Methods for the treatment of biological aging

Abstract

Disclosed herein are compositions and methods useful for reducing biological age measured by DNA methylation and the treatment or amelioration of various diseases, disorders, or conditions promoted by increased biological age. Some aspects pertain to a pharmaceutical composition 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 states thereof. Also presented herein is the surprising discovery that exposing a subject to the compositions disclosed herein can treat, reduce, or ameliorate a condition related to biological clocks.

Description

METHODS FOR THE TREATMENT OF BIOLOGICAL AGINGField of the Disclosure

[0001] The present disclosure relates to the field of chemistry and medicine. More particularly, the present disclosure relates generally to methods for the treatment of biological aging using 17-ethynyl-lOR, 13S-dimethyl 2, 3, 4, 7, 8R, 9S, 10, 11, 12, 13, 14S, 15, 16, 17- hexadecahydro-lH-cyclopenta[a]phenanthrene-3R, 7R, 17S-triol.BACKGROUND

[0002] While aging may be a complex, multifactorial process with no single cause or treatment, the issue of whether aging can be classified as a disease is widely debated. Many strategies for extending organismal life spans have been proposed, including replacing cells and organs, comprehensive strategies for repairing accumulated damage, using hormones to activate endogenous repair processes, and modulating aging processes through specific mutations, gene therapy, and small-molecule drugs. An animal's survival strongly depends on its ability to maintain homeostasis, achieved partly through intracellular and intercellular communication within and among different tissues.

[0003] Many biomarkers of aging have been proposed, including telomere length, intracellular and extracellular aggregates, racemization of amino acids, and genetic instability. Gene expression and DNA methylation profiles change during aging, and may also be used as aging biomarkers. DNA methylation algorithms are increasingly used to estimate biological aging; however, how these proposed measures of whole-organism biological aging relate to aging in the brain is not known or well understood.SUMMARY OF THE DISCLOSURE

[0004] Aspects of the disclosure relate to a method to treat, reduce, or ameliorate a disease or condition associated with biological clocks in a subject in need thereof. In some embodiments, the method includes administering to the subject 17a-ethynylandrost-5-ene- 30,70,170-triol. In some embodiments, the disease or condition associated with biological clocksin the subject in need thereof is based on modulation of DNA methylation of genes associated with biological clocks. In some embodiments, the disease or condition associated with a biological clock in the subject in need thereof is associated with genes or genomic regions hypermethylated with age. In some embodiments, the disease or condition associated with a biological clock in the subject in need thereof is associated with genes or genomic regions hypomethylated with age. In some embodiments, the disease or condition associated with a biological clock in a subject in need thereof is associated with Tau phosphorylation. In some embodiments, the disease or condition associated with a biological clock in a subject in need thereof is associated with hyperglycemia. In some embodiments, the disease or condition associated with a biological clock in a subject in need thereof is associated with insulin resistance. In some embodiments, the disease or condition associated with biological clock is mild cognitive impairment or late onset Alzheimer’s disease. In some embodiments, administering to the subject 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient decreases a subject’s Alzheimer’s Disease Composite Score. In some embodiments, the subject experiences a decrease between about a 5% to about a 100% reduction in symptoms connected to Alzheimer’s Disease Composite Score after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient. In some embodiments, administering to the subject 17a-ethynylandrost-5- ene-3p,7p,17p-triol and at least one pharmaceutically acceptable excipient decreases pTau in the subject. In some embodiments, the subject experiences a decrease between about a 5% to about a 100% reduction in conditions or symptoms connected to pTau after administration of 17a- ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient. In some embodiments, administering to the subject 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient increases leptin in the subject. In some embodiments, the subject experiences a decrease between about a 5% to about a 100% reduction in conditions or symptoms connected to leptin after administration of 17a-ethynylandrost-5-ene- 3p,7p,17p-triol and at least one pharmaceutically acceptable excipient. In some embodiments, administering to the subject 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient decreases DNA methylation in the subject. In some embodiments, the subject experiences a decrease between about a 5% to about a 100% reduction in conditions or symptoms connected to DNA methylation after administration of 17a- ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient. Insome embodiments, administering to the subject 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient decreases cardiovascular risk in the subject. In some embodiments, the subject experiences a decrease between about a 5% to about a 100% reduction in conditions or symptoms connected to cardiovascular risk after administration of 17a- ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient. In some embodiments, administering to the subject 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient decreases DNA methylation age in the subject. In some embodiments, the subject experiences a decrease between about a 5% to about a 100% reduction in conditions or symptoms connected to DNA methylation after administration of 17a- ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient. In some embodiments, administering to the subject 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient decreases DNA methylation phenoage in the subject. In some embodiments, the subject experiences a decrease between about a 5% to about a 100% reduction in conditions or symptoms connected to DNA methylation phenoage after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient. In some embodiments, administering to the subject 17a-ethynylandrost-5- ene-3p,7p,17p-triol and at least one pharmaceutically acceptable excipient decreases DNA methylation skin blood clock in the subject. In some embodiments, the subject experiences a decrease between about a 5% to about a 100% reduction in conditions or symptoms connected to DNA methylation skin blood clock after administration of 17a-ethynylandrost-5-ene-3P,7P,17P- triol and at least one pharmaceutically acceptable excipient. In some embodiments, the 17a- ethynylandrost-5-ene-3P,7P,17P-triol is administered orally. In some embodiments, the 17a- ethynylandrost-5-ene-3P,7P,17P-triol is administered intravenously. In some embodiments, the subject has a waist to hip ratio greater than or equal to approximately 0.90. In some embodiments, the subject has a waist to hip ratio greater than or equal to approximately 0.95. In some embodiments, the 17oc-ethynylandrost-5-ene-3p,7p,17P-triol is a solid state form of 17a- ethynylandrost-5-ene-3p,7p,17P-triol. In some embodiments, the solid state form of 17a- ethynylandrost-5-ene-3p,7p,17P-triol is crystalline solvate of 17a-ethynylandrost-5-ene- 3p,7p,17p-triol. In some embodiments, the crystalline solvate is crystalline methanolate 17a- ethynylandrost-5-ene-3p,7p,17P-triol. In some embodiments, the crystalline solvate is crystalline ethanolate 17a-ethynylandrost-5-ene-3p,7p,17P-triol. In some embodiments, the crystallinesolvate is crystalline hydrate 17a-ethynylandrost-5-ene-3p,7p,17P-triol. In some embodiments, the crystalline solvate is Form III 17oc-ethynylandrost-5-ene-3p,7p,17P-triol. In some embodiments, the crystalline solvate is Form IV 17oc-ethynylandrost-5-ene-3p,7p,17P-triol. In some embodiments, the crystalline solvate is Form V 17oc-ethynylandrost-5-ene-3p,7p,17P-triol. In some embodiments, the solid state form of 17oc-ethynylandrost-5-ene-3p,7p,17P-triol is amorphous 17oc-ethynylandrost-5-ene-3p,7p,17P-triol. In some embodiments, the pharmaceutical composition contains less than about 3% by weight of impurities.

[0005] Some embodiments relate to an in vitro screening method to identify a potential drug candidate or compound capable of treating, preventing, reducing, or ameliorating a disorder or disease. In some embodiments, the method includes (i) providing a sample for stimulation selected from the group consisting of a cell, tissue, blood, monocytes, microglia, macrophages, adipocytes, neuroblastoma, pheochromocytoma, and Lund human mesencephalic (LUHMES) cells, (ii) stimulating the sample with an agonist to induce a phenotype or phenotypic reaction, wherein the phenotype or phenotypic reaction substantially corresponds to a disease or condition associated with at least one DNA methylation at a CpG site in a region of DNA, (iii) contacting the one or more cells exhibiting the phenotype or phenotypic reaction with one or more potential drug candidate or compounds, (iv) determining a responsive change in the phenotype of the sample, and (v) providing the drug candidate or compound to a subject in need thereof to treat, reduce, prevent, or ameliorate a disease or condition associated with the DNA methylation 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, 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 phenotype or phenotypic reaction is selected from the group consisting of TNFa, GRC, CDR, MoCA, QDRS, GRC, ADCOMS, MoCA, QDRS-Cognition, ADAS-Cogl l, heart rate, frontal lobe, systolic blood pressure, grey 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 reaction is selected from the group consisting of ADAS-Cogl 1, ADCOMS, CDR, CSF glucose.CSF p Tau / Ab, frontal lobe volume, subcortical grey mater thickness, GRC, Heart rate, MoCA, PDQ-9, precuneus glutathione, QDRS, QDRS-behavior, QDRS-cognition, Systolic BP, Tau, TNFa, and weight. In some embodiments, the DNA methylation change is a decrease of > 50%. In some embodiments, the DNA methylation change is a decrease of > 55%. In some embodiments, the DNA methylation change is a decrease of > 60%. In some embodiments, the phenotype or phenotypic reaction is decreased in TNF, CDR, QRDS-cognition, wherein the subject QDRS-cognition is improved. In some embodiments, the responsive change is a decrease or loss in the phenotype and the decrease or loss is indicative that the potential drug candidate or compound is capable of preventing, reducing, or ameliorating 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), hippocampal sclerosis of aging (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 CpGs sites are interconnected with genes associated with Alzheimer’s disease and related dementias. In some embodiments, the disease or condition associated with inflammatory TNF signaling. In some embodiments, the disease or condition associated with DNA methylation associated with Tau phosphorylation. In some embodiments, the disease or condition associated with DNA methylation associated with hyperglycemia. In some embodiments, the disease or condition associated with DNA methylation associated with insulin resistance. In some embodiments, the disease or condition associated with DNA methylation associated with obesity, ADAS-Cogl l, ADCOMS, CDR, CSF glucose, CSF pTau / A0, frontal lobe volume, subcortical grey matter thickness, GRC, heart rate, MoCA, PDQ-9, precuneus glutathione, QDRS, QDRS- behavior, QDRS-cognition, systolic blood pressure, Tau, TNFa, and weight.

[0006] Some embodiments relate to a method of diagnose a patient with a disease or a condition. In some embodiments, the method includes (i) providing a patient with a potential drug candidate or compound capable of treating, preventing, reducing, or ameliorating a disorder or disease, (ii) identifying DNA methylation changes in the patient, and (iii) diagnosing the patient with a disease or condition associated with a biomarker associated with DNA methylation. In someembodiments, identifying DNA methylation changes in the patient identifies the CpGs decreased by more than 50%. In some embodiments, identifying DNA methylation changes is correlated with one or more clinical changes. In some embodiments, the DNA methylation changes at a 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, 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 biomarker associated with DNA methylation is selected from the group consisting of TNFa, GRC, CDR, MoCA, QDRS, GRC, ADCOMS, MoCA, QDRS-Cogmtion, ADAS-Cogl l, heart rate, frontal lobe, systolic blood pressure, grey matter, weight, MMSE, hippocampal volume, behavior, PDQ-9, CSF glucose, precuneus GLTH, CSF pTau / Ap, 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-Cogl l, ADCOMS, CDR, CSF glucose, CSF p'Tau / Ap, frontal lobe volume, subcortical grey matter thickness, GRC, Heart rate, MoCA, PDQ-9, precuneus glutathione, QDRS, QDRS-behavior, QDRS-cognition, Systolic BP, Tau, TNFa, and weight. In some embodiments, the biomarker associated with DNA methylation is decreased in TNF, CDR, QRDS-cognition, wherein the subject QDRS-cognition is improved. In some embodiments, the DNA methylation change is a decrease of > 50%. In some embodiments, the DNA methylation change is a decrease of > 55%. In some embodiments, the DNA methylation change is a decrease of > 60%. 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 (FID), hippocampal sclerosis of aging (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 CpGs sites are interconnected with genes associated with Alzheimer’s disease and related dementias. In some embodiments, the disease or condition associated with inflammatory TNF signaling. In some embodiments, the disease or condition associated with DNA methylationassociated with Tau phosphorylation. In some embodiments, the disease or condition associated with DNA methylation associated with hyperglycemia. In some embodiments, the disease or condition associated with DNA methylation associated with insulin resistance. In some embodiments, the disease or condition associated with DNA methylation associated with obesity, ADAS-Cogl l, ADCOMS, CDR, CSF glucose, CSF pTau / A P, frontal lobe volume, subcortical grey matter thickness, GRC, heart rate, MoCA, PDQ-9, precuneus glutathione, QDRS, QDRS- behavior, QDRS-cognition, systolic blood pressure, Tau, TNFa, and weight.

[0007] Not all objectives mentioned in this specification are necessarily achieved in all embodiments disclosed and / or claimed herein.BRIEF DESCRIPTION OF THE DRAWINGS

[0008] FIG. 1 illustrates a graph representing DNA methylation scores (pre- and posttreatment).

[0009] FIG. 2 illustrates a graph representing DNA methylation scores in relation to pack years.

[0010] FIG. 3 illustrates a graph estimating leptin scores.

[0011] FIG. 4 illustrates a graph measuring cardiovascular risk.

[0012] FIG. 5 illustrates a graph measuring changes in DNAmAGE.

[0013] FIG. 6 illustrates a graph measuring changes in DNAPhenoAge followingNE3107 treatment.

[0014] FIG. 7 illustrates a graph measuring changes in DNAmin te SkinBloodClock.

[0015] FIG. 8 illustrates a graph measuring DNAm changes following 14 weeks of NE3107 treatment.

[0016] FIG. 9 illustrates a graph representing changes in the Alzheimer’s Disease Composite Score (ADCOMS) change.

[0017] FIG. 10 illustrates a graph representing the relationship between pTau and ADCOMS.

[0018] FIG. 11 illustrates a graph representing CSF pTau in relation to ADCOMS for MMSE.

[0019] FIG. 12 illustrates a chart representing a Phase 3, randomized, placebo- controlled trial of NE3107 in subjects with mild to moderate probable Alzheimer’s disease.

[0020] FIGs. 13A-13G illustrate graphs for improvements in the blinded assessments from the Phase 3, Randomized, Placebo-Controlled Trials.

[0021] FIGs. 14A-14B illustrate graphs for imaging sub-studies of vMRI hippocampus volume and amygdala volume in the blinded assessments from the Phase 3, randomized, placebo- controlled trials.

[0022] FIG. 15 illustrates graphs representing imaging sub-studies using FDG-PET in blinded assessments from the Phase 3, randomized, placebo-controlled trials.

[0023] FIGs. 16A-16B illustrates graphs representing the neuropsychiatric inventory in blinded assessments from the Phase 3, randomized, placebo-controlled trials.

[0024] FIGs. 17A-17C illustrates graphs representing increased fasting insulin and H0MA2-%B with decreased HOMA2-%S without hypoglycemia.

[0025] FIGs. 18A-18G illustrate graphs representing placebo effects in various assessments.

[0026] FIGs. 19-20 illustrate graphs representing ADAS-Cogl2 Spearman correlations.

[0027] FIGs. 21-22 illustrate graphs representing clinician rating of global change Spearman correlations.

[0028] FIGs. 23-24 illustrate graphs representing Mini-Mental State Exam Spearman correlations.

[0029] FIG. 25 illustrate graphs representing ADCOMS Spearman correlations.

[0030] FIG. 26 illustrate graphs representing CDR sum of boxes Spearman correlations.

[0031] FIG. 27 illustrate graphs representing activities of daily living Spearman correlations.

[0032] FIG. 28 illustrate graphs representing improvement in ADAS-Cogl2 correlated with increased FDG-PET SUVR.

[0033] FIG. 29 illustrate a graph representing improvement in MMSE correlated with increased FDG-PET SUVR.

[0034] FIG. 30 illustrate graphs representing improvement in ADL correlated with increased FDG-PET SUVR.

[0035] FIG. 31 illustrate graphs representing improvement in HOMA2 insulin sensitivity correlated with increased FDG-PET SUVR.

[0036] FIG. 32 illustrate graphs representing an improvement in cholesterol correlated with increased FDG-PET SUVR.

[0037] FIG. 33 illustrate graphs representing Horvath skin blood clock scores.

[0038] FIG. 34 illustrate a graph representing a Clq score.

[0039] FIG. 35 illustrates graphs representing NE3107 MMSE increase proportional to biological age (dAge - Horvath Skin Blood Clock - Chronological Age).

[0040] FIG. 36 illustrate graphs representing NE3107 MMSE increase inversely proportional to baseline waist / hip ratio.

[0041] FIG. 37 illustrate graphs representing NE3107 MMSE increase proportional to RANTES increase.

[0042] FIG. 38 illustrate graphs representing NE3107 CDR SB decrease proportional to diastolic BP decrease.

[0043] FIG. 39 illustrate graphs representing NE3107 decrease CDR SB proportional to insulin sensitivity increase.

[0044] FIG. 40 illustrate graphs representing NE3107 decrease CDR SB proportional to cholesterol decrease.

[0045] FIG. 41 illustrate graphs representing NE3107 CDR SB decrease proportional to dAge (Biological Age [Horvath Skin Blood Clock] - Chronological Age) in pTau+ subjects.

[0046] FIG. 42 illustrate graphs representing placebo increased CDR SB proportional to increased GFAP.

[0047] FIG. 43 illustrate graphs representing placebo increased CDR SB proportional to increased NIL.

[0048] FIG. 44 illustrate graphs representing NE3107 decrease in ADCOMS proportional to increased RANTES.

[0049] FIG. 45 illustrate graphs representing placebo increased CDR SB proportional decreased Ab42 / 40 ratio.

[0050] FIG. 46 illustrate graphs representing NE3107 ADCOMS is inversely proportion to V10 leptin.

[0051] FIG. 47 illustrate graphs representing NE3107 decrease in ADCOMS proportional to cholesterol decrease.

[0052] FIG. 48 illustrate graphs representing NE3107 ADCOMS decrease proportional to dAge (Biological Age [Horvath Skin Blood Clock] - Chronological Age) inpTau+ subjects.

[0053] FIG. 49 illustrate graphs representing placebo ADCOMS increase proportional to GFAP increase.

[0054] FIG. 50 illustrate graphs representing placebo increased ADCOMS proportional to increased NfL.

[0055] FIG. 51 illustrate graphs representing placebo increased ADCOMS proportional to decreased Ab42 / 40 ratio. No outlier detected in placebo (Gibbs a=0.05, G = 2.42).

[0056] FIG. 52 illustrate graphs representing NE3107 decreased Cog 12 proportional to increased RANTES.

[0057] FIG. 53 illustrate graphs representing NE3107 decreased Cog 12 proportional to increased Adiponectin.

[0058] FIG. 54 illustrate graphs representing NE3107 decreased Cog 12 proportional to increased monocytes.

[0059] FIG. 55 illustrate graphs representing NE3107 decreased CGIC inversely proportional to baseline waist / hip ratio.

[0060] FIG. 56 illustrate graphs representing NE3107 decreased CGIC proportional to decreased dAge (Biological Age [Horvath Skin Blood Clock] - Chronological Age).

[0061] FIG. 57 illustrate graphs representing NE3107 decreased CGIC proportional to decreased NPI (Neuropsychiatric Inventory).

[0062] FIG. 58 illustrate graphs representing placebo increased CGIC proportional to decreased diastolic BP.

[0063] FIG. 59 illustrate graphs representing NE3107 decreased ADL proportional to decreased systolic BP.

[0064] FIG. 60 illustrate graphs representing decreased ADL proportional to decreased insulin.

[0065] FIG. 61 illustrate graphs representing decreased ADL proportional to decreased insulin resistance.

[0066] FIG. 62 illustrate graphs representing decreased ADL proportional to increased fructosamine.

[0067] FIG. 63 illustrate graphs representing decreased ADL proportional to increased C-reactive protein.

[0068] FIG. 64 illustrate graphs representing decreased ADL proportional to increased GFAP.

[0069] FIG. 65 illustrate graphs representing ADL decrease proportional to NfL increase.

[0070] FIG. 66 illustrate graphs representing NE3107 lowered WHR proportional to increased RANTES.

[0071] FIG. 67 illustrate graphs representing NE3107 lower WHR proportional to higher leptin. Outlier detected (Gibbs a = 0.05, G = 2.58).

[0072] FIG. 68 illustrate graphs representing NE3107 higher WHR proportional decreased triglycerides.

[0073] FIG. 69 illustrate graphs representing NE3107 lower WHR proportional to decreased dAge (Biological Age [Horvath Skin Blood Clock] - Chronological Age). Outlier detected (Gibbs a = 0.05, G = 2.77).

[0074] FIG. 70 illustrate graphs representing NE3107 lower WHR proportional to decreased NfL.

[0075] FIG. 71 illustrate graphs representing placebo higher WHR proportional to increased cholesterol.

[0076] FIG. 72 illustrate graphs representing NE3107 MCP1 cfb proportional to RANTES cfb.

[0077] FIG. 73 illustrate graphs representing NE3107 MCP1 cfb proportional to dAge (Biological Age [Horvath Skin Blood Clock] - Chronological Age) cfb.

[0078] FIG. 74 illustrate graphs representing NE3107 MCP1 cfb proportional to dAge (Biological Age [Horvath Skin Blood Clock] - Chronological Age) (pTau+).

[0079] FIG. 75 illustrate graphs representing MCP1 cfb proportional to NPI cfb.

[0080] FIG. 76 illustrate graphs representing placebo TNF cfb proportional to age.

[0081] FIG. 77 illustrate graphs representing NE3107 RABTES cfb proportional to fructoamine cfb.

[0082] FIG. 78 illustrate graphs representing placebo RANTES cfb proportional to diastolic BP cfb.

[0083] FIG. 79 illustrate graphs representing placebo systolic BP cfb proportional to triglycerides cfb.

[0084] FIG. 80 illustrate graphs representing placebo systolic BP cfb proportional to cholesterol cfb.

[0085] FIG. 81 illustrate graphs representing placebo systolic BP cfb proportional to dAge (Biological Age [Horvath Skin Blood Clock] - Chronological Age).

[0086] FIG. 82 illustrate graphs representing placebo diastolic BP cfb proportional to triglycerides cfb.

[0087] FIG. 83 illustrate graphs representing diastolic BP cfb proportional to cholesterol cfb.

[0088] FIG. 84 illustrate graphs representing placebo diastolic BP inversely proportional to C-reactive protein.

[0089] FIG. 85 illustrate graphs representing placebo diastolic BP proportional to dAge (Biological Age (Horvath Skin Blood Clock) - Chronological Age).

[0090] FIG. 86 illustrate graphs representing placebo diastolic BP cfb proportional to dAge (Biological Age (Horvath Skin Blood Clock) - Chronological Age) (A0+).

[0091] FIG. 87 illustrate graphs representing placebo diastolic BP cfb proportional to dAge (Biological Age (Horvath Skin Blood Clock) - Chronological Age) (pTau+).

[0092] FIG. 88 illustrate graphs representing glucose cholesterol, Clq and dAge (Biological Age (Horvath Skin Blood Clock) - Chronological Age).

[0093] FIG. 89 illustrate graphs representing that NE3107 insulin cfb proportional to triglycerides cfb.

[0094] FIG. 90 illustrate graphs representing that NE3107 insulin cfb proportional to Clq cfb.

[0095] FIG. 91 illustrate graphs representing placebo insulin cfb proportional to V10 leptin.

[0096] FIG. 92 illustrate graphs representing insulin resistance drives placebo weight gam.

[0097] FIG. 93 illustrate graphs representing that NE3107 pTau217 decrease proportional to biologic age decrease.

[0098] FIG. 94 illustrate graphs representing that NE3107 NfL decrease proportional to pTau217 decrease.

[0099] FIG. 95 illustrates a graph representing no significant change in weight from placebo and NE3107 (weight in kg).

[0100] FIG. 96 illustrates a graph representing a directional improvement in insulin sensitivity.

[0101] FIG. 97 illustrates a graph representing that NE3107 decreased dAge(Biological Age (Horvath Skin Blood Clock) - Chronological Age).

[0102] FIG. 98 illustrate graphs representing that NE3107 can alter MMSE dAge(Biological Age (Horvath Skin Blood Clock) - Chronological Age) axis.

[0103] FIG. 99 illustrates a graph representing no significant difference in MMSE medians.

[0104] FIG. 100 illustrate graphs representing that NE3107 may alter the MMSE WHR(waist / hip ratio) at screening axis.

[0105] FIG. 101 illustrate graphs representing that NE3107 may alter the MMSE- RANTES axis.

[0106] FIG. 102 illustrates a graph representing no significant difference in RANTES medians.

[0107] FIG. 103 illustrate graphs representing that NE3107 may alter CDR SB viaH0MA2 %S (H0MA2 %S = Homeostatic Model Assessment % Insulin Sensitivity).

[0108] FIG. 104 illustrates a graph no significant difference in CDR SB medians.

[0109] FIG. 105 illustrate graphs representing that NE3107 may alter the CDR SBCholesterol axis.

[0110] FIG. 106 illustrates a graph representing that NE3107 significantly decreased cholesterol.

[0111] FIG. 107 illustrate graphs representing that NE3107 may alter the CDR SB dAge (Biological Age (Horvath Skin Blood Clock) - Chronological Age) axis in pTau+ subjects.

[0112] FIG. 108 illustrate graphs representing that NE3107 significantly decreased dAge (Biological Age (Horvath Skin Blood Clock) - Chronological Age) pTau+ compared to placebo.

[0113] FIG. 109 illustrate graph representing that NE3107 may alter the CDR SB GFAP (glial fibrillary acid protein) axis.

[0114] FIG. 110 illustrates a graph representing no significant difference in GFAP (glial fibrillary acidic protein) medians (Hodges-Lehman difference = -1.49).

[0115] FIG. I l l illustrate graphs representing that NE3107 may alter the ADCOMS RANTES axis.

[0116] FIG. 112 illustrates a graph representing no significant differences in ADCOMS (Alzheimer’s Disease Composite Score) compared to placebo (Hodges-Lehmann difference = 0.033).

[0117] FIG. 113 illustrate graphs representing that NE3107 may alter the cholesterol ADCOMS axis.

[0118] FIG. 114 illustrate graphs representing that NE3107 may alter the ADCOMS GFAP axis.

[0119] FIG. 115 illustrate graphs representing that NE3107 may alter the ADCOMS NfL axis.

[0120] FIG. 116 illustrates a graph representing no significant difference in NfL (neurofilament light chain) between placebo and NE3107 (Hodges-Lehmann difference = 0.874).

[0121] FIG. 117 illustrate graphs representing that NE3107 may alter the Cogl2 RANTES axis.

[0122] FIG. 118 illustrates a graph representing no significant difference in Cog 12 (ADAS-Cogl2) in NE3107 compared to placebo (Hodges-Lehman difference = -1.16).

[0123] FIG. 119 illustrate graphs representing that NE3107 may alter the Cogl2 Adiponectin axis.

[0124] FIG. 120 illustrates a graph representing that NE3107 significantly increased adiponectin.

[0125] FIG. 121 illustrate graphs representing that NE3107 may alter Cogl2 monocytes axis.

[0126] FIG. 122 illustrates a graph representing NE3107 showing a directional change in monocytes compared to placebo.

[0127] FIG. 123 illustrate graphs representing that NE3107 may alter CGIC WHR axis.

[0128] FIG. 124 illustrates a graph representing no significant change in CGIC medians compared to placebo.

[0129] FIG. 125 illustrate graphs representing that NE3107 may alter the CGIC dAge (Biological Age [Horvath Skin Blood Clock] - Chronological Age) axis.

[0130] FIG. 126 illustrate graphs representing that NE3107 may alter the CGIC NPI (Neuropsychiatric Inventory) axis.

[0131] FIG. 127 illustrates a graph representing no significant difference in NPI total (total of Neuropsychiatric Inventory frequency times severity scores) (Hodges-Lehmann difference = -0.5).

[0132] FIG. 128 illustrates graphs representing placebo ADL cfb proportional to H0MA2 IR cfb.

[0133] FIG. 129 illustrates a graph representing directional change in H0MA2-IR compared to placebo (Homeostatic Model Assessment 2 Insulin Resistance).

[0134] FIG. 130 illustrate graphs representing NE3107 that may alter ADL-CRP axis.

[0135] FIG. 131 illustrates a graph representing ADL (activities of daily living) compared to placebo (Hodges-Lehman difference = +2.0).

[0136] FIG. 132 illustrates a graph representing no significant difference in CRP (C- reactive protein) medians compared to placebo.

[0137] FIG. 133 illustrate graphs representing that NE3107 may alter the ADL / GFAP axis.

[0138] FIG. 134 illustrate graphs representing that NE3107 may alter the obesity / leptin axis.

[0139] FIG. 135 illustrate a graph representing that NE3107 may alter the Leptin resistance.

[0140] FIG. 136 illustrate graphs representing that NE3107 may alter the obesity / triglyceride axis.

[0141] FIG. 137 illustrate a graph representing directional change in triglycerides compared to placebo.

[0142] FIG. 138 illustrate graphs representing that NE3107 may alter obesity effects on dAge (Biological Age [Horvath Skin Blood Clock] - Chronological Age).

[0143] FIG. 139 illustrate graphs representing that NE3107 may alter the obesity / NfL axis.

[0144] FIG. 140 illustrate graphs representing that NE3107 may alter the obesity / cholesterol axis.

[0145] FIG. 141 illustrate graphs representing that NE3107 may restore chemokin coregulation.

[0146] FIG. 142 illustrates a graph representing that NE3107 may decrease MCP1 (Monocyte Chemoattractant Protein 1).

[0147] FIG. 143 illustrate graphs representing that NE3107 may restore immune component coregulation.

[0148] FIG. 144 illustrates a graph representing Clq compared to placebo.

[0149] FIG. 145 illustrate graphs representing that NE3107 may alter the chemokine dAge (Biological Age [Horvath Skin Blood Clock] - Chronological Age) axis.

[0150] FIG. 146 illustrate graphs representing that NE3107 may alter the chemokine dAge (Biological Age [Horvath Skin Blood Clock] - Chronological Age) axis.

[0151] FIG. 147 illustrate graphs representing that NE3107 may alter the triglyceride / systolic BP axis.

[0152] FIG. 148 illustrates a graph representing a directional change in systolic BP.

[0153] FIG. 149 illustrate graphs representing that NE3107 may alter the cholesterol / systolic BP axis.

[0154] FIG. 150 illustrates graphs representing that NE3107 may alter systolic BP dAge (Biological Age [Horvath Skin Blood Clock] - Chronological Age) axis.

[0155] FIG. 151 illustrate graphs representing that NE3107 may alter the diastolic BP triglyceride axis.

[0156] FIG. 152 illustrate graphs representing directional change in diastolic BP compared to placebo.

[0157] FIG. 153 illustrate graphs representing that NE3107 may alter the diastolic BP cholesterol axis.

[0158] FIG. 154 illustrate graphs representing that NE3107 may alter the diastolic BP / dAge (Biological Age [Horvath Skin Blood Clock] - Chronological Age) axis.

[0159] FIG. 155 illustrate graphs representing that NE3107 may improve metabolic regulation.

[0160] FIG. 156 illustrates a graph representing that NE3107 decreased insulin levels compared to placebo.

[0161] FIG. 157 illustrate graphs representing that NE3107 may alter insulin the dAge (Biological Age [Horvath Skin Blood Clock] - Chronological Age) axis.

[0162] FIG. 158 illustrates a graph representing directional change in diastolic BP compared to placebo.

[0163] FIG. 159 illustrate graphs representing that NE3107 may alter metabolic regulation.

[0164] FIG. 160 illustrate graphs representing that NE3107 may alter the H0MA2 IR Clq axis.

[0165] FIG. 161 illustrate graphs representing that NE3107 may dissociate H0MA2 IR from H0MA2 %S.

[0166] FIG. 162 illustrate graphs representing that NE3107 may alter the cholesterol dAge (Biological Age (Horvath Skin Blood Clock) - Chronological Age) axis.

[0167] FIG. 163 illustrate graphs representing that NE3107 may alter dAge (Biological Age [Horvath Skin Blood Clock] - Chronological Age) pTau axis.

[0168] FIG. 164 illustrates a graph representing no significant change in pTau compared to placebo.

[0169] FIG. 165 illustrate graphs representing that NE3107 may alter the GFAP GST (global statistical test = CDR SB & ADAS-Cogl2) axis.

[0170] FIG. 166 illustrates a graph representing no significant difference in GST (global statistical test = CDR SB & ADAS-Cogl2) medians.

[0171] FIG. 167 illustrate graphs representing that NE3107 may alter the NfL GST axis.

[0172] FIG. 168 illustrate graphs representing NE3107 dAge pTau217 Z’ scores.

[0173] FIG. 169 illustrate graphs representing NE3107 dAge MMSE Z’ scores.

[0174] FIG. 170 illustrate graphs representing NE3107 dAge CGIC Z’ scores.

[0175] FIG. 171 illustrate graphs representing NE3107 dAge MCP1 Z’ scores.

[0176] FIG. 172 illustrate graphs representing NE3107 cAge TNF Z’ scores.

[0177] FIG. 173 illustrate graphs representing that GFAP is proportional to NfL.

[0178] FIG. 174 illustrate graphs representing that CDR SB is proportional to TNF.

[0179] FIG. 175 illustrate graphs representing that placebo CDR SB is proportional to CFAP.

[0180] FIG. 176 illustrate graphs representing that placebo CDR SB is proportional to NfL.

[0181] FIG. 177 illustrate graphs representing that placebo CDR SB is proportional to CRP.

[0182] FIG. 178 illustrate graphs representing that placebo ADCOMS is proportional to GFAP.

[0183] FIG. 179 illustrate graphs representing that placebo ADCOMS is proportional to NfL.

[0184] FIG. 180 illustrate graphs representing that placebo Cogl2 is proportional to GFAP.

[0185] FIG. 181 illustrate graphs representing that placebo ADL is inversely proportional to GFAP.

[0186] FIG. 182 illustrate graphs representing that placebo ADL is inversely proportional to NfL.

[0187] FIG. 183 illustrate graphs representing that placebo ADL is proportional to GST.

[0188] FIG. 184 illustrate graphs representing that MMSE is proportional to MCP1.

[0189] FIG. 185 illustrate graphs representing that MMSE is proportional to RANIES.

[0190] FIG. 186 illustrate graphs representing that MMSE is inversely proportional to fructosamine.

[0191] FIG. 187 illustrate graphs representing that placebo CDR SB is proportional to RANTES.

[0192] FIG. 188 illustrate graphs representing that CDR SB is inversely proportional to MCPl.

[0193] FIG. 189 illustrate graphs representing that Cogl2 is inversely proportional to MCP1.

[0194] FIG. 190 illustrate graphs representing that Cog 12 is proportional to Fructosamine.

[0195] FIG. 191 illustrate graphs representing that GST is inversely proportional to MCP1.

[0196] FIG. 192 illustrates a graph representing the relationship between placebo and NE3107 and dAge (dAge = Horvath Skin Blood Clock, Biological age - Chronological Age).

[0197] FIG. 193 illustrate graphs representing the relationship between placebo and NE3107 and metabolic biomarkers.

[0198] FIG. 194 illustrate graphs representing the relationship between placebo and NE3107 and homeostatic model assessment.

[0199] FIG. 195 illustrate graphs representing the relationship between placebo and NE3107 and adipokines.

[0200] FIG. 196 illustrate graphs representing the relationship between placebo and NE3107 and vital signs.

[0201] FIG. 197 illustrate graphs representing the relationship between placebo and NE3107 and inflammation biomarkers.

[0202] FIG. 198 illustrate graphs representing the relationship between placebo and NE3107 and AD biomarkers.

[0203] FIG. 199A illustrates an analysis of SkinBlood epigenetic aging clock results for the per-protocol population with evaluable, complete DNA methylation data, using two principal components identified for placebo correlations (n=16). FIG. 199B shows a similar analysis for the same population, but utilizing a single principal component identified for NE3107 correlations..

[0204] FIG. 200A illustrates a placebo Bayesian principal component analysis (PCA). FIG. 200B illustrates an enlarged version of the PCA from FIG. 200A.

[0205] FIG. 201 illustrates a graph representing the relationship between placebo (P) and NE3107 (B) in intrinsic epigenetic age acceleration (IEAA).

[0206] FIG. 202 illustrates a graph representing the relationship between placebo (P) and NE3107 (B) in grim age epigenetic age acceleration.

[0207] FIG. 203 illustrates a graph representing the relationship between placebo (P) and NE3107 (B) in component PackYears Clock.

[0208] FIG. 204 illustrates a graph representing the relationship between placebo (P) and NE3107 (B) in retroviral integration site epigenetic age acceleration.

[0209] FIG. 205 illustrates a graph representing the relationship between placebo (P) and NE3107 (B) in retroviral integration version 2 epigenetic age acceleration.

[0210] FIG. 206 illustrates a graph representing the relationship between placebo (P) and NE3107 (B) in stochastic Zhang epigenetic age acceleration.

[0211] FIG. 207 illustrates a graph representing the relationship between placebo (P) and NE3107 (B) in stochastic phenoage epigenetic acceleration.

[0212] FIG. 208 illustrates a graph representing the relationship between placebo (P) and NE3107 (B) in stochastic Horvath epigenetic age acceleration.

[0213] FIG. 209 illustrates a graph representing the relationship between placebo (P) and NE3107 (B) in systems age epigenetic age acceleration.

[0214] FIG. 210 illustrates a graph representing the relationship between placebo (P) and NE3107 (B) in heart epigenetic age acceleration.

[0215] FIG. 211 illustrates a graph representing the relationship between placebo (P) and NE3107 (B) in CD4 naive T cell.

[0216] FIG. 212 illustrates a graph representing the relationship between placebo (P) and NE3107 (B) in grimage based on predicted age epigenetic age acceleration.

[0217] FIG. 213 illustrates a graph representing the relationship between placebo (P) and NE3107 (B) in skinblood clock epigenetic age acceleration.DETAILED DESCRIPTION

[0218] The following description provides context and examples, but should not be interpreted to limit the scope of the disclosure covered by the claims that follow in this specification or in any other application that claims priority to this specification. No single component or collection of components is essential or indispensable. 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 that is described and / or illustrated in any embodiment in thisspecification can be used with or instead of any feature, structure, component, material, step, or method that is described and / or illustrated in any other embodiment in this specification.Definitions

[0219] As used herein and unless otherwise stated or implied by context, terms that are used herein have the meanings that are defined here. The descriptions of embodiments and examples that are described illustrate the disclosure and they are not intended to limit it in any way. Unless otherwise contraindicated or implied, e.g., by including mutually exclusive elements or options, in these definitions and throughout this specification, the terms “a” and “an” mean one or more and the term “or” means and / or.

[0220] The term “animal” is used herein to include all vertebrate animals, including transgenic animals. It also includes an individual animal in all stages of development, including embryonic and fetal stages. As used herein, the term “production animals” is used interchangeably with “livestock animals” and refers generally to animals raised primarily for food. For example, such animals include, but are not limited to, cattle (bovine), sheep (ovine), pigs (porcine or swine), poultry (avian), and the like. As used herein, the term “cow” or “cattle” is used generally to refer to an animal of bovine origin of any age. Interchangeable terms include “bovine”, “calf, “steer”, “bull”, “heifer”, “cow” and the like. As used herein, the term “pig” is used generally to refer to an animal of porcine origin of any age. Interchangeable terms include “piglet”, “sow” and the like. As used herein, the term “companion animals” is used herein to refers to a domestic animal. Companion animal is used generally to refer to a domestic dog, cat, rabbit, guinea pig, ferret, horse, or the like.

[0221] A “formulation” or the like means a composition that one can administer to a subject, e.g., human or animal. Formulations are suitable for human or veterinary applications and would typically have expected characteristics for the formulation, e.g., parenteral formulations for human use would usually be sterile solutions or suspensions.

[0222] An “excipient”, “carrier”, “pharmaceutically acceptable carrier” or similar terms mean one or more component(s) or ingredient(s) that is acceptable in the sense of being compatible with the other ingredients in the disclosed compositions or formulations and not overly deleterious to the patient, animal, tissues or cells to which the formulation is to be administered.

[0223] ‘Effective amount” refers to the amount required to produce a desired effect (e.g., enhancing the half-life, bioavailability or efficacy of a compound described herein, treating biological aging in a subject, reducing DNA methylation in a subject, etc.

[0224] As used herein, “subject,” “host,” “patient,” and “individual” are used interchangeably and shall be given their ordinary meaning in the art and shall also refer to an organism that has cancer and / or leukemia. This includes mammals, e.g., a human, a non-human primate, ungulates, canines, felines, equines, mice, rats, and the like. The term “mammal” includes both human and non-human mammals. The term “subject” includes animals.

[0225] “Preventing” in reference to a disease, disorder or condition refers to preventing a disease, disorder or condition, e.g., causing the clinical symptoms of the disease, disorder or condition not to develop. As used herein, the term “prevent,” “prevents,” or “prevention” (and grammatical equivalents thereof) may also refer to a delay in the onset of a disease or disorder or the lessening of symptoms upon onset of the disease or disorder. The terms are not meant to imply complete abolition of disease and encompass any type of prophylactic treatment that reduces the incidence of the condition or delays the onset and / or progression of the condition.

[0226] The terms “therapeutically effective amount” and “effective amount” refer to the amount of active pharmaceutical ingredient necessary to provide the desired pharmacologic result. In practice, the therapeutically effective amount will vary widely depending on the severity of the disease condition, age of the subject, and the desired therapeutic effect.

[0227] The terms “treatment,” “treating,” “treat,” and the like shall be given their ordinary meaning and shall also include herein to generally refer to obtaining a desired pharmacologic and / or physiologic effect. The effect may be prophylactic in terms of completely or partially preventing a disease or symptom thereof and / or may be therapeutic in terms of a partial or complete stabilization or cure for a disease and / or adverse effect attributable to the disease. The terms “treatment,” as used herein shall be given its ordinary meaning and shall also cover any treatment of a disease in a mammal, particularly a human, and includes: (a) preventing the disease or symptom from occurring in a subject which may be predisposed to the disease or symptom but has not yet been diagnosed as having it; (b) inhibiting the disease symptom, e.g., arresting its development; and / or (c) relieving the disease symptom, e.g., causing regression of the disease or symptom.T1

[0228] The term “about” or “approximately” means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend 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 deviations, per the practice in the art. Alternatively, “about” can mean a range of up to 20%, up to 10%, up to 5%, and up to 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. Where particular values are described in the application and claims, unless otherwise stated the term “about” meaning within an acceptable error range for the particular value should be assumed.

[0229] 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 any purpose. When definitions of terms in incorporated references appear to differ from the definitions provided in the present teachings, the definition provided in the present teachings shall control. It will be appreciated that there is an implied “about” prior to the temperatures, concentrations, times, etc. discussed in the present teachings, such that slight and insubstantial deviations are within the scope of the present teachings herein. In this application, the use of the singular includes the plural unless specifically stated otherwise. Also, the use of “comprise”, “comprises”, “comprising”, “contain”, “contains”, “containing”, “include”, “includes”, and “including” are not intended to be limiting. It is to be understood that both the general description and the following detailed description are exemplary and explanatory only and are not restrictive. The term “and / or” denotes that the provided possibilities can be used together or be used in the alternative. Thus, the term “and / or” denotes that both options exist for that set of possibilities.

[0230] Terms and phrases used in this application, and variations thereof, especially in the appended claims, unless otherwise expressly stated, should be construed as open ended as opposed to limiting. As examples of the foregoing, the term “including” should be read to mean “including, without limitation,” “including but not limited to,” or the like; the term “comprising” as used herein is synonymous with “including,” “containing,” or “characterized by,” and 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 “includes but is not limited to;” the term “example” is used to provide exemplary instances ofthe item in discussion, not an exhaustive or limiting list thereof; and use of terms like “preferably,” “preferred,” “desired,” or “desirable,” and words of similar meaning should not be understood as implying that certain features are critical, essential, or even important to the structure or function of the invention, but instead as merely intended to highlight alternative or additional features that may or may not be utilized in a particular embodiment of the disclosure. In addition, the term “comprising” is to 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. Likewise, a group of items linked with the conjunction “and” should not be read as requiring that each and every one of those items be present in the grouping, but rather should be read as “and / or” unless expressly stated otherwise. Similarly, a group of items linked with the conjunction “or” should not be read as requiring mutual exclusivity among that group, but rather should be read as “and / or” unless expressly stated otherwise.

[0231] With respect to the use of substantially any plural and / or singular terms herein, those having skill in the art can translate from the plural to the singular and / or from the singular to the plural as is appropriate to the context and / or application. The various singular / plural permutations may be expressly set forth herein for sake of 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 indicate 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.Methods of Treatment

[0232] Aspects of the present disclosure relate to methods to decrease biological age or DNA age and / or rate of biological aging through changes in patterns of DNA methylation (DNA methylome) resulting from treatment with at least one compound or a pharmaceutical composition described herein. In some embodiments, the treatment induced changes in the DNA methylome can be quantified by various “DNA methylation clocks” medical researchers have developed and published and which relate specific changes in the DNA methylome to a subject’s chronological age and / or susceptibility to various diseases or pathological conditions, including Alzheimer’sdisease, Parkinson’s disease, ALS, cardiovascular disease, rheumatoid arthritis, cancer, schizophrenia, type 2 diabetes, and life expectancy. Without wishing to be bound by theory, when a subject’s DNA methylome derived biological age is greater than the subject’s chronological age, the subject is experiencing age acceleration. Lower biological age than chronological age is age deceleration. Age acceleration is associated increased susceptibility to disease, poorer health, and shorter life expectancy. The reverse is true for age deceleration. In some embodiments, treatment of a subject, whose need for treatment in some instances may only be apparent from DNA methylome testing, while in other instances the need for treatment may be determined by known associations of the DNA methylome with a disease or condition, can be used to prevent, treat, reduce, or ameliorate a disease or condition associated with a biological clock in a subject in need thereof. The treatment effect on the DNA methylome may also be used to monitor the effectiveness of the treatment over time. In some embodiments, a method to prevent, treat, reduce, or ameliorate a disease or condition associated with a biological clock may include administering to a patient in need thereof an effective amount of a pharmaceutical composition. In some embodiments, the pharmaceutical composition includes a compound having the structure:wherein, one of R5and R6is — OH and the other R5and R6is — H, one of R12and R13is — OH and the other R12and R13is — H, R14and R15are — H, R16is — H, R17is — H or — OH, R18is — OH, R19is ethynyl, and R24and R25are — CH3.

[0233] In some embodiments, the pharmaceutical composition includes a compound having the structure:wherein, R1is — OH or an ester, R2is — OH or an ether, R3is — OH, =0, a halogen, an ester or =CH2, and R4is an optionally substituted amine, an amide, an N-linked amino acid, =N0H or — NHOH.|0234| In some embodiments, the pharmaceutical composition includes an effective amount of a compound having the structure:|0235| In some embodiments, the pharmaceutical composition includes an effective amount of a compound having the structure:|0236| In some embodiments, the pharmaceutical composition includes an effective amount of a compound having the structure:

[0237] In some embodiments, the pharmaceutical composition includes an effective amount of a compound having the structure:

[0238] In some embodiments, the pharmaceutical composition includes an effective amount of a compound having the structure:

[0239] In some embodiments, the pharmaceutical composition includes an effective amount of a compound having the structure:

[0240] In some embodiments, the pharmaceutical composition includes an effective amount of a compound having the structure:

[0241] In some embodiments, the pharmaceutical composition includes an effective amount of a compound having the structure:

[0242] In some embodiments, the pharmaceutical composition includes an effective amount of a compound having the structure:

[0243] In some embodiments, the pharmaceutical composition includes 17-ethynyl- 107?, 135-di methyl 2, 3, 4, 7, 87?, 95, 10, 11, 12, 13, 14S, 15, 16, 17-hexadecahydro-lH- cyclopenta[a]phenanthrene-3R, 77?, 175-triol, which is represented by Formula 1. The compound of Formula 1 may also be referred to as Compound 1 or 17a-ethynylandrost-5-ene-3p,7p,17P-triol and is represented by the structure below.

[0244] In some embodiments, the pharmaceutical composition includes(3S,5R,7S,8R,9S,10S,13S,14S,17R)-17-ethynyl-10,13-dimethylhexadecahydro-lH-cyclopenta[a]phenanthrene-3,7,17-triol, which is represented by Formula 2. The compound of Formula 2 may also be referred to as Compound 2, and is represented by the structure below.Formula 2

[0245] In some embodiments, a method is provided for to treat, reduce, prevent, or ameliorate a disease or condition in a subject. In some embodiments, the method to prevent a disease or condition in the subject may be by administration of a compound as described herein, or a pharmaceutical form thereof, and at least one pharmaceutically acceptable excipient. In some embodiments, the method to prevent a disease or condition in the subject may be by administration of Compound 1 or Compound 2, or a pharmaceutical form thereof, and at least one pharmaceutically acceptable excipient. In some embodiments, the method to prevent a disease or condition in the subject may be by administration of 17a-ethynylandrost-5-ene-30,70,170-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, the biological age of the subject is greater than the chronological age of the subject and reversing or decreasing the rate of increase in the biological age of the subject prevents or decreases a disease or condition in the subject related to the biological age of the subject being greater than the chronological age of the subject. In some embodiments, the disease or condition associated with a biological clock in a subject in need thereof is associated with DNA methylation levels in a subject. In some embodiments, the biological age may be assessed through epigenetic clocks, for example DNA methylation-based clocks (e.g., Horvath’s clock, Hannum’s clock, DNAmAge). In some embodiments, the disease or condition associated with the biological clock may include, but is not limited to, a neurodegenerative disease, a cardiovascular disease, a metabolic disorder, cancer,inflammatory and autoimmune diseases, chronic kidney disease, pulmonary diseases, infectious diseases, osteoporosis and musculoskeletal disorders, psychiatric and mood disorders. In some embodiments, the 17a-ethynylandrost-5-ene-3P,7P,17P-triol is administered at an amount sufficient to act as an epigenetic modulator. In some embodiments, the 17a-ethynylandrost-5-ene- 3p,7p,17p-triol is administered at an amount sufficient to slow or reverse the rate of biological aging as determined by DNA methylation-based epigenetic clocks. In some embodiments, the method further includes the use of combination therapies that target both the underlying disease and the biological aging process, thereby enhancing overall therapeutic efficacy. In some embodiments, the method further includes the use of epigenetic reprogramming agents that restore normal methylation levels in hypomethylated regions, thereby further improving age-related symptoms. In some embodiments, the method further comprises assessing additional biomarkers of biological aging, such as telomere length or senescence-associated markers, to customize the dosing regimen and optimize the reduction in DNA methylation. In some embodiments, the method further includes combining the composition with lifestyle interventions, such as dietary modifications and exercise, or co-administration of lipid-lowering agents to further enhance improvements in hyperlipidemia and associated obesity-related conditions. In some embodiments, the method further comprises assessing changes in metabolic biomarkers, for example, HOMA- IR, fasting insulin levels, and glucose tolerance, with epigenetic aging markers to optimize treatment efficacy. In some embodiments, the method further comprises monitoring epigenetic age markers, such as DNAmAge, to correlate reductions in neurodegenerative symptoms with a deceleration of biological aging. In some embodiments, the method further comprises assessing neuroendocrine markers and hippocampal volume, for example by neuroimaging, in conjunction with leptin signaling to provide a comprehensive evaluation of treatment effects on both cognitive function and biological aging. In some embodiments, the method further comprises tracking longitudinal changes in epigenetic age using DNA methylation clocks to evaluate the sustained impact of treatment on biological aging. In some embodiments, the subject is a mammal. In some embodiments, the subject is a livestock animal, e.g., sheep, cattle, pig, goat or poultry. In some embodiments, the subject is a warm-blooded animal, for example, cattle, sheep, pigs, cats, dogs, horses, llamas, deer, rabbits, skunks, raccoons, camels, etc., or birds. In some embodiments, the subject is a companion animal. In some embodiments, the companion animal is a cat, dog, or horse. In some embodiments, the subject is a human.

[0246] In some embodiments, the disease or condition associated with biological clocks in the subject in need thereof is based on modulation of DNA methylation of genes associated with biological clocks. In some embodiments, the disease or condition associated with a biological clock in a subject in need thereof is associated with genes or genomic regions hypermethylated with age. In some embodiments, the disease or condition associated with a biological clock in a subject in need thereof is associated with genes or genomic regions hypomethylated with age. In some embodiments, the disease or condition associated with a biological clock in a subject in need thereof is associated with Tau phosphorylation. In some embodiments, the disease or condition associated with a biological clock in a subject in need thereof is associated with hyperglycemia. In some embodiments, the disease or condition associated with a biological clock in a subject in need thereof is associated with insulin resistance. In some embodiments, the disease or condition may also be associated with chronic inflammation and oxidative stress. In some embodiments, the disease or condition may also be associated with mitochondrial dysfunction. In some embodiments, the disease or condition may also be associated with distinct DNA methylation signatures linked to advanced biological aging. In some embodiments, the disease or condition associated with biological clocks may be treated, reduced, ameliorated, or prevented by providing a therapeutically effective amount of at least one compound or composition as described herein to a subject in need thereof. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal.

[0247] In some embodiments, the present disclosure provides a method of reducing DNA methylation in a subject in need thereof. In some embodiments, the method includes administering to the subject an effective amount of a compound as 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.. In some embodiments, the compound is 17a-ethynylandrost-5-ene-3P,7P,17P- triol. The administering to the subject in need thereof has been shown to provide multiple beneficial responses to the subject. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. For example, in some embodiments, the administering 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 therein) as compared toa control measurement, e.g., as compared to DNA methylation in the subject prior to the administering (e.g., subject “baseline” DNA methylation). DNA methylation in the subject may be quantitatively and / or qualitatively evaluated by any standard technique in the art, e.g., as measured by a marker of relative global methylation as compared to a control, e.g., as measured by LINE-1 methylation as compared to a control. For example, in some embodiments, the administering 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) as compared to a control measurement, e.g., as compared to LINE-1 methylation in the subject prior to the administering (e.g., e.g., subject baseline LINE-1 methylation). For example, in some embodiments, the administering may reduce LINE-1 methylation in the subject by at least 5%, at least 8%, at least 10% or at least 15% or more. In some embodiments, the administering may reduce LINE-1 methylation in the subject by about 5% to about 20%, about 6% to about 15%, or by about 8% to about 10%. In some embodiments, the improvement in symptoms related to DNA methylation may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol. For example, subjects may experience an improvement in symptoms or conditions related DNA methylation from approximately 5% to objectively normal after administration of 17a- ethynylandrost-5-ene-3p,7p,17P-triol and at least one pharmaceutically acceptable excipient.

[0248] In some embodiments, the subject may experience an improvement in symptoms or conditions related to genes or genomic regions hypermethylated with age after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience an improvement in symptoms or conditions related to genes or genomic regions hypermethylated with age after administration of 17a-ethynylandrost- 5-ene-3P,7P,17P-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvement in symptoms related to genes or genomic regions hypermethylated with age may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of 17a-ethynylandrost-5-ene-3P,7P,17P- triol. For example, subjects may experience an improvement in symptoms or conditions related togenes or genomic regions hypermethylated with age ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0249] In some embodiments, the subject may experience an improvement in symptoms or conditions related to genes or genomic regions hypomethylated with age after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience an improvement in symptoms or conditions related to genes or genomic regions hypomethylated with age after administration of a composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvement in symptoms related to genes or genomic regions hypomethylated with age may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience an improvement in symptoms or conditions related to genes or genomic regions hypomethylated with age ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0250] In some embodiments, the subject may experience an improvement in symptoms or conditions related to hyperglycemia (which can lead to type I and type II diabetes) after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience an improvement in symptoms or conditions related to hyperglycemia (which can lead to type I and type II diabetes) after administration of 17a- ethynylandrost-5-ene-3P,7P,17P-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvement in symptoms related to hyperglycemia may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience an improvement in symptoms or conditions related to hyperglycemia31ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene- 30,70,170-triol and at least one pharmaceutically acceptable excipient.

[0251] In some embodiments, the subject may experience an improvement in symptoms or conditions related to hyperlipidemia (such as obesity-related conditions) after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience an improvement in symptoms or conditions related to hyperlipidemia (such as obesity-related conditions) after administration of 17a-ethynylandrost-5- ene-30,70,170-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvement in symptoms related to hyperlipidemia may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of 17a-ethynylandrost-5-ene-30,70,170-triol and at least one pharmaceutically acceptable excipient. For example, subjects may experience an improvement in symptoms or conditions related to hyperlipidemia ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-30,70,170-triol and at least one pharmaceutically acceptable excipient.

[0252] In some embodiments, the subject may experience an improvement in symptoms or conditions related to insulin resistance after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience an improvement in symptoms or conditions related to insulin resistance after administration of 17a- ethynylandrost-5-ene-30,70,170-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvement in symptoms related to insulin resistance may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience an improvement in symptoms or conditions related to insulin resistance ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene- 30,70,170-triol and at least one pharmaceutically acceptable excipient.

[0253] In some embodiments, the subject may experience a reduction or decrease in symptoms related to an Alzheimer’s Disease Composite Score after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a reduction or decrease in symptoms related to an Alzheimer’s Disease Composite Score after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to an Alzheimer’s Disease Composite Score may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience reduction in symptoms related to Alzheimer’s Disease Composite Score ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient. In some embodiments, reducing or decreasing symptoms related to Alzheimer’s Disease Composite Score may reduce other neuropsychological measures, including but not limited to, ADAS-Cog, MoCA, MMSE, CDR, QDRS, PDQ-9. In some embodiments, reducing or decreasing symptoms related to Alzheimer’s Disease Composite Score may improve neuroimaging, including but not limited to, ASL, BOLD, MRS, task-based or resting fMRI, vMRI and FDG-PET.

[0254] In some embodiments, the subject may experience a reduction or decrease in symptoms related to CSF phosphorylated tau (“pTau”) after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a reduction or decrease in symptoms related to pTau after administration of 17a-ethynylandrost- 5-ene-3P,7P,17P-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to pTau may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience reduction in symptoms related to pTau ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceuticallyacceptable excipient. In some embodiments, the method further includes evaluating changes in pTau in combination with improvements in epigenetic age indicators to tailor personalized treatment strategies.

[0255] In some embodiments, the subject may experience a reduction or decrease in symptoms related to leptin deficiency after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a reduction or decrease in symptoms related to leptin deficiency after administration of 17a-ethynylandrost-5- ene-3P,7P,17P-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to leptin deficiency may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience reduction in symptoms related to leptin deficiency ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient. In some embodiments, the method further comprises monitoring circulating leptin and adiponectin levels as biomarkers of metabolic and biological aging to optimize therapeutic outcomes.

[0256] In some embodiments, the subject may experience a reduction or decrease in symptoms related to leptin resistance after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a reduction or decrease in symptoms related to leptin resistance after administration of 17a-ethynylandrost-5- ene-3P,7P,17P-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to leptin resistance may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience reduction in symptoms related to leptin resistance ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient. In some embodiments, the method further includesevaluating changes in the leptin sensitivity index alongside epigenetic age assessments to personalize treatment for age-associated metabolic dysfunction.

[0257] In some embodiments, the subject may improve leptin / hypothalamic after administration of at least one compound or a composition as described herein. In some embodiments, the subject may improve leptin hippocampal after administration of a composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvement of leptin / hypothalamic and / or leptin hippocampal 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience an improvement related to leptin / hypothalamic and / or leptin hippocampal ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0258] In some embodiments, the subject may experience a reduction or decrease in symptoms related to DNA methylation after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a reduction or decrease in symptoms related to DNA methylation after administration of 17a-ethynylandrost-5- ene-3P,7P,17P-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to DNA methylation may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient. For example, subjects may experience reduction in symptoms related to DNA methylation ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient. In some embodiments, the method further includes evaluating both global and locus-specific methylation changes — such as alterations in LINE-1 methylation and DNAmAge — to monitor the therapeutic impact on biological aging.

[0259] In some embodiments, the subject may experience a reduction or decrease in symptoms related to cardiovascular risk after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a reduction or decrease in symptoms related to cardiovascular risk after administration of 17a-ethynylandrost-5- ene-3P,7P,17P-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to cardiovascular risk may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience reduction in symptoms related to cardiovascular risk ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient. In some embodiments, the method further comprises correlating improvements in cardiovascular parameters, for example, reduced arterial stiffness and improved endothelial function, with deceleration in epigenetic aging markers to guide treatment optimization.

[0260] In some embodiments, the subject may experience a reduction or decrease in symptoms related to a subject’s epigenetic age (“DNA methylation phenoage”) after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a reduction or decrease in symptoms related to DNA methylation phenoage after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to DNA methylation phenoage may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience reduction in symptoms related to DNA methylation phenoage ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0261] In some embodiments, the subject may experience a reduction or decrease in symptoms related to the subject’s epigenetic clock (“DNA methylation skin blood clock”) after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a reduction or decrease in symptoms related to the subject’s DNA methylation skin blood clock after administration of 17a-ethynylandrost-5-ene-30.70.170-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to DNA methylation skin blood clock may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience reduction in symptoms related to DNA methylation skin blood clock ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-30.70.170-triol and at least one pharmaceutically acceptable excipient.

[0262] In some embodiments, the subject may experience a reduction or decrease in symptoms related to the subject’s DunedinPACE clock after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a reduction or decrease in symptoms related to the subject’s DunedinPACE clock after administration of 17a-ethynylandrost-5-ene-30,70,170-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to DunedinPACE clock may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience reduction in symptoms related to DunedinPACE clock ranging from approximately 5% to 100% after administration of 17a- ethynylandrost-5-ene-30,70,170-triol and at least one pharmaceutically acceptable excipient. In some embodiments, the method further comprises correlating changes in DunedinPACE measurements with clinical outcomes to validate the treatment’s effect on decelerating biological aging.

[0263] In some embodiments, the subject may experience a prevention, reduction or decrease in conditions or symptoms related to a cancer or tumor after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a prevention, reduction or decrease in conditions or symptoms related to a cancer or tumor after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the prevention, reduction or decrease in conditions or symptoms related to a cancer or tumor may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience prevention, reduction or decrease in conditions or symptoms related to a cancer or tumor ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient. In some embodiments, the method further includes monitoring tumor-specific epigenetic markers alongside global DNA methylation profiles to assess the dual impact on cancer progression and biological aging.

[0264] In some embodiments, the subject may experience a reduction or decrease in symptoms related to atherosclerosis after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a reduction or decrease in symptoms related to atherosclerosis after administration of 17a-ethynylandrost-5-ene-3P,7P,17P- triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to atherosclerosis may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience reduction in symptoms related to atherosclerosis ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3p,7p,17P-triol and at least one pharmaceutically acceptable excipient. In some embodiments, the method further comprises assessing vascular function and arterial stiffness in conjunction with epigenetic aging markers to guide therapeutic interventions for atherosclerosis.

[0265] In some embodiments, the subject may experience a reduction or decrease in symptoms related to schizophrenia after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a reduction or decrease in symptoms related to schizophrenia after administration of 17a-ethynylandrost-5-ene-3P,7P,17P- triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to schizophrenia may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience reduction in symptoms related to schizophrenia ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3p,7p,17P-triol and at least one pharmaceutically acceptable excipient. In some embodiments, the method further includes evaluating neuroinflammatory markers and epigenetic age indicators to refine treatment strategies for schizophrenia associated with accelerated biological aging.

[0266] In some embodiments, the subject may experience a reduction or decrease in symptoms related to an autoimmune disease after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a reduction or decrease in symptoms related to an autoimmune disease after administration of 17a- ethynylandrost-5-ene-3P,7P,17P-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to an autoimmune disease may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience reduction in symptoms related to an autoimmune disease ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient. In some embodiments, the method further comprises monitoring immune cell profiles and epigenetic age signatures to adjust treatment regimens for autoimmune diseases in the context of overall biological aging.

[0267] In some embodiments, the subject may experience a reduction or decrease in symptoms related to rheumatoid arthritis after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a reduction or decrease in symptoms related to rheumatoid arthritis after administration of 17a-ethynylandrost- 5-ene-30,70,170-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to rheumatoid arthritis may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience reduction in symptoms related to rheumatoid arthritis ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-30,70,170-triol and at least one pharmaceutically acceptable excipient.

[0268] In some embodiments, the subject may experience a reduction or decrease in symptoms related to systemic lupus erythematosus after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a reduction or decrease in symptoms related to systemic lupus erythematosus after administration of 17a-ethynylandrost-5-ene-30,70,170-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to systemic lupus erythematosus may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience reduction in symptoms related to systemic lupus erythematosus ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene- 30,70,170-triol and at least one pharmaceutically acceptable excipient.

[0269] In some embodiments, the subject may experience a reduction or decrease in symptoms related to multiple sclerosis after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience a reduction or decrease in symptoms related to multiple sclerosis after administration of 17a-ethynylandrost-5- ene-30,70,170-triol. In some embodiments, the subject in need thereof is a human. In someembodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to multiple sclerosis may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience reduction in symptoms related to multiple sclerosis ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0270] In some embodiments, the subject may experience prevention or a reduction or decrease in the risk to ovum or sperm related to increased risks of offspring with autism spectrum disorder after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience prevention or a reduction or decrease in risk to ovum or sperm related to increased risks of offspring with symptoms related to autism spectrum disorder after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in ovum or sperm with increased risks of an offspring with symptoms related to autism spectrum disorder may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience prevention, reduction or decrease in ovum or sperm with increased risks of an offspring with symptoms related to autism spectrum disorder ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0271] In some embodiments, the subject may experience prevention or a reduction or decrease in DNA methylome changes in ovum or sperm associated with increased risks of an offspring with Down Syndrome after administration of at least one compound or a composition as described herein. In some embodiments, the subject may experience prevention or a reduction or decrease DNA methylome changes in ovum or sperm associated with increased risks of an offspring with Down Syndrome after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol. In some embodiments, the subject in need thereof is a human. In some embodiments, the subjectin need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to Down Syndrome may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of a composition as described herein. For example, subjects may experience prevention or reduction in ovum or sperm DNA methyome changes associated with increased risks of an offspring with Down Syndrome ranging from approximately 5% to 100% after administration of 17a-ethynylandrost-5-ene-3p,7p,17P-triol and at least one pharmaceutically acceptable excipient.

[0272] In some embodiments, the method to treat, reduce, improve, or ameliorate a disease or condition associated with biological clocks in a subject in need thereof further includes assessing the biological clock of a subject. In some embodiments, the method to treat, reduce, improve, or ameliorate a disease or condition associated with biological clocks in a subject in need thereof further includes measuring DNA methylation levels at a plurality of CpG sites using one or more techniques, for example, bisulfite sequencing, pyrosequencing, or methylation-specific PCR. In some embodiments, the measured DNA methylation levels are then compared to a reference database to determine the subject’s biological age relative to their chronological age. In some embodiments, the biological clock is assessed using one or more established epigenetic clocks, such as Horvath’s clock, Hannum’s clock, DNAmAge, or DunedinPACE. In some embodiments, the method further includes correlating the biological age with additional biomarkers, such as telomere length, oxidative stress markers, and senescence-associated secretory phenotype factors, to provide a comprehensive evaluation of the subject’s aging status. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal.

[0273] In some embodiments, the present disclosure provides a method for modulating the biological clock of a subject. The method comprises administering an effective amount of a compound or a combination of compounds of the disclosure that modulate DNA methylation, histone modifications, or non-coding RNA expression associated with aging. In some embodiments, the compounds or a combination of compounds are used in combination with at least one additional therapeutic agent, including, but are not limited to, sirtuin activators, mTOR inhibitors, DNMT inhibitors, or other epigenetic modulators, which are administered in a pharmaceutically acceptable formulation comprising at least one excipient. In some embodiments,the method further comprises administering adjunctive therapies, such as caloric restriction mimetics, antioxidants, or lifestyle interventions, to synergistically modulate the biological clock and slow the progression of biological aging. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal.

[0274] In some embodiments, the present disclosure provides a method for monitoring the efficacy of a treatment for biological aging. The method comprises obtaining a biological sample from the subject, for example, blood, saliva, or tissue, both before and after treatment. In some embodiments, the method further comprises measuring changes in DNA methylation at specific CpG sites and assessing global methylation patterns using markers such as LINE-1, thereby determining a change in the subject's biological age. In some embodiments, the measured change in biological age is compared to a predetermined threshold to evaluate treatment efficacy. In some embodiments, the method further includes utilizing machine learning algorithms to integrate multiple biomarkers. In some embodiments, the integrate multiple biomarkers may include, but are not limited to, epigenetic clock measurements, telomere length, and metabolic parameters — to predict long-term clinical outcomes and optimize treatment regimens. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal.

[0275] In some embodiments, the present disclosure provides a composition for use in modulating the biological clock in a subject. In some embodiments, the composition for use comprises an effective amount of one or more compounds of the disclosure capable of modulating epigenetic modifications associated with aging. In some embodiments, the one or more compounds of the disclosure consists of Compound 1, Compound 2, 17a-ethynylandrost-5-ene-3P,7P,17P- triol, or derivatives. In some embodiments, the composition further includes one or more pharmaceutically acceptable excipients to form a stable and effective formulation. In some embodiments, the composition further comprises a carrier or adjuvant selected to target the delivery of the compound to specific tissues, such as brain tissue, thereby modulating biological clocks in a tissue-specific manner. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal.

[0276] In several embodiments, the pharmaceutical composition includes a solid state form a compound as described herein. In several embodiments, the pharmaceutical compositions include a solid state form of 17oc-ethynylandrost-5-ene-3p,7p,17P-triol. In several embodiments,the solid state form is crystalline 17a-ethynylandrost-5-ene-3p,7p,17P-triol. In several embodiments, the solid state form is crystalline 17a-ethynylandrost-5-ene-3p,7p,17P-triol substantially free of 17a-ethynylandrost-5-ene-3p,7p,17P-triol in amorphous form.

[0277] In several embodiments, the solid state form is crystalline solvate 17a- ethynylandrost-5-ene-3p,7p,17P-triol. In several embodiments, the crystalline solvate is crystalline methanolate 17a-ethynylandrost-5-ene-3p,7p,17P-triol. In several embodiments, the crystalline solvate is crystalline ethanolate 17a-ethynylandrost-5-ene-3p,7p,17P-triol. In several embodiments, the crystalline solvate is crystalline hydrate 17a-ethynylandrost-5-ene-3p,7p,17P- triol.

[0278] In several embodiments, the crystalline solvate is Form III 17a-ethynylandrost- 5-ene-3p,7p,17P-triol. In several embodiments, the crystalline solvate is Form IV 17a- ethynylandrost-5-ene-3p,7p,17P-triol. In several embodiments, the crystalline solvate is Form V 17a-ethynylandrost-5-ene-3 P,7P, 17 P-triol.

[0279] In several embodiments, the solid-state form of 17a-ethynylandrost-5-ene- 3p,7p,17p-triol is amorphous 17a-ethynylandrost-5-ene-3p,7p,17P-triol. In several embodiments, the amorphous 17a-ethynylandrost-5-ene-3p,7p,17P-triol substantially free of 17a- ethynylandrost-5-ene-3p,7p,17P-triol in solid state form.

[0280] In some embodiments, the compound or pharmaceutical composition described herein is administered orally. In some embodiments, the compound or pharmaceutical composition described herein is administered intravenously. In some embodiments, the compound or pharmaceutical composition described herein is administered topically. In some embodiments, 17a-ethynylandrost-5-ene-3P,7P,17P-triol is administered orally. In other embodiments, 17a- ethynylandrost-5-ene-3P,7P,17P-triol is administered intravenously. In other embodiments, 17a- ethynylandrost-5-ene-3P,7P,17P-triol is administered topically.

[0281] In some embodiments, a compound as described herein is administered as a formulation or a composition with at least one pharmaceutically acceptable excipient. In some embodiments, a compound as described herein is administered as a formulation or a composition with at least one pharmaceutically acceptable excipient and at least one pharmaceutically acceptable carrier. In some embodiments, 17a-ethynylandrost-5-ene-3P,7P,17P-triol is administered as a formulation with at least one pharmaceutically acceptable excipient. In someembodiments, 17a-ethynylandrost-5-ene-3P,7P,17P-triol is administered as a formulation with at least one pharmaceutically acceptable excipient and at least one pharmaceutically acceptable carrier. In some embodiments, 17a-ethynylandrost-5-ene-3P,7P,17P-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 enhancing agents, acidifying agents, agents for modified release, alkalizing agents, antioxidants, buffering agents, chelating agents, coloring agents, complexing agents, emulsifying agents, flavoring agents, humectants, humidity-adjusting agents, pH-adjusting agents, preservatives, solubilizing agents, stabilizers, surface-active agents, suspending agents, sweetening agents, taste-masking agents, and wetting agents.

[0282] Formulations include compositions comprising 1, 2, 3, 4 or more pharmaceutically acceptable excipients or carriers. The compositions are used to prepare formulations suitable for human or animal use. Suitable administration routes 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). In general, aqueous and non-aqueous liquid or cream formulations are delivered by a parenteral, oral or topical route. In other embodiments, such as the disclosure intermittent dosing methods, 17a-ethynylandrost-5-ene-3P,7P,17P-triol may be present as an aqueous or a non-aqueous liquid formulation or a solid formulation suitable for administration by any of the routes disclosed herein, e.g., oral, topical, buccal, sublingual, parenteral, inhaled aerosol or a depot such as a subcutaneous depot or an intraperitoneal or intramuscular depot. It will be appreciated that the preferred route may vary with, for example, the subject’s pathological condition or weight or the subject’s response to therapy with 17a-ethynylandrost-5-ene-3P,7P,17P- triol or other therapy that is used or that is appropriate to the circumstances.

[0283] The formulations include those suitable for the foregoing administration routes. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods known in the art of pharmacy. Techniques, excipients and formulations generally are found in, e.g., Remington ’s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa. 2022, 23rdedition, 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, H. A. Lieberman, etal., editors, Pharmaceutical Dosage Forms, 19922ndrevised edition, volumes 1 and2, Marcel Dekker, ISBN 0824793870, J. T. Carstensen. Pharmaceutical Preformulation, 1998, pages 1-306, Technomic Publishing Co. ISBN 1566766907. Exemplary excipients for formulations include emulsifying wax, propyl gallate, citric acid, lactic acid, polysorbate 80, sodium chloride, isopropyl palmitate, glycerin, white petrolatum and other excipients disclosed herein.

[0284] Formulations, or compositions disclosed herein for use to make formulations suitable for administration by the routes disclosed herein optionally comprise an average particle size in the range of about 0.01 to about 500 microns, about 0.1 to about 100 microns or about 0.5 to about 75 microns. Average particle sizes include a range between 0.01 and 500 microns in 0.05 micron or in 0.1 micron or other increments, e.g., an 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, etc. microns). When 17a-ethynylandrost-5-ene-3P,7P,17P-triol or compositions that comprise 17a-ethynylandrost-5-ene-3P,7P,17P-triol are used as intermediates to make a formulation, they may comprise one, two, three or more of these average particle sizes, or size ranges. In preparing any of the compositions or formulations that are disclosed herein and that comprise 17a- ethynylandrost-5-ene-3P,7P,17P-triol (and optionally one or more excipients and / or one or more carriers), one may optionally mill, sieve or otherwise granulate the compound or composition to obtain a desired particle size.

[0285] Non-limiting examples of fillers suitable for use in the compositions include lactose, microcrystalline cellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose, methyl cellulose polymers hydroxyethylcellulose, sodium carboxymethylcellulose, carboxymethylene, carboxymethylhydroxyethylcellulose and other cellulose derivatives, sucrose, agarose, sorbitol, mannitol, dextrins, maltodextrins, starches or modified starches (including potato starch, maize starch and rice starch), calcium phosphate (e.g. basic calcium phosphate, calcium hydrogen phosphate, dicalcium phosphate hydrate), calcium sulfate, calcium carbonate, sodium alginate, and collagen.

[0286] Non-limiting examples of diluents suitable for use in the compositions include e.g. calcium carbonate, dibasic calcium phosphate, tribasic calcium phosphate, calcium sulfate, microcrystalline cellulose, powdered cellulose, dextrans, dextrin, dextrose, fructose, kaolin, lactose, mannitol, sorbitol, starch, pregelatinized starch, sucrose, and sugar.

[0287] 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, polacrillin potassium, sodium starch glycolate, starch, pregelatinized starch, and carboxymethyl starch.

[0288] 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 oxides, povidone, and pregelatinized starch.

[0289] 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 oils, corn starch, sodium stearyl fumarate, polyethylene glycols, alkyl sulfates, sodium benzoate, and sodium acetate.

[0290] 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 formaldehylde sulfoxylate, sodium metabisulfite, sodium thiosulfate, sulfur dioxide, tocopherol, tocopherol acetate, tocopherol hemisuccinate, and derivatives of tocopherol.

[0291] In several embodiments, the pharmaceutically acceptable excipient is selected from sodium dodecyl sulfate, microcrystalline cellulose, magnesium stearate, and any combination of the foregoing. In several embodiments, the pharmaceutically acceptable excipient is sodium dodecyl sulfate.

[0292] In several embodiments, the pharmaceutical compositions are formulated into oral dosage forms. In several embodiments, the dosage forms can include capsules and tablets. In some embodiments, the dosage forms can include one or more different types of delayed release layers selected from sealant and / or enteric layers. For example, delayed release layers having different release rate characteristics can provide the dosage form with different overall drug release characteristics. In some such embodiments, the pharmaceutically acceptable excipient is a surface active agent. In several embodiments, the surface active agent is present in an amount sufficient to provide 90% dissolution of the pharmaceutical composition in water at ambienttemperature after 30 min. In several embodiments, the surface active agent is sodium lauryl sulfate. In several embodiments, the pharmaceutical composition is a capsule or a tablet.

[0293] In several embodiments, the pharmaceutical compositions contain less than about 3% by weight of impurities.

[0294] In several embodiments, the pharmaceutical compositions contain less than about 5% by weight of 3P-hydroxy-androst-5-ene-7, 17-dione.

[0295] In several embodiments, the pharmaceutical compositions include a pharmaceutically acceptable formulation of 17a-ethynylandrost-5-ene-3p,7p,17P-triol.

[0296] In several embodiments, the use is concurrent with a use of at least one additional medicament. In several embodiments, the additional medicament is administered at a delay time after a first administration of the composition. In several embodiments, the first administration may occur using a dosage schedule that is daily, weekly, monthly, or any combination of the foregoing. In several embodiments, the dosage schedule of the first administration may include one, two, three or more daily dosages of the composition. In several embodiments, the dosage schedule of the first administration may include one, two, three or more weekly dosages of the composition. In several embodiments, the dosage schedule of the first administration may include one, two, three or more monthly dosages of the composition. In several embodiments, the delay time is equal to or greater than 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 years, or ranges including and / or spanning the aforementioned values. In several embodiments, the delay time is equal to or greater than 2 years. In some embodiments, the delay time is zero and the additional medicament is administered concurrently with the first administration of the composition. In several embodiments, the additional medicament is administered using a dosage schedule that is daily, weekly, monthly, or any combination of the foregoing. In several embodiments, the dosage schedule of the additional medicament may include one, two, three or more daily dosages of the composition. In several embodiments, the dosage schedule of the additional medicament may include one, two, three or more weekly dosages of the composition. In several embodiments, the dosage schedule of the additional medicament may include one, two, three or more monthly dosages of the composition.

[0297] Several embodiments of the present disclosure relate to the use of 17a- ethynylandrost-5-ene-3p,7p,17P-triol in the manufacture of a medicament for treating a neurodegenerative condition.

[0298] Aspects of the present disclosure relate to an in vitro screening method to identify a potential drug candidate. In some embodiments, the in vitro screening method may identify a potential drug candidate capable of treating, preventing, reducing, or ameliorating a disorder or disease. In some embodiments, the disorder or disease is a neurodegenerative disorder or disease. In some embodiments, the neurodegenerative disease or condition is dementia. As used herein, “drug candidate” refers to a specific molecule, compound, or therapeutic, whether of natural or synthetic origin, that has potential therapeutic effects and has been selected for further development and evaluation.

[0299] 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 tissue. In some embodiments, the sample is blood. In some embodiments, the sample includes monocytes. In some embodiments, the sample includes microglia. In some embodiments, the monocytes include, but are not limited to, CX3CR1!OW, CCR2pos, Ly6Ch!g!!, PD-Ll',sg, CD14++, CD16+, CD14dim, CD16+, CD16 CX3CRlhigh, CCR2”sg, Ly6Ciow, PD-Llpos. In some embodiments, the cells are T cells or granulocytes. In some embodiments, the cells are NK cells or granulocytes. In some embodiments, the cell for stimulation may be selected from the group consisting of, but not limited to, THP-1 human monocytes, RAW 264.7 macrophages, 3T3-L1 adipocytes, SH-S Y 5 Y neuroblastoma, PC- 12 pheochromocytoma and Lund human mesencephalic (LUHMES) cells.

[0300] In some embodiments, the in vitro screening method may include stimulating the cell with an agonist to induce a phenotype or a phenotypic change. In some embodiments, the phenotype may correspond to a phenotype of a cell or tissue 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), hippocampal sclerosis of aging (HS-Aging), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy, multiple system atrophy, corticobasal degeneration and vascular parkinsonism.

[0301] In some embodiments, the in vitro screening method may include contacting the one or more cells or tissue exhibiting the phenotype with the potential drug candidate. In some embodiments, the in vitro screening method may include contacting the one or more cells exhibiting the phenotype with the potential drug candidate in parallel in a high-throughputscreening method. In other embodiments, the in vitro screening method may include contacting the one or more cells exhibiting the phenotype with one or more potential drug candidates in parallel in a high throughput screening method. In still other embodiments, the in vitro screening method may include contacting the one or more cells exhibiting the phenotype with the potential drug candidate sequentially.

[0302] In some embodiments, the in vitro screening method may include determining a responsive change in the cell phenotype. In some embodiments, the responsive change may be a decrease, reduction, or loss in the cell phenotype. In some embodiments, the phenotype may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after contacting the cell exhibiting the phenotype with the potential drug candidate.

[0303] In some embodiments, the in vitro screening method further comprises exposing the sample to conditions that mimic accelerated aging, for example oxidative stress or inflammatory cytokines. In some embodiments, the in vitro screening method further comprises measuring changes in DNA methylation at age-associated CpG sites. In some embodiments, the in vitro screening method further comprises using reporter constructs linked to key regulators of the biological clock, for example, SIRT1, mTOR, or CLOCK genes, to provide a multi-parametric readout of a compound’s ability to modulate biological aging. In some embodiments, the in vitro screening method further comprises incorporating high-throughput imaging of senescence- associated markers (such as 0-galactosidase activity) to correlate epigenetic changes with cellular senescence reversal.

[0304] In some embodiments, the in vitro screening method is adapted for high- throughput screening to identify compounds that modulate the biological clock. In some embodiments, automated platforms are used to screen large libraries of small molecules, with readouts including changes in global and locus-specific DNA methylation, telomere length stabilization, and expression of age-regulated genes. In some embodiments, the hits from such screens may be further validated in cell-based assays that measure improvements in cellular senescence and metabolic function.

[0305] In some embodiments, the phenotype is associated with DNA methylation in the subject. In some embodiments, the DNA methylation is at a CpG site. In some embodiments,the CpG site is selected from, but not limited to, 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, DNA methylation may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after contacting the cell exhibiting the phenotype with the potential drug candidate. In some embodiments, the DNA methylation is decreased by more than 50%. In some embodiments, the DNA methylation is decreased by more than 55%. In some embodiments, the DNA methylation is decreased by more than 60%.

[0306] In some embodiments, the drug candidate, at least one compound or composition as described herein is provided to a subject to treat, prevent, reduce, or ameliorate a disease or condition associated with DNA methylation. In some embodiments, the disease or condition is associated with TNFa, GRC, CDR, MoCA, QDRS, GRC, ADCOMS, MoCA, QDRS- Cognition, ADAS-Cogl l, heart rate, frontal lobe, systolic blood pressure, grey matter, weight, MMSE, hippocampal volume, behavior, PDQ-9, CSF glucose, precuneus GLTH, CSF pTau / Ab. In some embodiments, treating, preventing, reducing or ameliorating a disease or condition associated with DNA methylation 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%, 100%, or ranges including and / or spanning the aforementioned values or establishing an objectively normal or disease- free condition after administration of the drug candidate or compound. For example, subjects may experience an improvement in symptoms or conditions related to TNFa, GRC, CDR, MoCA, QDRS, GRC, ADCOMS, MoCA, QDRS- Cognition, ADAS-Cogl l, heart rate, frontal lobe, systolic blood pressure, grey matter, weight, MMSE, hippocampal volume, behavior, PDQ-9, CSF glucose, precuneus GLTH, CSF pTau / Abregions hypermethylated with age ranging from approximately 5% to 100% or to an objectively normal condition after administration of the drug candidate, at least one compound or composition and at least one pharmaceutically acceptable excipient.

[0307] Aspects of the disclosure relate to a method of diagnosing a patient with a disease or a condition. In some embodiments, the method includes providing a patient with a potential drug candidate or compound as described herein capable of treating, preventing, reducing, or ameliorating a 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 with a disease or condition associated with a biomarker associated with DNA methylation. In some embodiments, the method of diagnosing a disease or condition further comprises determining the subject’s biological age by quantifying a panel of biomarkers, including DNA methylation at specific CpG sites, telomere length, and expression levels of age-related genes. In some embodiments, a composite biological clock score is generated by integrating these measurements using machine learning algorithms, which is then compared to the subject’s chronological age to predict susceptibility to age-associated disorders. In some embodiments, the drug candidate or compound is selected based on its capacity to modulate epigenetic clocks. The method comprises administering an effective amount of a compound that reverses age-associated DNA methylation changes and thereby reduces the subject’s biological age relative to their chronological age. In some embodiments, the compound is used in combination with lifestyle interventions, for example, caloric restriction mimetics, exercise regimens, or antioxidant therapies, to synergistically decelerate biological aging. Furthermore, the method may include longitudinal monitoring of the biological clock by repeated epigenetic assessments to adjust treatment regimens over time. In some embodiments, the method further includes formulating the drug candidate for targeted delivery to specific tissues known to exhibit accelerated biological aging, such as the brain, heart, or liver. In some embodiments, nanoparticle or liposomal delivery systems are employed to ensure the compound reaches the desired tissue, thereby providing localized modulation of DNA methylation patterns and improvement in tissue-specific function. In some embodiments, the present disclosure provides combinatorial therapeutic strategies that include a compound capable of modulating the biological clock in combination with standard-of- care treatments for neurodegenerative, metabolic, or cardiovascular diseases. In some embodiments, the combination therapy is optimized by correlating reductions in epigenetic age (as measured by DNA methylation clocks) with clinical improvements, thereby enabling personalized medicine approaches for patients with accelerated biological aging. In some embodiments, the in vitro screening method is adapted for high-throughput screening to identify compounds thatmodulate the biological clock. In some embodiments, automated platforms are used to screen large libraries of small molecules, with readouts including changes in global and locus-specific DNA methylation, telomere length stabilization, and expression of age-regulated genes. Hits from such screens may be further validated in cell-based assays that measure improvements in cellular senescence and metabolic function.

[0308] In some embodiments, identifying DNA methylation changes in the subject identifies the CpGs decreased by more than 50%. In some embodiments, identifying DNA methylation changes is correlated with one or more clinical changes. In some embodiments, the DNA methylation changes at a 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, 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.

[0309] In some embodiments, the disease or condition associated with a biomarker associated with DNA methylation is selected from the group consisting of TNFa, GRC, CDR, MoCA, QDRS, GRC, ADCOMS, MoCA, QDRS-Cogmtion, ADAS-Cogl l, heart rate, frontal lobe, systolic blood pressure, grey 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-Cogl 1, ADCOMS, CDR, CSF glucose, CSF pTau / Ab, frontal lobe volume, subcortical grey matter thickness, GRC, heart rate, MoCA, PDQ-9, precuneus glutathione, QDRS, QDRS-behavior, QDRS-cognition, Systolic BP, Tau, TNFa, and weight. In some embodiments, the biomarker associated with DNA methylation is decreased in TNF, CDR, QRDS-cognition, wherein the subject QDRS-cognition is improved. In some embodiments, the DNA methylation change is a decrease of > 50%. In some embodiments, the DNA methylation change is a decrease of > 55%. In some embodiments, the DNA methylation change is a decrease of > 60%.

[0310] In some embodiments, the subject may experience prevention or a reduction or decrease in obesity after administration of a drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience prevention or a reductionin obesity after administration of the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience an improvement to obesity - related metabolic inflammation after administration of a drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience an improvement to metabolic syndrome triglycerides after administration of a drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience an improvement in obesity and NfL after administration of a drug candidate, at least one compound or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the reduction or decrease in symptoms related to obesity may be reduced 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience prevention or reduction in obesity ranging from approximately 5% to 100% after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0311] In some embodiments, the subject may experience prevention or a reduction or decrease in ADCOMS after administration of a drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience improvements in ADCOMS after administration of the drug candidate, at least one compound, or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvements related to ADCOMS 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%, 100%, or ranges including and / or spanning the aforementioned values or objectively normal after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in ADCOMS ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition and at least one pharmaceutically acceptable excipient.

[0312] In some embodiments, the subject may experience prevention or a reduction or decrease in CDR after administration of a drug candidate, at least one compound or compositionas described herein. In some embodiments, the subject may experience improvements in CDR after administration of the drug candidate, at least one compound, or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvements related to CDR 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in CDR ranging from approximately 5% to 100% after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0313] In some embodiments, the subject may experience prevention or a reduction or decrease in CSF glucose after administration of a drug candidate, at least one compound, or composition as described herein. In some embodiments, the subject may experience improvements in CSF glucose after administration of the drug candidate, at least one compound, or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvements related to CSF glucose 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in CSF glucose ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0314] In some embodiments, the subject may experience prevention or a reduction or decrease in CSF pTau / AP after administration of a drug candidate at least one compound or composition as described herein. In some embodiments, the subject may experience improvements in CSF pTau / AP after administration of the drug candidate or compound. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvements related to CSF pTau / AP 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in CSF pTau / A0 ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0315] In some embodiments, the subject may experience prevention or a reduction or decrease in frontal lobe volume after administration of a drug candidate at least one compound or composition as described herein. In some embodiments, the subject may experience improvements in frontal lobe volume after administration of the drug candidate at least one compound or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvements 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in frontal lobe volume ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0316] In some embodiments, the subject may experience prevention or a reduction or decrease in subcortical grey matter thickness after administration of a drug candidate at least one compound or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience improvements in subcortical grey matter thickness after administration of the drug candidate, at least one compound or composition as described herein. In some embodiments, the improvements related to subcortical grey matter thickness 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in subcortical grey matter thickness ranging from approximately 5% to 100% or objectively normal after administration ofthe drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0317] In some embodiments, the subject may experience prevention or a reduction or decrease in GRC after administration of a drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience improvements in GRC after administration of the drug candidate or compound. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvements related to GRC 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in GRC ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0318] In some embodiments, the subject may experience prevention or a reduction or decrease in heart rate after administration of a drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience improvements in heart rate after administration of the drug candidate or compound. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvements related to heart rate 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in heart rate ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0319] In some embodiments, the subject may experience prevention or a reduction or decrease in MoCA after administration of a drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience improvements in MoCA after administration of the drug candidate or compound. In some embodiments, the subject in needthereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvements related to MoCA 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in MoCA ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0320] In some embodiments, the subject may experience prevention or a reduction or decrease in precuneus glutathione after administration of a drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience improvements in precuneus glutathione after administration of the drug candidate or compound. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvements related to precuneus glutathione 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in precuneus glutathione ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0321] In some embodiments, the subject may experience prevention or a reduction or decrease in QDRS after administration of a drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience improvements in QDRS after administration of the drug candidate or compound. In some embodiments, the improvements related to QDRS 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in QDRS ranging from approximately 5% to 100% or objectively normal afteradministration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0322] In some embodiments, the subject may experience prevention or a reduction or decrease in QDRS-behavior after administration of a drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience improvements in QDRS-behavior after administration of the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvements related to QDRS-behavior 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in QDRS-behavior ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0323] In some embodiments, the subject may experience prevention or a reduction or decrease in QDRS-behavior after administration of a drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience improvements in QDRS-behavior after administration of the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvements related to QDRS-behavior 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in QDRS-behavior ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0324] In some embodiments, the subject may experience prevention or a reduction or decrease in QDRS-cognition after administration of a drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience improvementsin QDRS-cognition after administration of the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvements related to QDRS-cognition 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in QDRS-cognition ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0325] In some embodiments, the subject may experience prevention or a reduction or decrease in systolic blood pressure after administration of a drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience improvements in systolic blood pressure after administration of the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvements related to systolic blood pressure 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in systolic blood pressure ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0326] In some embodiments, the subject may experience prevention or a reduction or decrease in pathological forms of Tau after administration of a drug candidate or compound as described herein. In some embodiments, the subject may experience improvements in Tau after administration of the drug candidate or compound. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvements related to pathological forms of Tau 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in pathological forms of Tau ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition and at least one pharmaceutically acceptable excipient.

[0327] In some embodiments, the subject may experience prevention or a reduction or decrease in TNFa after administration of a drug candidate or compound as described herein. In some embodiments, the subject may experience improvements in TNFa after administration of the drug candidate or compound. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the improvements related to TNFa 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in TNFa ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition and at least one pharmaceutically acceptable excipient.

[0328] In some embodiments, the subject may experience prevention or a reduction or decrease in weight after administration of a drug candidate or compound as described herein. In some embodiments, the subject may experience improvements in weight after administration of the drug candidate or compound. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience a reduction in adipose tissue accumulation, including a decrease in visceral fat. In some embodiments, the subject may experience stabilization or a reduction in body mass index (BMI) following administration of the drug candidate or compound. In some embodiments, the subject may experience a redistribution of body mass, characterized by a decrease in fat mass and an increase in lean muscle mass. In some embodiments, the subject may experience improved metabolic parameters, such as enhanced insulin sensitivity or increased energy expenditure, which correlates with improvements in weight. In some embodiments, the subject may experience a reduction in waist circumference or a decrease in the waist-to-hip ratio, further supporting overallimprovements in body composition. In some embodiments, the improvements related to weight may be improved by an amount equal to or greater than approximately 3%, 5%, 7.5%, 10%, 12.5%, 15%, 17.5%, 20%, 22.5%, 25%, 27.5%, 30%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience improvements in weight ranging from approximately 5% to 30% or objectively normal BMI after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0329] In some embodiments, the subject may experience a modification to their insulin levels after administration of the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience a modification that increases fasting insulin and H0MA2-%B in a subject. In some embodiments, the subject may experience an insulin modification that decreases HOMA2-%S without hypoglycemia. In some embodiments, the subject may experience an insulin modification that increases insulin and H0MA2-% B cell function and decreases H0MA2% insulin sensitivity. In some embodiments, the subject may experience improvements in insulin sensitivity. In some embodiments, the subject’s improvement in sensitivity is in an obese subject. In some embodiments, the obese subject has type-2 diabetes (“T2D”). In some embodiments, the subject improves clinical dementia rating (“CDR”) standard batter (“SB”) through Homeostatic Model Assessment 2 % Insulin Sensitivity (“H0MA2 %2”). In some embodiments, the subject’s CDR SB is decreased as insulin sensitivity is increased in the subject. In some embodiment, the subject’s insulin resistance is improved after administration of the drug candidate, at least one compound or composition as described herein. In some embodiments, improving insulin resistance increases central insulin to drive insulin receptor signaling. In some embodiments, the subject may decrease insulin levels after administration of the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may decrease insulin resistance after administration of the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject’s insulin dAge (age acceleration, biological age (Horvath Skin Blood Clock), and / or chronological age). In some embodiments, decreasing insulin resistance in the subject decreases Alzheimer’s Disease progression in the subject. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments,the modification to insulin may be in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience a modification in insulin levels ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0330] In some embodiments, a subject may experience one or more improvements to a neuropsychiatric condition after being administered to a compound as described herein. In some embodiments, the one or more neuropsychiatric conditions includes sleep and appetite. In some embodiments, an improvement in appetite is correlated with decreased Cog 12, CDR SB, and ADCOMS. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, an improvement in sleep is correlated with decreased CGIC and decreased TNFa. In some embodiments, the one or more improvements to a neuropsychiatric condition may be in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience one or more improvements to a neuropsychiatric condition ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0331] In some embodiments, a subject may experience an improvement in dAge. In some embodiments, an improvement in dAge includes, but is not limited to, improving biological age (as measured by the Hovarth Skin Blood Clock), and / or chronological age, after administration of the drug candidate or pharmaceutical composition as described herein. In some embodiments, a desired outcome may be either restoring dAge to match the subject's chronological age or reducing dAge below the chronological age. In some embodiments, the drug candidate or pharmaceutical composition as described herein may normalize or decrease disease parameters that are associated with an increased dAge. For example, the subject may experience a decrease or normalization in systolic blood pressure, cholesterol levels, or pTau levels followingadministration of the drug candidate or pharmaceutical composition as described herein. In some embodiments, a reduction in dAge is accompanied by a decrease in pTau217+ (pTau+) levels. In some embodiments, a reduction in dAge is accompanied by a decrease in pTau217+ (pTau+) levels demonstrates a correlation between improvements in dAge and improvements in at least one disease marker. In some embodiments, the drug candidate decelerates biological aging by reducing dAge as measured by the Hovarth Skin Blood Clock. In some embodiments, the drug candidate decelerates biological aging by reducing dAge as measured by the Hovarth Skin Blood Clock by at least 10% relative to baseline. In some embodiments, the deceleration correlates with an increase in life expectancy, as evidenced by normalization of age-associated biomarkers and improved survival outcomes compared to untreated subjects. In some embodiments, administration of the pharmaceutical composition results in a measurable decrease in the rate of biological aging as defined as a reduction of one biological year for each chronological year. In some embodiments, the reduction in dAge is associated with an extension of the subject’s projected lifespan. In some embodiments, an improvement in dAge is defined by a reduction in age acceleration markers (e.g., a decrease in pTau217+ levels) by a range of 5% to 100% relative to the subject’s initial dAge. In some embodiments, the biological age deceleration is directly linked to increased life expectancy, as subjects achieving such improvements may display a return to or improvement upon normal age-related biomarker levels, suggesting a slower progression of aging and enhanced longevity. In some embodiments, the administration of the drug candidate or the pharmaceutical composition may show beneficial effects on obesity-related parameters or chemokine profiles in response to a decrease in dAge. In some embodiments, the improvement in dAge may be one month, two months, three months, four months, five months, six months, seven months, eight months, nine months, ten months, eleven months, twelve months, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. In some embodiments, the improvement in dAge is dAge mediated. In some embodiments, the improvement in dAge may be 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, 12 years, 13 years, 14 years, 15 years, 16 years, 17 years, 18 years, 19 years, 20 years, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For instance, subjects may experience an improvement in dAge ranging from approximately 5% to 100% or achieve levels that are objectively normal or better (for example, lower) than the normal average for theirchronological age following administration with the drug candidate, at least one compound or composition as described herein, and at least one pharmaceutically acceptable excipient.

[0332] In some embodiments, a subject may experience an improvement in their cholesterol after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience an improvement to their CDR SB. In some embodiments, the subject may experience a decrease in their cholesterol after administration of the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience an improvement in ADCOMS after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience an improvement in obesity and cholesterol after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience an improvement to their diastolic blood pressure. In some embodiments, the subject may experience an improvement in their cholesterol in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience an improvement to their cholesterol ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0333] In some embodiments, a subject may experience an improvement in their MiniMental State Examination (“MMSE”) score after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject’s MMSE score decreases with age acceleration. In some embodiments, the subject’s waist-to-hip (“WHR”) ratio is high and the subject’s MMSE score is improved after being administered a compound as described herein. In some embodiments, the subject’s MMSE to Regulated on Activation, Normal T Cell Expressed and Secreted (RANTES) ratio is improved after being administered a compound as described herein. In some embodiments, the subject’s RANTES / neuroglia axis is realigned. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience an improvement intheir MMSE score in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience an improvement to their MMSE score ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0334] In some embodiments, a subject may experience an improvement in their metabolic regulation after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience an improvement in their metabolic regulation in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience an improvement to their metabolic regulation ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0335] In some embodiments, a subject may experience an improvement to CDR SB resulting from age deceleration mediated decrease of pathogenic pTau species after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience an improvement to CDR SB Glial Fibrillary Acid Protein (“GFAP”) ratio after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience a reduction in the pTau-associated age acceleration metric by at least approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 99%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience a decrease in the pTau-associated age acceleration metric from approximately 5%to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0336] In some embodiments, a subject may experience an improvement to Alzheimer’s Disease Composite Score (“ADCOMS”) to RANTES ratio after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience an improvement in the ADCOMS to RANTES ratio in the subject is a realignment of the RANTES mediated glial function after being administered a compound as described herein. In some embodiments, the subject may experience an improvement to their ADCOMS to GFAP ratio after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience an improvement in their ADCOMS to Neurofilament Light Chain (“NfL”) ratio after being administered a compound as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience an improvement in their ADCOMS to RANTES ratio in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience an improvement to their ADCOMS to RANTES ratio ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0337] In some embodiments, a method is provided to improve adiponectin in the subject by administering a therapeutically effective amount of the drug candidate, at least one compound or composition as described herein. In some embodiments, an improvement in adiponectin levels results in an actual increase in adiponectin. In some embodiments, the subject is obese. In some embodiments, the subject has impaired glucose intolerance. In some embodiments, a subject may experience an improvement in ratio of Cog 12 to RANTES. In some embodiments, a subject may experience an improvement in Cog 12 as a result of improved adiponectin in the subject. In some embodiments, the subject may experience an improvement in the Cog 12 to adiponectin ratio. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject13may experience an improvement in their adiponectin in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience an improvement to their adiponectin ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0338] In some embodiments, a subject may experience an improvement to triglycerides associated with increased systolic blood pressure after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, a subject may experience an increase in diastolic blood pressure mediated by triglycerides after being administered a compound as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience an improvement in their triglycerides associated with increased systolic blood pressure in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience an improvement to their triglycerides associated with increased systolic blood pressure ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0339] In some embodiments, a subject may experience an improvement in chemokine coregulation after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the chemokines MCP1 and RANTES are coregulated. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience an improvement cognition or neurodegenerative status as a result of improvements in their chemokine coregulation in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound asdescribed herein. For example, subjects may experience an improvement to their cognition or neurodeg enerative status ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0340] In some embodiments, a subject may experience an improvement in Cogl2 and an increase in M2 monocytes after being administered a compound as described herein. In some embodiments, improving M2 monocytes in the subject decreases (improves) Cog 12 in the subject. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience an M2 monocyte mediated improvement in their Cog 12 in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience an improvement to their Cogl2 ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0341] In some embodiments, a subject may improve CGIC and reduce WHR after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject has metabolic inflammation. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience an improvement in their CGIC resulting from decreased WHR in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience an improvement to their CGIC resulting from decreased WHR ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0342] In some embodiments, a subject may experience an improvement in CGIC after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience an improvement in CGIC after beingadministered the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience an improvement in their CGIC in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience an improvement to their CGIC ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0343] In some embodiments, a subject may experience a decrease in Monocyte Chemoattractant Protein 1 (“MCP1”) after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the MCP1 activates monocytes / microglia to Ml inflammatory Ml inflammatory cells. In some embodiments, administering to the subject the drug candidate, at least one compound or composition as described herein converts Ml inflammatory cells to M2 phagocytic cells. In some embodiments, MCP1 increases Clq. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience an improvement in their MCPlin 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience an improvement to their MCP1 ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0344] In some embodiments, a subject may experience an improvement in immune component coregulation after being administered a compound as described herein. In some embodiments, such improvements may be characterized as a partial or complete restoration of balanced immune regulation. In some embodiments, an improvement in immune component coregulation is a restoration of immune component coregulation. In embodiments wherequantitative measures are applicable, this improvement may be expressed as an increase from baseline of approximately 5% to 100%, or as restoration to values within the normal range.

[0345] In some embodiments, a subject may experience an increase to M2 monocytes / microglia after being administered the drug candidate, pharmaceutical composition, or at least one compound or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the treatment not only increases the number of M2 monocytes / microglia but also enhances their functional activity, as evidenced by increased expression of anti-inflammatory markers (e.g., IL- 10, CD206) and enhanced phagocytic capacity. In some embodiments, the drug candidate promotes a phenotypic shift from pro-inflammatory (Ml) to anti-inflammatory (M2) microglia, contributing to a more favorable neuroinflammatory environment and facilitating tissue repair. In some embodiments, the treatment leads to a sustained improvement in M2 monocyte / microglia levels over time, resulting in long-term modulation of the immune response in the subject. In some embodiments, the treatment may indirectly reduce levels of pro- inflammatory cytokines (e.g., TNF-a, IL-10) by promoting M2 polarization, further contributing to its therapeutic effects. In some embodiments, the subject may experience an improvement in their M2 monocytes / microglia to establish a functional presence that did not exist prior to treatment. This increase could be approximately a 2-fold, 5-fold, 10-fold, 15-fold, 20-fold, 25- fold, 50-fold, or 100-fold increase or more after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0346] In some embodiments, a subject may experience a decrease in TNF after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience an improvement in their TNF in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience an improvement to their TNF ranging from approximately 5% to 100% or objectively normal after administration of thedrug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0347] In some embodiments, the subject may experience an increase in HOMA2 IR after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject may experience a disassociation with H0MA2 IR from H0MA2 %S. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience an improvement in their H0MA2 IR in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience an improvement to their H0MA2 IR ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0348] In some embodiments, a method is provided herein to improve GFAP in a subject in need thereof after administering a drug candidate, pharmaceutical composition, or at least one compound as described herein. In some embodiments, the subject may experience a decrease in GFAP after being administered the drug candidate, at least one compound or composition as described herein. In some embodiments, the subject in need thereof is a human. In some embodiments, the subject in need thereof is an animal. In some embodiments, the subject may experience an improvement in their GFAP in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience an improvement to their GFAP ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient.

[0349] In some embodiments, a method is provided herein to improve NfL associated with GST in a subject in need thereof after administering a drug candidate, pharmaceutical composition, or at least one compound as described herein. In some embodiments, the subject may experience a decrease in NfL associated with improved GST after being administered the drugcandidate, at least one compound or composition as described herein. In some embodiments, the subject may experience an improvement in their NfL in 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%, 100%, or ranges including and / or spanning the aforementioned values after administration of the drug candidate or compound as described herein. For example, subjects may experience an improvement to their NfL associated with decreased GST ranging from approximately 5% to 100% or objectively normal after administration of the drug candidate, at least one compound or composition as described herein and at least one pharmaceutically acceptable excipient. Some embodiments relate to a method of decreasing dAge or biological clock age measured by DNA methylation in a subject. In some embodiments, the method is provided to the subject who exhibits an accelerated dAge relative to their chronological age. In some embodiments, the accelerated dAge relative to their chronological age is determined by DNA methylation profiling. In some embodiments, the method includes administering to the subject a therapeutically effective amount of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient. In some embodiments, the method includes administering to the subject a therapeutically effective amount of a pharmaceutical composition as described herein. In some embodiments, the administration of the pharmaceutical composition as describe herein may be provided in a range from several weeks to several months or longer, depending on the subject’s initial dAge and response to treatment.

[0350] Some embodiments of the disclosure relate to methods and uses of treating or preventing age-related conditions or diseases by administering a therapeutically effective amount of a compound of the disclosure or a composition as described herein. In some embodiments, the method includes measuring a subject’s intrinsic epigenetic age acceleration (IEAA) before treatment, administering a compound of the disclosure or a composition as described herein. In some embodiments, a composite marker, for example but not limited to, GrimAge clock, which integrates DNAm-based surrogate markers of plasma protein levels and smoking pack yerars, may be used to gauge the impact of a compound of the disclosure or a composition as described herein on predicted biological age and related health outcomes. In some embodiments, periodic dosing of a compound of the disclosure or a composition as described herein correspond to adjustments based on longitudinal measurements using stochastic clocks, retroelement-based clocks, or organ-specific clocks, for example but not limited to, systems age, heat, or Naive CD4 T cell clocks, to optimize the treatment regimen.

[0351] Some embodiments of the disclosure provide for a method or use to monitor or adjust a subject’s biological age. In some embodiments, the method includes administering to the subject a therapeutically effective amount of a pharmaceutical composition as described herein. In some embodiments, the administration of the pharmaceutical composition as describe herein may be provided in a range from several weeks to several months or longer, depending on the subject’s initial biological age and response to treatment. In some embodiments, the method includes utilizing at least one epigenetic biomarker to monitor, adjust, or validate the subject’s biological age.

[0352] Accordingly, some aspects described relate to the following numbered alternatives:

[0353] 1. A method to decrease biological age accelearation or biological age (“dAge”) as measured by a DNA methylation clock in a subject, the method comprising administering to the subject at least one compound or a pharmaceutical composition as described herein.

[0354] 2. A method to treat, reduce, or ameliorate a disease or condition associated with biological clocks in a subject in need thereof, the method comprising administering to the subject at least one compound or a pharmaceutical composition as described herein.

[0355] 3. The method of alternative 1 , wherein the DNA methylation clock is selected from the group consisting of Horvath Skin Blood Clock, universal pan mammalian clocks, Lu’s GrimAge, Youn and Wang Clock (MiAge), Levine’s DNAm PhenoAge, Bocklandt Clock, Hannum Clock, Zhang’s age predictor, Yang Clock [spiTOC]), or a combination thereof.

[0356] 4. The method of alternative 1 or 3, wherein decreasing dAge or biological age in the subject treat, reduces, improves, or ameliorates a disease or condition associated with biologiacal clocks.

[0357] 5. The method of any one of alternatives 1 to 4, wherein the at least one compound or pharmaceutical composition comprises 17a-ethynylandrost-5-ene-3P,7P,17P-triol.

[0358] 6. The method of any one of claims 1 to 5, wherein the improvement in the subject is decreased neuro inflammation

[0359] 7. The method of any one of claims 1 to 6, wherein the improvement in the subject is improved cognition.

[0360] 8. The method of any one of claims 1 to 7, wherein the improvement in the subject is decreased symptoms of a neurodegenerative disease.

[0361] 9. The method of any one of alternatives 1 to 5, wherein the disease or condition associated with biological clocks in the subject in need thereof is based on modulation of DNA methylation of genes associated with biological clocks.

[0362] 10. The method of alternative 1 to 6, wherein the disease or condition associated with a biological clock in the subject in need thereof is associated with genes or genomic regions hypermethylated with age.

[0363] 11. The method of any one of alternatives 1 to 10, wherein the disease or condition associated with a biological clock in the subject in need thereof is associated with genes or genomic regions hypomethylated with age.

[0364] 12. The method of any one of alternatives 1 to 11, wherein the disease or condition associated with a biological clock in a subject in need thereof is associated with Tau phosphorylation.

[0365] 13. The method of any one of alternatives 1 to 12, wherein the disease or condition associated with a biological clock in a subject in need thereof is associated with hyperglycemia.

[0366] 14. The method of any one of alternatives 1 to 13, wherein the disease or condition associated with a biological clock in a subject in need thereof is associated with insulin resistance.

[0367] 15. The method of any one of alternatives 1 to 14, wherein the disease or condition associate with biological clock is mild cognitive impairment or late onset Alzheimer’s disease.

[0368] 16. The method of any one of alternatives 5 to 15, wherein administering to the subject 17a-ethynylandrost-5-ene-3p,7p,17P-triol and at least one pharmaceutically acceptable excipient decreases a subject’s Alzheimer’s Disease Composite Score.

[0369] 17. The method of alternative 16, wherein the subject experiences a decrease between about a 5% to about a 100% reduction in symptoms connected to Alzheimer’s Disease Composite Score or objectively normal after administration of 17a-ethynylandrost-5-ene- 3p,7p,17p-triol and at least one pharmaceutically acceptable excipient.

[0370] 18. The method of any one of alternatives 5 to 17, wherein administering to the subject 17a-ethynylandrost-5-ene-3p,7p,17P-triol and at least one pharmaceutically acceptable excipient decreases pTau in the subject.

[0371] 19. The method of alternative 18, wherein the subject experiences a decrease between about a 5% to about a 100% reduction in conditions or symptoms connected to pTau or objectively normal after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0372] 20. The method of any one of alternatives 5 to 19, wherein administering to the subject 17a-ethynylandrost-5-ene-3p,7p,17P-triol and at least one pharmaceutically acceptable excipient decreases leptin in the subject.

[0373] 21. The method of alternative 20, wherein the subject experiences an increase between about a 5% to about a 100% in conditions or symptoms connected to leptin or objectively normal after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0374] 22. The method of any one of alternatives 1 to 21, wherein the subject experiences a decrease between about 5% to about 100% in conditions or symptoms connected to DNA methylation or objectively normal after administration of 17a-ethynylandrost-5-ene- 3p,7p,17p-triol and at least one pharmaceutically acceptable excipient.

[0375] 23. The method of any one of alternatives 5 to 22, wherein administering to the subject 17a-ethynylandrost-5-ene-3p,7p,17P-triol and at least one pharmaceutically acceptable excipient decreases cardiovascular risk in the subject.

[0376] 24. The method of alternative 23, wherein the subject experiences a decrease between about 5% to about 100%in conditions or symptoms connected to cardiovascular risk or objectively normal after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0377] 25. The method of any one of alternatives 5 to 24, wherein administering to the subject 17a-ethynylandrost-5-ene-3p,7p,17P-triol and at least one pharmaceutically acceptable excipient decreases DNA methylation age in the subject.

[0378] 26. The method of alternative 25, wherein the subject experiences a decrease between about 5% to about 100%in conditions or symptoms connected to DNA methylation orobjectively normal after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0379] 27. The method of any one of alternatives 5 to 26, wherein administering to the subject 17a-ethynylandrost-5-ene-3p,7p,17P-triol and at least one pharmaceutically acceptable excipient decreases DNA methylation phenoage in the subject.

[0380] 28. The method of alternative 27, wherein the subject experiences a decrease between about 5% to about 100%in conditions or symptoms connected to DNA methylation phenoage or objectively normal after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0381] 29. The method of any one of alternatives 5 to 28, wherein administering to the subject 17a-ethynylandrost-5-ene-3p,7p,17P-triol and at least one pharmaceutically acceptable excipient decreases DNA methylation skin blood clock in the subject.

[0382] 30. The method of alternative 29, wherein the subject experiences a decrease between about 5% to about 100%in conditions or symptoms connected to DNA methylation skin blood clock or objectively normal after administration of 17a-ethynylandrost-5-ene-3P,7P,17P- triol and at least one pharmaceutically acceptable excipient.

[0383] 31. The method of any one of alternatives 5 to 30, wherein administering to the subject 17a-ethynylandrost-5-ene-3p,7p,17P-triol and at least one pharmaceutically acceptable excipient decreases a subject’s CDR Score.

[0384] 32. The method of alternative 31, wherein the subject experiences a decrease between about 5% to about 100%in symptoms connected to CDR Score or objectively normal after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0385] 33. The method of any one of alternatives 5 to 32, wherein administering to the subject 17a-ethynylandrost-5-ene-3p,7p,17P-triol and at least one pharmaceutically acceptable excipient decreases a subject’s ADAS-Cog Score.

[0386] 34. The method of alternative 33, wherein the subject experiences a decrease between about 5% to about 100% in symptoms connected to ADAS-Cog Score or objectively normal after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0387] 35. The method of any one of alternatives 5 to 34, wherein administering to the subject 17a-ethynylandrost-5-ene-30,70,170-triol and at least one pharmaceutically acceptable excipient decreases a subject’s QDRS Score.

[0388] 36. The method of alternative 35, wherein the subject experiences between about a 5% to about a 100% reduction in symptoms connected to QDRS Score or objectively normal after administration of 17a-ethynylandrost-5-ene-30,70,170-triol and at least one pharmaceutically acceptable excipient.

[0389] 37. The method of any one of alternatives 5 to 36, wherein administering to the subject 17a-ethynylandrost-5-ene-30,70,170-triol and at least one pharmaceutically acceptable excipient increases a subject’s MMSE Score.

[0390] 38. The method of alternative 37, wherein the subject experiences about a 5% to about a 100% reduction in symptoms connected to MMSE Score or objectively normal after administration of 17a-ethynylandrost-5-ene-30,70,170-triol and at least one pharmaceutically acceptable excipient.

[0391] 39. The method of any one of alternatives 5 to 38, wherein administering to the subject 17a-ethynylandrost-5-ene-30,70,170-triol and at least one pharmaceutically acceptable excipient increases a subject’s MoCA Score.

[0392] 40. The method of alternative 39, wherein the subject experiences between about a 5% to about a 100% reduction in symptoms connected to MoCA Score or objectively normal after administration of 17a-ethynylandrost-5-ene-30,70,170-triol and at least one pharmaceutically acceptable excipient.

[0393] 41. The method of any one of alternatives 5 to 40, wherein administering to the subject 17a-ethynylandrost-5-ene-30,70,170-triol and at least one pharmaceutically acceptable excipient improves a subject’s GRC Score.

[0394] 42. The method of alternative 41, wherein the subject experiences between about a 5% to about a 100% reduction in symptoms connected to GRC Score or objectively normal after administration of 17a-ethynylandrost-5-ene-30,70,170-triol and at least one pharmaceutically acceptable excipient.

[0395] 43. The method of any one of alternatives 5 to 42, wherein administering to the subject 17a-ethynylandrost-5-ene-30,70,170-triol and at least one pharmaceutically acceptable excipient decreases a subject’s CGIC Score.

[0396] 44. The method of alternative 43, wherein the subject experiences between about a 5% to about a 100% reduction in symptoms connected to CGIC Score or objectively normal after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0397] 45. The method of any one of alternatives 5 to 44, wherein administering to the subject 17a-ethynylandrost-5-ene-3p,7p,17P-triol and at least one pharmaceutically acceptable excipient decreases a subject’s ADL Score.

[0398] 46. The method of alternative 45, wherein the subject experiences between about a 5% to about a 100% reduction in symptoms connected to ADL Score or objectively normal after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0399] 47. The method of any one of alternatives 5 to 46, wherein administering to the subject 17a-ethynylandrost-5-ene-3p,7p,17P-triol and at least one pharmaceutically acceptable excipient decreases a subject’s PDQ9 Score.

[0400] 48. The method of alternative 47, wherein the subject experiences between about a 5% to about a 100% reduction in symptoms connected to PDQ9 Score or objectively normal after administration of 17a-ethynylandrost-5-ene-3P,7P,17P-triol and at least one pharmaceutically acceptable excipient.

[0401] 49. The method of any one of alternatives 5 to 48, wherein the 17a- ethynylandrost-5-ene-3P,7P,17P-triol is administered orally.

[0402] 50. The method of any one of alternatives 5 to 48, wherein the 17a- ethynylandrost-5-ene-3P,7P,17P-triol is administered intravenously.

[0403] 51. The method of any one of alternatives 1 to 50, wherein the subject has a waist to hip ratio greater than or equal to approximately 0.90.

[0404] 52. The method of any one of alternatives 1 to 50, wherein the subject has a waist to hip ratio greater than or equal to approximately 0.95.

[0405] 53. The method of any one of Alternatives 5 to 52, wherein the 17a- ethynylandrost-5-ene-3b,7b,17b-triol is a solid state form of 17a-ethynylandrost-5-ene-3b,7b,17b- triol.

[0406] 54. The method of Alternative 53, wherein the solid state form of 17a- ethynylandrost-5-ene-3b,7b,17b-triol is crystalline solvate of 17a-ethynylandrost-5-ene- 3b,7b,17b-triol.

[0407] 55. The method of Alternative 54, wherein the crystalline solvate is crystalline methanolate 17a-ethynylandrost-5-ene-3b,7b, 17b-triol.

[0408] 56. The method of Alternative 55, wherein the crystalline solvate is crystalline ethanolate 17a-ethynylandrost-5-ene-3b,7b, 17b-triol.

[0409] 57. The method of Alternative 56, wherein the crystalline solvate is crystalline hydrate 17a-ethynylandrost-5-ene-3b,7b, 17b-triol.

[0410] 58. The method of Alternative 57, wherein the crystalline solvate is Form III17a-ethynylandrost-5-ene-3b,7b, 17b-triol.

[0411] 59. The method of Alternative 57, wherein the crystalline solvate is Form IV17a-ethynylandrost-5-ene-3b,7b, 17b-triol.

[0412] 60. The method of Alternative 57, wherein the crystalline solvate is Form V17a-ethynylandrost-5-ene-3b,7b, 17b-triol.

[0413] 61. The method of Alternative 55, wherein the solid state form of 17a- ethynylandrost-5-ene-3b,7b,17b-triol is amorphous 17a-ethynylandrost-5-ene-3b,7b,17b-triol.

[0414] 62. The method of any one of Alternatives 1 to 61, wherein the pharmaceutical composition contains less than about 3% by weight of impurities.

[0415] 63. An zn vitro screening method to identify a potential drug candidate or compound capable of treating, preventing, reducing, or ameliorating a disorder or disease, comprising: (i) providing a sample for stimulation selected from the group consisting of a cell, tissue, blood, monocytes, microglia, macrophages, adipocytes, neuroblastoma, pheochromocytoma, and Lund human mesencephalic (LUHMES) cells; (ii) stimulating the sample with an agonist to induce a phenotype or phenotypic reaction, wherein the phenotype or phenotypic reaction substantially corresponds to a disease or condition associated with at least one DNA methylation at a CpG site in a region of DNA; (iii) contacting the one or more cells exhibiting the phenotype or phenotypic reaction with one or more potential drug candidate or compounds; (iv) determining a responsive change in the phenotype of the sample; and (v) providing the drug candidate or compound to a subject in need thereof to treat, reduce, prevent, or ameliorate a disease or condition associated with the DNA methylation in the subject.

[0416] 64. The method of alternative 63, wherein 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, 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.

[0417] 65. The method of alternative 63 or 64, wherein the phenotype or phenotypic reaction is selected from the group consisting of increase or decrease of autocrine, paracrine, or endocrine signaling factors, NFkB p65 activation, iNOS activation, inflammasome activation, endoplasmic reticulum stress, unfolded protein response, cell death, mitochondrial stress, mitophagy, phospho proteome, lipidome, metabolome, DNA methylome, insulin signaling, adipokine increase or decrease, glial activation or change in Ml / Al to M2 / A2 character, RNA expression or stability.

[0418] 66. The method of any one of alternatives 63 to 64, wherein the DNA methylation change is a decrease of > 50%.

[0419] 67. The method of any one of alternatives 63 to 46, wherein the DNA methylation change is a decrease of > 55%.

[0420] 68. The method of any one of alternatives 63 to 46, wherein the DNA methylation change is a decrease of > 60%.

[0421] 69. The method of any one of alternatives 63 to 68, wherein the responsive change is a decrease or loss in the phenotype and the decrease or loss is indicative that the potential drug candidate or compound is capable of preventing, reducing, or ameliorating a neurodegenerative disorder or disease.

[0422] 70. The method of alternative 69, 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), hippocampal sclerosis of aging (HS-Aging), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy, multiple system atrophy, corticobasal degeneration and vascular parkinsonism.

[0423] 71. The method of alternative 69 or 70, wherein the neurodegenerative disorder or disease is Parkinson’s disease.

[0424] 72. The method of alternative 69 or 70, wherein the neurodegenerative disorder is Alzheimer’s disease.

[0425] 73. The method of any one of alternatives 63 to 72, wherein the CpGs sites are interconnected with genes associated with Alzheimer’s disease and related dementias.

[0426] 74. The method of alternative 73, wherein the disease or condition associated with inflammatory TNF signaling.

[0427] 75. The method of any one of alternatives 63 to 73, wherein the disease or condition associated with DNA methylation associated with Tau phosphorylation.

[0428] 76. The method of any one of alternatives 63 to 73, wherein the disease or condition associated with DNA methylation associated with hyperglycemia.

[0429] 77. The method of any one of alternatives 63 to 73, wherein the disease or condition associated with DNA methylation associated with insulin resistance.

[0430] 78. The method of any one of alternatives 63 to 73, wherein the disease or condition associated with DNA methylation associated with obesity.

[0431] 79. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s ADAS-Cogl 1.

[0432] 80. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s ADCOMS.

[0433] 81. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s CDR.

[0434] 82. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s CSF glucose.

[0435] 83. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s CSF pTau / A0.

[0436] 84. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s frontal lobe volume.

[0437] 85. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s subcortical grey matter thickness.

[0438] 86. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s GRC.

[0439] 87. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s heart rate.

[0440] 88. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s MoCA.

[0441] 89. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s PDQ-9.

[0442] 90. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s precuneus glutathione.

[0443] 91. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s QDRS.

[0444] 92. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s QDRS-behavior.

[0445] 93. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s QDRS-cognition.

[0446] 94. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s systolic blood pressure.

[0447] 95. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s function and homeostasis of the the protein tau and its phosphorylated and misfolded forms.

[0448] 96. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s TNFa.

[0449] 97. The method of any one of 63 to 73, wherein the disease or condition associated with DNA methylation is associated with a subject’s weight.

[0450] 98. The method of any one of alternatives 1 to 97, wherein the subject is a mammal.

[0451] 99. The method of any one of alternatives 1 to 98, wherein the subject is a human.

[0452] 100. The method of any one of alternatives 1 to 99, wherein the subject is an animal.

[0453] 101. The method of any one of alternatives 1 to 100, wherein the subject is a companion animal.EXAMPLES

[0454] The following examples are given for the purpose of illustrating various embodiments of the disclosure and are not meant to limit the present disclosure in any fashion. One skilled in the art will appreciate readily that the present disclosure is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those objects, ends and advantages inherent herein. Changes therein and other uses which are encompassed within the spirit of the disclosure as defined by the scope of the claims will occur to those skilled in the art.Example 1

[0455] An investigation study in early probable Alzheimer’s disease was performed.

[0456] In this study, 23 mild cognitive impaired (“MCI”) and Alzheimer’s disease (“AD”) subjects were enrolled based on Clinical Dementia Rating (“CDR”) from their Quick Dementia Rating System (“QDRS”) scale. Advanced MRI imaging (ASL, BOLD, MRS, taskbased fMRI) was also performed as well as cognition and memory tests were performed (Cog 12, MMSE, QDRS, MoCA). Biomarkers of the subjects were also taken (csf p-Tau, Abeta, and plasma TNF) and episome analyses were performed. Finally, a biological clock analysis was performed (DNA methylation profiling).

[0457] It was observed for in vitro samples receiving 17oc-ethynylandrost-5-ene- 3p,7p,170-triol lead to decreased activation of inflammatory p-IKKbeta, p-ERK, p-P38 and p-JNK (Tau phosphorylation) which resulted in decreased p-Tau.

[0458] Under normal circumstances, insulin binds to the insulin receptor, initiating tyrosine phosphorylation of IRS 1 / 2, leading to activation of Pi3K, Akt and inhibition of GSK3beta. Under inflammatory conditions, activation of the inflammatory kinases p-IKKbeta and p-JNK leads to serine phosphorylation of IRS- 1, inhibiting insulin signaling. Inflammation thus decreases insulin stimulated Akt inhibitory phosphorylation of GSK3beta, thus increasing p-Tau.

[0459] Without wishing to be bound by theory, it is believed that 17oc-ethynylandrost- 5-ene-3p,7p,17P-triol binds to ERK1 and ERK2, which decreases inflammatory activation of p- IKKbeta, p-JNK, p-ERK and p-P38, thereby decreasing serine IRS1 phosphorylation, restoringtyrosine IRS1 phosphorylation and insulin sensitivity. It was observed that this lead to increased Akt inhibition of GSK3beta and p-Tau.

[0460] Age related hyperglycemia results in insulin resistance, and hyperactivation of CDK5 / p25 kinase, leading not only to increased p-Tau, but also to inhibition of GSK3beta- induced degradation of beta catenin. This combination of effects leads to neuronal cell senescence and cell death (Chow, Herrup 2019).

[0461] The further results of this study are illustrated in FIGs. 1-11.Example 2

[0462] In this study, a follow up study from Example 1 was performed.

[0463] Illumina® 850K array DNA methylation changes following 14 weeks of NE3107 treatment were sorted and the top 400 decreased CpGs (decreases > 50%) were explored for correlations with changes in clinical results following treatment. Of these, 366 have CpGgene identifications. Changes for CpGs that showed Spearman correlations (p<0.05) with individual clinical changes (biomarker, cognition, function and imaging) were highly intercorrelated and were predominantly related to genes that are associated with Alzheimer's disease and related dementias.

[0464] Furthermore, many of these CpG decreases were correlated with more than one clinical change. As examples: KCNQ1DN, a potassium channel that is known to be decreased in AD brain showed a decrease in DNAm of 51% that was correlated with decreases (improvement) in TNF (inflammatory biomarker), CDR, QRDS-cognition, and an increase (improvement) in QDRS-cognition (clinical assessments). HLA-L, a protein that helps the immune system to clear amyloid plaques from the brain showed a 56% decrease in DNAm that was correlated with an improvement in subcortical grey matter thickness and frontal lobe volume (imaging). SLC26A1, decreases in this sulfate transporter is thought to contribute to cognitive decline m AD, showed a 55% decrease in DNAm That correlated with improvements in ADCOMS, QDRS (AD assessments) and CSF pTau / Ap ratio (biomarker).

[0465] Overall significant correlations between decreases in DNAm and individual clinical changes were observed in 157 of the 366 CpGs explored, and 45 of these correlated individually with multiple clinical changes. These 157 CpGs showed individual correlations with improvements in ADAS-Cogl I, ADCOMS, Ab42, CDR, CSF glucose, CSF pTau / Ab, frontal lobevolume, subcortical grey matter thickness, GRC, Heart rate, MoCA, PDQ-9, precuneus glutathione, QDRS, QDRS-behavior, QDRS-cognition, Systolic BP, I'au, TNFa, and weight. The results are of this study are illustrated in FIG. 8 and Table 1.Table 1Example 2

[0466] In this example, the clinical outcomes from a Phase 3, randomized, placebo- controlled trial of NE3107 (17a-ethynylandrost-5-ene-3p,7p,17P-triol) in subjects with mild to moderate probable Alzheimer’s disease.

[0467] Background

[0468] Recently, the roles of inflammation and insulin resistance in neurodegeneration have become better appreciated. NE3107, an oral small molecule, blood-brain permeable antiinflammatory insulin sensitizer that binds extracellular signal-regulated kinase, has been shown to selectively inhibit inflammation-driven ERK- and NF-KB-stimulated inflammatory mediators, including TNF-a, without inhibiting their homeostatic functions. We describe the rationale and design of NM101, the first randomized, multicenter Phase III clinical study to examine the safety and efficacy of 30-week treatment with NE3107 versus placebo in elderly adults with mild-to- moderate Alzheimer's disease. Patients (316) will be randomized in a 1 :1 ratio. The co-primary end points measure cognitive function (ADAS Cog 12), and functional and behavioral characteristics (ADCS CGIC). Trial registration number: NCT04669028 (Clinicaltrials.gov).

[0469] Trial Design

[0470] The trial design is illustrated in FIG. 12.

[0471] During the Phase 3 trials, the median improvement in various blinded assessments were measured. The results are described in Table 2 and FIGs. 13A-13G.Table 2

[0472] Similar distributions were observed for APOE4 + / -, mild / moderate Alzheimer’s disease, male / female, and older / younger participants.

[0473] Subjects metabolic and correlation changes were analyzed. Many of the genes associated with LOAD (late onset Alzheimer’s disease) are related to cholesterol metabolism, which is decreased in AD neuron membranes. Insulin and glycemic controls are known to be involved in neurodegeneration. Increased insulin and H0MA2-% 0 cell function, and decreased H0MA2% insulin sensitivity resulted in no cases of hypoglycemia. Increased mean amplitude of glycemic excursion (MAGE from CGM) increased risk of Alzheimer's disease progression. Leptin is an anti-inflammatory, neuroprotective and is decreased in AD. Leptin DNAm was observed to increase. The results are described in Tables 4 and 5.Table 4* P <0.0001Table 5* p <0.05, ** p<0.005, ***p<0.0001, #excludes obese subjects

[0474] Subjects were analyzed using imaging sub-studies vMRI on 23 subjects. It was observed that the volume was increased in the hippocampus and amygdala. In addition, it wasobserved a decreased CGIC was correlated with decreased hippocampi volume and increased MCP-1 was correlated with increased whole cortex. The results are described in FIGs. 14A-14B and Table 6.Table 6

[0475] Subjects were analyzed using a fluorodeoxyglucose (FDG)-positron emission tomography (PET) imaging scans. FDG-PET standardized uptake value ratios (“suvr”) were increased in 14 / 24 subjects with a baseline whole cortex suvr <1.29. This data correlates with Cog 12 and ADL with suvr improvements. Cholesterol increases trended with increased Cingulate suvr. The results are described in Table 7 and FIG. 15.Table 7* p<0.1, ** p<0.05, *** p<0.01

[0476] Subjects were analyzed for neuropsychiatric inventory. It was observed that overall improvement was seen in sleep (-1.0, p<0.0001) and appetite (-1.0, p<0.023). An improvement in appetite was also correlated with decreased Cog 12, CDR SB and ADCOMS, with increased Alzheimer’s disease. A decrease in anxiety was correlated with an increase ADL anddecreased CGIC. Sleep improvement was correlated with a decrease in CGIC and decreased in TNFa. The results are described in Table 8 and FIGs. 16A-16B.Table 8* p<0.10, ** p<0.05, *** p<0.01, ****p<0.00

[0477] The study overall had a very low rate of adverse effects (AEs) reported and only 10 subjects discontinued due to a reported AE (2.3%). Of the 439 subjects enrolled in the study, 156 experienced 1 or more AEs (35.5%). There were only 43 (9.8%) related AEs with the majority 42 ((.6%) categorized as non-serious AEs by the principal investigator. The study also had very low rate of serious AEs reported. There were 12 (2.7%) reported for the duration of the study. Only 1 of the serious AEs reported resulted in death. As this is a blinded analysis it is not known if the subjects was on IP or placebo.

[0478] Additional metrics were measured during the Phase 3 studies as represented in FIGs. 16A-33.

[0479] FIGs. 17A-17C illustrates graphs representing increased fasting insulin and H0MA2-%B with decreased HOMA2-%S w / o hypoglycemia. Table 9 provides additional information regarding the fasting insulin and H0MA2-%B with decreased HOMA2-%S w / o hypoglycemia.Table 9

[0480] FIGs. 18A-18G illustrate graphs representing placebo effects in various assessments. Medians showing improvement indicate that at least some placebo subjects showed improvement in neurocognitive and functional assessments. Similar findings in vMRI, NPI, HOMA, and APS suggest this is not just related to assessment inflation at baseline. FIGs. 19-20 illustrate graphs representing ADAS-Cogl2 spearman correlations. FIGs. 21-22 illustrate graphsrepresenting clinician rating of global change spearman correlations. FIGs. 23-24 illustrate graphs representing mini-mental state exam spearman correlations. FIG. 25 illustrate graphs representing ADCOMS spearman correlations. FIG. 26 illustrate graphs representing CDR sum of boxes spearman correlations. FIG. 27 illustrate graphs representing activities of daily living spearman correlations. A subset of subjects participated in FDG-PET analysis, and an increase in glucose uptake was observed in about half of the FDG-PET subjects. FIG. 28 illustrate graphs representing improvement in ADAS-Cogl2 correlated with increased FDG-PET SUVR. FIG. 29 illustrate a graph representing an improvement in MMSE correlated with increased FDG-PET SUVR. FIG. 30 illustrate graphs representing an improvement in ADL correlated with increased FDG-PET SUVR. FIG. 31 illustrate graphs representing an improvement in H0MA2 insulin sensitivity correlated with increased FDG-PET SUVR. FIG. 32 illustrate graphs representing an improvement in cholesterol correlated with increased FDG-PET SUVR.

[0481] Improvements in the blinded data for cognitive and accepted Alzheimer’s disease biomarkers (A0 and FDG-PET) suggests NE3107 is active in subjects with mild / moderate AD. The blinded analysis were also consistent with the hypothesis on NE3107’ s anti-inflammatory and insulin sensitizing activity in Alzheimer’s disease.

[0482] FIGs. 33-200B illustrate graphs representing NE3107 versus placebo effects in various assessments. FIG. 92 are graphs representing NE3107 may alter weight H0MA2 %S axis. One-tailed p; hypothesis is weight gain is known to be associated with decreased insulin sensitivity (metabolic information). FIG. 96 are graphs representing directional improvement in insulin sensitivity. One-tailed p; hypothesis is NE3107 should improve H0MA2 %S (H0MA2% = Homeostatic Model Assessment 2 % insulin sensitivity). NE3107 was seen to improve insulin sensitivity in obese and diabetic animals and in obese IGT and T2D human subjects. FIG. 97 is a graph representing NE3107 decreased dAge (dAge = Age acceleration = Biological age (Horvath Skin Blood Clock) - Chronological Age). One-tailed p; hypothesis is NE3107 should decrease dAge. Thus, it was believed and appears to be confirmed that NE3107 decreased biological age. FIG. 98 are graphs showing NE3107 may alter MMSE dAge axis. One-tailed p; hypothesis is MMSE decreases with Age acceleration. FIG. 100 are graphs representing that NE3107 may alter MMSE waist / hip ratio (WHR) axis. One-tailed p; hypothesis is higher WHR is associated with lower MMSE cfb (metabolic inflammation). FIG. 101 are graphs representing NE3107 may alter MMSE RANIES axis. Two-tailed p; hypothesis is positive correlation may suggest realignmentof RANTES / neuroglia axis. FIG. 103 are graphs representing NE107 may alter CDR SB via H0MA2 %S. One-tailed p; hypothesis: is CDR SB decrease is associated with insulin sensitivity increase. FIG. 105 are graphs representing NE3107 may alter CDR SB cholesterol axis. One-tailed p; hypothesis is CDR SB increase is associated with cholesterol increase. FIG. 106 illustrate a graph representing NE3107 significantly decreasing cholesterol. One-tailed p; hypothesis is NE3107 decreases cholesterol. FIG. 107 are graphs representing NE3107 may alter CDR SB dAge in pTau+ subjects. dAge (pTau+) = Age acceleration = Biological Agee (Horvath Skin Blood Clock) - Chronological Age in pTau217+ subjects. One-tailed p; hypothesis is increased CDR SB is associated with Age acceleration in pTau217+ NE3107. FIG. 108 is a graph representing NE3107 significantly decreased dAge (pTau+). One-tailed p; hypothesis is NE3107 should decrease dAge (pTau+). FIG. 109 are graphs representing NE3107 may alter CDR SB GFAP axis. One-tailed hypothesis is increased CDR SB is associated with increased GFAP (GFAP = Glial Fibrillary Acid Protein). FIG. I l l are graphs representing NE3107 may alter ADCOMS RANTES axis. Two-tailed p; hypothesis is negative correlation may suggest realignment of RANTES / neuroglia axis. FIG. 113 are graphs representing NE3107 may alter cholesterol ADCOMS axis. One-tailed p; hypothesis is increased cholesterol is associated with increased ADCOMS. FIG. 114 are graphs representing NE3107 may alter ADCOMS GFAP axis. One-tailed p; hypothesis is increased ADCOMS is associated with increased GFAP. FIG. 115 are graphs representing NE3107 may alter ADCOMS NfL axis. One-tailed p; hypothesis is increased ADCOMS is associated with increased NfL (neurofilament light chain). FIG. 117 are graphs representing NE3107 may alter Cogl2 RANTES axis. Two-tailed p; hypothesis is negative correlation may suggest realignment of RANTES / neuroglia axis. FIG. 119 are graphs representing NE3107 may alter Cogl2 adiponectin axis. One-tailed p; hypothesis is decreased Cogl2 is associated with decreased adiponectin. It was believed NE3107 increased adiponectin in obese IGT human subjects. FIG. 120 is a graph representing NE3107 significantly increased adiponectin. One-tailed p; hypothesis is NE3107 increases adiponectin. It was believed that NE3107 increased adiponectin in obese, IGT human subjects. FIG. 121 are graphs representing NE3107 may alter Cogl2 monocytes axis. One-tailed p; hypothesis is decreased Cogl2 is associated with increased M2 monocyte / microglia. FIG. 123 are graphs representing NE3107 may alter CGIC WHR axis. One-tailed p; hypothesis is increased CGIC is associated with higher WHR (metabolic inflammation). FIG. 125 are graphs representing NE3107 may alter CGIC dAge axis. One-tailedp; hypothesis is increased CGIC is associated with increased dAge. FIG. 126 are graphs representing NE3107 may alter CGIC NPI (neuropsychiatric index) axis. One-tailed p; hypothesis is increased CGIC is associated with Age acceleration. FIG. 128 are graphs representing placebo ADL cfb is proportional to H0MA2 IR cfb. One-tailed p; hypothesis is insulin resistance increases central insulin to drive insulin receptor signaling NE3107 outliers ROUT (Q=l%). FIG. 129 are graphs representing directional change in H0MA2 IR (Homeostatic Model Assessment 2 Insulin Resistance). One-tailed p; hypothesis is NE3107 decreases insulin resistance. NE3107 decreased insulin resistance in obese and diabetic animals, and in IGT and T2D human subjects. FIG. 130 are graphs representing NE3107 may alter ADL CRP axis. One-tailed p; hypothesis is inflammation is associated with decreased ADL. FIG. 133 are graphs representing NE3107 may alter ADL / GFAP axis. One-tailed p; hypothesis is increased GFAP is associated with decreased ADL. FIG. 134 are graphs representing NE3107 may alter obesity / I eptin axis. Two-tailed p; hypothesis NE3107 may alter leptin / hypothalamic and leptin hippocampal axes. FIG. 136 are graphs representing NE3107 may alter obesity / triglyceride axis. Two-tailed p; hypothesis is NE3107 may decrease obesity-related metabolic inflammation and metabolic syndrome triglycerides. FIG. 138 are graphs representing NE3107 may alter obesity effects on dAge. Two- tailed p; hypothesis is NE3107 may decrease obesity-related metabolic inflammaging. FIG. 139 are graphs representing NE3107 may alter obesity / NfL axis. One-tailed p; hypothesis is NfL increase associates with obesity. FIG. 140 are graphs representing NE3107 may alter obesity / cholesterol axis. One-tailed p; hypothesis is cholesterol increase associates with obesity. FIG. 141 are graphs representing NE3107 may restore chemokine coregulation. One-tailed hypothesis is chemokines MCP1 and RANTES are coregulated. FIG. 142 are graphs representing NE3107 may decrease MCP1 (Monocyte Chemoattractant Protein 1). One-sided p; hypothesis is MCP1 activates monocytes / microglia to Ml inflammatory cells, NE3107 converts them to M2 phagocytic cells. FIG. 143 are graphs representing NE3107 may restore immune component coregulation. One-tailed p; hyptoehsis is MCP1 increases Clq. FIG. 145 are graphs representing NE3107 may alter chemokine dAge axis. One-tailed p; hypothesis is MCP1 recruits M2 monocytes / microglia and decreases dAge. FIG. 146 are graphs representing N3107 may alter chemokine dAge axis. One-tailed p; hypothesis is MCP1 recruits M2 monocytes / microglia, decreases dAge. FIG. 147 are graphs representing NE3107 may alter the triglyceride / systolic BP axis. One-tailed p; hypothesis is increased triglycerides are associated with increased systolic BP.These results confirmed previous data demonstrating NE3107 decreases TNF in vitro and in animal models. FIG. 149 are graphs representing NE3107 may alter cholesterol / systolic BP axis. One-tailed hypothesis is increased cholesterol is associated with increased systolic BP. FIG. 150 are graphs representing NE3107 may alter systolic BP dAge axis. One-tailed p; hypothesis is systolic BP increases with dAge. FIG. 151 are graphs representing NE3107 may alter diastolic BP triglyceride axis. One-tailed p; hypothesis is systolic BP increase is associated with triglyceride increase (Metabolic Syndrome). FIG. 153 are graphs representing NE3107 may alter diastolic BP cholesterol axis. One-tailed p; hypothesis is systolic BP increase is associated with cholesterol increase (Metabolic Syndrome). FIG. 154 are graphs representing NE3107 may alter diastolic BP / dAge axis. FIG. 155 are graphs representing NE3107 may improve metabolic regulation. One- tailed p-hypothesis is triglyceride levels are known to be linked to insulin levels. FIG. 156 is a graph representing NE3107 decreased insulin levels. One-tailed p; hypothesis is NE3107 decreases insulin resistance. Insulin resistance is believed to be associated with Alzheimer’s Disease. FIG. 157 are graphs representing NE3107 may alter insulin dAge axis. A0+ = C2N APS (Amyloid Probability Score). One-tailed p; hypothesis insulin resistance increases Alzheimer's disease progression. FIG. 159 are graphs representing NE3107 may alter metabolic regulation. One-tailed p; hypothesis is triglyceride levels are known to be associated with insulin resistance. FIG. 160 are graphs representing NE3107 may alter H0MA2 IR Clq axis. One-tailed p; hypothesis is Clq levels are known to be associated with insulin resistance. FIG. 161 are graphs representing NE3107 may dissociate H0MA2 IR from H0MA2 %S. One-tailed p is hypothesis is H0MA2 IR and H0MA2 %S are associated in the model. FIG. 162 are graphs representing NE3107 may alter the cholesterol dAge axis. One-tailed p; hypothesis is increased cholesterol is associated with Age acceleration. FIG. 163 are graphs representing NE3107 may alter dAge pTau axis. One-tailed p; hypothesis is increased pTau is associated with Age acceleration. FIG. 165 are graphs representing NE3107 may alter FGAP GST (GST = Global Statistical test = CDR SB & ADAS-Cogl2) axis. One-tailed p; hypothesis is increased GFAP is associated with increased GST. FIG. 167 are graphs representing NE3107 may alter NfL GST axis. One-tailed p; hypothesis is increased NfL is associated with increased GST. The correlations of the SBC results and clinical measures were further examined using principal component analysis to reduce the data dimensionality (FIG. 199A and 199B). This analysis identified two principal components for correlations in placebo participants. The eigen vector clusters for metabolic, inflammatory, and dementia markers weredistinct from the clusters for neurological assessments, and all eigen vectors except for fructosamine shared PCI and PC2 contributions. The placebo dAge vector contributed least to the correlations. In distinction, there was only a single principal component identified for NE3107 correlations, combining major contributions from neurologic, metabolic, monocyte, regulated on activation, normal T-cell expressed and secreted (RANTES), and dAge vectors with minor contributions from inflammatory and dementia biomarkers, suggesting a potential homeostatic effect.

[0483] Examination of the NE3107 PCI loadings >|0.5| indicated that increases in fasting glucose, pancreatic beta cell function, and the chemokine RANTES were associated with increases (improvements) in MMSE and ADL; and that decreases in SBC dAge, diastolic BP, cholesterol, and insulin sensitivity were associated with improvements (decreases) in CGIC, ADAS-Cogl2, CDR-SB, ADCOMS, and GST. The placebo PCI loadings >|0.5| indicated that increased waist-to-hip ratio (WHR), insulin resistance, diastolic BP, TNF, C-reactive protein (CRP), triglycerides, and insulin were correlated; and that increased MCP1, GFAP, fructosamine, insulin sensitivity, and pTau217 were associated with increase (decline) in CGIC. Placebo PC2 loadings >|0.5| indicated that increased CRP was associated with increases (decline) in CDR-SB, GST, ADCOMS, and Cog 12; and that decreases (declines) in MMSE and ADL were correlated.

[0484] Tables 10 and 11 describe dAge correlations as determined from this example and FIGs.Table 10Table 11

[0485] FIGs. 201-211 illustrate differences in epigenetic age acceleration between NE3107 and placebo, with the clocks data adjusted for gender, age, and baseline CDR. In Figure 201, intrinsic epigenetic age acceleration (IEAA) is defined as the residual from a multivariable regression of Horvath DNAm age — estimated using 353 CpGs as specified in Horvath et al. (2013) — on chronological age and blood cell count estimates. This metric, independent of age- related changes in blood cell composition, captures cell-intrinsic aging properties preserved across different cell types and organs, suggesting a fundamental aging process. FIG. 202 presents the principal component GrimAge epigenetic age acceleration, a composite biomarker derived from DNAm-based surrogate markers of plasma protein levels and smoking pack-years; the resulting AgeAccelGrim is highly predictive of incident coronary heart disease and time-to-death, although the first three principal components were removed from the analysis due to technical differences. In FIG. 203, the PackYear clock quantifies smoking history by multiplying the number of cigarette packs smoked per day by the number of years a person has smoked, noting that changes in this clock may also occur due to metabolic factors unrelated to smoking, thereby providing additional insights into lung function and overall health. FIG. 204 introduces the retro-age clock, a composite retroelement-based epigenetic clock developed using DNA methylation patterns of retroelements — specifically, human endogenous retroviruses (HERVs) and long interspersed nuclear elements (LINEs). These retroelement clocks have been observed to reverse during transient epigenetic reprogramming, accelerate in individuals with HIV- 1, respond to antiretroviral therapy, and accurately estimate the long-term culture ages of human brain organoids. FIG. 205 details Retro-age clock version 2, an updated retroelement-based clock that incorporates CpGs covered by Illumina’s Infinium MethylationEPIC v2.0 Kit. FIGs 206-208 describe stochasticclocks derived from artificial DNAm datasets using a stochastic simulation model; unlike the original epigenetic clocks — developed from real DNAm datasets capturing both stochastic and nonstochastic elements — the stochastic clocks (StocH, StocZ, and StocP) were constructed from the corresponding CpGs of the Horvath, Zhang, and Pheno Age clocks, with FIG. 206 ’s clock measuring changes associated with disease-related DNAm alterations, while those in FIG. 207 and FIG. 208 capture changes related to normal aging. FIG. 209 features the systems age epigenetic acceleration clock, which correlates the aging of organ systems with biological age, and FIG. 210 shows the heat epigenetic age acceleration clock, correlating cardiac program activity with biological aging. FIG. 211 presents the Naive CD4 T cell clock, which correlates DNA methylation patterns with the functional status of naive CD4 T cells. FIG. 212 and FIG.213 report clocks data provided by the Epigenetic Clock Development Foundation; FIG. 212 displays the GrimAge clock analysis using predicted epigenetic age acceleration, and FIG. 213 illustrates the SkinBlood clock epigenetic age acceleration, currently considered the standard for accurately determining biological age.

[0486] While some embodiments have been illustrated and described, a person with ordinary skill in the art, after reading the foregoing specification, can effect changes, substitutions of equivalents and other types of alterations to the compounds of the present technology or salts, pharmaceutical compositions, derivatives, prodrugs, metabolites, tautomers or racemic mixtures thereof as set forth herein. Each aspect and embodiment described above can also have included or incorporated therewith such variations or aspects as disclosed in regard to any or all of the other aspects and embodiments.

[0487] The present technology is also not to be limited in terms of the particular aspects described herein, which are intended as single illustrations of individual aspects of the present technology. Many modifications and variations of this present 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 present technology, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the appended claims. It is to be understood that this present technology is not limited to particular methods, reagents, compounds, compositions, labeled compounds or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describingparticular aspects only, and is not intended to be limiting. Thus, it is intended that the specification be considered as exemplary only with the breadth, scope and spirit of the present technology indicated only by the appended claims, definitions therein and any equivalents thereof.

[0488] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby 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 generic disclosure also form part of the present technology. This includes the generic description of the present technology with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein.

[0489] All publications, patent applications, issued patents, and other documents (for example, journals, articles and / or textbooks) referred to in this specification are herein 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 that are contained in text incorporated by reference are excluded to the extent that they contradict definitions in this disclosure.

[0490] Other embodiments are set forth in the following claims, along with the full scope of equivalents to which such claims are entitled.

[0491] While the invention has been particularly shown and described with reference to a preferred embodiment and various alternate embodiments, it will be understood by persons skilled in the relevant art that various changes in form and details can be made therein without departing from the spirit and scope of the invention.

[0492] All references, issued patents and patent applications cited within the body of the instant specification are hereby incorporated by reference in their entirety, for all purposes.

[0493] Although 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 following claims.

Claims

WHAT IS CLAIMED IS:

1. A method to decrease biological age acceleration or biological age (“dAge”) as measured by a DNA methylation clock in a subject, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutically acceptable carrier and at least one compound selected from the group2. The method of claim 1 , wherein decreasing biological age acceleration or dAge in the subject treats, reduces, improves, or ameliorates a disease or condition associated with biological clocks.

3. The method of claim 1 or 2, wherein decreasing biological age acceleration or dAge in the subject decreases neuroinflammation.

4. The method of any one of claims 1 to 3, wherein decreasing biological age acceleration or dAge in the subject improves cognition.

5. The method any one of claims 1 to 4, wherein decreasing biological age acceleration or dAge in the subject decreases symptoms of a neurodegenerative disease.

6. The method of any one of claims 1 to 5, wherein decreasing biological age acceleration or dAge in the subject improves insulin sensitivity in an obese subject.

7. The method of any one of claims 1 to 6, wherein the subject is both obese and has impaired glucose tolerance.

8. The method of any one of claims 1 to 6, wherein the subject is both obese and has type- 2 diabetes (“T2D”).

9. The method of any one of claims 1 to 8, wherein administering to the subject the at least one compound decreases age acceleration (“dAge”) or biological age (Horvath Skin Blood Clock).

10. The method of any one of claims 1 to 9, wherein administering to the subject the at least one compound improves a subject’s Mini -Mental State Examination (MMSE) score.

11. The method of any one of claims 1 to 10, wherein administering to the subject the at least one compound improves clinical dementia rating (CDR) standard battery (SB) through Homeostatic Model Assessment 2 % Insulin Sensitivity (H0MA2 %2).

12. The method of any one of claims 1 to 11, wherein administering to the subject the at least one compound improves CDR SB cholesterol in the subject.

13. The method of any one of claims 1 to 12, wherein administering to the subject the at least one compound decreases cholesterol in the subject.

14. The method of any one of claims 1 to 13, wherein administering to the subject the at least one compound improves CDR SB age acceleration in pTau217+ (pTau+) subjects.

15. The method of any one of claims 1 to 14, wherein administering to the subject the at least one compound decreases age acceleration (dAge) in pTau217+ (pTau+) subjects.

16. The method of any one of claims 1 to 15, wherein administering to the subject the at least one compound improves CDR SB Glial Fibrillary Acid Protein (GFAP) in the subject.

17. The method of any one of claims 1 to 16, wherein administering to the subject the at least one compound improves ADCOMS (Alzheimer’s Disease Composite Score) RANIES in the subject.

18. The method of any one of claims 1 to 17, wherein administering to the subject the at least one compound improves adiponectin in the subject.

19. The method of any one of claims 1 to 18, wherein administering to the subject the at least one compound improves Cog 12 through an increase in M2 monocytes in the subject.

20. The method of any one of claims 1 to 19, wherein administering to the subject the at least one compound improves CGIC through a decrease in WHR in the subject.

21. The method of any one of claims 1 to 20, wherein administering to the subject the at least one compound improves CGIC through age deceleration in the subject.

22. The method of any one of claims 1 to 21, administering to the subject the at least one compound improves insulin sensitivity.

23. The method of any one of claims 1 to 22, wherein administering to the subject the at least one compound improves leptin concentrations or activity in the hypothalamus or hippocampus of the subject.

24. The method of any one of claims to 1 to 23, wherein administering to the subject the at least one compound improves obesity-related metabolic inflammation and / or metabolic syndrome triglycerides.

25. The method of any one of claims to 1 to 24, wherein administering to the subject the at least one compound improves obesity effects on age acceleration or biological age (Horvath Skin Blood Clock).

26. The method of any one of claims 1 to 25, wherein administering to the subject the at least one compound improves obesity and NfL in the subject.

27. The method of any one of claims 1 to 26, wherein administering to the subject the at least one compound improves obesity and cholesterol in the subject.

28. The method of any one of claims 1 to 27, wherein administering to the subject the at least one compound improves chemokine regulation in the subject.

29. The method of claim 28, wherein the chemokines MCP1 and RANTES are coregulated.

30. The method of any one of claims 1 to 29, wherein administering to the subject the at least one compound decreases Monocyte Chemoattractant Protein 1 (MCP1).

31. The method of any one of claims 1 to 30, wherein administering to the subject the at least one compound restores immune component coregulation.

32. The method of any one of claims 1 to 31, wherein administering to the subject the at least one compound improves chemokine age acceleration or biological age (Horvath Skin Blood Clock), in the subject.

33. The method of claim 32, wherein improving chemokine age acceleration or biological age (Horvath Skin Blood Clockjin the subject decreases age acceleration or biological age (Horvath Skin Blood Clock).

34. The method of claim 33, wherein administering to the subject the at least one compound increases M2 monocytes / microglia in the subject.

35. The method of any one of claims 1 to 34, wherein administering to the subject the at least one compound improves diastolic blood pressure.

36. The method of any one of claims 1 to 35, wherein administering to the subject the at least one compound improves metabolic regulation in the subject.

37. The method of any one of claims 1 to 36, wherein administering to the subject the at least one compound decreases insulin levels in the subject.

38. The method of any one of claims 1 to 37, wherein administering to the subject the at least one compound decreases insulin resistance.

39. The method of any one of claims 1 to 38, wherein administering to the subject the at least one compound decreases insulin age acceleration, biological age (Horvath Skin Blood Clock), and / or biological age in the subject.

40. The method of any one of claims 1 to 39, wherein administering to the subject the at least one compound increases H0MA2 IR by decreasing Clq in the subject.

41. The method of any one of claims 1 to 40, wherein administering to the subject the at least one compound dissociates H0MA2 IR from H0MA2 %S.

42. The method of any one of claims 1 to 41, wherein the subject is a mammal.

43. The method of any one of claims 1 to 42, wherein the subject is a human.

44. The method of any one of claims 1 to 42, wherein the subject is an animal.

45. The method of any one of claims 1 to 44, wherein the at least one compound is 17a- ethyny landrost- 5 -ene-30, 70, 170-tr iol .

46. The method of any one of claims 1 to 44, wherein the at least one compound is47. The method of any one of claims 1 to 44, wherein the at least one compound is48. The method of any one of claims 1 to 44, wherein the at least one compound is

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