Treatment of biological aging

Abstract

Disclosed herein are compositions and methods useful for treating or ameliorating various diseases, disorders, or conditions. Some embodiments relate to pharmaceutical compositions comprising 17-ethynyl-10R,13S-dimethyl-2,3,4,7,8R,9S,10,11,12,13,14S,15,16,17-hexadecahydro-1H-cyclopenta[a]phenanthrene-3R,7R,17S-triol (including solid forms thereof). Also presented herein is the surprising discovery that exposing a subject to the compositions disclosed herein can treat, inhibit, or ameliorate conditions associated with the circadian clock.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 63 / 381,521, filed October 28, 2022, and U.S. Provisional Application No. 63 / 508,856, filed June 16, 2023. All of the foregoing applications are incorporated herein by reference in their entirety for all purposes.

[0002] The present disclosure relates generally to the fields of chemistry and medicine. More particularly, the present disclosure relates to methods for treating biological aging using 17-ethynyl-10R,13S-dimethyl-2,3,4,7,8R,9S,10,11,12,13,14S,15,16,17-hexadecahydro-1H-cyclopenta[a]phenanthrene-3R,7R,17S-triol. [Background technology]

[0003] Aging is a complex, multifactorial process with no single cause or cure, and the question of whether aging can be classified as a disease is widely debated. Many strategies have been proposed to extend an organism's lifespan, including cell and organ replacement, comprehensive strategies to repair accumulated damage, the use of hormones to activate endogenous repair processes, and modulation of the aging process with specific mutations, gene therapy, and small molecule drugs. Animal survival strongly depends on the ability to maintain homeostasis, which is achieved in part by intracellular and intercellular communication within and between different tissues.

[0004] Many biomarkers of aging have been proposed, including telomere length, intracellular and extracellular aggregates, amino acid racemization, and genetic instability. Gene expression and DNA methylation profiles change during the aging process and may also be used as aging biomarkers. DNA methylation algorithms are increasingly being used to estimate biological aging. However, how these proposed measures of whole-organism biological aging relate to brain aging is unknown and poorly understood. Summary of the Invention

[0005] Aspects of the present disclosure relate to methods for treating, inhibiting, or ameliorating a disease or condition associated with the circadian clock in a subject in need thereof. In some embodiments, the method comprises administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol to the subject. In some embodiments, the disease or condition associated with the circadian clock in the subject in need thereof is due to modulation of DNA methylation of a gene associated with the circadian clock. In some embodiments, the disease or condition associated with the circadian clock in the subject in need thereof is associated with a gene or genomic region that becomes hypermethylated with age. In some embodiments, the disease or condition associated with the circadian clock in the subject in need thereof is associated with a gene or genomic region that becomes hypomethylated with age. In some embodiments, the disease or condition associated with the circadian clock in the subject in need thereof is associated with tau phosphorylation. In some embodiments, the disease or condition associated with the circadian clock in the subject in need thereof is associated with hyperglycemia. In some embodiments, the disease or condition associated with the circadian clock in the subject in need thereof is associated with insulin resistance. In some embodiments, the disease or condition associated with the circadian clock in the subject in need thereof is mild cognitive impairment or late-onset Alzheimer's disease. In some embodiments, administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces the subject's Alzheimer's disease composite score. In some embodiments, the subject experiences about a 5% to about 100% reduction in symptoms associated with Alzheimer's disease composite score after administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient. In some embodiments, administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces pTau (phosphorylated tau protein) in the subject.In some embodiments, the subject experiences about a 5% to about 100% reduction in a condition or symptom associated with pTau after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient. In some embodiments, administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject increases leptin in the subject. In some embodiments, the subject experiences about a 5% to about 100% reduction in a condition or symptom associated with leptin after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient. In some embodiments, administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces DNA methylation in the subject. In some embodiments, the subject experiences about a 5% to about 100% reduction in a condition or symptom associated with DNA methylation after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient. In some embodiments, administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces cardiovascular risk in the subject. In some embodiments, the subject experiences about a 5% to about 100% reduction in a condition or symptom associated with cardiovascular risk after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient. In some embodiments, administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces DNA methylation age in the subject.In some embodiments, the subject experiences about a 5% to about 100% reduction in a condition or symptom associated with DNA methylation after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient. In some embodiments, administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces DNA methylation phenoage in the subject. In some embodiments, the subject experiences about a 5% to about 100% reduction in a condition or symptom associated with DNA methylation phenoage after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient. In some embodiments, administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces DNA methylation skin blood clock in the subject. In some embodiments, the subject experiences about a 5% to about 100% reduction in conditions or symptoms associated with DNA methylation skin blood clock after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient. In some embodiments, the 17α-ethynylandrost-5-ene-3β,7β,17β-triol is administered orally. In some embodiments, the 17α-ethynylandrost-5-ene-3β,7β,17β-triol is administered intravenously. In some embodiments, the subject's waist-to-hip ratio is greater than or equal to approximately 0.90. In some embodiments, the subject has a waist-to-hip ratio of greater than or equal to approximately 0.95. In some embodiments, the 17α-ethynylandrost-5-ene-3β,7β,17β-triol is a solid-state form of 17α-ethynylandrost-5-ene-3β,7β,17β-triol.In some embodiments, the solid-state form of 17α-ethynylandrost-5-ene-3β,7β,17β-triol is a crystalline solvate of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the crystalline solvate is crystalline methanolate 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the crystalline solvate is crystalline ethanolate 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the crystalline solvate is crystalline hydrate 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the crystalline solvate is Form III 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the crystalline solvate is Form IV 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the crystalline solvate is Form V 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the solid-state form of 17α-ethynylandrost-5-ene-3β,7β,17β-triol is amorphous 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the pharmaceutical composition contains less than about 3% by weight of impurities.

[0006] Some embodiments relate to an in vitro screening method for identifying potential drug candidates or compounds capable of treating, preventing, suppressing, or ameliorating a disorder or disease, the method comprising: (i) providing a stimulation sample selected from the group consisting of cells, tissues, blood, monocytes, microglia, macrophages, adipocytes, neuroblastoma, pheochromocytoma, and Lund human mesencephalon (LUHMES) cells; (ii) stimulating the sample with an agonist to induce a phenotype or phenotypic response, wherein the phenotype or phenotypic response substantially corresponds to a disease or condition associated with at least one DNA methylation at a CpG site within a region of DNA; (iii) contacting the one or more cells exhibiting the phenotype or phenotypic response with one or more potential drug candidates or compounds; (iv) determining a change in the phenotypic responsiveness of the sample; and (v) providing the drug candidates or compounds to a subject in need thereof to treat, suppress, prevent, or ameliorate the DNA methylation-associated disease or condition in the subject. In some embodiments, the at least one DNA methylation at the CpG site is selected from the group consisting of AC073869.20, SP100, KCNQ1DN, DBNDD2, CEP112, CEP85L, SPDYE4, ZNF211, NR3C1, HLA-L, TPP2, SLC26A1, SLC37A1, CAB39L, ILKAP, NPHP4, PATE4, ARHGEF12, CELA1, OR10G7, PFN2, W DR59, snoU13, ANXA3, SVIL-AS1, PPHLN1, AP000442.1, FA, KIAA0319L, ZNF509, DLEU2L, ABL2, SGK1, TMEM245, SRSF4, DAP, GRAMD1C, FABP5P1, MCM10, ANP32E, ZNF268, ESPN, DHFR, U6, MTUS1, ATP1B3, or a combination thereof.In some embodiments, the phenotype or phenotypic response is selected from the group consisting of TNFα, GRC, CDR, MoCA, QDRS, GRC, ADCOMS, MoCA, QDRS-Cognition, ADAS-Cogl 1, heart rate, frontal lobe, systolic blood pressure, gray matter, weight, MMSE, hippocampal volume, behavior, PDQ-9, CSF glucose, precuneus GLTH, CSF pTau / Ab, or a combination thereof. In some embodiments, the phenotype or phenotypic response is selected from the group consisting of ADAS-Cog11, ADCOMS, CDR, CSF glucose, CSF pTau / Ab, frontal lobe volume, subcortical gray matter thickness, GRC, heart rate, MoCA, PDQ-9, precuneus glutathione, QDRS, QDRS-behavior, QDRS-cognition, systolic BP, tau, TNFα, and body weight. In some embodiments, the DNA methylation change is a reduction of more than 50%. In some embodiments, the DNA methylation change is a reduction of more than 55%. In some embodiments, the DNA methylation change is a reduction of more than 60%. In some embodiments, the phenotype or phenotypic response is a reduction in TNF, CDR, or QDRS-cognition, and the subject's QDRS-cognition improves. In some embodiments, the change in responsiveness is a reduction or elimination of the phenotype, and the reduction or elimination indicates that the potential drug candidate or compound can prevent, inhibit, or ameliorate a neurodegenerative disorder or disease. In some embodiments, the neurodegenerative disorder or disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, levodopa-induced dyskinesia (LID), amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), age-related hippocampal sclerosis (HS-Aging), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy, multiple system atrophy, corticobasal degeneration, and vascular parkinsonism. In some embodiments, the neurodegenerative disorder or disease is Parkinson's disease. In some embodiments, the neurodegenerative disorder is Alzheimer's disease. In some embodiments, the CpG site is correlated with genes associated with Alzheimer's disease and related dementias.In some embodiments, the disease or condition is associated with inflammatory TNF signaling. In some embodiments, the DNA methylation-associated disease or condition is associated with tau phosphorylation. In some embodiments, the DNA methylation-associated disease or condition is associated with hyperglycemia. In some embodiments, the DNA methylation-associated disease or condition is associated with insulin resistance. In some embodiments, the DNA methylation-associated disease or condition is associated with obesity, ADAS-Cog11, ADCOMS, CDR, CSF glucose, CSF pTau / Aβ, frontal lobe volume, subcortical gray matter thickness, GRC, heart rate, MoCA, PDQ-9, precuneus glutathione, QDRS, QDRS-behavior, QDRS-cognition, systolic blood pressure, tau, TNFα, and weight.

[0007] Some embodiments relate to methods for diagnosing a patient with a disease or condition. In some embodiments, the method includes: (i) providing a patient with a potential drug candidate or compound that can treat, prevent, inhibit, or ameliorate a disorder or disease; (ii) identifying DNA methylation changes in the patient; and (iii) diagnosing the patient for a disease or condition associated with a DNA methylation-related biomarker. In some embodiments, identifying DNA methylation changes in the patient identifies CpGs that are reduced by more than 50%. In some embodiments, identifying DNA methylation changes is correlated with one or more clinical changes. In some embodiments, DNA methylation changes at CpG sites are detected by AC073869.20, SP100, KCNQ1DN, DBNDD2, CEP112, CEP85L, SPDYE4, ZNF211, NR3C1, HLA-L, TPP2, SLC26A1, SLC37A1, CAB39L, ILKAP, NPHP4, PATE4, ARHGEF12, CELA1, OR10G7, PFN2, WDR59, snoU13, ANXA3, SVIL-AS1, PPHLN1, AP000442.1, FA, KIAA0319L, ZNF509, DLEU2L, ABL2, SGK1, TMEM245, SRSF4, DAP, GRAMD1C, FABP5P1, MCM10, ANP32E, ZNF268, ESPN, DHFR, U6, MTUS1, ATP1B3, or a combination thereof. In some embodiments, the disease or condition associated with a DNA methylation related biomarker is selected from the group consisting of TNFα, GRC, CDR, MoCA, QDRS, GRC, ADCOMS, MoCA, QDRS-Cognition, ADAS-Cogl 1, heart rate, frontal lobe, systolic blood pressure, gray matter, weight, MMSE, hippocampal volume, behavior, PDQ-9, CSF glucose, precuneus GLTH, CSF pTau / Ab, or a combination thereof.In some embodiments, the disease or condition associated with a biomarker associated with DNA methylation is selected from the group consisting of ADAS-Cog11, ADCOMS, CDR, CSF glucose, CSF pTau / Ab, frontal lobe volume, subcortical gray matter thickness, GRC, heart rate, MoCA, PDQ-9, precuneus glutathione, QDRS, QDRS-behavior, QDRS-cognition, systolic BP, tau, TNFα, and body weight. In some embodiments, the biomarker associated with DNA methylation is decreased in TNF, CDR, or QDRS-cognition, and the subject's QDRS-cognition improves. In some embodiments, the DNA methylation change is a greater than 50% decrease. In some embodiments, the DNA methylation change is a greater than 55% decrease. In some embodiments, the DNA methylation change is a greater than 60% decrease. In some embodiments, the disease or condition is selected from the group consisting of Alzheimer's disease, Parkinson's disease, levodopa-induced dyskinesia (LID), amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), age-related hippocampal sclerosis (HS-Aging), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy, multiple system atrophy, corticobasal degeneration, and vascular parkinsonism. In some embodiments, the neurodegenerative disorder or disease is Parkinson's disease. In some embodiments, the neurodegenerative disorder is Alzheimer's disease. In some embodiments, the CpG sites are correlated with genes associated with Alzheimer's disease and related dementias. In some embodiments, the disease or condition is associated with inflammatory TNF signaling. In some embodiments, the DNA methylation-associated disease or condition is associated with tau phosphorylation. In some embodiments, the DNA methylation-associated disease or condition is associated with hyperglycemia. In some embodiments, the DNA methylation-associated disease or condition is associated with insulin resistance.In some embodiments, the DNA methylation-associated disease or condition is associated with obesity, ADAS-Cog11, ADCOMS, CDR, CSF glucose, CSF pTau / Aβ, frontal lobe volume, subcortical gray matter thickness, GRC, heart rate, MoCA, PDQ-9, precuneus glutathione, QDRS, QDRS-behavior, QDRS-cognition, systolic blood pressure, tau, TNFα, and weight.

[0008] Not every objective set forth herein is necessarily achieved in every embodiment disclosed and / or claimed herein. [Brief explanation of the drawings]

[0009] [Figure 1] 1 is a graph showing the change in Alzheimer's Disease Composite Score (ADCOMS). [Figure 2] Graph of pTAU vs. ADCOMS. [Figure 3] The relationship between pTAU in CSF and ADCOMS is shown for MMSE. [Figure 4] Graph of DNA methylation scores (before and after). [Figure 5] A graph of DNA methylation scores against pack-years is shown. [Figure 6] 1 is a graph showing an estimated leptin score. [Figure 7] 1 is a graph measuring cardiovascular risk. [Figure 8] This is a graph measuring changes in DNAmAGE. [Figure 9] 1 is a graph measuring changes in DNA PhenoAge. [Figure 10] This is a graph measuring changes in DNAm. [Figure 11] This is a graph measuring changes in DNAm after 14 weeks of Vegistellim treatment. [Figure 12]1 is a graph depicting a Phase 3, randomized, placebo-controlled study of NE3107 (17α-ethynylandrost-5-ene-3β,7β,17β-triol) in subjects with possible mild to moderate Alzheimer's disease. [Figure 13A] 1 is a graph showing improvement in blinded assessments of a phase 3, randomized, placebo-controlled trial. [Figure 13B] 1 is a graph showing improvement in blinded assessments of a phase 3, randomized, placebo-controlled trial. [Figure 13C] 1 is a graph showing improvement in blinded assessments of a phase 3, randomized, placebo-controlled trial. [Figure 13D] 1 is a graph showing improvement in blinded assessments of a phase 3, randomized, placebo-controlled trial. [Figure 13E] 1 is a graph showing improvement in blinded assessments of a phase 3, randomized, placebo-controlled trial. [Figure 13F] 1 is a graph showing improvement in blinded assessments of a phase 3, randomized, placebo-controlled trial. [Figure 13G] 1 is a graph showing improvement in blinded assessments of a phase 3, randomized, placebo-controlled trial. [Figure 14A] Graph of vMRI hippocampal volume and amygdala volume imaging substudy in a blinded evaluation of a phase 3, randomized, placebo-controlled trial. [Figure 14B] Graph of vMRI hippocampal volume and amygdala volume imaging substudy in a blinded evaluation of a phase 3, randomized, placebo-controlled trial. [Figure 15] 1 shows a graph of the FDG-PET imaging substudy in a blinded evaluation of a phase 3, randomized, placebo-controlled trial. [Figure 16A] 1 is a graph of the neuropsychiatric inventory in the blinded evaluation of a phase 3, randomized, placebo-controlled trial. [Figure 16B] 1 is a graph of the neuropsychiatric inventory in the blinded evaluation of a phase 3, randomized, placebo-controlled trial. [Figure 17A] This graph shows that increases in fasting insulin and HOMA2-%B and decreases in HOMA2-%S occur without hypoglycemia. [Figure 17B] This graph shows that increases in fasting insulin and HOMA2-%B and decreases in HOMA2-%S occur without hypoglycemia. [Figure 17C] This graph shows that increases in fasting insulin and HOMA2-%B and decreases in HOMA2-%S occur without hypoglycemia. [Figure 18A] 1 is a graph showing the placebo effect on various endpoints. [Figure 18B] 1 is a graph showing the placebo effect on various endpoints. [Figure 18C] 1 is a graph showing the placebo effect on various endpoints. [Figure 18D] 1 is a graph showing the placebo effect on various endpoints. [Figure 18E] 1 is a graph showing the placebo effect on various endpoints. [Figure 18F] 1 is a graph showing the placebo effect on various endpoints. [Figure 18G] 1 is a graph showing the placebo effect on various endpoints. [Figure 19] 1 is a graph showing Spearman correlation for ADAS-Cog12. [Figure 20] 1 is a graph showing Spearman correlation for ADAS-Cog12. [Figure 21] 13 is a graph showing Spearman correlation for clinician global assessment of change. [Figure 22] 13 is a graph showing Spearman correlation for clinician global assessment of change. [Figure 23] 1 is a graph showing Spearman correlation for the Mini-Mental State Examination. [Figure 24] 1 is a graph showing Spearman correlation for the Mini-Mental State Examination. [Figure 25] 1 is a graph showing Spearman correlation for ADCOMS. [Figure 26] Graph showing Spearman correlation for CDR box scores. [Figure 27] 1 is a graph showing Spearman correlation for activities of daily living. [Figure 28] 1 is a graph showing that improvement in ADAS-Cog12 correlates with an increase in FDG-PET suvr. [Figure 29] 1 is a graph showing that improvement in MMSE correlates with an increase in FDG-PET suvr. [Figure 30] 1 is a graph showing that improvement in ADL correlates with an increase in SUVR on FDG-PET. [Figure 31] FIG. 10 is a graph showing that improvement in HOMA2 insulin sensitivity correlates with increased FDG-PET SUVR. [Figure 32] 1 is a graph showing that cholesterol improvement correlates with increased FDG-PET SUVR. DETAILED DESCRIPTION OF THE INVENTION

[0010] The following description provides context and examples, but should not be construed as limiting the scope of the disclosure covered by the claims that follow this specification or any other application claiming priority to this specification. No single element or group of elements is essential. For example, in some embodiments, one or more variables such as Y or Y and Q may be omitted. Any feature, structure, component, material, step, or method described and / or illustrated in any embodiment herein can be used in conjunction with or in place of any feature, structure, component, material, step, or method described and / or illustrated in any other embodiment herein.

[0011] definition Terms used herein have the meanings defined herein unless otherwise stated or implied by context. The descriptions of the described embodiments and examples are illustrative of the disclosure and are not intended to limit the disclosure in any way. In these definitions and throughout this specification, the terms "a" and "an" mean one or more, and the term "or" means and / or, unless otherwise contraindicated or implied, e.g., including mutually exclusive elements or alternatives.

[0012] "Formulation" and the like means a composition that can be administered to a subject, e.g., a human or an animal. Formulations are suitable for human or veterinary applications and typically have the properties expected of a pharmaceutical; e.g., parenteral formulations for humans typically are sterile solutions or suspensions.

[0013] "Excipient," "carrier," "pharmaceutically acceptable carrier" or similar terms means one or more component(s) or ingredient(s) that are acceptable in the sense of being compatible with the other ingredients in the disclosed composition or formulation and not overly toxic to the patient, animal, tissue or cell to which the formulation is administered.

[0014] An "effective amount" refers to the amount necessary to produce a desired effect (e.g., increasing the half-life, bioavailability or efficacy of a compound described herein, treating biological aging in a subject, inhibiting DNA methylation in a subject, etc.).

[0015] As used herein, the terms "subject," "host," "patient," and "individual" are used interchangeably and are given their ordinary meaning in the art and refer to organisms afflicted with cancer and / or leukemia, including mammals such as humans, non-human primates, ungulates, dogs, cats, horses, mice, rats, etc. The term "mammal" includes both human and non-human mammals.

[0016] "Preventing," in reference to a disease, disorder, or condition, refers to preventing the disease, disorder, or condition, e.g., preventing the clinical symptoms of the disease, disorder, or condition from developing. As used herein, the terms "prevent," "prevents," or "prevention" (and grammatical equivalents) can also refer to delaying the onset of a disease or disorder or alleviating symptoms of a disease or disorder once it has developed. This term does not imply complete eradication of the disease, but encompasses any type of prophylactic treatment that reduces the occurrence of the condition or delays the onset and / or progression of the condition.

[0017] The terms "therapeutically effective amount" and "effective amount" refer to the amount of an active pharmaceutical ingredient required to produce the desired pharmacological result. In practice, the therapeutically effective amount will vary widely depending on the severity of the condition, the age of the subject, and the desired therapeutic effect.

[0018] The terms "treatment," "treating," "treat," and the like shall be given their ordinary meaning and, as used herein, shall generally include reference to obtaining a desired pharmacological and / or physiological effect. The effect may be prophylactic, in that a disease or its symptoms are completely or partially prevented, and / or therapeutic, in that a disease and / or side effects resulting from the disease are partially or completely stabilized or cured. As used herein, the term "treatment" shall have its ordinary meaning and shall cover any treatment of disease in mammals, particularly humans, and shall include: (a) preventing a disease or condition from occurring in a subject who may be predisposed to, but has not yet been diagnosed as having, the disease or condition; (b) inhibiting a disease symptom, e.g., preventing its onset; and / or (c) alleviating a disease symptom, e.g., causing regression of the disease or condition.

[0019] The term "about" or "approximately" means within an acceptable error range for a particular value as determined by one of ordinary skill in the art, which depends in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example, "about" can mean within 1 or more than 1 standard deviation, according to practice in the art. Alternatively, "about" can mean within 20%, 10%, 5%, and 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, within 5-fold, and within 2-fold of a value. When a particular value is described in this application and claims, unless otherwise stated, the term "about" meaning within an acceptable error range for the particular value should be assumed.

[0020] All literature and similar materials cited in this application (including, but not limited to, patents, patent applications, articles, books, treatises, and internet web pages) are expressly incorporated by reference in their entirety for all purposes. Where definitions of terms in incorporated references appear to differ from those of the present teachings, the definitions of the present teachings shall control. There is an implicit "about" before temperatures, concentrations, times, etc. discussed in the present teachings, and it will be understood that minor, insubstantial deviations are within the scope of the teachings herein. In this application, unless specifically stated otherwise, the use of the singular includes the plural. Furthermore, the use of "comprise," "comprises," "comprising," "contain," "contains," "containing," "include," "includes," and "including" is not intended to be limiting. It should be understood that both the Summary of the Invention and the following Detailed Description are merely exemplary and explanatory, and not limiting. The term "and / or" indicates that the provided options can be used together or alternatively. Thus, the term "and / or" means that both possibilities exist in the combination of options.

[0021] Terms and phrases used in this application, and variations thereof, particularly in the appended claims, unless expressly stated otherwise, should be construed as open-ended rather than limiting. As examples of the foregoing, the term "including" should be read to mean "including, but not limited to," etc., and the term "comprising," as used herein, is synonymous with "including," "containing," or "characterized by," is inclusive or open-ended, and does not exclude additional, unrecited elements or method steps. The term "having" should be interpreted as "having at least." The term "includes" should be interpreted as "including, but not limited to." The term "examples" is used to provide illustrative examples of the item under discussion, rather than an exhaustive or exclusive enumeration thereof. The use of terms such as "preferably," "preferred," "desired," or "desirable," and words of similar import, should not be understood to imply that a particular feature is critical, essential, or even important to structure or function, but instead should be understood as merely intended to highlight alternative or additional features that may or may not be utilized in a particular embodiment. Additionally, the term "comprising" should be interpreted synonymously with the phrases "having at least" or "including at least." When used in the context of a process, the term "comprising" means that the process includes at least the recited steps, but may include additional steps. When used in the context of a compound, composition, or device, the term "comprising" means that the compound, composition, or device includes at least the recited features or components, but may also include additional features or components.Similarly, a group of items associated with the conjunction "and" should not be read as requiring each and every item to be present in the group, but should be read as "and / or" unless expressly stated otherwise. Similarly, a group of items associated with the conjunction "or" should not be read as requiring mutual exclusivity between the group, but should be read as "and / or" unless expressly stated otherwise.

[0022] With respect to the use of virtually any plural and / or singular term herein, those skilled in the art can convert from the plural to the singular and / or from the singular to the plural as appropriate to the context and / or application. Various singular / plural permutations may be expressly set forth herein for clarity. The indefinite article "a" or "an" does not exclude a plurality. The mere fact that certain measures are recited in mutually different dependent claims does not suggest that a combination of these measures cannot be used to advantage. Any reference signs in the claims should not be construed as limiting the scope.

[0023] Treatment method Aspects of the present disclosure relate to methods for preventing, treating, suppressing, or ameliorating a disease or condition associated with the circadian clock in a subject in need thereof. In some embodiments, the method for preventing, treating, suppressing, or ameliorating a disease or condition associated with the circadian clock may comprise administering to a patient in need thereof an effective amount of a pharmaceutical composition. In some embodiments, the pharmaceutical composition comprises a compound having the following structure: [ka] where R 5 and R 6 One of them is - OH and the other is R 5 and R 6 - H and R 12 and R 13 One of them is -OH and the other is R12 and R 13 - H and R 14 and R 15 - H and R 16 - H and R 17 is -H or -OH, and R 18 is -OH and R 19 is ethynyl and R 24 and R 25 is - CH3.

[0024] In some embodiments, the pharmaceutical composition comprises a compound having the structure [ka] where R 1 is -OH or an ester, and R 2 is -OH or ether, and R 3 -OH, =O, halogen, ester, or =CH2, R 4 is an optionally substituted amine, amide, N-linked amino acid, = NOH, or - NHOH.

[0025] In some embodiments, the pharmaceutical composition comprises an effective amount of a compound having the following structure: [ka]

[0026] In some embodiments, the pharmaceutical composition comprises an effective amount of a compound having the following structure: [ka]

[0027] In some embodiments, the pharmaceutical composition comprises an effective amount of a compound having the following structure: [ka]

[0028] In some embodiments, the pharmaceutical composition comprises an effective amount of a compound having the following structure: [ka]

[0029] In some embodiments, the pharmaceutical composition comprises an effective amount of a compound having the following structure: [ka]

[0030] In some embodiments, the pharmaceutical composition comprises an effective amount of a compound having the following structure: [ka]

[0031] In some embodiments, the pharmaceutical composition comprises an effective amount of a compound having the following structure: [ka]

[0032] In some embodiments, the pharmaceutical composition comprises 17-ethynyl-10R,13S-dimethyl-2,3,4,7,8R,9S,10,11,12,13,14S,15,16,17-hexadecahydro-1H-cyclopenta[a]phenanthrene-3R,7R,17S-triol, represented by Formula 1. The compound of Formula 1 may also be referred to as Compound 1 or 17α-ethynylandrost-5-ene-3β,7β,17β-triol, and is represented by the following structure: [ka]

[0033] In some embodiments, the pharmaceutical composition comprises (3S,5R,7S,8R,9S,10S,13S,14S,17R)-17-ethynyl-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthrene-3,7,17-triol, represented by Formula 2. The compound of Formula 2 may also be referred to as Compound 2 and is represented by the following structure: [ka]

[0034] In some embodiments, methods are provided for treating, suppressing, preventing, or ameliorating a disease or condition in a subject. In some embodiments, the method for preventing a disease or condition in a subject may involve administering a compound described herein, or a pharmaceutical form thereof, and at least one pharmaceutically acceptable excipient. In some embodiments, the method for preventing a disease or condition in a subject may involve administering Compound 1 or Compound 2, or a pharmaceutical form thereof, and at least one pharmaceutically acceptable excipient. In some embodiments, the method for preventing a disease or condition in a subject may involve administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient. In some embodiments, the method further includes measuring the biological age of the subject. In some embodiments, measuring the biological age of the subject involves determining the subject's chronological age and reversing or reducing the rate of increase in the subject's biological age, thereby preventing the disease or condition in the subject. In some embodiments, the disease or condition associated with the circadian clock in the subject in need thereof is associated with DNA methylation levels in the subject.

[0035] In some embodiments, the circadian clock-related disease or condition in the subject in need is due to modulation of DNA methylation of circadian clock-related genes. In some embodiments, the circadian clock-related disease or condition in the subject in need is associated with genes or genomic regions that become hypermethylated with age. In some embodiments, the circadian clock-related disease or condition in the subject in need is associated with genes or genomic regions that become hypomethylated with age. In some embodiments, the circadian clock-related disease or condition in the subject in need is associated with tau phosphorylation. In some embodiments, the circadian clock-related disease or condition in the subject in need is associated with hyperglycemia. In some embodiments, the circadian clock-related disease or condition in the subject in need is associated with insulin resistance.

[0036] In some embodiments, the present disclosure provides a method for suppressing DNA methylation in a subject in need thereof. In some embodiments, the method comprises administering to the subject an effective amount of a compound described herein, or a pharmaceutical form thereof, and at least one pharmaceutically acceptable excipient, thereby reducing DNA methylation in the subject. In some embodiments, the compound is Compound 1 or Compound 2, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is 17α-ethynylandrost-5-ene-3β,7β,17β-triol, which, when administered to a subject in need thereof, has been shown to produce multiple beneficial responses. For example, in some embodiments, administration reduces DNA methylation in the subject by at least 5% (e.g., at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15% or more, or any value or range thereof) compared to a control measurement, e.g., compared to DNA methylation in the subject before administration (e.g., subject "baseline" DNA methylation). DNA methylation in a subject can be assessed quantitatively and / or qualitatively by any standard technique in the art, e.g., measured by a marker of relative global methylation compared to a control, or measured by LINE-1 methylation compared to a control. For example, in some embodiments, administration reduces LINE-1 methylation in the subject by at least 5% (e.g., at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15% or more) compared to a control measurement, e.g., compared to LINE-1 methylation in the subject prior to administration (e.g., subject baseline LINE-1 methylation). For example, in some embodiments, administration may suppress LINE-1 methylation in the subject by at least 5%, at least 8%, at least 10%, or at least 15%. In some embodiments, administration may reduce LINE-1 methylation in the subject by about 5% to about 20%, about 6% to about 15%, or about 8% to about 10%.In some embodiments, improvement of symptoms associated with DNA methylation may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol, or by a range including and / or spanning the aforementioned values. For example, a subject may experience an improvement in symptoms or conditions associated with DNA methylation from approximately 5% to objectively normal after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0037] In some embodiments, a subject may experience an improvement in symptoms or conditions associated with genes or genomic regions that become hypermethylated with age after administration of a composition described herein. In some embodiments, a subject may experience an improvement in symptoms or conditions associated with genes or genomic regions that become hypermethylated with age after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the improvement in symptoms associated with genes or genomic regions that become hypermethylated with age after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values. For example, a subject may experience an improvement ranging from approximately 5% to 100% in symptoms or conditions associated with genes or genomic regions that become hypermethylated with age after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0038] In some embodiments, a subject may experience an improvement in symptoms or conditions associated with genes or genomic regions that become hypomethylated with age after administration of a composition described herein. In some embodiments, a subject may experience an improvement in symptoms or conditions associated with genes or genomic regions that become hypomethylated with age after administration of a composition described herein. In some embodiments, the improvement in symptoms associated with genes or genomic regions that become hypomethylated with age may be inhibited by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values, after administration of a composition described herein. For example, a subject may experience an improvement ranging from approximately 5% to 100% in symptoms or conditions associated with genes or genomic regions that become hypomethylated with age after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0039] In some embodiments, a subject may experience an improvement in symptoms or conditions related to hyperglycemia (which may lead to type I and type II diabetes) after administration of a composition described herein. In some embodiments, a subject may experience an improvement in symptoms or conditions related to hyperglycemia (which may lead to type I and type II diabetes) after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the improvement in symptoms related to hyperglycemia may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values, after administration of a composition described herein. For example, a subject may experience an improvement in symptoms or conditions related to hyperglycemia ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0040] In some embodiments, a subject may experience an improvement in symptoms or conditions related to hyperlipidemia (such as obesity-related conditions) after administration of a composition described herein. In some embodiments, a subject may experience an improvement in symptoms or conditions related to hyperlipidemia (such as obesity-related conditions) after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the improvement in symptoms related to hyperlipidemia may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, or by a range including and / or spanning the aforementioned values. For example, a subject may experience an improvement in symptoms or conditions related to hyperlipidemia ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0041] In some embodiments, a subject may experience an improvement in symptoms or conditions related to insulin resistance after administration of a composition described herein. In some embodiments, a subject may experience an improvement in symptoms or conditions related to insulin resistance after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the improvement in symptoms related to insulin resistance after administration of a composition described herein may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values. For example, a subject may experience an improvement in symptoms or conditions related to insulin resistance ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0042] In some embodiments, a subject may experience a suppression or reduction in symptoms related to Alzheimer's disease composite score after administration of a composition described herein. In some embodiments, a subject may experience a suppression or reduction in symptoms related to Alzheimer's disease composite score after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the suppression or reduction in symptoms related to Alzheimer's disease composite score after administration of a composition described herein may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values. For example, a subject may experience a reduction in symptoms related to the Alzheimer's Disease Composite Score ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient. In some embodiments, reducing or suppressing symptoms related to the Alzheimer's Disease Composite Score may also reduce other neuropsychological measures (including, but not limited to, ADAS-Cog, MoCA, MMSE, CDR, QDRS, and PDQ-9). In some embodiments, reducing or suppressing symptoms related to the Alzheimer's Disease Composite Score may also improve neuroimaging results (including, but not limited to, ASL, BOLD, MRS, task- or resting-state fMRI, vMRI, and FDG-PET).

[0043] In some embodiments, a subject may experience a suppression or reduction in symptoms associated with CSF phosphorylated tau ("pTau") after administration of a composition described herein. In some embodiments, a subject may experience a suppression or reduction in symptoms associated with pTau after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the suppression or reduction in symptoms associated with pTau may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values, after administration of a composition described herein. For example, a subject may experience a suppression of pTau-related symptoms ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0044] In some embodiments, a subject may experience a suppression or reduction in symptoms associated with leptin deficiency after administration of a composition described herein. In some embodiments, a subject may experience a suppression or reduction in symptoms associated with leptin deficiency after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the suppression or reduction in symptoms associated with leptin deficiency may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values, after administration of a composition described herein. For example, a subject may experience a suppression of symptoms associated with leptin deficiency ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0045] In some embodiments, a subject may experience a suppression or reduction in symptoms associated with leptin resistance after administration of a composition described herein. In some embodiments, a subject may experience a suppression or reduction in symptoms associated with leptin resistance after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the suppression or reduction in symptoms associated with leptin resistance may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values ​​after administration of a composition described herein. For example, a subject may experience a suppression of symptoms associated with leptin resistance ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0046] In some embodiments, a subject may experience a suppression or reduction in symptoms associated with DNA methylation after administration of a composition described herein. In some embodiments, a subject may experience a suppression or reduction in symptoms associated with DNA methylation after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the suppression or reduction in symptoms associated with DNA methylation may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values, after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient. For example, a subject may experience a suppression of symptoms associated with DNA methylation ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0047] In some embodiments, after administering the compositions described herein, subjects may experience a suppression or reduction in symptoms related to cardiovascular risk. In some embodiments, after administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol, subjects may experience a suppression or reduction in symptoms related to cardiovascular risk. In some embodiments, after administering the compositions described herein, the suppression or reduction in symptoms related to cardiovascular risk may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values. For example, a subject may experience a reduction in symptoms related to cardiovascular risk ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0048] In some embodiments, a subject may experience a suppression or reduction in symptoms associated with the subject's epigenetic age ("DNA methylation phenoage") after administration of a composition described herein. In some embodiments, a subject may experience a suppression or reduction in symptoms associated with DNA methylation phenoage after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the suppression or reduction in symptoms associated with DNA methylation phenoage may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% after administration of a composition described herein, or by a range including and / or spanning the aforementioned values. For example, a subject may experience a suppression of symptoms associated with DNA methylation phenoaging ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0049] In some embodiments, a subject may experience a suppression or reduction in symptoms related to the subject's epigenetic clock ("DNA methylation skin blood clock") after administration of a composition described herein. In some embodiments, a subject may experience a suppression or reduction in symptoms related to the subject's DNA methylation skin blood clock after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the suppression or reduction in symptoms related to the DNA methylation skin blood clock may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% after administration of a composition described herein, or by a range including and / or spanning the aforementioned values. For example, a subject may experience a suppression of symptoms associated with DNA methylation skin blood clock ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0050] In some embodiments, a subject may experience a suppression or reduction in symptoms related to the DunedinPACE clock after administration of a composition described herein. In some embodiments, a subject may experience a suppression or reduction in symptoms related to the DunedinPACE clock after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the suppression or reduction in symptoms related to the DunedinPACE clock after administration of a composition described herein may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values. For example, a subject may experience a suppression of symptoms associated with the DunedinPACE clock ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0051] In some embodiments, a subject may experience prevention, suppression, or reduction of cancer- or tumor-related conditions or symptoms after administration of a composition described herein. In some embodiments, a subject may experience prevention, suppression, or reduction of cancer- or tumor-related conditions or symptoms after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the prevention, suppression, or reduction of cancer- or tumor-related conditions or symptoms after administration of a composition described herein may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values. For example, a subject may experience a cancer or tumor-related condition or symptom being prevented, suppressed, or reduced by approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0052] In some embodiments, a subject may experience a reduction or suppression of symptoms associated with atherosclerosis after administration of a composition described herein. In some embodiments, a subject may experience a reduction or suppression of symptoms associated with atherosclerosis after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, a reduction or suppression of symptoms associated with atherosclerosis may be approximately equal to or greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% after administration of a composition described herein, or within a range including and / or spanning the aforementioned values. For example, a subject may experience a reduction in symptoms associated with atherosclerosis ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0053] In some embodiments, a subject may experience a suppression or reduction in symptoms associated with schizophrenia after administration of a composition described herein. In some embodiments, a subject may experience a suppression or reduction in symptoms associated with schizophrenia after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the suppression or reduction in symptoms associated with schizophrenia after administration of a composition described herein may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values. For example, a subject may experience a suppression of symptoms associated with schizophrenia ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0054] In some embodiments, a subject may experience a suppression or reduction in symptoms associated with autoimmune disease after administration of a composition described herein. In some embodiments, a subject may experience a suppression or reduction in symptoms associated with autoimmune disease after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the suppression or reduction in symptoms associated with autoimmune disease may be suppressed by approximately or greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% after administration of a composition described herein, or by a range including and / or spanning the aforementioned values. For example, a subject may experience a suppression of symptoms associated with an autoimmune disease ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0055] In some embodiments, a subject may experience a suppression or reduction in symptoms associated with rheumatoid arthritis after administration of a composition described herein. In some embodiments, a subject may experience a suppression or reduction in symptoms associated with rheumatoid arthritis after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the suppression or reduction in symptoms associated with rheumatoid arthritis may be suppressed by approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% or by a range including and / or spanning the aforementioned values ​​after administration of a composition described herein. For example, a subject may experience a suppression of symptoms associated with rheumatoid arthritis ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0056] In some embodiments, a subject may experience a suppression or reduction in symptoms associated with systemic lupus erythematosus after administration of a composition described herein. In some embodiments, a subject may experience a suppression or reduction in symptoms associated with systemic lupus erythematosus after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the suppression or reduction in symptoms associated with systemic lupus erythematosus after administration of a composition described herein may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values. For example, a subject may experience a suppression of symptoms associated with systemic lupus erythematosus ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0057] In some embodiments, after administration of the compositions described herein, subjects may experience suppression or reduction of symptoms associated with multiple sclerosis. In some embodiments, after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol, subjects may experience suppression or reduction of symptoms associated with multiple sclerosis. In some embodiments, after administration of the compositions described herein, the suppression or reduction of symptoms associated with multiple sclerosis may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values. For example, a subject may experience a suppression of symptoms associated with multiple sclerosis ranging from approximately 5% to 100% after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0058] In some embodiments, after administration of a composition described herein, a subject may experience prevention, suppression, or reduction of risks to eggs or sperm associated with an increased risk of having a child with an autism spectrum disorder. In some embodiments, after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol, a subject may experience prevention, suppression, or reduction of risks to eggs or sperm associated with an increased risk of having a child with symptoms associated with autism spectrum disorder. In some embodiments, the suppression or reduction of eggs or sperm at increased risk of producing a child with symptoms associated with autism spectrum disorder may be suppressed by approximately or greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% after administration of a composition described herein. For example, a subject may experience approximately 5% to 100% prevention, suppression, or reduction of eggs or sperm at increased risk of producing a child with symptoms associated with autism spectrum disorder after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0059] In some embodiments, a subject may experience prevention, suppression, or reduction in eggs or sperm at increased risk of having a baby with Down syndrome after administration of a composition described herein. In some embodiments, a subject may experience prevention, suppression, or reduction in eggs or sperm at increased risk of having a baby with Down syndrome after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, suppression or reduction of symptoms associated with Down syndrome may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values, after administration of a composition described herein. For example, a subject may experience approximately 5% to 100% prevention or suppression of eggs or sperm that are at increased risk of giving birth to a child with Down syndrome after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0060] In some embodiments, the pharmaceutical composition comprises a solid-state form of a compound described herein. In some embodiments, the pharmaceutical composition comprises a solid-state form of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the solid-state form is crystalline 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the solid-state form is crystalline 17α-ethynylandrost-5-ene-3β,7β,17β-triol that is substantially free of amorphous 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

[0061] In some embodiments, the solid-state form is a crystalline solvate of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the crystalline solvate is a crystalline methanolate of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the crystalline solvate is a crystalline ethanolate of 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the crystalline solvate is a crystalline hydrate of 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

[0062] In some embodiments, the crystalline solvate is Form III 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the crystalline solvate is Form IV 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the crystalline solvate is Form V 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

[0063] In some embodiments, the solid-state form of 17α-ethynylandrost-5-ene-3β,7β,17β-triol is amorphous 17α-ethynylandrost-5-ene-3β,7β,17β-triol. In some embodiments, the amorphous 17α-ethynylandrost-5-ene-3β,7β,17β-triol is substantially free of solid-state form of 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

[0064] In some embodiments, the compounds or pharmaceutical compositions described herein are administered orally. In some embodiments, the compounds or pharmaceutical compositions described herein are administered intravenously. In some embodiments, the compounds or pharmaceutical compositions described herein are administered topically. In some embodiments, 17α-ethynylandrost-5-ene-3β,7β,17β-triol is administered orally. In other embodiments, 17α-ethynylandrost-5-ene-3β,7β,17β-triol is administered intravenously. In other embodiments, 17α-ethynylandrost-5-ene-3β,7β,17β-triol is administered topically.

[0065] In some embodiments, the compounds described herein are administered as a formulation or composition with at least one pharmaceutically acceptable excipient.In some embodiments, the compounds described herein are administered as a formulation or composition with at least one pharmaceutically acceptable excipient and at least one pharmaceutically acceptable carrier.In some embodiments, 17α-ethynylandrost-5-ene-3β,7β,17β-triol is administered as a formulation with at least one pharmaceutically acceptable excipient.In some embodiments, 17α-ethynylandrost-5-ene-3β,7β,17β-triol is administered as a formulation with at least one pharmaceutically acceptable excipient and at least one pharmaceutically acceptable carrier.In some embodiments, 17α-ethynylandrost-5-ene-3β,7β,17β-triol is administered as a formulation with at least one pharmaceutically acceptable excipient and at least one pharmaceutically acceptable carrier.In some embodiments, 17α-ethynylandrost-5-ene-3β,7β,17β-triol is administered as a formulation with at least one pharmaceutically acceptable carrier. Other pharmaceutically acceptable excipients suitable for use in the compositions include absorption enhancers, acidifying agents, sustained-release agents, alkalizing agents, antioxidants, buffers, chelating agents, coloring agents, complexing agents, emulsifying agents, flavoring agents, humectants, humidity adjusting agents, pH adjusting agents, preservatives, solubilizing agents, stabilizers, surfactants, suspending agents, sweetening agents, flavoring agents, and wetting agents.

[0066] Formulations include compositions containing one, two, three, four, or more pharmaceutically acceptable excipients or carriers. The compositions are used to prepare formulations suitable for human or animal use. Suitable routes of administration for formulations include oral, rectal, nasal, transmucosal, topical (including buccal and sublingual), vaginal, rectal, and parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal, intraocular, and epidural). Generally, aqueous and non-aqueous liquid or cream formulations are delivered parenterally, orally, or topically. In other embodiments, such as the disclosed intermittent administration methods, 17α-ethynylandrost-5-ene-3β,7β,17β-triol may be present as an aqueous or non-aqueous liquid or solid formulation suitable for administration via any of the routes disclosed herein, for example, oral, topical, buccal, sublingual, parenteral, inhalation aerosol, or depot formulations such as subcutaneous, intraperitoneal, or intramuscular depots. It will be appreciated that the preferred route may vary depending, for example, on the condition or weight of the subject, or the subject's response to treatment with 17α-ethynylandrost-5-ene-3β,7β,17β-triol, or other treatments used or appropriate for the situation.

[0067] The formulations include those suitable for the above-mentioned administration routes.The formulations may conveniently be presented in unit dosage form and may be prepared by any method well known in the art of pharmacy.Techniques and formulations are generally described in, for example, Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa. 2022, 23 rd edition, Adeboye et al.,PDA J.Pharm.Sci.Tech.1997 51:166-171, G.Cole,et al.,editors,Pharmaceutical Coating Technology,1995,Taylor &Francis,ISBN 0 136628915,HALieberman,et al.,editors,Pharmaceutical Dosage Forms,1992 2 ndrevised edition, volumes 1 and 2, Marcel Dekker, ISBN 0824793870, JT Carstensen. Pharmaceutical Preformulation, 1998, pages 1-306, Technomic Publishing Co. ISBN 1566766907. Exemplary excipients for the formulation include emulsifying wax, propyl gallate, citric acid, lactic acid, polysorbate 80, sodium chloride, isopropyl palmitate, glycerin, white petrolatum, and other excipients disclosed herein.

[0068] Formulations or compositions disclosed herein for use in preparing formulations suitable for administration by the routes disclosed herein optionally comprise an average particle size ranging from about 0.01 to about 500 microns, about 0.1 to about 100 microns, or about 0.5 to about 75 microns, including the range of 0.01 to 500 microns in increments of 0.05 microns or 0.1 microns or other increments, e.g., average particle size of about 0.05, 0.1, 0.5, 1, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 50, 60, 75, 85, 100, 120 microns, etc. When 17α-ethynylandrost-5-ene-3β,7β,17β-triol or a composition comprising 17α-ethynylandrost-5-ene-3β,7β,17β-triol is used as an intermediate for producing a formulation, it may comprise one, two, three, or more of these average particle sizes or particle size ranges. When preparing any of the compositions or formulations disclosed herein comprising 17α-ethynylandrost-5-ene-3β,7β,17β-triol (and optionally one or more excipients and / or one or more carriers), the compound or composition can optionally be milled, sieved, or otherwise granulated to obtain the desired particle size.

[0069] Non-limiting examples of fillers suitable for use in the compositions include lactose, microcrystalline cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, methylcellulose polymers, hydroxyethyl cellulose, sodium carboxymethyl cellulose, carboxymethylene, carboxymethylhydroxyethyl cellulose and other cellulose derivatives, sucrose, agarose, sorbitol, mannitol, dextrin, maltodextrin, starch or modified starch (including potato starch, corn starch, rice starch), calcium phosphate (e.g., basic calcium phosphate, calcium hydrogen phosphate, dicalcium phosphate hydrate), calcium sulfate, calcium carbonate, sodium alginate, and collagen.

[0070] Non-limiting examples of diluents suitable for use in the compositions include, for example, calcium carbonate, dibasic calcium phosphate, tribasic calcium phosphate, calcium sulfate, microcrystalline cellulose, powdered cellulose, dextran, dextrin, dextrose, fructose, kaolin, lactose, mannitol, sorbitol, starch, pregelatinized starch, sucrose, and sugar.

[0071] Non-limiting examples of disintegrants suitable for use in the compositions include alginic acid or alginates, microcrystalline cellulose, low-substituted hydroxypropyl cellulose and other cellulose derivatives, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, starch, pregelatinized starch, and carboxymethyl starch.

[0072] Non-limiting examples of binders suitable for use in the compositions include acacia, alginic acid, agar, calcium carrageenan, sodium carboxymethylcellulose, microcrystalline cellulose, dextrin, ethylcellulose, gelatin, liquid glucose, guar gum, hydroxypropyl methylcellulose, methylcellulose, pectin, PEG, polyethylene oxide, povidone, and pregelatinized starch.

[0073] Non-limiting examples of glidants and / or lubricants suitable for use in the compositions include stearic acid, magnesium stearate, calcium stearate, or other metallic stearates, talc, waxes, and glycerides, light mineral oil, PEG, glyceryl behenate, colloidal silica, hydrogenated vegetable oil, corn starch, sodium stearyl fumarate, polyethylene glycol, alkyl sulfates, sodium benzoate, and sodium acetate.

[0074] Non-limiting examples of antioxidants suitable for use in the compositions include ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, hypophosphorous acid, monothioglycerol, potassium metabisulfite, propyl gallate, sodium formaldehyde sulfoxylate, sodium metabisulfite, sodium thiosulfate, sulfur dioxide, tocopherol, tocopherol acetate, tocopherol hemisuccinate, and derivatives of tocopherol.

[0075] In some embodiments, the pharmaceutically acceptable excipient is selected from sodium dodecyl sulfate, microcrystalline cellulose, magnesium stearate, and any combination of the foregoing, hi some embodiments, the pharmaceutically acceptable excipient is sodium dodecyl sulfate.

[0076] In some embodiments, the pharmaceutical composition is formulated into an oral dosage form. In some embodiments, the dosage form may include a capsule or tablet. In some embodiments, the dosage form may include one or more different types of delayed-release layers selected from a sealant layer and / or an enteric layer. For example, delayed-release layers with different release rate characteristics can provide dosage forms with different overall drug release characteristics. In some such embodiments, the pharmaceutically acceptable excipient is a surfactant. In some embodiments, the surfactant is present in an amount sufficient to provide 90% dissolution of the pharmaceutical composition in water at ambient temperature after 30 minutes. In some embodiments, the surfactant is sodium lauryl sulfate. In some embodiments, the pharmaceutical composition is a capsule or tablet.

[0077] In some embodiments, the pharmaceutical composition contains less than about 3% by weight of impurities.

[0078] In some embodiments, the pharmaceutical composition contains less than about 5% by weight of 3β-hydroxy-androst-5-ene-7,17-dione.

[0079] In some embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable formulation of 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

[0080] In some embodiments, the use is concurrent with the use of at least one additional pharmaceutical agent. In some embodiments, the additional pharmaceutical agent is administered with a delay following the first administration of the composition. In some embodiments, the first administration can be administered using a dosing regimen that is daily, weekly, monthly, or any combination of the foregoing. In some embodiments, the first administration regimen can include one, two, three, or more daily doses of the composition. In some embodiments, the first administration regimen can include one, two, three, or more weekly doses of the composition. In some embodiments, the first administration regimen can include one, two, three, or more monthly doses of the composition. In some embodiments, the delay period is about 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more years, or a range including and / or spanning the aforementioned values. In some embodiments, the delay period is two years or more. In some embodiments, the delay time is zero, and the additional pharmaceutical agent is administered simultaneously with the first administration of the composition. In some embodiments, the additional pharmaceutical agent is administered using a dosing regimen that is daily, weekly, monthly, or any combination of the foregoing. In some embodiments, the dosing regimen of the additional pharmaceutical agent may include one, two, three, or more daily doses of the composition. In some embodiments, the dosing regimen of the additional pharmaceutical agent may include one, two, three, or more weekly doses of the composition. In some embodiments, the dosing regimen of the additional pharmaceutical agent may include one, two, three, or more monthly doses of the composition.

[0081] Some embodiments of the present disclosure relate to the use of 17α-ethynylandrost-5-ene-3β,7β,17β-triol in the manufacture of a medicament for treating a neurodegenerative condition.

[0082] Aspects of the present disclosure relate to in vitro screening methods for identifying potential drug candidates. In some embodiments, the in vitro screening methods may identify potential drug candidates that can treat, prevent, suppress, or ameliorate a disorder or disease. In some embodiments, the disorder or disease is a neurodegenerative disorder or condition. In some embodiments, the neurodegenerative disease or condition is dementia. As used herein, "drug candidate" refers to a specific molecule, compound, or therapeutic agent, whether natural or synthetic, that has potential therapeutic benefit and is selected for further development and evaluation.

[0083] In some embodiments, the in vitro screening method may include providing a sample for stimulation. In some embodiments, the sample is a cell. In some embodiments, the sample is a tissue. In some embodiments, the sample is blood. In some embodiments, the sample comprises monocytes. In some embodiments, the sample comprises microglia. In some embodiments, the monocytes are CX3CR1 low (low expression), CCR2 pos (positive), Ly6C high (high expression), PD-L1 neg (negative), CD14 ++ , CD16 + , CD14 dim (weakly positive), CD16 + , CD16 - , CX3CR1 high , CCR2 neg , Ly6C low , PD-L1 pos In some embodiments, the cells are T cells or granulocytes, including but not limited to, THP-1 human monocytes, RAW264.7 macrophages, 3T3-L1 adipocytes, SH-SY5Y neuroblastoma, PC-12 pheochromocytoma, and Lund human mesencephalon (LUHMES) cells. In some embodiments, the cells are NK cells or granulocytes. In some embodiments, the cells for stimulation may be selected from the group consisting of, but not limited to, THP-1 human monocytes, RAW264.7 macrophages, 3T3-L1 adipocytes, SH-SY5Y neuroblastoma, PC-12 pheochromocytoma, and Lund human mesencephalon (LUHMES) cells.

[0084] In some embodiments, the in vitro screening method may include stimulating cells with an agonist to induce a phenotype or phenotypic change. In some embodiments, the phenotype may correspond to the phenotype of cells or tissues affected by a neurodegenerative disease or disorder. In some embodiments, the neurodegenerative disorder or disease may be selected from the group consisting of, but not limited to, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), age-related hippocampal sclerosis (HS-Aging), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy, multiple system atrophy, corticobasal degeneration, and vascular parkinsonism.

[0085] In some embodiments, in vitro screening methods may involve contacting one or more cells or tissues exhibiting the phenotype with the potential drug candidate. In some embodiments, in vitro screening methods may involve contacting one or more cells exhibiting the phenotype with the potential drug candidate in parallel in a high-throughput screening method. In other embodiments, in vitro screening methods may involve contacting one or more cells exhibiting the phenotype with one or more potential drug candidates in parallel in a high-throughput screening method. In yet other embodiments, in vitro screening methods may involve sequentially contacting one or more cells exhibiting the phenotype with the potential drug candidate.

[0086] In some embodiments, the in vitro screening method may include determining a change in responsiveness in a cellular phenotype. In some embodiments, the change in responsiveness may be a reduction, suppression, or elimination of the cellular phenotype. In some embodiments, the phenotype may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values, after contacting cells exhibiting the phenotype with a potential drug candidate.

[0087] In some embodiments, the phenotype is associated with DNA methylation in the subject. In some embodiments, the DNA methylation occurs at CpG sites. In some embodiments, the CpG sites include AC073869.20, SP100, KCNQ1DN, DBNDD2, CEP112, CEP85L, SPDYE4, ZNF211, NR3C1, HLA-L, TPP2, SLC26A1, SLC37A1, CAB39L, ILKAP, NPHP4, PATE4, ARHGEF12, CELA1, OR10G7, PFN2, WDR59, snoU13, AN XA3, SVIL-AS1, PPHLN1, AP000442.1, FA, KIAA0319L, ZNF509, DLEU2L, ABL2, SGK1, TMEM245, SRSF4, DAP, GRAMD1C, FABP5P1, MCM10, ANP32E, ZNF268, ESPN, DHFR, U6, MTUS1, ATP1B3, or a combination thereof. In some embodiments, DNA methylation may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or a range including and / or spanning the aforementioned values, after contacting cells exhibiting the phenotype with a potential drug candidate. In some embodiments, DNA methylation is reduced by more than 50%. In some embodiments, DNA methylation is reduced by more than 55%. In some embodiments, DNA methylation is reduced by more than 60%.

[0088] In some embodiments, the drug candidates or compounds described herein are provided to a subject to treat, prevent, inhibit, or ameliorate a disease or condition associated with DNA methylation. In some embodiments, the disease or condition is associated with TNFα, GRC, CDR, MoCA, QDRS, GRC, ADCOMS, MoCA, QDRS-Cognition, ADAS-Cogl 1, heart rate, frontal lobe, systolic blood pressure, gray matter, body weight, MMSE, hippocampal volume, behavior, PDQ-9, CSF glucose, precuneus GLTH, or CSF pTau / Ab. In some embodiments, treating, preventing, inhibiting, or ameliorating a DNA methylation-related disease or condition may be improved by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% following administration of the drug candidate or compound, or may be improved in a range including and / or spanning the aforementioned values, or may be improved to the extent that an objectively normal or disease-free state is established. For example, a subject may experience an improvement ranging from approximately 5% to 100%, or improvement to objectively normal, in a symptom or condition related to TNFα, GRC, CDR, MoCA, QDRS, GRC, ADCOMS, MoCA, QDRS-Cognition, ADAS-Cogl 1, heart rate, frontal lobe, systolic blood pressure, gray matter, weight, MMSE, hippocampal volume, behavior, PDQ-9, CSF glucose, precuneus GLTH, CSF pTau / Ab, regions hypermethylated with aging, or a symptom or condition related to regions hypermethylated with aging, following administration of the drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0089] Aspects of the present disclosure relate to methods for diagnosing a patient with a disease or condition. In some embodiments, the method includes providing a patient with a potential drug candidate or compound described herein that can treat, prevent, suppress, or ameliorate the disorder or disease. In some embodiments, the method includes identifying DNA methylation changes in the patient. In some embodiments, the method includes diagnosing the patient for a disease or condition associated with a DNA methylation-related biomarker.

[0090] In some embodiments, identifying DNA methylation changes in a patient identifies CpGs that are reduced by more than 50%. In some embodiments, identifying DNA methylation changes is correlated with one or more clinical changes. In some embodiments, DNA methylation changes at CpG sites include AC073869.20, SP100, KCNQ1DN, DBNDD2, CEP112, CEP85L, SPDYE4, ZNF211, NR3C1, HLA-L, TPP2, SLC26A1, SLC37A1, CAB39L, ILKAP, NPHP4, PATE4, ARHGEF12, CELA1, OR10G7, PFN2, WDR59, snoU13, ANXA3, SVIL-AS1, PPHLN1, AP000442.1, FA, KIAA0319L, ZNF509, DLEU2L, ABL2, SGK1, TMEM245, SRSF4, DAP, GRAMD1C, FABP5P1, MCM10, ANP32E, ZNF268, ESPN, DHFR, U6, MTUS1, ATP1B3, or a combination thereof.

[0091] In some embodiments, the disease or condition associated with a biomarker related to DNA methylation is selected from the group consisting of TNFα, GRC, CDR, MoCA, QDRS, GRC, ADCOMS, MoCA, QDRS-Cognition, ADAS-Cogl 1, heart rate, frontal lobe, systolic blood pressure, gray matter, body weight, MMSE, hippocampal volume, behavior, PDQ-9, CSF glucose, precuneus GLTH, CSF pTau / Ab, frontal lobe volume, subcortical gray matter thickness, GRC, heart rate, MoCA, PDQ-9, precuneus glutathione, QDRS, QDRS-behavior, QDRS-cognition, systolic BP, tau, TNFα, and body weight. In some embodiments, DNA methylation-related biomarkers such as TNF, CDR, and QDRS-cognition are decreased, and the subject's QDRS-cognition is improved. In some embodiments, the DNA methylation change is a decrease of more than 50%. In some embodiments, the DNA methylation change is a decrease of more than 55%. In some embodiments, the DNA methylation change is a decrease of more than 60%.

[0092] In some embodiments, a subject may experience prevention, or suppression or reduction of obesity after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience prevention or suppression of obesity after administration of a drug candidate or compound. In some embodiments, suppression or reduction of obesity-related symptoms may be suppressed by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% after administration of a drug candidate or compound described herein, or in a range including and / or spanning the aforementioned values. For example, a subject may experience prevention or suppression of obesity in the range of approximately 5% to 100% after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0093] In some embodiments, a subject may experience prevention, suppression, or reduction of ADCOMS after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience improvement in ADCOMS after administration of a drug candidate or compound. In some embodiments, the improvement related to ADCOMS may be equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or within a range including and / or spanning the aforementioned values, or to an objectively normal state, after administration of a drug candidate or compound described herein. For example, a subject may experience an improvement in ADCOMS ranging from approximately 5% to 100%, or to an objectively normal state, after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0094] In some embodiments, a subject may experience prevention, suppression, or reduction of CDR after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience an improvement in CDR after administration of a drug candidate or compound. In some embodiments, the improvement in CDR may be equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% after administration of a drug candidate or compound described herein, or within a range including and / or spanning the aforementioned values. For example, a subject may experience an improvement in CDR ranging from approximately 5% to 100% after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0095] In some embodiments, a subject may experience prevention, suppression, or reduction of CSF glucose after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience improvement in CSF glucose after administration of a drug candidate or compound. In some embodiments, the improvement in CSF glucose may be equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or a range including and / or spanning the aforementioned values, after administration of a drug candidate or compound described herein. For example, a subject may experience an improvement in CSF glucose ranging from approximately 5% to 100%, or improvement to objectively normal, after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0096] In some embodiments, a subject may experience prevention, or suppression or reduction of CSF pTau / Aβ after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience improvement in CSF pTau / Aβ after administration of a drug candidate or compound. In some embodiments, the improvement in CSF pTau / Aβ may be improved by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values, after administration of a drug candidate or compound described herein. For example, a subject may experience an improvement ranging from approximately 5% to 100%, or improvement to objectively normal, in CSF pTau / Aβ following administration of the drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0097] In some embodiments, a subject may experience prevention, suppression, or reduction of frontal lobe volume after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience improvement in frontal lobe volume after administration of a drug candidate or compound. In some embodiments, the improvement related to frontal lobe volume may be improved by an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or by a range including and / or spanning the aforementioned values, after administration of a drug candidate or compound described herein. For example, a subject may experience an improvement in frontal lobe volume ranging from approximately 5% to 100%, or improvement to objectively normal, after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0098] In some embodiments, a subject may experience prevention, suppression, or reduction of subcortical gray matter thickness after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience improvement in subcortical gray matter thickness after administration of a drug candidate or compound. In some embodiments, the improvement in subcortical gray matter thickness may be approximately equal to or greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or a range including and / or spanning the aforementioned values, after administration of a drug candidate or compound described herein. For example, a subject may experience an improvement in subcortical gray matter thickness ranging from approximately 5% to 100%, or improvement to objectively normal, after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0099] In some embodiments, a subject may experience prevention, suppression, or reduction of GRC after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience improvement in GRC after administration of a drug candidate or compound. In some embodiments, the improvement in GRC may be equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or a range including and / or spanning the aforementioned values, after administration of a drug candidate or compound described herein. For example, a subject may experience an improvement in GRC ranging from approximately 5% to 100%, or improvement to objectively normal, after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0100] In some embodiments, a subject may experience prevention, suppression, or reduction of heart rate after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience improvement in heart rate after administration of a drug candidate or compound. In some embodiments, the improvement in heart rate may be equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% improvement, or a range including and / or spanning the aforementioned values, after administration of a drug candidate or compound described herein. For example, a subject may experience an improvement in heart rate ranging from approximately 5% to 100%, or improvement to objectively normal, after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0101] In some embodiments, a subject may experience prevention, suppression, or reduction of MoCA after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience improvement in MoCA after administration of a drug candidate or compound. In some embodiments, the improvement related to MoCA may be an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or a range including and / or spanning the aforementioned values, after administration of a drug candidate or compound described herein. For example, a subject may experience an improvement ranging from approximately 5% to 100%, or improvement to objectively normal, in MoCA after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0102] In some embodiments, a subject may experience prevention, suppression, or reduction of precuneus glutathione after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience improvement in precuneus glutathione after administration of a drug candidate or compound described herein. In some embodiments, the improvement related to precuneus glutathione may be an improvement of approximately or greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or a range including and / or spanning the aforementioned values, after administration of a drug candidate or compound described herein. For example, a subject may experience an improvement ranging from approximately 5% to 100%, or improvement to objectively normal, in precuneus glutathione after administration of the drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0103] In some embodiments, a subject may experience prevention, suppression, or reduction of QDRS after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience improvement in QDRS after administration of a drug candidate or compound. In some embodiments, the improvement related to QDRS may be an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% improvement, or a range including and / or spanning the aforementioned values, after administration of a drug candidate or compound described herein. For example, a subject may experience an improvement ranging from approximately 5% to 100% in QDRS, or improvement to objectively normal, after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0104] In some embodiments, a subject may experience prevention, suppression, or reduction of QDRS-behavior after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience improvement in QDRS-behavior after administration of a drug candidate or compound. In some embodiments, the improvement in QDRS-behavior may be equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% after administration of a drug candidate or compound described herein, or within a range including and / or spanning the aforementioned values. For example, a subject may experience an improvement in QDRS-behavior ranging from approximately 5% to 100%, or improvement to objectively normal, after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0105] In some embodiments, a subject may experience prevention, suppression, or reduction of QDRS-behavior after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience improvement in QDRS-behavior after administration of a drug candidate or compound. In some embodiments, the improvement in QDRS-behavior may be equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% after administration of a drug candidate or compound described herein, or within a range including and / or spanning the aforementioned values. For example, a subject may experience an improvement in QDRS-behavior ranging from approximately 5% to 100%, or improvement to objectively normal, after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0106] In some embodiments, a subject may experience prevention, suppression, or reduction of QDRS-cognition after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience improvement in QDRS-cognition after administration of a drug candidate or compound. In some embodiments, the improvement in QDRS-cognition may be equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or a range including and / or spanning the aforementioned values, after administration of a drug candidate or compound described herein. For example, a subject may experience an improvement in QDRS-cognition ranging from approximately 5% to 100%, or improvement to objectively normal, after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0107] In some embodiments, a subject may experience prevention, suppression, or reduction of systolic blood pressure after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience improvement in systolic blood pressure after administration of a drug candidate or compound. In some embodiments, the improvement in systolic blood pressure may be equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% after administration of a drug candidate or compound described herein, or within a range including and / or spanning the aforementioned values. For example, a subject may experience an improvement in systolic blood pressure ranging from approximately 5% to 100%, or improvement to objectively normal, after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0108] In some embodiments, a subject may experience tau prevention, or suppression or reduction, after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience tau improvement after administration of a drug candidate or compound. In some embodiments, the tau-related improvement may be an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% improvement, or a range including and / or spanning the aforementioned values, after administration of a drug candidate or compound described herein. For example, a subject may experience an improvement in tau ranging from approximately 5% to 100%, or improvement to objectively normal, after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0109] In some embodiments, a subject may experience prevention, or suppression or reduction of TNFα after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience improvement in TNFα after administration of a drug candidate or compound. In some embodiments, the improvement in TNFα may be an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or a range including and / or spanning the aforementioned values, after administration of a drug candidate or compound described herein. For example, a subject may experience an improvement in TNFα ranging from approximately 5% to 100%, or improvement to objectively normal, after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0110] In some embodiments, a subject may experience prevention, suppression, or reduction in body weight after administration of a drug candidate or compound described herein. In some embodiments, a subject may experience an improvement in body weight after administration of a drug candidate or compound. In some embodiments, the weight-related improvement may be an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% improvement after administration of a drug candidate or compound described herein, or a range including and / or spanning the aforementioned values. For example, a subject may experience an improvement in body weight ranging from approximately 5% to 100%, or improvement to objectively normal, after administration of a drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0111] In some embodiments, a subject may experience a modulation of their insulin levels after administration of a compound described herein. In some embodiments, a subject may experience a modulation that increases fasting insulin and HOMA2-%B in the subject. In some embodiments, a subject may experience an insulin modulation that decreases HOMA2-%S without hypoglycemia. In some embodiments, a subject may experience an insulin modulation that increases insulin and HOMA2% B cell function and decreases HOMA2% insulin sensitivity. In some embodiments, the modulation of insulin after administration of a drug candidate or compound described herein may be an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100%, or a range including and / or spanning the aforementioned values. For example, a subject may experience a modulation in insulin levels ranging from approximately 5% to 100%, or to objectively normal, after administration of the drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0112] In some embodiments, a subject may experience improvement in one or more neuropsychiatric symptoms after administration of a compound described herein. In some embodiments, the one or more neuropsychiatric symptoms include sleep and appetite. In some embodiments, improved appetite is correlated with decreases in Cog12, CDR SB, and ADCOMS. In some embodiments, improved sleep is correlated with decreases in CGIC and TNFα. In some embodiments, the improvement in one or more neuropsychiatric symptoms may be an amount equal to or greater than approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% after administration of a drug candidate or compound described herein, or a range including and / or spanning the aforementioned values. For example, a subject may experience one or more improvements in neuropsychiatric symptoms ranging from approximately 5% to 100%, or improvement to objectively normal, after administration of the drug candidate or compound and at least one pharmaceutically acceptable excipient.

[0113] Accordingly, some aspects described relate to the following numbered example configurations:

[0114] 1. A method for treating, suppressing, or ameliorating a disease or condition associated with the circadian clock in a subject in need thereof, comprising administering to the subject 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

[0115] 2. The method of embodiment 1, wherein the disease or condition associated with the circadian clock in the subject in need thereof is based on modulation of DNA methylation of genes associated with the circadian clock.

[0116] 3. The method of configuration example 1 or 2, wherein the disease or condition associated with the circadian clock in the subject in need thereof is associated with a gene or genomic region that becomes hypermethylated with age.

[0117] 4. The method according to any one of configuration examples 1 to 3, wherein the disease or condition associated with the circadian clock in the subject in need thereof is associated with a gene or genomic region that becomes hypomethylated with age.

[0118] 5. The method according to any one of configurations 1 to 4, wherein the disease or condition associated with the circadian clock in the subject in need thereof is associated with tau phosphorylation.

[0119] 6. The method according to any one of configuration examples 1 to 5, wherein the disease or condition associated with the circadian clock in the subject in need thereof is associated with hyperglycemia.

[0120] 7. The method according to any one of configurations 1 to 6, wherein the disease or condition associated with the circadian clock in the subject in need thereof is associated with insulin resistance.

[0121] 8. A method according to any one of configuration examples 1 to 7, wherein the disease or condition associated with the circadian clock is mild cognitive impairment or late-onset Alzheimer's disease.

[0122] 9. The method of any one of configurations 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces the subject's Alzheimer's disease composite score.

[0123] 10. The method of embodiment 9, wherein the subject experiences about a 5% to about 100% reduction in symptoms associated with Alzheimer's disease composite score, or a reduction to objectively normal, after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

[0124] 11. The method of any one of configurations 1 to 8, wherein administering to the subject 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient reduces pTau in the subject.

[0125] 12. The method of embodiment 11, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences about a 5% to about 100% reduction in a condition or symptom associated with pTau, or a reduction to an objectively normal state.

[0126] 13. The method of any one of configurations 1 to 8, wherein administering to the subject 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient reduces leptin in the subject.

[0127] 14. The method of embodiment 13, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences an increase of about 5% to about 100% in the leptin-related condition or symptom, or an increase to objectively normal.

[0128] 15. The method of any one of configurations 1 to 8, wherein administering to the subject 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient reduces DNA methylation in the subject.

[0129] 16. The method of embodiment 15, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences about a 5% to about 100% reduction in a condition or symptom associated with DNA methylation, or a reduction to an objectively normal state.

[0130] 17. The method of any one of configurations 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces cardiovascular risk in the subject.

[0131] 18. The method of embodiment 17, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction of about 5% to about 100% in a condition or symptom associated with cardiovascular risk, or a reduction to an objectively normal state.

[0132] 19. The method of any one of configurations 1 to 8, wherein administering to the subject 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient reduces DNA methylation age in the subject.

[0133] 20. The method of embodiment 19, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences about a 5% to about 100% reduction in a condition or symptom associated with DNA methylation, or a reduction to an objectively normal state.

[0134] 21. The method of any one of configurations 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces DNA methylation phenoage in the subject.

[0135] 22. The method of embodiment 21, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences about a 5% to about 100% reduction in a condition or symptom associated with DNA methylation phenoaging, or a reduction to an objectively normal state.

[0136] 23. The method of any one of configurations 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces DNA methylation skin blood clock in the subject.

[0137] 24. The method described in Example 23, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction of about 5% to about 100% in a condition or symptom associated with DNA methylation skin blood clock, or a reduction to an objectively normal state.

[0138] 25. The method of any one of configurations 1 to 8, wherein administering to the subject 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient reduces the subject's CDR score.

[0139] 26. The method of embodiment 25, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction in symptoms associated with a CDR score of about 5% to about 100%, or a reduction to an objectively normal state.

[0140] 27. The method of any one of configurations 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces the subject's ADAS-Cog score.

[0141] 28. The method of embodiment 27, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences about a 5% to about 100% reduction in symptoms associated with ADAS-Cog score, or a reduction to objectively normal.

[0142] 29. The method of any one of configurations 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces the subject's QDRS score.

[0143] 30. The method described in Example 29, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction of symptoms associated with QDRS score of about 5% to about 100%, or a reduction to an objectively normal state.

[0144] 31. The method of any one of configurations 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject increases the subject's MMSE score.

[0145] 32. The method of embodiment 31, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction of symptoms associated with MMSE score of about 5% to about 100%, or a reduction to objectively normal.

[0146] 33. The method of any one of configurations 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces the subject's MoCA score.

[0147] 34. The method of embodiment 33, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction in symptoms associated with MoCA score of about 5% to about 100%, or a reduction to an objectively normal state.

[0148] 35. The method of any one of configurations 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces the subject's GRC score.

[0149] 36. The method of embodiment 35, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction of symptoms associated with a GRC score of about 5% to about 100%, or a reduction to an objectively normal state.

[0150] 37. The method of any one of configurations 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces the subject's CGIC score.

[0151] 38. The method of embodiment 37, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction in symptoms associated with CGIC score of about 5% to about 100%, or a reduction to an objectively normal state.

[0152] 39. The method of any one of configurations 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces the subject's ADL score.

[0153] 40. The method described in Example 39, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction of symptoms related to ADL score of about 5% to about 100%, or a reduction to an objectively normal state.

[0154] 41. The method of any one of configurations 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces the subject's PDQ9 score.

[0155] 42. The method of embodiment 41, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction in symptoms associated with PDQ9 score of about 5% to about 100%, or a reduction to an objectively normal state.

[0156] 43. The method of any one of embodiments 1 to 42, wherein 17α-ethynylandrost-5-ene-3β,7β,17β-triol is administered orally.

[0157] 44. The method of any one of embodiments 1 to 42, wherein 17α-ethynylandrost-5-ene-3β,7β,17β-triol is administered intravenously.

[0158] 45. The method of any one of configurations 1 to 44, wherein the subject has a waist-to-hip ratio greater than or equal to approximately 0.90.

[0159] 46. ​​The method of any one of configuration examples 1 to 44, wherein the subject's waist-to-hip ratio is greater than or equal to approximately 0.95.

[0160] 47. The method of any one of configurations 1 to 46, wherein the 17a-ethynylandrost-5-ene-3b,7b,17b-triol is a solid-state form of 17a-ethynylandrost-5-ene-3b,7b,17b-triol.

[0161] 48. The method of embodiment 47, wherein the solid-state form of 17a-ethynylandrost-5-ene-3b,7b,17b-triol is a crystalline solvate of 17a-ethynylandrost-5-ene-3b,7b,17b-triol.

[0162] 49. The method of embodiment 48, wherein the crystalline solvate is crystalline methanolate 17a-ethynylandrost-5-ene-3b,7b,17b-triol.

[0163] 50. The method of embodiment 49, wherein the crystalline solvate is crystalline ethanolate 17a-ethynylandrost-5-ene-3b,7b,17b-triol.

[0164] 51. The method of embodiment 50, wherein the crystalline solvate is crystalline hydrate 17a-ethynylandrost-5-ene-3b,7b,17b-triol.

[0165] 52. The method described in configuration example 50, wherein the crystalline solvate is Form III 17a-ethynylandrost-5-ene-3b,7b,17b-triol.

[0166] 53. The method of embodiment 50, wherein the crystalline solvate is Form IV 17a-ethynylandrost-5-ene-3b,7b,17b-triol.

[0167] 54. The method of embodiment 50, wherein the crystalline solvate is Form V 17a-ethynylandrost-5-ene-3b,7b,17b-triol.

[0168] 55. The method of embodiment 49, wherein the solid state form of 17a-ethynylandrost-5-ene-3b,7b,17b-triol is amorphous 17a-ethynylandrost-5-ene-3b,7b,17b-triol.

[0169] 56. The method of any one of embodiments 1 to 55, wherein the pharmaceutical composition contains less than about 3% by weight of impurities.

[0170] 57. An in vitro screening method for identifying potential drug candidates or compounds capable of treating, preventing, suppressing, or ameliorating a disorder or disease, comprising: (i) providing a stimulation sample selected from the group consisting of cells, tissues, blood, monocytes, microglia, macrophages, adipocytes, neuroblastoma, pheochromocytoma, and Lund human mesencephalon (LUHMES) cells; (ii) stimulating the sample with an agonist to induce a phenotype or phenotypic response, wherein the phenotype or phenotypic response substantially corresponds to a disease or condition associated with at least one DNA methylation at a CpG site within a region of DNA; (iii) contacting the one or more cells exhibiting the phenotype or phenotypic response with one or more potential drug candidates or compounds; (iv) determining a change in responsiveness in the phenotype of the sample; and (v) providing the drug candidates or compounds to a subject in need thereof, thereby treating, suppressing, preventing, or ameliorating the DNA methylation-associated disease or condition in the subject.

[0171] 58. At least one DNA methylation at the CpG site is selected from the group consisting of AC073869.20, SP100, KCNQ1DN, DBNDD2, CEP112, CEP85L, SPDYE4, ZNF211, NR3C1, HLA-L, TPP2, SLC26A1, SLC37A1, CAB39L, ILKAP, NPHP4, PATE4, ARHGEF12, CELA1, OR10G7, PFN2, WDR59, and snoU1. 3, ANXA3, SVIL-AS1, PPHLN1, AP000442.1, FA, KIAA0319L, ZNF509, DLEU2L, ABL2, SGK1, TMEM245, SRSF4, DAP, GRAMD1C, FABP5P1, MCM10, ANP32E, ZNF268, ESPN, DHFR, U6, MTUS1, ATP1B3, or a combination thereof.

[0172] 59. The method of embodiment 57 or 58, wherein the phenotype or phenotypic response is selected from the group consisting of TNFα, GRC, CDR, MoCA, QDRS, GRC, ADCOMS, MoCA, QDRS-Cognition, ADAS-Cogl 1, heart rate, frontal lobe, systolic blood pressure, gray matter, body weight, MMSE, hippocampal volume, behavior, PDQ-9, CSF glucose, precuneus GLTH, CSF pTau / Ab, or a combination thereof.

[0173] 60. The method of embodiment 59, wherein the phenotype or phenotypic response is selected from the group consisting of ADAS-Cog11, ADCOMS, CDR, CSF glucose, CSF pTau / Ab, frontal lobe volume, subcortical gray matter thickness, GRC, heart rate, MoCA, PDQ-9, precuneus glutathione, QDRS, QDRS-behavior, QDRS-cognition, systolic BP, tau, TNFa, and body weight.

[0174] 61. The method of any one of configurations 57 to 60, wherein the DNA methylation change is a decrease of more than 50%.

[0175] 62. The method of any one of embodiments 57 to 60, wherein the DNA methylation change is a decrease of more than 55%.

[0176] 63. The method of any one of configurations 57 to 60, wherein the DNA methylation change is a decrease of more than 60%.

[0177] 64. The method of embodiment 59, wherein the phenotype or phenotypic response is reduced in TNF, CDR, or QDRS-cognition, and the subject's QDRS-cognition is improved.

[0178] 65. The method of any one of configuration examples 57 to 64, wherein the change in responsiveness is a reduction or elimination of the phenotype, and the reduction or elimination indicates that the potential drug candidate or compound is capable of preventing, suppressing, or ameliorating a neurodegenerative disorder or disease.

[0179] 66. The method of embodiment 65, wherein the neurodegenerative disorder or disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, levodopa-induced dyskinesia (LID), amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), age-related hippocampal sclerosis (HS-Aging), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy, multiple system atrophy, corticobasal degeneration, and vascular parkinsonism.

[0180] 67. The method of any one of embodiments 65 to 66, wherein the neurodegenerative disorder or disease is Parkinson's disease.

[0181] 68. The method of any one of embodiments 65 and 66, wherein the neurodegenerative disorder is Alzheimer's disease.

[0182] 69. The method of any one of embodiments 57 to 68, wherein the CpG sites correlate with genes associated with Alzheimer's disease and related dementias.

[0183] 70. The method of embodiment 69, wherein the disease or condition is associated with inflammatory TNF signaling.

[0184] 71. The method of any one of embodiments 57 to 69, wherein the DNA methylation-associated disease or condition is associated with tau phosphorylation.

[0185] 72. The method of any one of configurations 57 to 69, wherein the DNA methylation-related disease or condition is related to hyperglycemia.

[0186] 73. The method of any one of embodiments 57 to 69, wherein the DNA methylation-associated disease or condition is associated with insulin resistance.

[0187] 74. The method of any one of embodiments 57 to 69, wherein the DNA methylation-associated disease or condition is associated with obesity.

[0188] 75. The method of any one of configurations 57 to 69, wherein the DNA methylation-associated disease or condition is associated with ADAS-Cog11 in the subject.

[0189] 76. The method of any one of configurations 57 to 69, wherein the DNA methylation-associated disease or condition is associated with ADCOMS in the subject.

[0190] 77. The method of any one of embodiments 57 to 69, wherein the DNA methylation-associated disease or condition is associated with the CDR of interest.

[0191] 78. The method of any one of embodiments 57 to 69, wherein the DNA methylation-associated disease or condition is associated with CSF glucose in the subject.

[0192] 79. The method of any one of embodiments 57 to 69, wherein the DNA methylation-associated disease or condition is associated with pTau / Aβ in the subject's CSF.

[0193] 80. The method of any one of configuration examples 57 to 69, wherein the DNA methylation-related disease or condition is related to the subject's frontal lobe volume.

[0194] 81. The method of any one of configuration examples 57 to 69, wherein the DNA methylation-associated disease or condition is associated with subcortical gray matter thickness in the subject.

[0195] 82. The method of any one of configurations 57 to 69, wherein the DNA methylation-associated disease or condition is associated with GRC in the subject.

[0196] 83. The method of any one of configurations 57 to 69, wherein the DNA methylation-related disease or condition is related to the subject's heart rate.

[0197] 84. The method of any one of configurations 57 to 69, wherein the DNA methylation-associated disease or condition is associated with MoCA in the subject.

[0198] 85. The method of any one of configurations 57 to 69, wherein the DNA methylation-associated disease or condition is associated with MoCA in the subject.

[0199] 86. The method of any one of embodiments 57 to 69, wherein the DNA methylation-related disease or condition is related to the subject's PDQ-9.

[0200] 87. The method of any one of embodiments 57 to 69, wherein the DNA methylation-associated disease or condition is associated with precuneus glutathione in the subject.

[0201] 88. The method of any one of configurations 57 to 69, wherein the DNA methylation-related disease or condition is associated with the subject's QDRS.

[0202] 89. The method of any one of configurations 57 to 69, wherein the DNA methylation-related disease or condition is related to the subject's QDRS-behavior.

[0203] 90. The method of any one of configurations 57 to 69, wherein the DNA methylation-related disease or condition is related to QDRS-cognition of the subject.

[0204] 91. The method of any one of configurations 57 to 69, wherein the DNA methylation-associated disease or condition is associated with the subject's systolic blood pressure.

[0205] 92. The method of any one of embodiments 57 to 69, wherein the DNA methylation-associated disease or condition is associated with tau in the subject.

[0206] 93. The method of any one of configurations 57 to 69, wherein the DNA methylation-associated disease or condition is associated with TNFα in the subject.

[0207] 94. The method of any one of embodiments 57 to 69, wherein the DNA methylation-associated disease or condition is associated with the subject's body weight.

[0208] 95. A method for diagnosing a patient having a disease or condition, comprising: (i) providing the patient with a potential drug candidate or compound that can treat, prevent, inhibit, or ameliorate the disorder or disease; (ii) identifying DNA methylation changes in the patient; and (iii) diagnosing the patient for a disease or condition associated with a biomarker related to DNA methylation.

[0209] 96. The method described in Example 95, wherein DNA methylation changes in the patient are identified to identify CpGs that are reduced by more than 50%.

[0210] 97. The method of any one of embodiments 95 to 96, wherein identifying DNA methylation changes is correlated with one or more clinical changes.

[0211] 98. The DNA methylation changes at CpG sites are AC073869.20, SP100, KCNQ1DN, DBNDD2, CEP112, CEP85L, SPDYE4, ZNF211, NR3C1, HLA-L, TPP2, SLC26A1, SLC37A1, CAB39L, ILKAP, NPHP4, PATE4, ARHGEF12, CELA1, OR10G7, PFN2, WDR59, snoU13, ANXA3, SV 98. The method of any one of configurations 95 to 97, wherein the gene is selected from the group consisting of IL-AS1, PPHLN1, AP000442.1, FA, KIAA0319L, ZNF509, DLEU2L, ABL2, SGK1, TMEM245, SRSF4, DAP, GRAMD1C, FABP5P1, MCM10, ANP32E, ZNF268, ESPN, DHFR, U6, MTUS1, ATP1B3, or a combination thereof.

[0212] 99. The method of any one of configuration examples 95 to 98, wherein the disease or condition associated with a biomarker related to DNA methylation is selected from the group consisting of TNFα, GRC, CDR, MoCA, QDRS, GRC, ADCOMS, MoCA, QDRS-Cognition, ADAS-Cog11, heart rate, frontal lobe, systolic blood pressure, gray matter, body weight, MMSE, hippocampal volume, behavior, PDQ-9, CSF glucose, precuneus GLTH, CSF pTau / Ab, or a combination thereof.

[0213] 100. The method of claim 99, wherein the disease or condition associated with a biomarker related to DNA methylation is selected from the group consisting of ADAS-Cog11, ADCOMS, CDR, CSF glucose, CSF pTau / Ab, frontal lobe volume, subcortical gray matter thickness, GRC, heart rate, MoCA, PDQ-9, precuneus glutathione, QDRS, QDRS-behavior, QDRS-cognition, systolic BP, tau, TNFa, and body weight.

[0214] 101. The method of embodiment 99, wherein biomarkers related to DNA methylation are reduced in TNF, CDR, and QDRS-cognition, and QDRS-cognition of the subject is improved.

[0215] 102. The method of any one of embodiments 95 to 101, wherein the DNA methylation change is a decrease of more than 50%.

[0216] 103. The method of any one of embodiments 95 to 101, wherein the DNA methylation change is a decrease of more than 55%.

[0217] 104. The method of any one of embodiments 95 to 101, wherein the DNA methylation change is a decrease of more than 60%.

[0218] 105. The method of any one of configuration examples 95 to 104, wherein the disease or condition is selected from the group consisting of Alzheimer's disease, Parkinson's disease, levodopa-induced dyskinesia (LID), amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), age-related hippocampal sclerosis (HS-Aging), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy, multiple system atrophy, corticobasal degeneration, and vascular parkinsonism.

[0219] 106. The method of embodiment 105, wherein the neurodegenerative disorder or disease is Parkinson's disease.

[0220] 107. The method of embodiment 105, wherein the neurodegenerative disorder is Alzheimer's disease.

[0221] 108. The method of any one of embodiments 95 to 107, wherein the CpG sites correlate with genes associated with Alzheimer's disease and related dementias.

[0222] 109. The method of any one of embodiments 95 to 108, wherein the disease or condition is associated with inflammatory TNF signaling.

[0223] 110. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with tau phosphorylation.

[0224] 111. The method of any one of embodiments 95 to 108, wherein the DNA methylation-related disease or condition is related to hyperglycemia.

[0225] 112. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with insulin resistance.

[0226] 113. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with obesity.

[0227] 114. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with ADAS-Cog11 in the subject.

[0228] 115. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with ADCOMS in the subject.

[0229] 116. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with the CDR of interest.

[0230] 117. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with CSF glucose in the subject.

[0231] 118. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with pTau / Aβ in the subject's CSF.

[0232] 119. The method of any one of embodiments 95 to 108, wherein the DNA methylation-related disease or condition is related to the subject's frontal lobe volume.

[0233] 120. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with subcortical gray matter thickness in the subject.

[0234] 121. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with GRC in the subject.

[0235] 122. The method of any one of embodiments 95 to 108, wherein the DNA methylation-related disease or condition is related to the subject's heart rate.

[0236] 123. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with MoCA in the subject.

[0237] 124. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with MoCA in the subject.

[0238] 125. The method of any one of embodiments 95 to 108, wherein the DNA methylation-related disease or condition is related to the subject's PDQ-9.

[0239] 126. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with precuneus glutathione in the subject.

[0240] 127. The method of any one of embodiments 95 to 108, wherein the DNA methylation-related disease or condition is associated with a QDRS of the subject.

[0241] 128. The method of any one of embodiments 95 to 108, wherein the DNA methylation-related disease or condition is related to the subject's QDRS-behavior.

[0242] 129. The method of any one of embodiments 95 to 108, wherein the DNA methylation-related disease or condition is related to QDRS-cognition of the subject.

[0243] 130. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with the subject's systolic blood pressure.

[0244] 131. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with tau in the subject.

[0245] 132. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with TNFα in the subject.

[0246] 132. The method of any one of embodiments 95 to 108, wherein the DNA methylation-associated disease or condition is associated with the subject's body weight.

[0247] 133. The method of any one of embodiments 95 to 108, wherein administering the drug candidate or compound to the subject reduces the subject's CDR score.

[0248] 134. The method of embodiment 133, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction in symptoms associated with CDR score of about 5% to about 100%.

[0249] 135. The method of any one of embodiments 95 to 108, wherein administering the drug candidate or compound to the subject reduces the subject's ADAS-Cog score.

[0250] 136. The method of embodiment 135, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences about a 5% to about 100% reduction in symptoms associated with ADAS-Cog score.

[0251] 137. The method of any one of embodiments 95 to 108, wherein administering the drug candidate or compound to the subject reduces the subject's QDRS score.

[0252] 138. The method of embodiment 137, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction in symptoms associated with QDRS score of about 5% to about 100%.

[0253] 139. The method of any one of embodiments 95 to 108, wherein administering the drug candidate or compound to the subject increases the subject's MMSE score.

[0254] 140. The method of embodiment 139, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction of symptoms associated with MMSE score of about 5% to about 100%, or a reduction to an objectively normal state.

[0255] 141. The method of any one of configurations 95 to 108, wherein administering the drug candidate or compound to the subject increases the subject's MoCA score.

[0256] 142. The method of embodiment 141, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction in symptoms associated with MoCA score of about 5% to about 100%.

[0257] 143. The method of any one of embodiments 95 to 108, wherein administering the drug candidate or compound to the subject increases the subject's GRC score.

[0258] 144. The method of embodiment 143, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction in symptoms associated with a GRC score of about 5% to about 100%, or a reduction to an objectively normal state.

[0259] 145. The method of any one of embodiments 95 to 108, wherein administering the drug candidate or compound to the subject reduces the subject's CGIC score.

[0260] 146. The method of embodiment 145, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction of symptoms associated with CGIC score of about 5% to about 100%, or a reduction to an objectively normal state.

[0261] 147. The method of any one of embodiments 95 to 108, wherein administering the drug candidate or compound to the subject increases the subject's ADL score.

[0262] 148. The method of embodiment 147, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction of symptoms related to ADL score of about 5% to about 100%, or a reduction to an objectively normal state.

[0263] 149. The method of any one of embodiments 95 to 108, wherein administering the drug candidate or compound to the subject reduces the subject's PDQ9 score.

[0264] 150. The method of embodiment 149, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences a reduction in symptoms associated with PDQ9 score of about 5% to about 100%, or a reduction to an objectively normal state. [Example]

[0265] The following examples are given for the purpose of illustrating various embodiments of the present disclosure and are not intended to limit the disclosure in any manner. Those skilled in the art will readily appreciate that the present disclosure is well adapted to carry out the objects and obtain the objects and advantages set forth above, as well as those inherent therein. Modifications therein and other uses will occur to those skilled in the art which are encompassed within the spirit of the disclosure as defined by the scope of the claims.

[0266] Example 1 Research studies were conducted into possible early Alzheimer's disease.

[0267] This study enrolled 23 subjects with mild cognitive impairment (MCI) and Alzheimer's disease (AD) as assessed by the Clinical Dementia Rating (CDR) based on the Quick Dementia Rating System (QDRS) scale. Advanced MRI imaging (ASL, BOLD, MRS, task-based fMRI) was also performed, as were cognitive and memory tests (Cog12, MMSE, QDRS, MoCA). Subject biomarkers (cerebrospinal fluid (CSF) p-Tau, Aβ, plasma TNF) were also collected, and episomal analysis was performed. Finally, circadian clock analysis (DNA methylation profiling) was performed.

[0268] In vitro samples treated with 17α-ethynylandrost-5-ene-3β,7β,17β-triol showed reduced activation of inflammatory p-IKKβ, p-ERK, p-P38, and p-JNK (phosphorylation of tau), resulting in a decrease in p-Tau.

[0269] Under normal conditions, insulin binds to the insulin receptor, initiating tyrosine phosphorylation of IRS1 / 2, leading to activation of Pi3K and Akt and inhibition of GSK3β. Under inflammatory conditions, activation of the inflammatory kinases p-IKKβ and p-JNK leads to serine phosphorylation of IRS-1, which inhibits insulin signaling. Thus, inflammation reduces insulin-stimulated Akt-inhibitory phosphorylation of GSK3β and increases p-Tau.

[0270] Without wishing to be bound by theory, 17α-ethynylandrost-5-ene-3β,7β,17β-triol is thought to bind to ERK1 and ERK2 and reduce the inflammatory activation of p-IKKβ, p-JNK, p-ERK, and p-P38, thereby decreasing the serine phosphorylation of IRS1 and restoring the tyrosine phosphorylation of IRS1 and insulin sensitivity. As a result, we observed an increased inhibitory effect of Akt on GSK3β and p-Tau.

[0271] Age-related hyperglycemia induces insulin resistance, leading to overactivation of CDK5 / p25 kinase, which not only increases p-Tau but also inhibits the degradation of β-catenin by GSK3β, a combination of which contributes to neuronal aging and cell death (Chow and Herrup 2019).

[0272] Further results of this study are shown in Figures 1-10.

[0273] Example 2 In this study, a follow-up study to Example 1 was carried out.

[0274] DNA methylation changes after 14 weeks of bezisterim treatment were sorted using an Illumina® 850K array, and the top 400 CpGs showing a >50% decrease were examined for correlation with changes in clinical outcomes after treatment. Genes were identified for 366 of these CpGs. CpG changes that showed Spearman correlations (p<0.05) with individual clinical changes (biomarkers, cognition, function, imaging) were highly correlated with each other and were primarily associated with genes associated with Alzheimer's disease and related dementias.

[0275] Furthermore, many of these CpG reductions correlated with multiple clinical changes. For example, KCNQ1DN (a potassium channel known to be reduced in AD brains) showed a 51% reduction in DNAm (DNA methylation), which correlated with decreases (improvements) in TNF (an inflammatory biomarker), CDRs, and QRDS-cognition, and increases (improvements) in QDRS-cognition (clinical assessment). HLA-L (a protein that helps the immune system remove amyloid plaques from the brain) showed a 56% reduction in DNAm, which correlated with improvements in subcortical gray matter thickness and frontal lobe volume (imaging). SLC26A1 (reductions in this sulfate transporter are thought to contribute to cognitive decline in AD) showed a 55% reduction in DNAm, which correlated with improvements in ADCOMS, QDRS (AD assessment), and CSF pTau / Aβ ratio (biomarker).

[0276] Of the 366 CpGs examined, 157 showed significant overall correlations between DNAm reduction and individual clinical changes, 45 of which were individually correlated with multiple clinical changes. These 157 CpGs showed individual correlations with improvements in ADAS-Cog11, ADCOMS, Ab42, CDR, CSF glucose, CSF pTau / Ab, frontal lobe volume, subcortical gray matter thickness, GRC, heart rate, MoCA, PDQ-9, precuneus glutathione, QDRS, QDRS-behavior, QDRS-cognition, systolic blood pressure, tau, TNFα, and weight. The results of this study are shown in Figure 11 and Table 1. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5]

[0277] Example 2 This example presents the clinical outcomes of a phase 3, randomized, placebo-controlled trial of NE3107 (17α-ethynylandrost-5-ene-3β,7β,17β-triol) in subjects with possible mild to moderate Alzheimer's disease.

[0278] background Recently, the roles of inflammation and insulin resistance in neurodegeneration have become better understood. NE3107 is an orally available, small-molecule, anti-inflammatory insulin sensitizer that binds to extracellular signal-regulated kinase (ESK) and can cross the blood-brain barrier. It has been shown to selectively inhibit inflammatory mediators (including TNF-α) stimulated by inflammation-activated ERK and NF-κB without interfering with homeostatic functions. The rationale and design of NM101 are described below. NM101 is the first randomized, multicenter, Phase 3 clinical trial to evaluate the safety and efficacy of 30 weeks of treatment with NE3107 compared with placebo in elderly patients with mild to moderate Alzheimer's disease. Patients (316) were randomized 1:1. Co-primary endpoints measured cognitive function (ADAS Cog12) and functional and behavioral characteristics (ADCS CGIC). Trial registration number: NCT04669028 (Clinicaltrials.gov).

[0279] Study design The study design is shown in Figure 12.

[0280] The phase 3 study measured median improvement in various blinded assessments, and the results are shown in Table 2 and Figures 13A-13G. [Table 2]

[0281] Similar distributions were observed for APOE4 presence / absence (+ / -), mild / moderate Alzheimer's disease, male / female, and elderly / young.

[0282] Metabolic and correlated changes in subjects were analyzed. Many of the genes associated with late-onset Alzheimer's disease (LOAD) are involved in cholesterol metabolism, which is reduced in AD neuronal membranes. Insulin and blood glucose control are known to be involved in neurodegeneration. Increased insulin and HOMA2-% beta cell function and decreased HOMA2% insulin sensitivity resulted in no cases of hypoglycemia. Increased mean amplitude of blood glucose excursions (MAGE from CGM) increased the risk of progression to Alzheimer's disease. Leptin has anti-inflammatory and neuroprotective effects and is reduced in AD. An increase in leptin DNAm was observed. The results are shown in Tables 4 and 5. [Table 3] [Table 4]

[0283] A substudy of 23 subjects was analyzed using vMRI imaging. Increased volume was observed in the hippocampus and amygdala. Furthermore, decreases in CGIC correlated with decreases in hippocampal volume, and increases in MCP-1 correlated with increases throughout the cerebral cortex. Results are shown in Figures 14A-B and Table 6. [Table 5]

[0284] Subjects were analyzed using fluorodeoxyglucose (FDG)-positron emission tomography (PET) imaging scans. Fourteen of 24 subjects with a baseline FDG-PET standardized uptake value ratio ("SUVR") of less than 1.29 in the whole cerebral cortex experienced an increase in SUVR. This data correlated with improved SUVR in Cog12 and ADL. Increased cholesterol levels tended to be associated with increased SUVR in the cingulate gyrus. Results are shown in Table 7 and Figure 15. [Table 6]

[0285] Subjects were analyzed for neuropsychiatric assessment. Overall improvements were seen in sleep (-1.0, p<0.0001) and appetite (-1.0, p<0.023). Improved appetite also correlated with declines in Cog12, CDR SB, and ADCOMS, and correlated with Alzheimer's disease progression. Decreased anxiety correlated with increased ADL and decreased CGIC. Improved sleep correlated with decreased CGIC and decreased TNFα. Results are shown in Table 8 and Figures 16A-16B. [Table 7]

[0286] The overall incidence of reported adverse events (AEs) in this study was very low, with only 10 subjects (2.3%) discontinuing due to a reported AE. Of the 439 subjects enrolled in the study, 156 (35.5%) experienced one or more AEs. Only 43 (9.8%) were relevant AEs, the majority of which, 42 (0.6%), were classified by the investigator as non-serious. The study also had a very low rate of reported serious adverse events; 12 cases (2.7%) were reported during the study period. Of the reported serious adverse events, only one resulted in death. Because this analysis was blinded, it is unclear whether subjects received the investigational drug (IP) or placebo.

[0287] Additional metrics were measured during the Phase 3 study, as shown in Figures 16A-33.

[0288] Figures 17A-17C are graphs showing increases in fasting insulin and HOMA2-%B and decreases in HOMA2-%S occurring without hypoglycemia. Table 10 provides additional information regarding increases in fasting insulin and HOMA2-%B and decreases in HOMA2-%S without hypoglycemia. [Table 8]

[0289] Figures 18A-18G show graphs of the placebo effect on various endpoints. Median improvements are shown, indicating that at least some placebo subjects demonstrated improvement in neurocognitive and functional assessments. Similar findings were observed for vMRI, NPI, HOMA, and APS, suggesting that this is not simply related to baseline overestimation (assessment inflation). Figures 19-20 show Spearman correlations for the ADAS-Cog12. Figures 21-22 show Spearman correlations for the Clinician Global Assessment of Change. Figures 23-24 show Spearman correlations for the Mini-Mental State Examination. Figure 25 shows Spearman correlations for the ADCOMS. Figure 26 shows Spearman correlations for the CDR box score (CDR sum of boxes). Figure 27 shows Spearman correlations for activities of daily living. Among the subjects participating in the FDG-PET analysis, increased glucose uptake was observed in approximately half of the FDG-PET subjects. Figure 28 is a graph showing that improvement in ADAS-Cog12 correlates with an increase in FDG-PET suvr. Figure 29 is a graph showing that improvement in MMSE correlates with an increase in FDG-PET suvr. Figure 30 is a graph showing that improvement in ADL correlates with an increase in FDG-PET suvr. Figure 31 is a graph showing that improvement in HOMA2 insulin sensitivity correlates with an increase in FDG-PET suvr. Figure 32 is a graph showing that improvement in cholesterol correlates with an increase in FDG-PET suvr.

[0290] Improvements in cognitive function and recognized Alzheimer's disease biomarkers (Aβ and FDG-PET) in blinded data suggest that NE3107 is effective in subjects with mild / moderate AD. Blinded analyses were also consistent with the hypothesis that NE3107 has anti-inflammatory and insulin-sensitizing effects in Alzheimer's disease.

[0291] Although several embodiments have been illustrated and described, one of ordinary skill in the art, after reading the foregoing specification, may affect the modifications, equivalent substitutions, and other types of alterations described herein to the compounds of the present technology or their salts, pharmaceutical compositions, derivatives, prodrugs, metabolites, tautomers, or racemic mixtures. Each of the above-described aspects and embodiments may also include or incorporate variations or aspects as disclosed with respect to any or all of the other aspects and embodiments.

[0292] The present technology is also not limited in terms of the specific embodiments described herein, which are intended as single examples of individual embodiments of the technology. Many modifications and variations of the technology can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods within the scope of the technology, in addition to those recited herein, will be apparent to those skilled in the art from the foregoing description. Such modifications and variations are intended to fall within the scope of the appended claims. It is understood that the present technology is not limited to particular methods, reagents, compounds, compositions, labeled compounds, or biological systems, which may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments and is not intended to be limiting. Therefore, the specification is intended to be considered exemplary only, with the breadth, scope, and spirit of the technology being indicated only by the appended claims, the definitions therein, and their equivalents.

[0293] The embodiments illustratively described herein may suitably be practiced in the absence of any element or elements, limitation, or limitations not specifically disclosed herein. Thus, for example, terms such as "comprising," "including," and "containing" are intended to be broadly understood and not limiting. Furthermore, the terms and expressions used herein are used as terms of description rather than limitation, and the use of such terms and expressions is not intended to exclude any equivalents of the shown and described features or portions thereof, recognizing that various modifications are possible within the scope of the claimed technology. Additionally, the phrase "consisting essentially of" will be understood to include the elements specifically recited and additional elements that do not materially affect the basic and novel characteristics of the claimed technology. The phrase "consisting of" excludes any elements not specified.

[0294] Additionally, when features or aspects of the present disclosure are described in terms of Markush groups, those skilled in the art will recognize that the present disclosure is also described in terms of any individual member or subgroup of members of the Markush group. Each of the narrower species and subgeneric groupings falling within the general disclosure also constitutes part of the present invention. This includes the general description of the present technology with a condition or negative limitation removing any subject matter from the genus, regardless of whether the omitted material is specifically set forth herein.

[0295] All publications, patent applications, issued patents, and other documents (e.g., journals, articles, and / or textbooks) mentioned herein are incorporated by reference as if each individual publication, patent application, issued patent, or other document was specifically and individually indicated to be incorporated by reference in its entirety. Definitions contained in text incorporated by reference are excluded to the extent they conflict with definitions in this disclosure.

[0296] Other embodiments are set forth in the following claims, along with the full scope of equivalents to which such claims are entitled.

[0297] While the present invention has been particularly shown and described with reference to preferred and various alternative embodiments, it will be understood by those skilled in the art that various changes in form and detail can be made therein without departing from the spirit and scope of the invention.

[0298] All publications, issued patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.

[0299] While the invention has been described with reference to embodiments and examples, it should be understood that numerous and various modifications can be made without departing from the spirit of the invention. Accordingly, the invention is limited only by the scope of the following claims.

Claims

1. 1. A method for treating, inhibiting, or ameliorating a disease or condition associated with the circadian clock in a subject in need thereof, comprising: administering to the subject 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

2. 10. The method of claim 1, wherein the disease or condition associated with the circadian clock in the subject in need thereof is based on the regulation of DNA methylation of genes associated with the circadian clock.

3. 3. The method of claim 1 or 2, wherein the disease or condition associated with the circadian clock in the subject in need thereof is associated with a gene or genomic region that becomes hypermethylated with age.

4. 4. The method of any one of claims 1 to 3, wherein the disease or condition associated with the circadian clock in the subject in need thereof is associated with a gene or genomic region that becomes hypomethylated with age.

5. 5. The method of any one of claims 1 to 4, wherein the disease or condition associated with the circadian clock in the subject in need thereof is associated with tau phosphorylation.

6. 6. The method of any one of claims 1 to 5, wherein the disease or condition associated with the circadian clock in the subject in need thereof is associated with hyperglycemia.

7. 7. The method of any one of claims 1 to 6, wherein the disease or condition associated with the circadian clock in the subject in need thereof is associated with insulin resistance.

8. 8. The method of any one of claims 1 to 7, wherein the disease or condition associated with the circadian clock is mild cognitive impairment or late-onset Alzheimer's disease.

9. 9. The method of any one of claims 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces the subject's Alzheimer's disease composite score.

10. 9. The method of any one of claims 1 to 8, wherein administering to the subject 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient reduces leptin in the subject.

11. 11. The method of claim 10, wherein the subject experiences about a 5% to about 100% increase in the leptin-related condition or symptom after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient.

12. 9. The method of any one of claims 1 to 8, wherein administering to the subject 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient reduces DNA methylation in the subject.

13. 13. The method of claim 12, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences about a 5% to about 100% reduction in the condition or symptom associated with DNA methylation.

14. 9. The method of any one of claims 1 to 8, wherein administering to the subject 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient reduces cardiovascular risk in the subject.

15. 15. The method of claim 14, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences about a 5% to about 100% reduction in the cardiovascular risk condition or symptom.

16. 9. The method of any one of claims 1 to 8, wherein administering to the subject 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient decreases DNA methylation age in the subject.

17. 17. The method of claim 16, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences about a 5% to about 100% reduction in a condition or symptom involving DNA methylation.

18. 9. The method of any one of claims 1 to 8, wherein administering to the subject 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient reduces DNA methylation phenoage in the subject.

19. 19. The method of claim 18, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences about a 5% to about 100% reduction in a condition or symptom associated with DNA methylation phenoaging.

20. 9. The method of any one of claims 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces DNA methylation skin blood clock in the subject.

21. 21. The method of claim 20, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences about a 5% to about 100% reduction in a condition or symptom associated with the DNA methylation Skin Blood Clock.

22. 9. The method of any one of claims 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces the subject's CDR score.

23. 23. The method of claim 22, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences about a 5% to about 100% reduction in symptoms associated with CDR scores.

24. 9. The method of any one of claims 1 to 8, wherein administering to the subject 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient reduces the subject's ADAS-Cog score.

25. 25. The method of claim 24, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences about a 5% to about 100% reduction in symptom associated ADAS-Cog score.

26. 9. The method of any one of claims 1 to 8, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences about a 5% to about 100% reduction in symptom related MMSE score.

27. 9. The method of any one of claims 1 to 8, wherein administering 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient to the subject reduces the subject's MoCA score.

28. 9. The method of any one of claims 1 to 8, wherein after administration of 17α-ethynylandrost-5-ene-3β,7β,17β-triol and at least one pharmaceutically acceptable excipient, the subject experiences about a 5% to about 100% reduction in symptom-related ADL score.

29. 29. The method of any one of claims 1 to 28, wherein 17α-ethynylandrost-5-ene-3β,7β,17β-triol is administered orally.

30. 29. The method of any one of claims 1 to 28, wherein 17α-ethynylandrost-5-ene-3β,7β,17β-triol is administered intravenously.

31. 31. The method of any one of claims 1 to 30, wherein the subject has a waist-to-hip ratio of greater than or equal to approximately 0.

90.

32. 31. The method of any one of claims 1 to 30, wherein the subject has a waist-to-hip ratio of greater than or equal to approximately 0.

95.

33. 33. The method of any one of claims 1 to 32, wherein the 17α-ethynylandrost-5-ene-3β,7β,17β-triol is a solid-state form of 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

34. 34. The method of claim 33, wherein the solid state form of 17α-ethynylandrost-5-ene-3β,7β,17β-triol is a crystalline solvate of 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

35. 35. The method of claim 34, wherein the crystalline solvate is crystalline methanolate 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

36. 36. The method of claim 35, wherein the crystalline solvate is crystalline ethanolate 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

37. 37. The method of claim 36, wherein the crystalline solvate is crystalline hydrate 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

38. 37. The method of claim 36, wherein the crystalline solvate is Form III 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

39. 37. The method of claim 36, wherein the crystalline solvate is Form IV 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

40. 37. The method of claim 36, wherein the crystalline solvate is Form V 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

41. 36. The method of claim 35, wherein the solid state form of 17α-ethynylandrost-5-ene-3β,7β,17β-triol is amorphous 17α-ethynylandrost-5-ene-3β,7β,17β-triol.

42. 42. The method of any one of claims 1 to 41, wherein the 17α-ethynylandrost-5-ene-3β,7β,17β-triol is in a pharmaceutical composition, and the pharmaceutical composition contains less than about 3% by weight of impurities.

43. 1. An in vitro screening method for identifying potential drug candidates or compounds capable of treating, preventing, inhibiting, or ameliorating a disorder or disease, comprising: (i) providing a sample containing one or more stimulator cells selected from the group consisting of cells, tissue, blood, monocytes, microglia, macrophages, adipocytes, neuroblastoma, pheochromocytoma, and Lund human mesencephalon (LUHMES) cells; (ii) stimulating the sample with an agonist to induce a phenotype or phenotypic response, wherein said phenotype or phenotypic response substantially corresponds to a disease or condition associated with at least one DNA methylation at a CpG site within the region of DNA; (iii) contacting said one or more cells exhibiting said phenotype or phenotypic response with one or more potential drug candidates; (iv) determining a change in the phenotype or responsiveness of the sample; and (v) providing the drug candidate to a subject in need thereof to treat, inhibit, prevent, or ameliorate the DNA methylation-associated disease or condition in the subject.

44. At least one DNA methylation at the CpG site is selected from AC073869.20, SP100, KCNQ1DN, DBNDD2, CEP112, CEP85L, SPDYE4, ZNF211, NR3C1, HLA-L, TPP2, SLC26A1, SLC37A1, CAB39L, ILKAP, NPHP4, PATE4, ARHGEF12, CELA1, OR10G7, PFN2, WDR59, snoU13, 44. The method of claim 43, wherein the gene is selected from the group consisting of ANXA3, SVIL-AS1, PPHLN1, AP000442.1, FA, KIAA0319L, ZNF509, DLEU2L, ABL2, SGK1, TMEM245, SRSF4, DAP, GRAMD1C, FABP5P1, MCM10, ANP32E, ZNF268, ESPN, DHFR, U6, MTUS1, ATP1B3, or a combination thereof.

45. 45. The method of any one of claims 43 to 44, wherein the change in responsiveness is a reduction or elimination of the phenotype, and wherein the reduction or elimination indicates that the potential drug candidate is capable of preventing, inhibiting, or ameliorating a neurodegenerative disorder or disease.

46. 46. ​​The method of claim 45, wherein the neurodegenerative disorder or disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, levodopa-induced dyskinesia (LID), amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), age-related hippocampal sclerosis (HS-Aging), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy, multiple system atrophy, corticobasal degeneration, and vascular parkinsonism.

47. 47. The method of claim 46, wherein identifying DNA methylation changes in the subject identifies CpGs that are reduced by more than 50%.

48. 48. The method of claim 47, wherein identifying DNA methylation changes is correlated with one or more clinical changes.

49. The DNA methylation changes at CpG sites include AC073869.20, SP100, KCNQ1DN, DBNDD2, CEP112, CEP85L, SPDYE4, ZNF211, NR3C1, HLA-L, TPP2, SLC26A1, SLC37A1, CAB39L, ILKAP, NPHP4, PATE4, ARHGEF12, CELA1, OR10G7, PFN2, WDR59, snoU13, ANXA3, SV 49. The method of any of claims 44 to 48, wherein the target gene is selected from the group consisting of IL-AS1, PPHLN1, AP000442.1, FA, KIAA0319L, ZNF509, DLEU2L, ABL2, SGK1, TMEM245, SRSF4, DAP, GRAMD1C, FABP5P1, MCM10, ANP32E, ZNF268, ESPN, DHFR, U6, MTUS1, ATP1B3, or a combination thereof.

50. 50. The method of any of claims 44 to 49, wherein the disease or condition associated with a DNA methylation related biomarker is selected from the group consisting of TNFα, GRC, CDR, MoCA, QDRS, GRC, ADCOMS, MoCA, QDRS-Cognition, ADAS-Cogl 1, heart rate, frontal lobe, systolic blood pressure, grey matter, body weight, MMSE, hippocampal volume, behavior, PDQ-9, CSF glucose, precuneus GLTH, CSF pTau / Ab, or a combination thereof.

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