wetting with a binder such as syrup, starch paste, or solutions of cellulosic or polymeric materials and forcing through a screen. As an alternative to granulating, the powder mixture can be run through the tablet machine and the result is imperfectly formed slugs broken into granules. The granules can be lubricated to prevent sticking to the tablet forming dies by means of the addition of stearic acid, a stearate salt, talc or mineral oil. The lubricated mixture is then compressed into tablets. The compounds disclosed herein can also be combined with a free- flowing inert carrier and compressed into tablets directly without going through the granulating or slugging steps. A clear or opaque protective coating consisting of a sealing coat of shellac, a coating of sugar or polymeric material and a polish coating of wax can be provided. Dyestuffs can be added to these coatings to distinguish different unit dosages.

[0154] Oral fluids such as solutions, syrups and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of the compound. Solutions and syrups can be prepared by dissolving the compound in a suitably flavored aqueous solution, while elixirs are prepared through the use of a pharmaceutically acceptable alcoholic vehicle. Suspensions can be formulated by dispersing the compound in a pharmaceutically acceptable vehicle. Solubilizers and emulsifiers such as ethoxylated isostearyl alcohols and polyoxy ethylene sorbitol ethers, preservatives, flavor additive such as peppermint oil or natural sweeteners or saccharin or other artificial sweeteners, and the like can also be added.

[0155] Where appropriate, unit dosage formulations for oral administration can be microencapsulated. The formulation can also be prepared to prolong or sustain the release as for example by coating or embedding particulate material in polymers, wax or the like.

[0156] The compounds of the disclosure can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearyl amine, or phosphatidylcholines.

[0157] Pharmaceutical formulations adapted for topical administration in the mouth include lozenges, pastilles, and mouth washes.

[0158] Pharmaceutical formulations adapted for rectal administration may be presented as suppositories or as enemas.

[0159] Pharmaceutical formulations adapted for nasal administration wherein the carrier is a solid include a coarse powder having a particle size for example in the range 20 to 500 microns which is administered in the manner in which snuff is taken, i.e. by rapid inhalation through the nasal passage from a container of the powder held close up to the nose. Suitable formulations wherein the carrier is a liquid, for administration as a nasal spray or as nasal drops, include aqueous or oil solutions of the active ingredient.

[0160] Pharmaceutical formulations adapted for administration by inhalation include fine particle dusts or mists, which may be generated by means of various types of metered dose pressurized aerosols, metered dose inhalers, dry powder inhalers, nebulizers, or insufflators.

[0161] Pharmaceutical formulations adapted for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams, or spray formulations.

[0162] Pharmaceutical formulations adapted for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain antioxidants, buffers, bacteriostats and solutes which render the formulation of pharmaceutically acceptable tonicity with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The formulations may be presented in unitdose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injection, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets. It should be understood that in addition to the ingredients particularly mentioned above, the formulations may include other agents conventional in the art having regard to the type of formulation in question, for example, those suitable for oral administration may include flavoring agents.

[0163] V. Methods of Use

[0164] In certain examples, this disclosure relates to methods of treating or preventing diseases or conditions associated with LRH-1 such as diabetes, cancer, or cardiovascular disease by administering a therapeutically effective amount of a hexahydropentalene derivative disclosed herein to a subject in need thereof.

[0165] In certain examples, the disclosure relates to methods of treating or preventing diabetes comprising administering a therapeutically effective amount of a pharmaceutical composition comprising compounds disclosed herein to a subject in need thereof. In certain examples, the subject is at risk of, exhibiting symptoms of, or diagnosed with diabetes, insulin-dependent diabetes mellitus, non-insulin-dependent diabetes mellitus, or gestational diabetes.

[0166] In certain examples, this disclosure relates to compounds disclosed herein that are LRH- 1 agonists for use in the prevention of progressive loss of pancreatic beta-cells. It also relates to an LRH-1 agonist for use in the preservation or restoration of pancreatic beta-cells. Further, it relates to an LRH- 1 agonist for use in the prevention or treatment of type I diabetes or insulindependent diabetes mellitus, the increment of survival of pancreatic beta-cells, the increment of the performance of pancreatic beta-cells, the increment of the survival of a beta-cell graft, the in vitro preservation of pancreatic beta-cells, maintaining insulin secretion and / or in a method of transplanting pancreatic islet cells.

[0167] In some cases, the disclosure relates to the treatment or prevention of pancreatitis. In certain examples, the methods of treating or preventing pancreatitis comprises administering a therapeutically effective amount of a pharmaceutical composition comprising a compound disclosed herein to a subject in need thereof.

[0168] Diabetes mellitus (DM) is often simply referred to as diabetes. Diabetes is a condition in which a person has a high blood sugar (glucose) level as a result of the body either not producing enough insulin, or because body cells do not properly respond to the insulin that is produced.

[0169] In healthy persons, blood glucose levels are maintained within a narrow range, primarily by the actions of the hormone insulin. Insulin is released by pancreatic beta-cells at an appropriate rate in response to circulating glucose concentrations, the response being modulated by other factors including other circulating nutrients, islet innervation and incretin hormones. Insulin maintains glucose concentrations by constraining the rate of hepatic glucose release to match the rate of glucose clearance.

[0170] Insulin thus enables body cells to absorb glucose, to turn into energy. If the body cells do not absorb the glucose, the glucose accumulates in the blood (hyperglycemia), leading to various potential medical complications. Accordingly, diabetes is characterized by increased blood glucose resulting in secondary complications such as cardiovascular diseases, kidney failure, retinopathy and neuropathy if not properly controlled. Two major pathophysiologies are related to increase glycemia. The first is an autoimmune attack against the pancreatic insulin-producing beta-cells (Type 1 diabetes or insulin-dependent diabetes) whilst the second is associated to poor beta-cell function and increased peripheral insulin resistance (Type 2 diabetes or non-insulin dependent diabetes). Similar to Type 1, beta-cell death is also observed in Type 2 diabetes. Type 1 and often Type 2 diabetes requires the person to inject insulin.

[0171] Type 1 DM is typically characterized by loss of the insulin-producing beta-cells of the islets of Langerhans in the pancreas leading to insulin deficiency. This type of diabetes can be further classified as immune-mediated or idiopathic. The majority of Type 1 diabetes is of the immune-mediated nature, where beta-cell loss is a T-cell mediated autoimmune attack. Sensitivity and responsiveness to insulin are usually normal, especially in the early stages. Type 1 diabetes can affect children or adults but was traditionally termed "juvenile diabetes" because it represents a majority of the diabetes cases in children.

[0172] Type 2 DM is characterized by beta-cell dysfunction in combination with insulin resistance. The defective responsiveness of body tissues to insulin is believed to involve the insulin receptor. Similar to Type 1 diabetes, an insufficient beta cell mass is also a pathogenic factor in many Type 2 diabetic patients. In the early stage of Type 2 diabetes, hyperglycemia can be reversed by a variety of measures and medications that improve insulin secretion and reduce glucose production by the liver. As the disease progresses, the impairment of insulin secretion occurs, and therapeutic replacement of insulin may sometimes become necessary in certain patients. In certain examples, the treatment of diabetes by administering compounds disclosed herein is in combination with the administration of insulin.

[0173] Diabetes without proper treatments can cause many complications. Acute complications include hyperglycemia, diabetic ketoacidosis, or nonketotic hyperosmolar coma. Serious long-term complications include cardiovascular disease, chronic renal failure, retinal damage. In some examples, this disclosure relates to methods of treating or preventing central nervous system (CNS) disorders, such as diabetic neuropathy. In some cases, this disclosure relates to improved mitochondrial dysfunction and a decreased level of apoptosis in podocytes. In certain examples, this disclosure relates to methods of treating or preventing cardiovascular disease comprising administering a therapeutically effective amount of a pharmaceutical composition comprising a compound disclosed herein to a subject in need thereof.

[0174] In certain examples, the cardiovascular disease is coronary artery diseases (CAD), angina, myocardial infarction, stroke, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, heart arrhythmia, congenital heart disease, valvular heart disease, carditis, aortic aneurysms, peripheral artery disease, and venous thrombosis.

[0175] In some examples, this disclosure relates to methods of treating or preventing diabetic nephropathy (i.e., diabetic kidney disease). In some cases, this disclosure relates to improved mitochondrial dysfunction, a decreased level of apoptosis in podocytes, and improved podocyte functi on / survival .

[0176] In certain examples, this disclosure relates to methods of treating or managing cancer. “Cancer” refers any of various cellular diseases with malignant neoplasms characterized by the proliferation of cells. It is not intended that the diseased cells must actually invade surrounding tissue and metastasize to new body sites. Cancer can involve any tissue of the body and have many different forms in each body area. Within the context of certain examples, whether "cancer is reduced" may be identified by a variety of diagnostic manners known to one skill in the art including, but not limited to, observation the reduction in size or number of tumor masses or if an increase of apoptosis of cancer cells observed, e.g., if more than a 5 % increase in apoptosis of cancer cells is observed for a sample compound compared to a control without the compound. It may also be identified by a change in relevant biomarker or gene expression profile, such as PSA for prostate cancer, HER2 for breast cancer, or others.

[0177] In certain examples, this disclosure relates to methods of treating cancer comprising administering a therapeutically effective amount of an agent to a subject in need thereof. Optionally, the cancer is bladder cancer, brain cancer, breast cancer (e.g., triple negative breast cancer), bronchus cancer, colorectal cancer (e.g., colon cancer, rectal cancer), cervical cancer, chondrosarcoma, endometrial cancer, gastrointestinal cancer, gastric cancer, genitourinary cancer, glioblastoma, head and neck cancer, hepatic cancer, hepatocellular carcinoma, leukemia, liver cancer, lung cancer, lymphoma, melanoma of the skin, ovarian cancer, pancreatic cancer, prostate cancer, renal cancer, skin cancer, testicular cancer, thyroid cancer, or uterine cancer. Optionally, the cancer is a cancer that affects one or more of the following sites: oral cavity and pharynx (e.g., tongue, mouth, pharynx, or other oral cavity); digestive system (e.g., esophagus, stomach, small intestine, colon, rectum, anus, anal canal, anorectum, liver and intrahepatic bile duct, gallbladder and other biliary, pancreas, or other digestive organs); respiratory system (e.g., larynx, lung and bronchus, or other respiratory organs); bones and joints; soft tissue (e.g., heart); skin (e.g., melanoma of the skin or other nonepithelial skin); breast; genital system (e.g., uterine cervix, uterine corpus, ovary, vulva, vagina and other female genital areas, prostate, testis, penis and other male genital areas); urinary system (e.g., urinary bladder, kidney and renal pelvis, and ureter and other urinary organs); eye and orbit; brain and other nervous system; endocrine system (e.g., thyroid and other endocrine); lymphoma (e.g., Hodgkin lymphoma and non-Hodgkin lymphoma); myeloma; or leukemia (e.g., acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, or other leukemia). Optionally, the cancer is a cancer that has an increased expression of LRH-1 as compared to non-cancerous cells of the same cell type.

[0178] In certain examples, the cancer is pancreatic cancer, breast cancer, liver cancer, colon cancer, or gastrointestinal tumors.

[0179] Benod et al. report LRH-1 regulates pancreatic cancer cell growth and proliferation. Proc Natl Acad Sci U S A, 2011, 108(41): 16927-31. Pan et al. report LRH-1 -dependent programming of mitochondrial glutamine processing drives liver cancer. Genes Dev, 2016, 30(11): 1255-1260. Holly et al. LRH-1 drives colon cancer cell growth by repressing the expression of the CDKN1 A gene in a p53-dependent manner. Nucleic Acids Res, 2016, 44(2): 582-594.

[0180] In certain examples, the cancer is selected from bladder cancer, breast cancer, colon cancer, rectal cancer, endometrial cancer, kidney cancer, leukemia, lung cancer, melanoma, non-Hodgkin lymphoma, pancreatic cancer, prostate cancer, and thyroid cancer.

[0181] The compounds disclosed herein can be used alone in the treatment of each of the foregoing conditions or can be used to provide additive or potentially synergistic effects with certain existing chemotherapies, radiation, biological or immunotherapeutics (including monoclonal antibodies) and vaccines. The compounds disclosed herein may be useful for restoring effectiveness of certain existing chemotherapies and radiation and or increasing sensitivity to certain existing chemotherapies and / or radiation.

[0182] Coste et al. report LRH-1 -mediated glucocorticoid synthesis in enterocytes protects against inflammatory bowel disease. PNAS, 2007. 104 (32) 13098-13103. See also Fernandez- Marcos et al. Emerging actions of the nuclear receptor LRH-1 in the gut, Biochim Biophys Acta. 2011 August; 1812(8): 947-955. Mueller et al. The nuclear receptor LRH-1 critically regulates extra-adrenal glucocorticoid synthesis in the intestine, Journal of Experimental Medicine Sep 2006, 203 (9) 2057-2062. Thus, in certain examples, this disclosure relates to methods to prevent or treat gut, intestinal, and colonic inflammation, comprising administrating a compound disclosed herein in a therapeutically effective amount to a subject in need thereof. In certain examples, the subject is at risk of, exhibiting symptoms of, or diagnosed with intestinal and colonic inflammation.

[0183] In another aspect, the disclosure relates to methods to prevent or treat inflammatory bowel diseases (IBD), comprising administrating a compound disclosed herein in a therapeutically effective amount to a subject in need thereof. In certain examples, the subject is at risk of, exhibiting symptoms of, or diagnosed with inflammatory bowel diseases (IBD). As outlined in Mays et al., Cell Chemical Biology, 29: 1174-1186 (2022), agonists of LRH-1 improve outcomes in T- cell transfer mouse model of colitis, resulting in heightened expression of genes involved in steroidogenesis and lower expression of inflammatory cytokine genes in the gut.

[0184] Optionally, the disclosure relates to methods to prevent or treat diseases characterized by inflammation of the bowel, including celiac disease, immune checkpoint inhibitor colitis, IgA nephropathy, Crohn's disease, ulcerative colitis, regional enteritis, granulomatous enteritis, distal ileitis, regional ileitis, terminal ileitis, and others. In some cases, the disclosure relates to methods to protect tissues from inflammatory bowel disease, including ulcerative colitis and Crohn’s disease.

[0185] In another aspect, the disclosure relates to methods to prevent or treat Crohn's disease, comprising administrating a compound disclosed herein in a therapeutically effective amount to a subject in need thereof. In certain examples, the subject is at risk of, exhibiting symptoms of, or diagnosed with Crohn's disease.

[0186] In another aspect, the disclosure relates to methods to prevent or treat colitis or ulcerative colitis, comprising administrating a compound disclosed herein in a therapeutically effective amount to a subject in need thereof. In certain examples, the subject is at risk of, exhibiting symptoms of, or diagnosed with colitis or ulcerative colitis.

[0187] Overweight and obesity are increasingly common conditions in the world. Doctors measure body mass index (BMI) to screen for obesity. Obesity is a serious medical condition that can cause complications such as metabolic syndrome, high blood pressure, atherosclerosis, heart disease, diabetes, high serum cholesterol, cancers, and sleep disorders. Thus, there is a need to reduce obesity.

[0188] Fatty liver, or hepatic steatosis, is a term that describes the buildup of fat in the liver. Excessive alcohol use causes fat to accumulate, damages the liver, and cirrhosis may develop. Nonalcoholic fatty liver disease (NAFLD) is a fatty liver disease associated with obesity- related disorders, such as type-2 diabetes and metabolic syndrome, occurring in people who drink little or no alcohol. Nonalcoholic steatohepatitis (NASH) is a more advanced and severe subtype of NAFLD where steatosis is complicated by liver-cell injury and inflammation, with or without fibrosis. NASH can be severe and can lead to cirrhosis, in which the liver is permanently damaged and scarred and no longer able to work properly. Insulin resistance altered lipid storage and metabolism, accumulation of cholesterol within the liver, oxidative stress resulting in increased hepatic injury, and bacterial translocation secondary to disruption of gut microbiota have all been implicated as important co-factors contributing to progression of NASH. Due to the growing epidemic of obesity and diabetes, NASH is projected to become the most common cause of advanced liver disease and the most common indication for liver transplantation.

[0189] Lee et al. report dilauroyl phosphatidylcholine (DLPC) is an LRH-1 agonist ligand in vitro. DLPC treatment induces bile acid biosynthetic enzymes in mouse liver, increases bile acid levels, and lowers hepatic triglycerides and serum glucose. DLPC treatment also decreases hepatic steatosis and improves glucose homeostasis in two mouse models of insulin resistance. Nature volume 474, pages 506-510 (2011). Sahini et al. report differentially expressed genes (DEGs) were identified which are mechanistically linked to lipid droplet (LD) formation in hepatocytes. LD-associated DEGs frequently regulated in patient samples were identified. Liver-receptor homolog- 1 (NR5A2), was commonly repressed among patients examined. Translational Research, 177: 41- 69 (2016).

[0190] In certain examples, this disclosure relates to methods to prevent or treat hepatic steatosis or metabolic syndrome, comprising administrating compounds disclosed herein in a therapeutically effective amount to a subject in need thereof. In certain examples, the subject is at risk of, exhibiting symptoms of, or diagnosed with nonalcoholic fatty liver disease (NAFLD). In certain examples, the subject is at risk of, exhibiting symptoms of, or diagnosed with nonalcoholic steatohepatitis (NASH). In certain examples, the subject is at risk of, exhibiting symptoms of, or diagnosed with alcoholic liver disease (ALD). In certain examples, the subject is at risk of, exhibiting symptoms of, or diagnosed with alcoholic steatohepatitis (ASH).

[0191] In certain examples, a subject is at risk of NAFLD due to obesity, insulin resistance, an enlarged liver, signs of cirrhosis, or abnormal levels of liver enzymes, triglycerides and / or cholesterol. Signs of insulin resistance include darkened skin patches over your knuckles, elbows, and knees. Signs of cirrhosis include jaundice, a condition that causes your skin and whites of your eyes to turn yellow. A sign of NAFLD or NASH includes blood test showing increased levels of the liver enzymes alanine aminotransferase (ALT) and aspartate aminotransferase (AST). An enlarged liver or an abnormal amount of fat in a liver may be identified by ultrasound, computerized tomography (CT) scans, magnetic resonance imaging or combinations thereof. A liver biopsy may be used to detect liver inflammation and damage to diagnose NASH.

[0192] Metabolic syndrome is typically diagnosed in the presence of three or more of the following medical issues: large waist size, e.g., 40 inches or more, high triglycerides e.g., triglyceride level of 150 mg / dL or higher, low levels of HDL cholesterol less than 50 mg / dL, high blood pressure, e.g., 130 / 85 mmHg or higher, and high blood glucose (or blood sugar) levels, a fasting blood sugar level of 100 mg / dL or higher.

[0193] In another aspect, the disclosure relates to methods to control or reduce the serum cholesterol level, comprising administrating compounds disclosed herein in a therapeutically effective amount to a subject in need thereof. In certain examples, the subject has a borderline high serum cholesterol level, 200-239 mg / dL. In certain examples, the subject has a high serum cholesterol level, >240 mg / dL. In certain examples, the subject is at risk of, exhibiting symptoms, or diagnosed with hypercholesterolemia.

[0194] In another aspect, the disclosure relates to methods to prevent or treat hepatic steatosis, comprising administrating a compound disclosed herein in a therapeutically effective amount to a subject in need thereof. In certain examples, the subject is at risk of, exhibiting symptoms of, or diagnosed with alcoholic liver disease (ALD). In certain examples, the subject is at risk of, exhibiting symptoms of, or diagnosed with alcoholic steatohepatitis (ASH). In certain examples, the subject is at risk of, exhibiting symptoms of, or diagnosed with nonalcoholic fatty liver disease (NAFLD). In certain examples, the subject is at risk of, exhibiting symptoms of, or diagnosed with nonalcoholic steatohepatitis (NASH). In certain examples, a subject is at risk of NAFLD due to obesity, insulin resistance, an enlarged liver, signs of cirrhosis, or abnormal levels of liver enzymes, triglycerides and / or cholesterol. Signs of insulin resistance include darkened skin patches over your knuckles, elbows, and knees. Signs of cirrhosis include jaundice, a condition that causes your skin and whites of your eyes to turn yellow. A sign of NAFLD or NASH includes blood test showing increased levels of the liver enzymes alanine aminotransferase (ALT) and aspartate aminotransferase (AST). An enlarged liver or an abnormal amount of fat in a liver may be identified by ultrasound, computerized tomography (CT) scans, magnetic resonance imaging or combinations thereof. A liver biopsy may be used to detect liver inflammation and damage to diagnose NASH. The precise therapeutically effective amount of the compounds of this disclosure will depend on a number of factors. There are variables inherent to the compounds including, but not limited to, the following: molecular weight, absorption, bioavailability, distribution in the body, tissue penetration, half-life, metabolism, protein binding, and excretion. These variables determine what dose of compound needs to be administered in a sufficient percentage and for a sufficient amount of time to have the desired effect on the condition being treated (e.g., neoplasm). The duration of drug exposure will be limited only by the compound half-life, and side effects from treatment requiring cessation of dosing. The amount of compound administered will also depend on factors related to patients and disease including, but not limited to, the following: the age, weight, concomitant medications, and medical condition of the subject being treated, the precise condition requiring treatment and its severity, the nature of the formulation, and the route of administration. Ultimately the dose will be at the discretion of the attendant physician or veterinarian. Typically, the compound disclosed herein will be given for treatment in the range of 0.01 to 30 mg / kg body weight of recipient (mammal) per day or per dose or per cycle of treatment and more usually in the range of 0.1 to 10 mg / kg body weight per day or per dose or per cycle of treatment. Thus, for an adult human being treated for a condition, the actual amount per day or per dose or per cycle of treatment would usually be from 1 to 2000 mg and this amount may be given in a single or multiple doses per day or per dose or per cycle of treatment. Dosing regimens may vary significantly and will be determined and altered based on clinical experience with the compound. The full spectrum of dosing regimens may be employed ranging from continuous dosing (with daily doses) to intermittent dosing. A therapeutically effective amount of a pharmaceutically acceptable salt of a compound disclosed herein may be determined as a proportion of the therapeutically effective amount of the compound as the free base.

[0195] The methods of treating or preventing diseases or conditions associated with LRH-1 (e.g., diabetes, cancer, or cardiovascular disease) in a subject can further comprise administering to the subject one or more additional agents. The one or more additional agents and the compounds described herein or pharmaceutically acceptable salts or prodrugs thereof can be administered in any order, including concomitant, simultaneous, or sequential administration. Sequential administration can be administration in a temporally spaced order of up to several days apart. The methods can also include more than a single administration of the one or more additional agents and / or the compounds described herein or pharmaceutically acceptable salts or prodrugs thereof. The administration of the one or more additional agents and the compounds described herein or pharmaceutically acceptable salts or prodrugs thereof can be by the same or different routes and concurrently or sequentially.

[0196] Additional therapeutic agents include, but are not limited to, chemotherapeutic agents, antibodies, antivirals, steroidal and non-steroidal anti-inflammatories, conventional immunotherapeutic agents, cytokines, chemokines, and / or growth factors. The additional therapeutic agents can be biomolecules.

[0197] A chemotherapeutic agent, chemotherapy agent, chemotherapeutic, anti-cancer agent, or the like is a compound or composition effective in inhibiting or arresting the growth of an abnormally growing cell. Such agents can be recognized to aid in the treatment of a cancer. Thus, such an agent may be used therapeutically to treat cancer as well as other diseases marked by abnormal cell growth. Illustrative examples of chemotherapeutic compounds include, but are not limited to, bexarotene, gefitinib, erlotinib, gemcitabine, paclitaxel, docetaxel, topotecan, irinotecan, temozolomide, carmustine, vinorelbine, capecitabine, leucovorin, oxaliplatin, bevacizumab, cetuximab, panitumumab, bortezomib, oblimersen, hexamethylmelamine, ifosfamide, CPT-11, deflunomide, cycloheximide, dicarbazine, asparaginase, mitotant, vinblastine sulfate, carboplatin, colchicine, etoposide, melphalan, 6- mercaptopurine, teniposide, vinblastine, antibiotic derivatives (e.g. anthracyclines such as doxorubicin, liposomal doxorubicin, and diethylstilbestrol doxorubicin, bleomycin, daunorubicin, and dactinomycin); antiestrogens (e.g., tamoxifen); antimetabolites (e.g., fluorouracil (FU), 5-FU, methotrexate, floxuridine, interferon alpha-2B, glutamic acid, plicamycin, mercaptopurine, and 6-thioguanine); cytotoxic agents (e.g., carmustine, BCNU, lomustine, CCNU, cytosine arabinoside, cyclophosphamide, estramustine, hydroxyurea, procarbazine, mitomycin, busulfan, cisplatin, vincristine and vincristine sulfate); hormones (e.g., medroxyprogesterone, estramustine phosphate sodium, ethinyl estradiol, estradiol, megestrol acetate, methyltestosterone, diethylstilbestrol diphosphate, chlorotrianisene, and testolactone); nitrogen mustard derivatives (e.g., mephalen, chlorambucil, mechlorethamine (nitrogen mustard) and thiotepa); and steroids (e.g., bethamethasone sodium phosphate).

[0198] In some cases, contemplated examples of chemotherapeutic agents include the following molecules or derivatives such as temozolomide, carmustine, bevacizumab, procarbazine, lomustine, vincristine, gefitinib, erlotinib, cisplatin, carboplatin, oxaliplatin, 5- fluorouracil, gemcitabine, tegafur, raltitrexed, methotrexate, cytosine arabinoside, hydroxyurea, adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin, mithramycin, vinblastine, vindesine, vinorelbine, paclitaxel, taxol, docetaxel, etoposide, teniposide, amsacrine, topotecan, camptothecin, bortezomib, anagrelide, tamoxifen, toremifene, raloxifene, droloxifene, idoxifene, fulvestrant, bicalutamide, flutamide, nilutamide, cyproterone, goserelin, leuprorelin, buserelin, megestrol, anastrozole, letrozole, vorozole, exemestane, finasteride, marimastat, trastuzumab, cetuximab, dasatinib, imatinib, combretastatin, thalidomide, azacitidine, azathioprine, capecitabine, chlorambucil, cyclophosphamide, cytarabine, daunorubicin, doxifluridine, epothilone, irinotecan, mechlorethamine, mercaptopurine, mitoxantrone, pemetrexed, tioguanine, valrubicin and / or lenalidomide or combinations thereof such as cyclophosphamide, methotrexate, 5 -fluorouracil (CMF); doxorubicin, cyclophosphamide (AC); mustine, vincristine, procarbazine, prednisolone (MOPP); adriamycin, bleomycin, vinblastine, dacarbazine (ABVD); cyclophosphamide, doxorubicin, vincristine, prednisolone (CHOP); bleomycin, etoposide, cisplatin (BEP); epirubicin, cisplatin, 5 -fluorouracil (ECF); epirubicin, cisplatin, capecitabine (ECX); methotrexate, vincristine, doxorubicin, cisplatin (MV AC).

[0199] Any of the aforementioned therapeutic agents can be used in any combination with the compositions described herein. Combinations are administered either concomitantly (e.g., as an admixture), separately but simultaneously (e.g., via separate intravenous lines into the same subject), or sequentially (e.g., one of the compounds or agents is given first followed by the second). Thus, the term combination is used to refer to concomitant, simultaneous, or sequential administration of two or more agents. The term “combination with” when used to describe administration with an additional treatment means that the agent may be administered prior to, together with, or after the additional treatment, or a combination thereof.

[0200] Optionally, a compound or therapeutic agent as described herein may be administered in combination with a radiation therapy, an immunotherapy, a gene therapy, or a surgery.

[0201] The methods and compounds as described herein are useful for both prophylactic and therapeutic treatment. For prophylactic use, a therapeutically effective amount of the compounds and compositions or pharmaceutically acceptable salts thereof as described herein are administered to a subject prior to onset (e.g., before obvious signs of an LRH-l-related disease), during early onset (e.g., upon initial signs and symptoms of an LRH-l-related disease), or after the development of an LRH-l-related disease. Prophylactic administration can occur for several days to years prior to the manifestation of symptoms of an LRH-l-related disease. Therapeutic treatment involves administering to a subject a therapeutically effective amount of the compounds and compositions or pharmaceutically acceptable salts thereof as described herein after an LRH-l-related disease is diagnosed.

[0202] The compounds described herein are also useful in modulating LRH-1 in a cell. The methods for modulating LRH-1 activity in a cell includes contacting a cell with a therapeutically effective amount of one or more of the compounds as described herein. Optionally, the contacting is performed in vivo. Optionally, the contacting is performed in vitro.

[0203] The methods herein for prophylactic and therapeutic treatment optionally comprise selecting a subject with or at risk of developing an LRH-1 -related disease. A skilled artisan can make such a determination using, for example, a variety of prognostic and diagnostic methods, including, for example, a personal or family history of the disease or condition, clinical tests (e.g., imaging, biopsy, genetic tests), and the like. Optionally, the methods herein can be used for preventing relapse of cancer in a subject in remission (e.g., a subject that previously had cancer).

[0204] VI. Kits

[0205] Also provided herein are kits for treating or preventing an LRH-l-related disease (e.g., diabetes, cancer, and / or cardiovascular disease) in a subject. A kit can include any of the compounds or compositions described herein. For example, a kit can include one or more compounds of Formula I. A kit can further include one or more additional agents, such as one or more anti-inflammatory agents and / or chemotherapeutic agents. A kit can include an oral formulation of any of the compounds or compositions described herein. A kit can include an intravenous formulation of any of the compounds or compositions described herein. A kit can additionally include directions for use of the kit (e.g., instructions for treating a subject), a container, a means for administering the compounds or compositions (e.g., a syringe), and / or a carrier. The kits of this disclosure can optionally include a carrier container being compartmentalized to receive in close confinement one or more containers such as vials, tubes, and the like, each of the containers comprising one of the separate elements to be used in the method.

[0206] The compounds as described in this disclosure for use in treating patients may be delivered in a pharmaceutical package or kit to doctors and patients. Such packaging can be designed and manufactured to improve patient convenience and compliance with the treatment plan. Optionally, the packaging comprises paper (cardboard) or plastic. In some cases, the kit or pharmaceutical package further comprises instructions for use (e.g., for administering according to a method as described herein).

[0207] In some examples, a pharmaceutical package or kit includes unit dose forms of a compound described herein. In some embodiments, the pharmaceutical package or kit further comprises unit dose forms of one or more of a chemotherapeutic agent, a cytotoxic agent, a radiotherapeutic agent, or an immunotherapeutic agent.

[0208] Optionally, kits are provided for producing a single-dose administration unit. In certain embodiments, kits containing one or more containers of a formulation described in this disclosure are included. Optionally, the kit or pharmaceutical package includes doses suitable for multiple days of administration, such as one week, one month, or three months.

[0209] As used herein the terms treatment, treat, or treating refer to a method of reducing one or more symptoms of a disease or condition. Thus, in the disclosed method, treatment can refer to a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% reduction in the severity of one or more symptoms of the disease or condition. For example, a method for treating a disease is considered to be a treatment if there is a 10% reduction in one or more symptoms or signs (e.g., size of the tumor or rate of tumor growth) of the disease in a subject as compared to a control. As used herein, control refers to the untreated condition (e.g., the tumor cells not treated with the compounds and compositions described herein). Thus, the reduction can be a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any percent reduction in between 10% and 100% as compared to native or control levels. It is understood that treatment does not necessarily refer to a cure or complete ablation of the disease, condition, or symptoms of the disease or condition.

[0210] As used herein, the terms prevent, preventing, and prevention of a disease or disorder refer to an action, for example, administration of a composition or therapeutic agent, that occurs before or at about the same time a subject begins to show one or more symptoms of the disease or disorder, which inhibits or delays onset or severity of one or more symptoms of the disease or disorder. It is not intended that the present disclosure be limited to complete prevention. In some examples, the onset is delayed, or the severity of the disease is reduced.

[0211] As used herein, references to decreasing, reducing, or inhibiting include a change of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater as compared to a control level. Such terms can include, but do not necessarily include, complete elimination.

[0212] As used herein, subject means any animal, including mammals and non-mammals. Mammals include, for example, humans; non-human primates, e.g., apes and monkeys; cattle; horses; sheep; rats; mice; pigs; and goats. Non-mammals include, for example, fish and birds. In some examples, subject refers to a human patient, livestock, rodent, monkey, or domestic pet.

[0213] Throughout this application, various publications are referenced. The disclosures of these publications in their entireties are hereby incorporated by reference into this application. The examples below are intended to further illustrate certain aspects of the methods and compositions described herein, and are not intended to limit the scope of the claims.

[0214] EXAMPLES

[0215] Example 1: Synthesis

[0216] All reactions were carried out in flame-dried glassware, equipped with a stir bar and under a nitrogen atmosphere with dry solvents under anhydrous conditions, unless otherwise noted. Solvents used in anhydrous reactions were purified by passing over activated alumina and storing under argon. Yields refer to chromatographically and spectroscopically (JH NMR) homogenous materials, unless otherwise stated. Reagents were purchased at the highest commercial quality and used without further purification, unless otherwise stated, n- Butyllithium (n-BuLi) was used as a 2.5 M solution in hexanes (Aldrich), was stored at 4 °C and titrated prior to use. Organic solutions were concentrated under reduced pressure on a rotary evaporator using a water bath. Chromatographic purification of products was accomplished using forced-flow chromatography on 230-400 mesh silica gel. Thin-layer chromatography (TLC) was performed on 250 pm SiliCycle silica gel F-254 plates. Visualization of the developed chromatogram was performed by fluorescence quenching or by staining using KMnO4, / >-anisaldehyde, or ninhydrin stains.

[0217] JH and13C NMR spectra were recorded on a Bruker Avance III HD 600 equipped with cryo-probe (600 MHz), INOVA 600 (600 MHz), INOVA 500 (500 MHz), INOVA 400 (400 MHz), VNMR 400 (400 MHz), and are internally referenced to residual protio solvent signals. Data for 'H NMR are reported as follows: chemical shift (ppm), multiplicity (s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, dd = doublet of doublets, dt = doublet of triplets, ddd= doublet of doublet of doublets, dtd= doublet of triplet of doublets, b = broad, etc.), coupling constant (Hz), integration, and assignment, when applicable. Data for decoupled13C NMR are reported in terms of chemical shift and multiplicity when applicable. Liquid Chromatography Mass Spectrometry (LC-MS) was performed on an Agilent 6120 mass spectrometer with an Agilent 1220 Infinity liquid chromatography inlet. Preparative High- Pressure Liquid chromatography (Prep-HPLC) was performed on an Agilent 1200 Infinity Series chromatograph using an Agilent Prep-C18 30 x 250 mm 10 pm column, or an Agilent Prep-C18 21.2 x 100 mm, 5 pm column.

[0218] Synthesis of 6N-10CA has been previously reported (Cato et al., J Med Chem 65(9): 6888-6902 (2022)). Synthetic methods for RJW100 have been previously published (Mays et al., J Med Chem 62(24): 11022-11034 (2019)). 10CA was synthesized as previously described (Flynn et al., ACS Med Chem Let 9(10): 1051-1056 (2018)).

[0219] 10-(6-(methoxymethoxy)-3-phenyl-3a-(l-phenylvinyl)-l, 3a, 4,5,6, 6a-hexahydropentalen-

[0220] 2-yl)decan-l-ol (SI):

[0221] The title compound was prepared according to the reported procedure and the NMR data were consistent with those previously reported (Flynn et al., ACS Med Chem Let 9(10): 1051-1056 (2018)).

[0222] 10-(6-(methoxymethoxy)-3-phenyl-3a-(l-phenylvinyl)-l, 3a, 4,5,6, 6a-hexahydropentalen-

[0223] 2-yl)decanoic acid (S2):

[0224] The title compound was prepared according to the reported procedure and the NMR data were consistent with those previously reported (Flynn et al., ACS Med Chem Let 9(10): 1051-1056 (2018)).

[0225] 10-(6-(methoxymethoxy)-3-phenyl-3a-(l-phenylvinyl)-l, 3a, 4,5,6, 6a-hexahydropentalen-

[0226] 2-yl)-l-(piperazin-l-yl)decan-l-one (S3):

[0227] A flame-dried vial was charged with a stir bar and l,l’-carbinyliimidazole (CDI). The vial was placed under vacuum and backfilled 3x with nitrogen before the addition of S2 (27.6 mg, 0.05 mmol, 1.0 equiv) in dry THF. The reaction stirred at room temperature for 1.5 hours. Piperizine (94.7 mg, 1.1 mmol, 20 equiv) was added to the solution and the reaction proceeded for 16 hours. The reaction mixture was then diluted in ethyl acetate and washed with water twice. The organic layer was then dried with magnesium sulfate, filtered, and concentrated via rotary evaporation. The resulting residue was purified by flash chromatography (5-10% MeOH / DCM) to afford the title compound (10.9 mg, 35% yield) as a clear oil.1H NMR (600 MHz, CDCh) 5 7.34 - 7.32 (m, 3H), 7.29 (dd, J= 7.1, 1.2 Hz, 2H), 7.26 - 7.23 (m, 3H), 7.22 - 7.19 (m, 2H), 5.05 (d, J= 1.4 Hz, 1H), 5.00 (d, J= 1.5 Hz, 1H), 4.60 - 4.57 (m, 2H), 3.80 - 3.75 (m, 3H), 3.62 (s, 2H), 3.30 (s, 3H), 3.01 (t, J= 8.9 Hz, 3H), 2.42 (dd, J= 9.3, 1.7 Hz, 1H), 2.32 (dt, J= 15.6, 8.4 Hz, 4H), 2.08 - 2.06 (m, 1H), 2.06 - 1.98 (m, 4H), 1.77 - 1.71 (m, 1H), 1.68 - 1.57 (m, 6H), 1.34 - 1.26 (m, 10H).13C NMR (151 MHz, CDCh) 5 171.74, 154.55, 144.14, 141.34, 139.25, 137.50, 129.65, 127.85, 127.65, 126.63, 126.58, 114.89, 94.76, 86.75, 69.10, 55.18, 52.79, 49.46, 40.54, 33.17, 32.44, 31.46, 29.74, 29.71, 29.44, 29.43, 29.41, 27.86, 26.99, 25.22. LRMS (ESI, APCI) m / z: calculated for C38H53N2O3 [M+H]+ 585.8, found 584.8.

[0228] 10-(6-hydroxy-3-phenyl-3a-(l-phenylvinyl)-l, 3a, 4,5,6, 6a-hexahydropentalen-2-yl)-l- (piperazin-l-yl)decan-l-one (Pip):

[0229] In a reaction vial, S3 (5.5 mg, 0.01 mmol, 1.0 equiv) was dissolved in acetonitrile. A few drops of hydrochloric acid were added at room temperature and the reaction proceeded until complete by TLC (30 minutes- 2 h). The crude mixture was concentrated and purified by flash chromatography (10% MeOH / DCM) to afford the product 2.1 mg, 42% yield). ’H NMR (600 MHz, CDCh) 5 7.37 (ddd, J= 5.4, 3.0, 1.0 Hz, 3H), 7.34 - 7.31 (m, 2H), 7.28 - 7.27 (m, 3H), 7.21 (dt, J= 7.1, 1.4 Hz, 2H), 5.09 (d, J= 1.5 Hz, 1H), 5.01 (d, J= 1.4 Hz, 1H), 3.97 (s, 3H), 3.85 (m, 2H), 3.78 - 3.62 (m, 5H), 3.57 - 3.48 (m, 3H), 3.31 (s, 1H), 3.29 - 3.18 (m, 2H), 2.40 - 2.27 (m, 6H), 2.15 (d, J= 4.8 Hz, 1H), 2.13 - 2.08 (m, 4H), 2.08 - 2.02 (m, 2H), 1.76 - 1.66 (m, 2H), 1.37 - 1.29 (m, 5H).13C NMR (151 MHz, CDCh) 5 154.55, 144.21, 141.14, 139.19, 137.37, 129.72, 127.77, 127.67, 126.70, 126.63, 115.09, 82.08, 69.35, 55.75, 53.49, 40.25, 34.03, 33.37, 32.13, 31.96, 29.74, 29.70, 29.67, 29.63, 29.61, 29.54, 29.41, 29.39, 29.33, 29.30, 29.28, 27.82, 27.76, 22.74. LRMS (ESI, APCI) m / z: calculated for C36H50N2O2 [M+H]+ 542.4, found 542.8. 10-(6-hydroxy-3-phenyl-3a-(l-phenylvinyl)-l, 3a, 4,5,6, 6a-hexahydropentalen-2- yl)decanamide (Am):

[0230] MOMO-protected 10CA was prepared as described before (S2) and added to a scintillation vial (52 mg, 0.1 mmol, 1.0 equiv), equipped with a magnetic stir bar and dissolved in THF. Carbonyldiimidazole (35 mg, 0.2 mmol, 2.0 equiv) was added in small portions (effervesces) and stirred until completion was detected by LCMS (small portions of the activated ester was quenched with methanol to visualize reaction progress). Ammonia (as a 0.5 M solution in dioxane, 100 pL, excess) was added to the reaction mixture, which was stirred at room temperature until completion as detected by TLC and LCMS. The reaction mixture was concentrated and the crude residue was passed through silica (50-100% EtOAc / Hex eluent), concentrated, and dissolved in acetonitrile. A magnetic stirrer was added, and two drops of concentrated HC1 was added to the solution. After deprotection was complete (approximately 1 hour), the reaction mixture was concentrated and purified on silica (50-100% EtOAc / Hex eluent) to give the title compound (10.1 mg, 54% yield).1H NMR (600 MHz, CDC13) 5 7.40 - 7.15 (m, 10H), 5.35 (d, J = 36.9 Hz, 2H), 5.06 (s, 1H), 4.99 (s, 1H), 3.95 (s, 1H), 2.35 (dd, J = 16.8, 9.3 Hz, 1H), 2.29 (d, J = 8.8 Hz, 1H), 2.21 (t, J = 7.6 Hz, 2H), 2.12 - 1.95 (m, 4H), 1.74 - 1.60 (m, 6H), 1.42 - 1.14 (m, 11H).13C NMR (126 MHz, CDC13) 5 175.2, 155.0, 143.7, 141.9, 138.9, 137.8, 129.7, 127.8, 127.7, 127.6, 126.7, 126.6, 116.4, 86.0, 70.1, 54.0, 40.2, 35.9, 34.0, 32.1, 29.6, 29.6, 29.3, 29.2, 29.2, 27.3, 25.5. LRMS (ESI, APCI) m / z: calculated for C32H41NO2 [M+H]+ 472.3, found 471.8.

[0231] (10-(6-hydroxy-3-phenyl-3a-(l-phenylvinyl)-l,3a,4,5,6,6a-hexahydropentalen-2- yl)decanoyl)serine (Ser): A flame-dried vial was charged with a stir bar and l,l’-carbinyliimidazole (CDI) (30 mg, 0.14 mmol, 2.5 equiv). The vial was placed under vacuum and backfilled 3 times with nitrogen before the addition of the S2 (35 mg, 0.07 mmol, 1.0 equiv) dissolved in dry THF (0.1 M). The reaction was stirred at room temperature for 1.5 hours. The L-Serine methyl ester (16 mg, 0.13 mmol, 2.0 equiv) was added to the solution and the reaction proceeded for 16 hours. The reaction mixture was then diluted in ethyl acetate and washed with water twice. The organic layer was then dried with magnesium sulfate, filtered, and concentrated via rotary evaporation. The resulting oil was dissolved in acetonitrile (0.1 M). A few drops of hydrochloric acid was added at room temperature and the reaction proceeded until complete by TLC (30 minutes- 2 h). The crude mixture was concentrated and purified by flash chromatography (10% MeOH / DCM) to afford the product (2.1 mg, 42% yield). * H NMR (500 MHz, CDC13) 8 7.42 - 7.07 (m, 10H), 6.76 (s, 1H), 5.05 (s, 1H), 4.99 (s, 1H), 4.57 (s, 1H), 4.10 - 3.77 (m, 5H), 2.39 - 2.18 (m, 4H), 2.14 - 1.94 (m, 6H), 1.77 - 1.55 (m, 5H), 1.38 - 1.09 (m, 11H).13C NMR (151 MHz, CDCh) 5 154.58, 144.10, 141.15, 139.31, 137.36, 129.71, 127.82, 127.78, 127.69, 126.75, 126.66, 115.00, 44.80, 40.20, 32.11, 31.94, 29.72, 29.57, 29.37, 29.21, 27.66, 23.13, 22.71, 14.13. LRMS (ESI, APCI) m / z: calculated for C35H44NO5 [M-H]- 558.3, found 558.3.

[0232] A- hydroxy- 10-(6-hydroxy-3-phenyl-3a-(l-phenylvinyl)- 1 , 3a, 4, 5, 6,6a- hexahydropentalen-2-yl)decanamide (HA) :

[0233] A flame-dried vial was charged with a stir bar and l,l’-carbinyliimidazole (CDI) (42 mg, 0.18 mmol, 2 equiv). The vial was placed under vacuum and backfilled 3 time with nitrogen before the addition of the S2 (49 mg, 0.09 mmol, 1.0 equiv) dissolved in dry THF (0.1 M). The reaction stirred at room temperature for 1.5 hours. The hydroxylamine hydrochloride (32 mg, 0.4 mmol, 5.0 equiv) was added to the solution and the reaction proceeded for 16 hours. The reaction mixture was then diluted in ethyl acetate and washed with water twice. The organic layer was then dried with magnesium sulfate, filtered, and concentrated via rotary evaporation. The resulting oil was dissolved in acetonitrile (0.1 M). A few drops of hydrochloric acid were added at room temperature and the reaction proceeded until complete by TLC (30 minutes- 2 h). The crude mixture was concentrated and purified by flash chromatography (10% MeOH / DCM) to afford the product (12.9 mg, 66% yield).1H NMR (600 MHz, CDCh) 5 7.38 - 7.30 (m, 4H), 7.28 - 7.26 (m, 3H), 7.22 (dt, J= 6.4, 1.4 Hz, 3H), 5.09 (d, J= 1.5 Hz, 1H), 5.02 (d, J= 1.5 Hz, 1H), 3.99 - 3.95 (m, 1H), 2.54 - 2.45 (m, 2H), 2.40 - 2.30 (m, 3H), 2.22 - 2.16 (m, 2H), 2.13 - 1.99 (m, 6H), 1.76 - 1.62 (m, 4H), 1.62 - 1.54 (m, 2H), 1.39 - 1.27 (m, 10H).13C NMR (151 MHz, CDCh) 5 154.59, 144.16, 141.13, 139.26, 137.39, 129.71, 127.79, 127.74, 127.66, 126.69, 126.63, 115.02, 82.16, 69.35, 55.83, 40.21, 34.05, 32.09, 29.98, 29.71, 29.65, 29.60, 29.45, 29.23, 29.18, 29.10, 29.07, 29.04, 27.71. LRMS (ESI, APCI) m / z: calculated for C32H42NO3 [M+H]+ 488.3, found 488.6.

[0234] 9-(6-(methoxymethoxy)-3-phenyl-3a-(l-phenylvinyl)-l, 3a, 4,5,6, 6a-hexahydropentalen-2- yl)nonan-l-ol (S4):

[0235] Hexahydropentalene formation was accomplished through slight modification of Whibty’s procedure. Prior to cyclization, all non-volatile reagents were dried by azeotropic removal of water using benzene. A dry three-neck round bottom flask backfilled with nitrogen three times (3x) containing bis(cyclopentadienyl)zirconium(IV) dichloride (1.403 g, 4.8 mmol, 1.2 equiv) was dissolved in anhydrous, degassed tetrahydrofuran (THF, 16 mL) and cooled to -78 °C. The resulting solution was treated with n-BuLi (2.5 M in hexanes, 3.8 mL, 9.6 mmol, 2.4 equiv). The light yellow solution stirred for 45 minutes at -78 °C. A solution of te / 7- butyldimethyl((7-phenylhept-l-en-6-yn-3-yl)oxy)silane (921.2 mg, 4.0 mmol, 1 equiv) was added dropwise to the solution in anhydrous THF (16 mL). The solution stirred at -78 °C for 45 minutes before the bath was removed and the solution stirred at room temperature for 2.5 hours turning a salmon-colored mixture. The mixture was cooled to -78 °C before the addition of / c / 7-butyl(( I O, I O-dibromodecyl)oxy)diphenylsilane (2.45 g, 4.4 mmol, 1.1 equiv) in anhydrous THF (16 mL). followed immediately by freshly prepared lithium diisopropylamide (LD A, 4.4 mmol, 1.1 equiv) in anhydrous THF (16 mL). The red-solution stirred for 15 minutes before the addition of freshly prepared lithium phenylacetylide (1.5 mL, 14.4 mmol, 3.6 equiv) in anhydrous THF (16 mL). The reaction mixture continued at -78 °C for 1.5 h before being quenched with methanol (24 mL) and saturated aqueous sodium bicarbonate (24 mL). The resulting slurry was allowed to warm to room temperature before being diluted with ethyl acetate and being washed with water. The aqueous layer was reextracted with ethyl acetate two times (2x) before the combined organic layers were then washed with brine (2x). The resulting organic layer was dried with magnesium sulfate, filtered and concentrated via rotary evaporation. The resulting yellow slurry was then pushed through a short plug of silica (100% ethyl acetate eluent) and concentrated in vacuo. The crude product was then dissolved in THF and treated with tetrabutylammonium fluoride hydrate (3.0 equiv) overnight. The reaction mixture was concentrated and the diastereomers were purified and separated by careful silica gel chromatography (20-40% EtOAc / Hexanes eluent) to afford the exo diastereomer in a RATIO (Flynn et al., 2018). ’H NMR (400 MHz, CDCh) 5 7.39 - 7.29 (m, 6H), 7.27 - 7.21 (m, 2H), 5.07 (d, J= 1.5 Hz, 1H), 5.02 (d, J= 1.5 Hz, 1H), 4.61 (d, J= 1.5 Hz, 1H), 3.66 (td, J = 6.6, 3.6 Hz, 3H), 3.33 (s, 3H), 2.46 - 2.41 (m, 1H), 2.34 (dd, J= 16.8, 9.2 Hz, 1H), 2.21 (td, J= 7.1, 2.7 Hz, 1H), 2.10 (d, J= 1.7 Hz, 1H), 2.08 - 2.01 (m, 4H), 1.72 - 1.63 (m, 2H), 1.61 - 1.48 (m, 8H), 1.40 - 1.20 (m, 10H).13C NMR (101 MHz, CDCh) 5 154.32, 143.95, 141.19, 139.03, 138.09, 137.30, 134.62, 128.92, 127.65, 127.47, 126.45, 126.39, 114.73, 94.52, 86.52, 68.90, 62.90, 55.38, 52.08, 39.45, 32.62, 32.58, 32.24, 31.25, 29.53, 29.45, 29.29, 29.20, 29.16, 29.13, 27.62, 25.52.

[0236] 9-(6-(methoxymethoxy)-3-phenyl-3a-(l-phenylvinyl)-l, 3a, 4,5,6, 6a-hexahydropentalen-2- yl)nonyl methanesulfonate (S5):

[0237] A flame-dried round bottom flask equipped with a stir bar was backfilled three times with nitrogen gas before the addition of RMS-IV-162 (504.9 mg, 1.0 mmol, 1 equiv) in DCM (10 mL), methanesulfonyl chloride (160 pL, 2.0 mmol, 2 equiv), and triethylamine (290 pL, 2.0 mmol, 2 equiv). The reaction stirred for 60 minutes before the crude mixture was washed with water. The aqueous layer was then extracted with DCM two times before the combined organic layers were dried with MgSCh, filtered, and concentrated via rotary evaporation. The crude mixture was purified via flash chromatography (10% EtOAc / Hex) to give the title compound (288.4 mg, 0.51 mmol, 51% yield) as a clear oil. ’H NMR (600 MHz, Chloroform- d) 5 7.37 - 7.27 (m, 5H), 7.25 (t, J= 3.3 Hz, 3H), 7.23 - 7.17 (m, 2H), 5.05 (d, J= 1.5 Hz, 1H), 5.00 (d, J= 1.5 Hz, 1H), 4.60 - 4.57 (m, 2H), 4.21 (t, J= 6.6 Hz, 2H), 3.79 (p, J= 2.1 Hz, 1H), 3.31 (s, 3H), 2.99 (s, 3H), 2.43 - 2.41 (m, 1H), 2.33 (dd, J = 17.0, 9.2 Hz, 1H), 2.19 (td, J = 7.1, 2.6 Hz, 1H), 2.08 - 2.04 (m, 2H), 2.02 (ddd, J = 10.0, 7.2, 4.6 Hz, 2H), 1.73 (p, J = 6.8 Hz, 5H), 1.66 - 1.62 (m, 2H), 1.44 - 1.18 (m, 9H).13C NMR (151 MHz, CDCh) 5 154.46, 144.05, 141.20, 139.23, 137.41, 129.57, 127.77, 127.58, 127.50, 126.55, 126.51, 114.82, 94.68, 86.66, 70.08, 69.03, 55.09, 52.71, 40.47, 37.31, 32.36, 31.38, 29.63, 29.56, 29.24, 29.21, 29.06, 28.94, 27.73, 25.32. LRMS (ESI, APCI) m / z: calculated for C34H47O5S [M+H]+ 567.8, found 536.3 (-OCH3).

[0238] 9-(6-(methoxymethoxy)-3-phenyl-3a-(l-phenylvinyl)-l, 3a, 4,5,6, 6a-hexahydropentalen-2- yl)nonyl sulfamate (S6):

[0239] An oven-dried vial backfilled with nitrogen was cooled to 0 °C before the addition of chlorosulfonyl isocyanate (0.5 mL, 5.744 mmol) and formic acid (216 pL, 5.744 mmol). The resulting solution (28 pL) was added to a solution of S5 (70.6 mg, 0.144 mmol, 1 equiv) was dissolved in DMA. The reaction stirred for 24 hours before being diluted in EtOAc and washed with water. The organic layer was dried with magnesium sulfate, filtered, and concentrated via rotary evaporation. The crude mixture was purified via flash chromatography (5-30% EtOAc / Hexanes and then flushed with 100% EtOAc) to collect impure product taken to the next step without further purification.

[0240] 9-(6-hydroxy-3-phenyl-3a-(l-phenylvinyl)-l,3a,4,5,6,6a-hexahydropentalen-2-yl)nonyl sulfamate (Sulf): In a reaction vial the S6 (37.9 mg, 0.07 mmol, 1.0 equiv) was dissolved in acetonitrile. A few drops of hydrochloric acid was added at room temperature and the reaction proceeded until complete by TLC (30 minutes- 2 hours). The crude mixture was concentrated and purified by reverse phase liquid chromatography using a 50-99% MeCN / IfcO gradient over 35 minutes to afford the title compound.1H NMR (500 MHz, Chloroform -t / ) 5 7.36 - 7.27 (m, 5H), 7.25 (d, J= 3.1 Hz, 3H), 7.21 - 7.17 (m, 2H), 5.07 (d, J= 1.4 Hz, 1H), 4.98 (d, J = 1.4 Hz, 1H), 4.79 (s, 2H), 4.19 (t, J= 6.6 Hz, 2H), 3.95 (d, J= 3.9 Hz, 1H), 2.36 (dd, J= 16.7, 9.3 Hz, 1H), 2.31 - 2.25 (m, 1H), 2.12 - 1.98 (m, 4H), 1.76 - 1.64 (m, 4H), 1.36 (dt, J= 19.7, 7.0 Hz, 3H), 1.30 - 1.19 (m, 11H).13C NMR (126 MHz, CDC13) 5 154.50, 144.07, 140.98, 139.13, 137.28, 129.61, 127.65, 127.63, 127.54, 126.58, 126.52, 114.93, 82.00, 71.43, 69.26, 55.73, 40.13, 33.90, 32.00, 29.51, 29.38, 29.13, 29.08, 28.84, 28.68, 27.62, 25.29. LRMS (ESI, APCI) m / z: calculated for C31H42NO4S [M+H]+ 524.7, found 522.2.

[0241] 10-(6-(methoxymethoxy)-3-phenyl-3a-(l-phenylvinyl)-l, 3a, 4,5,6, 6a-hexahydropentalen- 2-yl)decanenitrile (S7):

[0242] A flame-dried round bottom flask equipped with a stir bar was backfilled three times with nitrogen. Sodium cyanide (250 mg, 5.1 mmol, 10 equiv) was quickly added before the addition of S6 (288.4 mg, 0.51 mmol, 1 equiv) in DMF (5.1 mL). A vent needle to a saturated KOH solution was added before heating the reaction to 110 °C. After 10 minutes the reaction showed completion by TLC and was cooled to room temperature before being diluted with EtOAc. The solution was washed with saturated KOH two times, water one time, and brine three times. The resulting organic layer was dried with MgSO4, filtered, and concentrated via rotary evaporation. The crude mixture was purified by flash chromatography to give the title compound (224.8 mg, 0.45 mmol, 89% yield) as a clear oil. ’H NMR (600 MHz, Chloroform- d) 5 7.35 - 7.29 (m, 6H), 7.26 - 7.24 (m, 2H), 7.22 - 7.20 (m, 2H), 5.05 (d, J= 1.4 Hz, 1H), 5.01 (d, J= 1.5 Hz, 1H), 4.59 (d, J= 2.6 Hz, 2H), 3.80 - 3.78 (m, 1H), 3.31 (s, 3H), 2.43 (dt, J = 9.2, 1.7 Hz, 1H), 2.32 (t, J= 7.1 Hz, 3H), 2.03 (dtd, J= 12.6, 7.2, 6.2, 2.9 Hz, 4H), 1.74 (ddt, J = 6.8, 4.7, 2.3 Hz, 1H), 1.67 - 1.62 (m, 4H), 1.45 - 1.39 (m, 3H), 1.33 (t, J= 7.0 Hz, 2H), 1.30 - 1.18 (m, 9H).13C NMR (151 MHz, CDCI3) 5 154.45, 144.05, 141.17, 139.24, 137.40, 129.56, 127.76, 127.57, 127.56, 126.54, 126.50, 119.75, 114.81, 94.67, 86.64, 69.02, 55.08, 52.70, 40.46, 32.35, 31.37, 29.61, 29.52, 29.17, 29.11, 28.66, 28.55, 27.71, 25.28, 17.04. LRMS (ESI, APCI) m / z: calculated for C34H44NO2 [M+H]+ 498.7, found 498.4.

[0243] 5-(9-(6-(methoxymethoxy)-3-phenyl-3a-(l-phenylvinyl)-l,3a,4,5,6,6a- hexahydropentalen-2-yl)nonyl)- 1 / / -let r azole (S8) :

[0244] A round bottom flask was charged with a stir bar and S7 (112.4 mg, 0.23 mmol, 1.0 equiv) in toluene (500 pL). Trimethyl silyl azide (81 pL, 0.46 mmol, 2 equiv) and dibutyltin oxide (5.7 mg, 0.023 mmol, 0.1 equiv) were added to the reaction vial before heating to reflux. After 48 hours the reaction was cooled to room temperature and concentrated before being redissolved in methanol (10 mL). The reaction was reconcentrated before being dissolved in EtOAc and washed with saturated sodium bicarbonate two times. The organic layer was dried with magnesium sulfate, filtered, and concentrated. The crude reaction mixture was purified by flash chromatography (50% EtOAc / Hex then 10% MeOH / DCM) to give the title compound (53.3 mg, 0.1 mmol, 43% yield) as a clear oil.1H NMR (400 MHz, Chloroform-t / ) 5 7.27 (m, 8H), 7.18 - 7.13 (m, 2H), 5.01 (d, J= 1.4 Hz, 1H), 4.96 (d, J = 1.6 Hz, 1H), 4.63 - 4.56 (m, 2H), 3.76 (t, J= 3.1 Hz, 1H), 3.31 (s, 3H), 2.97 (t, J= 7.7 Hz, 2H), 2.38 (d, J = 9.1 Hz, 1H), 2.27 (dd, J= 17.1, 8.9 Hz, 1H), 2.01 - 1.93 (m, 5H), 1.81 - 1.74 (m, 2H), 1.67 - 1.56 (m, 2H), 1.19 - 1.13 (m, 12H).13C NMR (101 MHz, CDCI3) 5 154.20, 143.91, 141.04, 139.29, 137.19, 129.47, 127.61, 127.56, 127.52, 127.49, 126.50, 126.46, 125.59, 114.83, 94.48, 86.89, 68.96, 54.97, 52.60, 40.24, 32.22, 31.22, 29.53, 29.28, 28.87, 28.82, 28.76, 28.66, 28.59, 27.46, 27.29, 23.40. LRMS (ESI, APCI) m / z: calculated for C34H45N4O2 [M+H]+ 541.8, found 542.4.

[0245] 5-(9-( l / / -tetr:izol-5-yl)nonyl)-4-phenyl-3a-(l-phenylvinyl)-1.2.3.3a.6.6a- hexahydropentalen-l-ol (Tet): In a reaction vial the S8 (37.6 mg, 0.07 mmol, 1.0 equiv) was dissolved in acetonitrile (500 pL). A few drops of hydrochloric acid was added at room temperature and the reaction proceeded until complete by TLC (30 minutes- 2 hours). The crude mixture was concentrated and purified by flash chromatography (0-2% MeOH / DCM) to afford the product as a clear oil (20.0 mg, 57% yield). ’H NMR (400 MHz, Chloroforms / ) 5 7.27 (t, J= 7.7 Hz, 5H), 7.21 (t, J = 3.8 Hz, 3H), 7.16 (d, J= 7.4 Hz, 2H), 5.03 (d, J= 7.8 Hz, 1H), 4.95 (d, J= 4.3 Hz, 1H), 3.95 (d, J = 4.2 Hz, 1H), 2.95 (t, J = 7.7 Hz, 1H), 2.31 - 2.23 (m, 1H), 2.10 - 1.89 (m, 2H), 1.84 - 1.59 (m, 2H), 1.38 - 1.14 (m, 12H).13C NMR (151 MHz, CDCh) 5 154.46, 144.04, 141.08, 139.38, 137.30, 129.68, 127.79, 127.75, 127.69, 126.73, 126.68, 115.04, 82.34, 72.97, 69.31, 55.89, 49.45, 40.10, 34.12, 32.07, 29.46, 29.18, 29.09, 28.99, 28.93, 28.87, 27.73, 27.57. LRMS (ESI, APCI) m / z: calculated for C32H41N4O [M+H]+ 497.3, found 497.9.

[0246] 5-(9-( l / / -tetr:izol-5-yl)nonyl)-4-phenyl-3:i-(l-phenylvinyl)-3.3:i.6.6:i-tetr:ihydropentalen- l(2 / / )-one (S9):

[0247] In a round bottom flask Tet (18.9 mg, 0.04 mmol, 1 equiv) was dissolved in DCM (760 pL). Dess-Martin Periodinate (21.2 mg, 0.05 mg, 1.2 equiv) was added and the reaction proceeded open to air. After completion the mixture was diluted in MTBE pushed through a plug of celite before being concentrated via rotary evaporation to give the title compound as a yellow oil (21.1 mg, quant.). ’H NMR (400 MHz, CDCh) 5 7.35 - 7.29 (m, 6H), 7.28 - 7.22 (m, 4H), 5.23 (d, J= 1.4 Hz, 1H), 5.14 (d, J= 1.3 Hz, 1H), 3.04 (t, J= 7.6 Hz, 2H), 2.54 (d, J = 7.6 Hz, 1H), 2.51 - 2.27 (m, 3H), 2.24 - 2.10 (m, 3H), 1.96 (dd, J= 13.8, 6.8 Hz, 1H), 1.85 (tt, J= 15.1, 7.6 Hz, 4H), 1.43 - 1.29 (m, 12H).13C NMR (101 MHz, CDCh) 5 152.68, 144.54, 142.17, 137.68, 136.22, 134.61, 128.77, 128.08, 128.03, 128.01, 127.50, 127.45, 127.02, 126.91, 115.30, 65.34, 55.63, 38.31, 37.60, 29.52, 29.33, 28.80, 28.60, 28.38, 28.28, 28.22, 28.11, 27.13, 27.02, 23.15. LRMS (ESI, APCI) m / z: calculated for C32H39N4O [M+H]+ 495.7, found 496.3. 5-(9-( l / / -tetrazol-5-yl)nonyl)-4-phenyl-3a-(l-phenylvinyl)-1.2.3.3a.6.6a- hexahydropentalen-l-amine (S10):

[0248] To a flame-dried round bottom equipped with a stir bar and backfilled (3x) with nitrogen S9 (18 mg, 0.04 mmol, 1 equiv) dissolved in dry ethanol (400 pL) was added. Titanium isopropoxide (18 pL, 0.06 mmol, 1.5 equiv) was added to the reaction followed by ammonia (7N in methanol) (120 pL, 0.8 mmol, 20 equiv). The reaction stirred overnight before the addition of sodium borohydride (4.5 mg, 0.12 mmol, 3 equiv). The reaction mixture stirred for another hour before being diluted in ethyl acetate and extracted (2x) with sat. Rochelle’s salt. The organic layer was dried with magnesium sulfate, filtered, and concentrated via rotary evaporation. The crude mixture was then purified via flash chromatography (10-20% MeOH / DCM) to give the title compound (7.8 mg, 39% yield) as a yellow oil.1H NMR (600 MHz, MeOD) 5 7.35 - 7.31 (m, 5H), 7.29 - 7.26 (m, 3H), 7.21 - 7.17 (m, 2H), 5.10 (d, J = 1.2 Hz, 1H), 4.93 (d, J = 1.2 Hz, 1H), 3.51 (ddd, J= 11.9, 9.0, 5.8 Hz, 1H), 2.87 (t, J= 7.5 Hz, 3H), 2.73 (dd, J= 9.1, 2.7 Hz, 1H), 2.30 (dd, J= 17.8, 9.1 Hz, 1H), 2.24 (d, J= 2.8 Hz, 1H), 2.14 (ddd, J= 13.7, 9.2, 6.8 Hz, 1H), 2.08 - 1.97 (m, 4H), 1.91 - 1.78 (m, 2H), 1.76 - 1.71 (m, 3H), 1.27 - 1.16 (m, 11H).13C NMR (151 MHz, MeOD) 5 155.35, 144.86, 143.56, 140.73, 137.50, 130.96, 129.07, 128.92, 128.74, 128.26, 116.61, 70.88, 54.94, 47.17, 35.92, 34.13, 30.60, 30.53, 30.40, 30.18, 30.12, 30.07, 29.94, 29.20, 28.44, 24.89. LRMS (ESI, APCI) m / z: calculated for C32H42N5 [M+H]+ 496.7, found 498.9.

[0249] \-(5-(9-( l / / -tetrazol-5-yl)nonyl)-4-phenyl-3a-( l-phenylvinyl)-l .2.3.3a.6.6a- hexahydropentalen-l-yl)-sulfanoylamine (6N-Tet):

[0250] S10 was added to a reaction vial in EtOH (314 pL). H2O (160 pL) was added followed by sulfamide (7.6 mg, 0.09 mmol, 5 equiv) and triethylamine (6.5 pL, 0.05 mmol, 3 equiv). The reaction proceeded for 16 hours at reflux before being cooled to room temperature. The reaction mixture was concentrated via rotatory evaporation. The crude mixture was then purified via flash chromatography (10-20% MeOH / DCM) to give the title compound (2.1 mg, 35% yield) as a yellow oil. ’H NMR (400 MHz, cdcl3) 5 7.27 (d, J= 10.6 Hz, 7H), 7.20 - 7.15 (m, 3H), 5.07 (s, 1H), 4.96 (s, 1H), 4.70 (br. s, 2H), 3.81 (s, 1H), 2.96 (s, 2H), 2.60 (t, J= 8.6 Hz, 1H), 2.45 (d, J = 17.6 Hz, 1H), 2.19 - 1.90 (m, 7H), 1.73 (dd, J= 20.7, 7.6 Hz, 7H), 1.40 (t, J= 7.1 Hz, 10H).13C NMR (151 MHz, CDC13) 5 155.35, 144.86, 143.56, 140.73, 137.50, 130.96, 129.07, 128.92, 128.74, 128.26, 116.61, 82.34, 72.97, 69.31, 35.92, 34.13, 30.60, 30.53, 30.40, 30.18, 30.12, 30.07, 29.94, 29.20, 28.44, 24.89. LRMS (ESI, APCI) m / z: calculated for C32H43N6O2S [M+H]+ 575.8, found 574.9.

[0251] Example 2: Biological Evaluation of Compounds

[0252] Nuclear receptors (NRs) are ligand-regulated transcription factors that allow biological systems to sense and respond to lipophilic molecules by altering gene expression. Their functions are driven by a modular structure consisting of a ligand binding domain (LBD) and DNA-binding domain (DBD). Ligand binding to the LBD promotes NR association with coactivators, which drive target gene expression through recruitment of transcriptional machinery and remodeling of chromatin. Liver receptor homolog-1 (LRH-1; NR5A2) is a nuclear receptor that is primarily expressed in tissues of endodermal origin and has been thoroughly characterized in the liver, where it regulates lipid and glucose homeostasis. This receptor also plays a key role in cell renewal and local glucocorticoid biosynthesis in the gut, making it an attractive target for inflammatory bowel disease.

[0253] While the endogenous ligand of LRH-1 remains unknown, phosphatidylcholines of medium chain length directly bind the receptor (Figure 1A, B) and increase coactivator association. In a diabetic mouse model, the exogenous phospholipid (PL) 1,2-dilauroyl-sn- glycero-3 -phosphocholine (DLPC) suppresses genes involved in lipogenesis and alleviates glucose intolerance and liver steatosis in an LRH-1 -dependent manner (Figure 1C). However, due to the poor solubility of PLs, synthetic agonists are more favorable tools for targeting LRH- 1 in the clinic and laboratory. The first class of LRH-1 small molecule agonists (including “RJW100”) bound deep in the ligand binding pocket, away from PL-coordinating residues at the pocket mouth, through a conserved network of water molecules (Figure IB, C).

[0254] A series of compounds was previously designed to mimic PL binding by engaging the PL-coordinating residues at the pocket mouth. A carboxylic acid attached to the RJW100 scaffold through a 10-carbon linker (“10CA”; Figure 1C) exhibited improved potency and effect in reporter assays. Structural studies confirmed that the carboxylic acid engaged in hydrogen bonding with residues contacted by activating PLs (G421, Y516, and K520) at the pocket mouth (Figure IB).

[0255] Described herein is a series of 10CA isosteres to explore the effects of different polar moieties that target the same pocket mouth residues. As shown herein in the present example, a tetrazole-containing isostere improves potency in reporter assays and drives greater target gene expression in HepG2 cells. Structural studies paired with molecular dynamics (MD) simulations reveal that this isostere improves hydrogen bond contacts at the pocket mouth and preserves lOCA-like allostery. Additionally developed small molecules described herein exhibit improved binding and potency and show specificity for NR5 As, providing a useful tool for targeting LRH-1 for clinical and laboratory purposes.

[0256] Materials and Methods

[0257] Cell culture. Cells were cultured under standard conditions (5% CO2, 37 °C). HepG2 cells were cultured in phenol red-free DMEM + 10% fetal bovine serum (FBS). HeLa cells were cultured in phenol red-free MEMa + 10% FBS - charcoal / dextran treated (FBS-S).

[0258] Data analysis and visualization. Average values from technical replicates were used for all data analyses. These values represented either biological replicates (in-cell work) or independent experiments (in vitro work) that were combined for data analyses. Bar charts and curves were constructed with GraphPad Prism (version 9), structural figures were constructed with either PyMol or VMD, and difference distance matrices were constructed with Bio3D. GraphPad Prism (version 9) was used for all data analyses. All figures were constructed using Adobe Illustrator 2021 (Adobe Inc.). Values were consistently reported with two significant figures.

[0259] Protein purification. The LRH-1 LBD was expressed and purified as described previously. Briefly, BL21(DE3) Escherichia coli cells were transformed with the human LRH- 1 LBD (residues 299-541) with an N-terminal six-His tag in a pMCSG7 vector. Cells were grown at 37 °C in lysogeny broth until ODeoo 0.6. Protein expression was induced with 1 mM isopropyl P-D-l -thiogalactopyranoside for 4 hours at 30 °C. Cells were centrifuged and stored at -80 °C. The cell pellet was resuspended in lysis buffer [20 mM Tris-HCl, 150 mMNaCl, 5% glycerol, 25 mM imidazole, 0.2 mM phenylmethyl sulfonyl fluoride, DNase, and lysozyme (pH 7.4)] and lysed via sonication. Protein was isolated with Ni2+affinity chromatography. The human LRH-1 LBD purifies bound to bacterial phospholipids when expressed in Escherichia coli. Co-purified bacterial lipids for the LRH-1 LBD used in FP competition assays were removed by incubating the protein with 4-fold molar excess of DLPC overnight at 4 °C. The LBD was then purified with size-exclusion chromatography (SEC) into assay buffer [150 mM NaCl, 20 mM Tris-HCl, and 5 % glycerol (pH 7.4)]. The LRH-1 LBD used for thermal stability assays was purified in a similar manner but was not complexed with DLPC prior to SEC purification. LRH-1 LBD used for crystallography was incubated with TEV protease to remove the six-His tag and subjected to a second round of Ni2+affinity chromatography before being complexed with Tet (see below). All protein was stored at -80 °C until use.

[0260] Fluorescence polarization competition assays. Forward binding assays were run as described previously. Briefly, binding affinity for 6N conjugated to fluorescein amidite (FAM) was determined using 10 nM 6N-FAM and protein concentrations indicated in the figure (Figure 2). Plates were incubated overnight at 4 °C. Polarization was monitored on a Neo plate reader (Biotek, Winooski, VT) at an excitation / emission wavelength of 485 / 528 nm. Eight independent experiments were conducted, each with three technical replicates. Data were baseline-corrected and fit with a one-site binding (total) curve in GraphPad Prism (version 9). The resulting curve is provided in supplemental information (Figure 2) and the Kd was 8.1 nM.

[0261] FP competition assays were conducted as described previously. Briefly, experiments were conducted in 30 pL of assay buffer [150 mM NaCl, 20 mM Tris-HCl, and 5% glycerol (pH 7.4)]. 6N-FAM (10 nM / well) was incubated with LRH-1 LBD (5 nM / well). Unlabeled compounds were added at concentrations indicated in figures, with DMSO in each well held constant at 6.7% v / v. Data were excluded from analysis from wells with le'4M Am and HA, as the corresponding polarization values distorted the curve fit because they were abnormally high, potentially because of compound insolubility. Each experiment was performed two times with four technical replicates each. Data from each independent experiment were first normalized so that the highest and lowest values corresponded to 100 and zero, respectively. Data were then fit to a one-site, fit Ki curve, assuming a final probe concentration of 10 nM and probe affinity (Kd = 8.1 nM) determined with forward binding assays (Figure 2).

[0262] Thermal stability assays. Thermal stability of the LRH-1 LBD complexed with ligands was determined using a Tycho NT.6 Nanotemper. The LRH-1 LBD was incubated with 5-fold molar excess of ligand (final DMSO concentration was 1.4%) overnight at 4 °C in assay buffer [150 mM NaCl, 20 mM Tris-HCl, and 5% glycerol (pH 7.4)]. Complexes were centrifuged at high speed for five minutes and then loaded into capillaries. Tryptophan / tyrosine fluorescence was monitored at wavelengths 330 and 350 nm over a 30 °C / min gradient (35 °C - 95 °C). The inflection point was determined with Tycho NT.6 software. Two separate experiments were conducted in triplicate. Calculated inflection point values were displayed as bar graphs constructed with Prism (version 9). Luciferase reporter assays. LRH-1 reporter assays were conducted. Briefly, HeLa cells were seeded at -7,500 cells per well in 96-well plates (white-walled, clear bottom) in MEMa + 10% FBS-S. Once cells reached 70-90% confluence, they were transfected with LRH-1 (in pCI vector, 5 ng / well), a reporter plasmid with an NR5A response element derived from the NR0B2 promoter cloned upstream of firefly luciferase (in pGL3 -Basic vector, 50 ng / well), and a plasmid expressing Renilla luciferase constitutively from a CMV promoter (1 ng / well). Cells were transfected with FuGENE at a ratio of 2.5: 1 (FuGENE:DNA). Twenty-four hours after transfection, compounds were diluted in Opti-MEM and introduced to cells at final concentrations indicated in figures (final DMSO concentration was 0.37%). Luciferase signal was measured after 24 hours using the DualGlo kit (Promega). Experiments were conducted with three biological replicates, each with three technical replicates. Each well’s Firefly luciferase signal intensity was divided by the well’s Renilla signal intensity and then normalized relative to the DMSO control. Data were analyzed with GraphPad Prism (version 9) using a stimulating dose-response curve (Hill slope = 1). The fold change was determined to be the calculated span + 1. Relative efficacies reported were determined by dividing the small molecule’s fold change by that of 10CA. Data were excluded from analysis for cells treated with 3e'5M of Tet and Sul as low Renilla signal and cell morphology suggested cytotoxicity. Data were included for this concentration in reporter assays comparing Tet and 6N-Tet, as there was no observable cell death during these experiments.

[0263] RT-qPCR. RT-qPCR was performed as described previously. HepG2 cells were seeded at 400,000 cells per well in 24-well plates in DMEM + 10% FBS. When cells reached -90% confluence, the medium was exchanged with medium containing DMSO or compound at the desired concentration (final DMSO concentration: 0.3%). Small molecules were added concentrations indicated in figure legends. After 24 hours, the medium was decanted, cells were washed with phosphate buffered saline, and cells were collected in RET lysis buffer (+ 1% 2- mercaptoethanol). Cells were stored at -80 °C prior to RNA extraction. RNA was extracted from cells using the RNeasy Mini Kit (QIAGEN), with on-column DNase digestion. RNA was reverse transcribed with the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). cDNA was quantified using Power SYBR Green PCR Master Mix (Applied Biosystems), using human ACTB (actin beta) as a housekeeping gene. Ct values were calculated by resident software on the StepOne Plus thermocycler. Data were normalized using the AACt method (Livak and Schmittgen, 2001). Each experiment was conducted with two or four biological replicates that were normalized independently for data analysis. Data were analyzed with GraphPad Prism, using a Brown-Forsythe and Welch one-way ANOVA and Dunnett T3 multiple comparisons test. Primers used for RT-qPCR were as follows: hACTB forward 5’-AGGCACCAGGGCGTGAT-3’ (SEQ ID NO:1) reverse 5’-GCCCACATAGGAATCCTTCTGAC-3’ (SEQ ID NO:2) hNR0B2 forward 5’-GCTTAGCCCCAAGGAATATGC-3’ (SEQ ID NO:3) reverse 5’-GTTCCAGGACTTCACACAGC-3’ (SEQ ID NO:4) hCYP7Al forward 5’-GAGAAGGCAAACGGGTGAAC-3’ (SEQ ID NO: 5) reverse 5’-GGATTGGCACCAAATTGCAGA-3’ (SEQ ID NO: 6)

[0264] X-ray crystallography. LRH-1 LBD-Tet crystals were generated. Briefly, cleaved (six- His tag removed) LRH-1 LBD was incubated with Tet at 4-fold molar excess overnight at 4 °C. The complex was then purified via SEC into crystallization buffer [150 mMNaCl, 100 mM ammonium acetate, 1 mM EDTA, 2 mM CHAPS, and 1 mM DTT (pH 7.4)] and incubated with a peptide corresponding to human TIF2 NR box 3 ( HiN-KENALLRYLLDKDD-CO?-, SEQ ID NO:7) at 4-fold molar excess, along with an additional 2-fold molar excess of Tet, for two hours at room temperature. The complex was then concentrated to ~7 mg / mL and crystals were generated via hanging drop vapor diffusion in crystallant containing 0.1 M tri-Na citrate - pH 4.6, 10-14% tert-butanol, and 0-7.5% glycerol at 4 °C. Crystals were flash frozen in liquid N2 using cryoprotectant consisting of crystallant supplemented with 30% glycerol. Data were collected remotely from the Southeast Regional Collaborative Access Team (SER-CAT) at the Advanced Photon Source (Argonne National Laboratories, Chicago, IL). Data were processed using HKL2000 and phased with molecular replacement, using PDB entry 4DOS as the search model. Structure refinement was performed with Phenix and Coot. Additional refinement was performed with PDB-REDO (Joosten et al., 2014). Twinning was detected with xtriage in Phenix, and we used the recommended twin law (-h,-k,l) during refinement. Final figures were constructed with PyMOL (Schrodinger, LLC), which was also used to predict hydrogen bonds between the ligand and LRH-1.

[0265] Model construction. Complexes for MD simulations with LRH-1 LBD were constructed as described previously (Cato et al., 2022). Three complexes were included: i) apo LRH-1 -TIF2; ii) LRH-1 -TIF2-Tet; and iii) LRH-1-TIF2-10CA. Ligand-bound (or apo) LRH- 1 LBD complexes were generated by using the structure of LRH-1 -TIF2-Tet (PDB entry 8F8M; currently unpublished) as a starting model. PDB entry 8F8M underwent further refinement before being uploaded to PDB but was nearly identical to that used for MD studies. 1 OCA was modeled into the complex using the positioning of the ligand from the previous crystal structure (PDB entry 7JYD). Complexes included LRH-1 residues 299-540 and residues 742-752 of TIF2 CH3N-NALLRYLLDKD-CO2; SEQ ID NO:8). Residues 539 and 540 were added using the positioning from PDB entry 7JYE.

[0266] Maestro (Schrodinger, LLC) was used to optimize hydrogen bond assignments (pH 7.0), add N- and C-terminal caps to LRH-1 and TIF2, and run initial minimization on the structure. The complexes were solvated in an octahedral box of TIP3P water with a 10 A buffer around the protein complex. Na+and CL ions were added to neutralize the protein and achieve physiological buffer conditions (150 mM NaCl). Systems were set up using the xleap tool in AmberTools20 of Amber 2020 (Case et al., 2020), with ffl4SB (protein), GAFF2 (ligand), and TIP3P (water) forcefields. Parameters for 10CA and Tet were obtained using Antechamber in AmberTools20. Note that both ligands were deprotonated in simulations, giving each a net charge of -1.

[0267] Molecular dynamics simulations. MD simulations were performed as described previously. For minimization, 5000 steps of steepest descent were used, followed by 5000 steps of conjugate gradient minimization. Minimizations were first performed with 500 kcal / mol A2 restraints on all protein and ligand atoms. Restraints were then removed on all atoms except the ligand and TIF2 peptide, and the protocol was repeated. Restraints were then removed on all atoms except the ligand, and the protocol was repeated. Restraints were subsequently lowered to 100 kcal / mol A2 and then finally removed from all atoms for two final rounds of minimization. Minimized systems were heated from 0 to 300 K with a 100 ps MD run, with constant volume periodic boundaries and 10 kcal / mol A2 restraints on all protein and ligand atoms. A 10 ns equilibration was performed for all complexes with 10 kcal / mol A2 restraints on all protein and ligand atoms using the NPT ensemble. Restraints were then removed on all atoms except the ligand, and the protocol was repeated. The protocol was repeated with a 1 kcal / mol A2 restraint on the ligand. Note that the water molecules critical for ligand engagement deep within the pocket (four waters) and at the mouth (one water) were restrained along with ligand.

[0268] Production trajectories of 500 ns were obtained for unrestrained complexes in the NPT ensemble. All bonds between heavy atoms and hydrogens were fixed with the SHAKE algorithm. A cutoff distance of 10 A was used to evaluate long-range electrostatics with particle mesh Ewald and for van der Waals forces. Structural averaging was performed using the CPPTRAJ module of AmberTools. Four 500 ns simulations were run and concatenated with CPPTRAJ, with every fifth frame (total of 40,000 frames) used for data analysis. Water, Na+, Cl’, along with N- and C-terminal caps, were removed for data analysis.

[0269] CPPTRAJ was used to for hydrogen bond analyses, as well as the construction of average structures. Note that hydrogen bond analysis was conducted with the default angle cutoff of 135° and distance cutoff of 3.5 A. Bio3D was used to create difference distance matrices comparing average structures between apo and ligand-bound complexes. Dynamic networks were constructed from trajectories using the Network View plugin in VMD and the Carma program. Networks were constructed by defining all protein Ca atoms as nodes, using Cartesian covariance (calculated in Carma) to measure communication within the network. Hydrogen atoms were excluded from network construction, and edges between neighboring residues were disallowed. Pairs of nodes that reside within a 4.5 A cutoff for 75% of the simulation are connected via an edge. Communities are constructed using the Girvan-Newman algorithm, and the minimum number of communities possible were generated while maintaining at least 98% maximum modularity. Communities were visualized with VMD.

[0270] Cross-reactivity studies. Reporter assays comparing LRH-1 and SF-1 activity were conducted as above. However, for experiments testing SF-1 activity, cells were transfected with SF-1 in a pcDNA vector (5 ng / well). Cells were transfected with FuGENE at a ratio of 4: 1 (FuGENE:DNA). Cells were treated with 10 pM of compound for 24 hours (final DMSO concentration was 0.37%). Reporter assays assessing cross-reactivity with non-NR5A receptors were conducted by INDIGO Biosciences, Inc. Reporter cells expressed either the native receptor (AhR, AR, ERa, GR, and MR) or a receptor hybrid in which the native N- terminal DBD has been replaced with that of the yeast Gal4 DBD (RORy, CAR3, FXR, PPARa, PPAR5, PPARy, and PXR). Huh7 (AhR), CV-1 (AR), CHO (CAR3, ERa, FXR, GR, PPARa, PPAR5, and PPARy), or HEK293 (RORy, MR, and PXR) cells were used in studies. A gene encoding Firefly luciferase was downstream of a receptor-specific genetic response element or the GAL4 upstream activation sequence. All reference compounds used, aside from ursolic acid (RORy inverse agonist), were agonists and were as follows: ursolic acid (RORy), MeBio (AhR), 5a-Dihydro-ll- ketoTestosterone (AR), CITCO (CAR3), 17P-estradiol (ERa), GW4064 (FXR), dexamethasone (GR), aldosterone (MR), GW7647 (PPARa), GW0742 (PPAR5), rosiglitazone (PPARy), and rifampicin (PXR). Experiments were run in triplicate in 96-well plates (medium = cell recovery medium). Assay plates were incubated for 24 hours and then the treatment medium was discarded. Luciferase detection reagent was added and relative bioluminescence was measured. All graphical manipulations were performed using GraphPad Prism (version 9). Results

[0271] Isosteres as described above were synthesized including the following moi eties: a tetrazole (“Tet”), amide (“Am”), hydroxamic acid (“HA”), piperazine (“Pip”), sulfamate (“Sul”), and serine (“Ser”) (Figure 3). Initial studies were performed to assess the compound binding to the LRH-1 LBD utilizing an internally developed fluorescence polarization (FP) competition assay. The fluorescence polarization competition assay assesses binding of the synthesized 10CA isosteres to the ligand binding pocket. The Tet analog demonstrated the greatest affinity (Ki = 23 nM) (Figure 4, Panel A; Figure 2). Surprisingly, the inhibition constant of the Tet analog was measured and found to be similar to that of 10CA ( = 26 nM). Thermal stability testing revealed that 10CA(+ 6.65 °C) and Tet (+ 6.4 °C) were similar, driving the greatest improvement to LRH-1 LBD stability (Figure 4, Panel B). To assess effects on LRH-1 activity, luciferase reporter assays were performed. Tet (ECso = 710 nM) and Sul (ECso = 470 nM) demonstrated improved potency relative to 10CA. (Figure 4, Panel C; Figure 5). Interestingly, other analogs exhibited decreased affinity, thermal stability, and / or potency relative to 10CA.

[0272] To further explore divergent effects on LRH-1 activity, ligand-driven modulation of key liver target genes NR0B2 (small heterodimer partner; SHP) and CYP7A1 (cytochrome P450 7A1; also known as cholesterol 7-a-hydroxylase) was assessed in HepG2 cells derived from the liver. Driving the greatest increase in mRNA steady-state levels for both genes was the Tet analog (Figure 4, Panel D). Taken collectively, the tetrazole appeared to best recapitulate carboxylic acid-mediated effects on binding and activity while improving potency and target gene expression in cultured cells.

[0273] Crystal structures of the LRH-1 LBD bound to Tet and a fragment of coactivator transcriptional intermediate factor 2 (TIF2) were evaluated to assess the molecular mechanisms of Tet-mediated activity (Figure 6, Panel A; Table 1).

[0274] Table !:

[0275] Observed was visible electron density corresponding to the core, tail, and tetrazole head (Figure 6, Panel B), and the ligand orientation of Tet was similar to that of 10CA (Figure 6, Panel C). Like RJW100 and 10CA, the hydroxyl group of Tet engaged indirectly with polar residues deep in the pocket through a conserved network of water molecules visible in previous structures presented in the literature (Figure 6, Panel D). Surprisingly, while the tetrazole permitted extensive contact with residues at the mouth of the pocket, K520 was slightly out of hydrogen bond distance (Figure 6, Panel E), which appears to be a consequence of reorientation of K520 rather than the tetrazole (Figure 6, Panel E). Crystals were generated at pH 4.6, which may have protonated the tetrazole tail group. This may decrease the electrostatic interaction with K520, which may consequently be repositioned further away from the ligand.

[0276] Subsequently, MD simulations (4 x 500 ns) were performed to compare pocket mouth interactions made by the deprotonated tetrazole (tetrozolate) and carboxylic acid (carboxylate), which may better reflect their ionized states at physiological pH. Tet appeared to make considerably stronger contacts with pocket mouth residues than 10CA (Figure 7, Panel A). The tetrazole interacted with G421 twenty times longer and with Y516 and K520 for twice as long. Additional simulations were performed to explore ligand-driven effects on protein dynamics. Comparison of average structures revealed that both 10CA and Tet mobilized H6- H7 (Figure 7, Panel B). This region may correspond with the alternate activation function surface (AF-B), which may communicate ligand-binding status to the coactivator binding interface on LRH-1. Community analysis, which reveals “communities” of residues with highly correlated motions, was utilized to compare residue motions driven by apo and ligandbound. Differences were observed between ligand-bound and apo states, including that 10CA and Tet promoted interhelical communication between H6 and H7, suggesting that both ligands drive correlated motion in the AF-B (Figure 7, Panel C). However, correlated motion with the TIF2 coactivator peptide differed between 10CA- and Tet-bound LBD, suggesting that these ligands may differ in coactivator communication in certain contexts. Altogether, molecular dynamic (MD) simulations suggest that Tet makes stronger contacts at the pocket mouth and drives similar allosteric motions in the AF-B as 10CA.

[0277] Previous reports suggest that replacement of the hydroxyl group on RJW100 with a sulfamide moiety may improve compound binding and agonism (the resulting compound is “6N”) (data not shown). Recently it was uncovered that appending the sulfamide moiety to 10CA (“6N-10CA”) may radically improve compound binding, while preserving carboxylic acid-mediated efficacy (Figure 8, Panel A). Thus, the effects of combining Tet with the 6N sulfamide were explored (Figure 8, Panel B). Addition of the sulfamide (“6N-Tet”) demonstrated improved binding (Figure 8, Panel C) and potency (Figure 8, Panel D) and enhanced thermal stability by ~ 5 °C (Figure 8, Panel E). 6N-Tet also increased NR 0B2 mRNA levels to the same extent as Tet (Figure 8, Panel F). This demonstrates the utility of the sulfamide in improving compound binding while maintaining the agonistic effects driven by pocket mouth contacts.

[0278] The specificity of 6N-Tet and isosteres with the strongest binding isosteres (Sul, Ser, Tet, and 6N-Tet) for LRH-1 over its close homolog steroidogenic factor-1 (SF-1; NR5A1) was evaluated. It was observed that the small molecules described herein were cross-reactive with SF-1 (Figure 9, Panel A). This is unsurprising, as the two receptors have nearly identical ligand binding pockets and previous studies have noted the cross-reactivity of most LRH-1 small molecule agonists with SF-1. Aside from minimal activation of PXR, 6N-Tet showed no modulation of non-NR5 A receptors PPARy, PPAR5, PPARa, MR, GR, FXR, ERa, CAR3, AR, AhR, and RORy (Figure 9, Panel B). This provides compelling evidence that the small molecules described herein are NR5 A specific.

[0279] The compounds and methods of the appended claims are not limited in scope by the specific compounds and methods described herein, which are intended as illustrations of a few aspects of the claims and any compounds and methods that are functionally equivalent are within the scope of this disclosure. Various modifications of the compounds and methods in addition to those shown and described herein are intended to fall within the scope of the appended claims. Further, while only certain representative compounds, methods, and aspects of these compounds and methods are specifically described, other compounds and methods are intended to fall within the scope of the appended claims. Thus, a combination of steps, elements, components, or constituents can be explicitly mentioned herein; however, all other combinations of steps, elements, components, and constituents are included, even though not explicitly stated.

Claims

WHAT IS CLAIMED IS:

1. A compound of the following formula:Formula I or a prodrug, salt, or stereoisomer thereof, wherein: n is 1 to 10;X is heteroaryl, amido, or sulfamate, wherein X is optionally substituted with one or more, the same or different, of hydrogen, alkyl, hydroxy, hydroxyalkyl, alkanoyl, carbocyclyl, benzyl, aryl, or heterocyclyl;R1is halogen, nitro, cyano, hydroxy, amino, sulfamoylamino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl, wherein R1is optionally substituted with one or more, the same or different, R10;R2is hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl, wherein R2is optionally substituted with one or more, the same or different, R10;R3is hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl, wherein R3is optionally substituted with one or more, the same or different, R10;R4is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl, wherein R4is optionally substituted with one or more, the same or different, R10;R5is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino,alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl, wherein R5is optionally substituted with one or more, the same or different, R10;R10is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, hydroxyalkyl, alkylthio, thioalkyl, alkylamino, aminoalkyl, (alkyl)2amino, alkanoyl, alkoxycarbonyl, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, sulfamoyl, carbocyclyl, benzoyl, benzyl, aryl, or heterocyclyl, wherein R10is optionally substituted with one or more, the same or different, R11; andR11is halogen, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy, isopropoxy, tert-butoxy, hydroxymethyl, hydroxyethyl, thiomethyl, thioethyl, aminomethyl, aminoethyl, acetyl, acetoxy, methylamino, ethylamino, dimethylamino, diethylamino, N-methyl-N- ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl, tert-butoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N- diethylsulfamoyl, N-methyl-N-ethylsulfamoyl, benzoyl, benzyl, carbocyclyl, aryl, or heterocyclyl.

2. The compound of claim 1, wherein X is -C(O)NR6R7, -O-S(=O)2NR8R9, or a tetrazole, wherein:R6, R7, R8, and R9are each independently hydrogen, alkyl, hydroxy, hydroxyalkyl, alkanoyl, carbocyclyl, benzyl, aryl, or heterocyclyl, orR6and R7or R8and R9, together with the nitrogen atom to which they are connected, form a heterocyclyl or heteroaryl, wherein R6, R7, R8, and / or R9is optionally substituted with one or more, the same or different, R10.

3. The compound of claim 1 or 2, wherein the compound has the following formula:Formula I-Aor a prodrug, salt, or stereoisomer thereof, wherein:R6and R7are each independently hydrogen, alkyl, hydroxy, hydroxyalkyl, alkanoyl, carbocyclyl, benzyl, aryl, or heterocyclyl, or R6and R7together with the nitrogen atom to which they are connected form a heterocyclyl or heteroaryl, wherein R6and / or R7is optionally substituted with one or more, the same or different, R10.

4. The compound of claim 3, wherein the compound has the following formula:Formula I-Al or a prodrug, salt, or stereoisomer thereof.

5. The compound of claim 3, wherein the compound has the following formula:Formula I-A2 or a prodrug, salt, or stereoisomer thereof.

6. The compound of claim 3, wherein the compound has the following formula:Formula I- A3 or a prodrug, salt, or stereoisomer thereof.

7. The compound of claim 3, wherein the compound has the following formula:Formula I-A4 or a prodrug, salt, or stereoisomer thereof.

8. The compound of claim 1 or 2, wherein the compound has the following formula:Formula I-B or a prodrug, salt, or stereoisomer thereof, wherein:R8and R9are each independently hydrogen, alkyl, hydroxy, hydroxyalkyl, alkanoyl, carbocyclyl, benzyl, aryl, or heterocyclyl, or R8and R9together with the nitrogen atom to which they are connected form a heterocyclyl or heteroaryl, wherein R8and / or R9is optionally substituted with one or more, the same or different,R10.

9. The compound of claim 8, wherein the compound has the following formula:Formula I-Bl or a prodrug, salt, or stereoisomer thereof.

10. The compound of claim 1 or 2, wherein the compound has the following formula:Formula I-Cor a prodrug, salt, or stereoisomer thereof, wherein:R8is hydrogen, alkyl, hydroxy, hydroxyalkyl, alkanoyl, carbocyclyl, benzyl, aryl, or heterocyclyl, wherein R8is optionally substituted with one or more, the same or different, R10.

11. The compound of claim 10, wherein the compound has the following formula:Formula I-Cl or a prodrug, salt, or stereoisomer thereof.

12. The compound of any one of claims 1-11, wherein R4is 1-phenylvinyl.

13. The compound of any one of claims 1-12, wherein n is 2 to 7.

14. The compound of any one of claims 1-13, wherein R1is hydroxy or sulfamoylamino.

15. The compound of any one of claims 1-14, wherein R2and R3are hydrogen.

16. The compound of any one of claims 1-15, wherein R5is phenyl.

17. A compound of the following structure:

18. A compound of the following structure:

19. A compound of the following structure:

20. A compound of the following structure:

21. A compound of the following structure:

22. A compound of the following structure:

23. A compound of the following structure:

24. A compound of the following structure:

25. A compound of the following structure:

26. A compound of the following structure:

27. A compound of the following structure:

28. A compound of the following structure:

29. A pharmaceutical composition comprising a compound of any one of claims 1-28 or pharmaceutically acceptable salt and a pharmaceutically acceptable excipient.

30. A method of treating or preventing cancer comprising administering a therapeutically effective amount of a pharmaceutical composition of claim 29 to a subject in need thereof.

31. The method of claim 30, wherein the cancer is selected from the group consisting of bladder cancer, breast cancer, colon cancer, rectal cancer, endometrial cancer, kidney cancer, leukemia, lung cancer, melanoma, non-Hodgkin lymphoma, pancreatic cancer, prostate cancer, and thyroid cancer.

32. A method of treating or preventing diabetes comprising administering a therapeutically effective amount of a pharmaceutical composition of claim 29 to a subject in need thereof.

33. The method of claim 32, wherein the diabetes is insulin-dependent diabetes mellitus, non-insulin-dependent diabetes mellitus, or gestational diabetes.

34. A method of treating or preventing cardiovascular disease comprising administering a therapeutically effective amount of a pharmaceutical composition of claim 29 to a subject in need thereof.

35. A method of treating or preventing inflammatory bowel diseases (IBD), comprising administering a therapeutically effective amount of a pharmaceutical composition of claim 29 to a subject in need thereof.

36. A method of treating or preventing colitis or ulcerative colitis, comprising administering a therapeutically effective amount of a pharmaceutical composition of claim 29 to a subject in need thereof.-n-37. A method of treating or preventing diabetic nephropathy, comprising administering a therapeutically effective amount of a pharmaceutical composition of claim 29 to a subject in need thereof.

38. A method of modulating steroidogenic factor-1 (SF-1) activity in a cell, comprising: contacting the cell with a therapeutically effective amount of a compound of any one of claims 1-28.

39. The method of claim 38, wherein the contacting is performed in vitro.

40. The method of claim 38, wherein the contacting is performed in vivo.

41. A method of modulating liver receptor homolog- 1 (LRH-1) activity in a cell, comprising: contacting the cell with a therapeutically effective amount of a compound of any one of claims 1-28.

42. The method of claim 41, wherein the contacting is performed in vitro.

43. The method of claim 41, wherein the contacting is performed in vivo.

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