Compounds and methods for promoting longevity and treating senopathies

Abstract

Senescent cells are implicated in aspects of age-related decline in health and may contribute to certain diseases. Embodiments include formulations and methods for promoting longevity and treating a senescence associated disease or disorder. Applicants present three distinct classes and multiple associated therapeutic molecules that can function alone or in combination with the other molecules. The classes include transcription regulators, lipid intermediates and microRNAs. Additional embodiments include methods of promoting longevity by administering GRAS substances.

Description

COMPOUNDS AND METHODS FOR PROMOTING LONGEVITY AND TREATING SENOPATHIESRELATED APPLICATIONS

[0001] The present application claims priority to U.S. provisional patent application No. 63 / 675,257 filed July 24, 2024, the contents of which are incorporated herein by reference.FIELD OF THE INVENTION

[0002] The invention relates to therapeutics, and more specifically, it relates to compounds and methods for promoting longevity and treating senescence-associated diseases and disorders.BACKGROUND

[0003] Aging can be defined as the process of becoming older. In humans, aging represents the accumulation of changes over time and can encompass physical, psychological, and social changes. Advanced age is the greatest risk factor for many chronic diseases. More than 90% of adults aged 65 or older experience at least one chronic disease such as cancer, diabetes, or cardiovascular disease. Aging phenotypes and pathologies, including diverse age-associated diseases and disorders, are causally linked to the accumulation of senescent cell burden with age.

[0004] While aging may be a complex multifactorial process with no single cause or treatment, the issue whether aging can be classified as the disease is debated. Many strategies for extending organismal life spans have been proposed including replacing cells and organs, comprehensive strategies for repairing the accumulated damage, using hormetins to activate endogenous repair processes, modulating the aging processes through specific mutations, gene therapy and small molecule drugs.

[0005] Senescent cells are characterized by irreversible cell-cycle arrest of proliferation-competent cells, morphological and metabolic changes, altered gene expression, chromatin reorganization, and a unique pro-inflammatory senescence-associated secretory phenotype (SASP). Senescent cells in older adults contribute to chronic inflammation and damage to surrounding tissues. It has been demonstrated that removal of senescent cells via genetic manipulation in transgenic mouse models can prevent or delay tissue dysfunction, improve age-related pathologies, and extend health span. This suggests that removal of senescent cell burden in aging adults, merits further study as a therapeutic target of interest for the treatment and prevention of the disease of aging. Senolytic drugs have also shown promising results in mice and human cell culture models.

[0006] Treatments and therapies for senescence reversal (i.e., aging reversal) have been generally unsuccessful because of the complexity of the underlying mechanisms of senescence and the lack of tools for understanding and treating senescence. Recent developments in bioinformatics (e.g., deep neural networks) have presented opportunities for developing highly-personalized senescence reversal treatments, based in part on gene expression of senescent tissues versus non-senescent tissues. The present invention includes compounds and methods of promoting longevity and treating age-related diseases and conditions.SUMMARY OF THE INVENTION

[0007] The inventions described and claimed herein have many attributes and embodiments including, but not limited to, those set forth or described or referenced in this brief summary. The inventions described and claimed herein are not limited to, or by, the features or embodiments identified in this summary, which is included for purposes of illustration only and not restriction.

[0008] Embodiments also include therapies for promoting longevity and preventing or treating senescence-associated diseases and disorders using multiple agents.

[0009] Embodiments also include methods of increasing life expectancy and / or reducing the probability of suffering from senescence-associated diseases and disorders.

[0010] Embodiments also include the formulations and methods for promotinglongevity, for anti-aging and for the treating, preventing or improving the prognosis of senescence-associated diseases and disorders.

[0011] Embodiments include a therapeutic formulation that includes (a) one or more transcription regulators, one or more lipid intermediates and one or more microRNAs.

[0012] In aspects, the one or more transcription regulators are proteins, transcription factors, long non-coding RNA and / or protein transcriptional complexes. In aspects, they include one or more of MEG3, IRISIN, MOTS-C and HUMANIN.

[0013] In aspects, the one or more lipid intermediates are selected from lipoxin A4 (LXA4), maresins (MaR1 / MaR2), neuroprotection D1 . In aspects, the one or more microRNAs are selected from MIR-145 3p, Let-7c-3p, MIR-548aj-3p, MIR-548aj-4p and hsa-MIR-548x-3p.

[0014] In aspects, the formulation also includes a senolytic agent (e.g., Navitoclax, Venetoclax, Dasatinib / Quercetin, Fisetin, 17-DMAG / IPI504, UBX0101 / FGX04-DRI peptide, cardiotonic steroids, GMD / SSK1 , erastin, vacuolin-1 or apilimod). In aspects, the formulation also includes a senomorphic agent (e.g., Fisetin, Metformin, Rapamycin, Resveratrol, Quercetin, Dasatinib, Kinase inhibitors, an inhibitor of IkB kinase (IKK), an inhibitor of nuclear factor (NF)-kB5, a free radical scavenger or a Janus kinase (JAK) pathway inhibitor).

[0015] In embodiments, the formulations described herein are administered to slow the aging process or reduce signs of aging in a subject.

[0016] Embodiments also include methods to slow the aging process or reduce signs of aging. The methods can include administering (a) one or more transcription regulators, (b) one or more lipid intermediates and (c) one or more microRNAs. In aspects, the one or more transcription regulators target one or more genes selected from MEG3, IRISIN, MOTS-C and HUMANIN. In aspects, the one or more lipid intermediates include lipoxin A4 (LXA4), maresins (MaR1 / MaR2), neuroprotection D1. In aspects, the one or more microRNAs include MIR-145 3p, Let-7c-3p, MIR-548aj-3p, MIR-548aj-4p and / or hsa-MIR-548x-3p. In aspects, the methods also includeadministering a senolytic agent and / or a senomorphic agent. In aspects, one or more of the above compounds work synergistically with one another.

[0017] In aspects, the miRNAs are administered using a lipid-based delivery (e.g., liposomes). In aspects, the miRNAs are administered in a Bound Nucleic Acid (BNA) formation, for example, an RNA-DNA hybrid, RNA-RNA hybrid, miRNA-siRNA or IncRNA etc. In aspects, the miRNA agent is an isolated miRNA that is 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of a miRNA listed above. In aspects, the miRNA agent is a seed sequence of a miRNA listed above.

[0018] In embodiments, the miRNAs are non-coding and act as repressors on one or more sites. In aspects, the miRNAs are packaged into nanoparticles (e.g., exosome or liposomes) for administration. In aspects, the miRNAs are conjugated (e.g., with locked nucleic acids (LNAs), 5'-(E)-vinylphosphonate modification or pyrimidine methylation) for administration.

[0019] In aspects, the miRNAs are modified to improve one or more of activity, bioavailability, solubility, stability, etc. In aspects, the miRNA nucleic acids are modified miRNAs, for example, an miRNA nucleic acid including one or more sequence modifications, modified nucleotides, and / or a 5'-end and / or 3'-end modification. The use of agomirs, mimics or mimetics are also contemplated.

[0020] In embodiments, the formulations described herein are useful to reduce signs / symptoms of a senescence-associated disease or disorder. In aspects the formulations can promote longevity by, for example, improving energy levels, increasing strength / endurance / athletic performance, decreasing frailty, etc.

[0021] Another embodiment is a formulation that includes (a) nicotinamide adenine dinucleotide (NAD), (b) urolithin A, (c) docosahexaenoic acid (DHA), and (d) trimethylglycine (TMG). The formulation can also include (e) resveratrol, (f) spermidine, (g) quercetin and (h) fisetin. In aspects, the formulation can be administered to promote longevity and / or treat a senescence-associated disease or disorder.

[0022] Additional embodiments include methods for treating, preventing or reducingthe risk of a senescence-associated disease or disorder associated with aberrant expression of one or more miRNAs. The methods can include administering a composition that includes an oligonucleotide that down-regulates the over-expression of at least one miRNA. The oligonucleotide can be a) complementary to a nucleotide sequence or b) hybridize to a nucleotide sequence. In aspects, the composition includes one or of nicotinamide adenine dinucleotide (NAD), urolithin A, docosahexaenoic acid (DHA), trimethylglycine (TMG), resveratrol, spermidine, quercetin, fisetin.

[0023] In embodiments, the formulations and methods described herein include an lnterleukin-11 (IL-11 ) antagonist (e.g., anakinra, canakinumab or rilonacept) or an IL-11 antibody.

[0024] In aspects, encapsulation and drug delivery modalities can be used such as synthetic or natural exosomes, lipid bilayers, liposomal and alginate formulation or other synthetic or natural polymer encapsulation techniques. In aspects, the miRNAs and / or small molecules described herein are administered by sublingual, acid-stable capsules, and / or transdermal methods.

[0025] Aspects also include methods of abrogating or reducing senescence that can delay the onset of senescence symptoms and / or extend longevity. In aspects, the methods include administering one or more of NAD (or one or more precursors of NAD), urolithin A, DHA, TMG, resveratrol, spermidine, quercetin and fisetin. In aspects, the NAD, urolithin A, DHA and TMG are administered early in the day (e.g., within one hour after awakening). In aspects, the resveratrol is administered after exercise. In aspects, the resveratrol, spermidine, quercetin and fisetin are administered late in the day (e.g., within two hours of going to sleep).

[0026] Embodiments include a formulation that includes NAD (or one or more precursors of NAD), urolithin A, DHA, TMG, resveratrol, spermidine, quercetin and / or fisetin.

[0027] Embodiments include a morning (“AM”) formulation that includes NAD (orone or more precursors of NAD), urolithin A, DHA, TMG, resveratrol, spermidine, quercetin and / or fisetin.

[0028] Embodiments include an evening (“PM”) formulation that includes resveratrol, spermidine, quercetin and / or fisetin.

[0029] In embodiments, the methods described herein include one or more steps of theranosis. In aspects, the methods include a step of administering a nociceptive stimulus (e.g., immersion in an ice-bath or cold water). In aspects, the methods include a step of intermittent fasting.

[0030] In aspects, the formulation includes about 0.5 to 5 wt.% of each of each of the above compounds for either oral administration or injection.

[0031] In another aspect, the specification provides a use of the pharmaceutical composition including the same in the preparation of drugs for the prevention or treatment of a neurodegenerative disease.

[0032] The methods described herein can delay onset or progression of an age- related disease or condition by, for example, identifying and selectively removing senescent cells over non-senescent cells. In aspects, the formulations and methods described herein reduce the level of one or more biomarkers associated with senescence.BRIEF DESCRIPTION OF THE DRAWINGS

[0033] The accompanying drawings illustrate aspects of the present invention. In such drawings:

[0034] FIG. 1 is a flowchart that depicts compounds and methods of promoting longevity and preventing senescence-associated diseases and disorders

[0035] FIG. 2 is a flowchart that depicts a GRAS treatment to promote longevity and prevent senescence-associated diseases and disorders.Definitions

[0036] Reference in this specification to "one embodiment / aspect" or "an embodiment / aspect" means that a particular feature, structure, or characteristic described in connection with the embodiment / aspect is included in at least one embodiment / aspect of the disclosure. The use of the phrase "in one embodiment / aspect" or "in another embodiment / aspect" in various places in the specification are not necessarily all referring to the same embodiment / aspect, nor are separate or alternative embodiments / aspects mutually exclusive of other embodiments / aspects. Moreover, various features are described which may be exhibited by some embodiments / aspects and not by others. Similarly, various requirements are described which may be requirements for some embodiments / aspects but not other embodiments / aspects. Embodiment and aspect can be in certain instances be used interchangeably.

[0037] The terms used in this specification generally have their ordinary meanings in the art, within the context of the disclosure, and in the specific context where each term is used. Certain terms that are used to describe the disclosure are discussed below, or elsewhere in the specification, to provide additional guidance to the practitioner regarding the description of the disclosure. It will be appreciated that the same thing can be said in more than one way.

[0038] Consequently, alternative language and synonyms may be used for any one or more of the terms discussed herein. Nor is any special significance to be placed upon whether or not a term is elaborated or discussed herein. Synonyms for certain terms are provided. A recital of one or more synonyms does not exclude the use of other synonyms. The use of examples anywhere in this specification including examples of any terms discussed herein is illustrative only and is not intended to further limit the scope and meaning of the disclosure or of any exemplified term. Likewise, the disclosure is not limited to various embodiments given in this specification.

[0039] Without intent to further limit the scope of the disclosure, examples of instruments, apparatus, methods and their related results according to the embodimentsof the present disclosure are given below. Note that titles or subtitles may be used in the examples for convenience of a reader, which in no way should limit the scope of the disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. In the case of conflict, the present document, including definitions, will control.

[0040] The term “senescence” refers to gradual deterioration of functional characteristics in living organisms. Cellular senescence is often defined as a stress- induced, durable cell cycle arrest of previously replication-competent cells. The effects of senescent cells can be thought of as beneficial or detrimental with regard to host physiology and disease, although in some contexts, senescent cells affect a disease state in a complex manner both promoting and opposing certain conditions.

[0041] The term “senescence-associated disease or disorders” or “senopathies” refer to ailments that can be associated with age and can include, for example, atherosclerosis, osteoarthritis, osteoporosis, hypertension, arthritis, cataracts, cancer, Alzheimer’s disease, chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis. Other ailments (including age-related conditions) associated with age or senescence include hair graying, sarcopenia, adiposity, neurogenesis, fibrosis and glaucoma.

[0042] Still other ailments associated with age or senescence include cardiovascular disease (e.g., atherosclerosis, angina, arrhythmia, cardiomyopathy, congestive heart failure, coronary artery disease, carotid artery disease, endocarditis, coronary thrombosis, myocardial infarction, hypertension, aortic aneurysm, cardiac diastolic dysfunction, hypercholesterolemia, hyperlipidemia, mitral valve prolapsed, peripheral vascular disease, cardiac stress resistance, cardiac fibrosis, brain aneurysm, and stroke). A senescence-associated disease or disorder can also be an inflammatory or autoimmune disease or disorder (e.g., osteoarthritis, osteoporosis, oral mucositis, inflammatory bowel disease or kyphosis). A senescence-associated disease or disorder can also be a neurodegenerative disease (e.g., Alzheimer's disease,Parkinson's disease, Huntington's disease, dementia, mild cognitive impairment or motor neuron dysfunction). A senescence-associated disease or disorder can also be a metabolic disease (e.g., diabetes, diabetic ulcer, metabolic syndrome or obesity). A senescence-associated disease or disorder can also be a pulmonary disease (e.g., pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, cystic fibrosis, emphysema, bronchiectasis or age-related loss of pulmonary function). A senescence- associated disease or disorder can also be an eye disease or disorder (e.g., macular degeneration, vitreous degeneration, glaucoma, cataracts, presbyopia or vision loss). A senescence-associated disease or disorder is an age-related disorder that can also be renal disease, renal failure, frailty, hearing loss, muscle fatigue, skin conditions, skin wound healing, liver fibrosis, pancreatic fibrosis, oral submucosa fibrosis or sarcopenia. A senescence-associated disease or disorder can also be a dermatological disease or disorder (e.g., eczema, psoriasis, hyperpigmentation, nevi, rashes, atopic dermatitis, urticaria, diseases or disorders related to photosensitivity or photoaging).

[0043] The term “senescence-associated B-galactosidase,” “SA-[3-gal” or “SABG” is a hypothetical hydrolase enzyme that catalyzes the hydrolysis of [3-galactosides into monosaccharides only in senescent cells. Senescence-associated beta-galactosidase, along with p16lnk4A, can be used as a biomarker of cellular senescence.

[0044] The term “senolytic” or “senolytic agent” refers to a therapeutic such as a small molecule that can selectively or preferentially induce death of senescent cells. A senolytic agent may kill senescent cells by inducing (i.e. , activating, stimulating or removing inhibition of) an apoptotic pathway that leads to cell death. Senolytic agents may be useful for treatment of senescence-associated diseases or disorders. For example, the drugs dasatinib, quercetin, fisetin and navitoclax have potential senolytic activities.

[0045] The term “theranosis” or “theranostics” generally refers to processes used to tailor therapy for a patient. It is the use of diagnostic tests to identify those patients who are better-suited for a drug, drugs, medicaments or therapeutics or to determine how well a drug(s), medicament(s) or therapeutic(s) is / are working.

[0046] The term "biomarker" refers generally to a DNA, RNA, protein, carbohydrate, or glycolipid-based molecular marker, the expression or presence of which in a subject's sample can be detected by standard methods (or methods disclosed herein) and is predictive or prognostic of the effective responsiveness or sensitivity of a mammalian subject with an ailment. Biomarkers may be present in a test sample but absent in a control sample, absent in a test sample but present in a control sample, or the amount of biomarker can differ between a test sample and a control sample. For example, protein biomarkers can be present in such a sample, but not in a control sample, or certain biomarkers are seropositive in the sample, but seronegative in a control sample. Also, expression of such a biomarker may be determined to be higher than that observed from a control sample. The terms "marker" and "biomarker" are used herein interchangeably.

[0047] A biomarker that is “upregulated” generally refers to an increase in the level of expression in response to a given treatment or condition. A biomarker that is “downregulated” generally refers to a decrease in the level of expression of the biomarker in response to a given treatment or condition. In some situations, the biomarker level can remain unchanged upon a given treatment or condition. A biomarker from a patient sample can be “upregulated,” i.e., the level can be increased, for example, by about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 200%, about 300%, about 500%, about 1 ,000%, about 5,000% or more compared to a reference level. Alternatively, a biomarker can be “downregulated,” i.e., the level can be decreased, for example, by about 99%, about 95%, about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 30%, about 20%, about 10%, about 5%, about 2%, about 1 % or less compared to a reference level.

[0048] The term "detecting" or "determining" with respect to a biomarker value includes the use of both the instrument required to observe and record a signal corresponding to a biomarker value and the material(s) required to generate that signal. In various embodiments, the biomarker value is detected using any suitable method, including fluorescence, chemiluminescence, surface plasmon resonance, surfaceacoustic waves, mass spectrometry, infrared spectroscopy, Raman spectroscopy, atomic force microscopy, scanning tunneling microscopy, electrochemical detection methods, nuclear magnetic resonance, quantum dots, and the like.

[0049] The term “fingerprint,” “disease fingerprint,” or “biomarker signature” refers to a plurality or pattern of biomarkers that have elevated or reduced levels in a subject with disease. A fingerprint can be generated by comparing subjects with the disease to healthy subjects and used for screening / diagnosis of the disease.

[0050] The term “nanoparticle” refers to a particle of matter, generally between about 1 and 100 nanometers (nm) in diameter. As used herein, a nanoparticle can refer to a liposome, virus, viral vector or other viral particle.

[0051] The term “liposome” refers to a spherical vesicle having at least one lipid bilayer (i.e. an aqueous solution core surrounded by a hydrophobic membrane). Liposomes can be prepared by disrupting biological membranes (such as by sonication). Liposomes are formed when phospholipids and their derivatives are dispersed in water. Upon dispersion in water the phospholipids form closed vesicles called “liposomes,” which are characterized by lipid bilayers encapsulating an aqueous core. Various liposomes have been used as carriers for entrapped therapeutic agents, such as drugs, enzymes and genetic sequences for use in medical science, in pharmaceutical science and in biochemistry. Specific uses include delivery of nutrients and pharmaceutical drugs, such as lipid nanoparticles in mRNA vaccines and DNA vaccines. Liposomes can be modified by the incorporation of polyethylene glycol or other hydrophilic polymers (e.g., a PEG liposome where one or more of the constituent lipids is modified by attachment of PEG). Liposomes can also be modified to target particular cell types by incorporating targeting factors (e.g., “targeting ligands”) for particular cell types. Examples include asialoglycoprotein, folate, transferrin, antibodies, etc.

[0052] The term “exosome” refers to a membrane-bound extracellular vesicles (EVs) that are produced in the endosomal compartment of most eukaryotic cells. In multicellular organisms, exosomes and other EVs are found in biological fluids includingsaliva, blood, urine and cerebrospinal fluid. Exosomes are similar to liposomes in terms of consisting of bilayered phospholipids, but the biogenesis of exosomes ensures their biocompatibility and low toxicity. It also significantly complicates pharmaceutical development, production and safety profiling (immunogenicity, and potential biological impurities).

[0053] The term “nicotinamide adenine dinucleotide” or “NAD” refers to a coenzyme central to metabolism. Found in all living cells, NAD is called a dinucleotide because it consists of two nucleotides joined through their phosphate groups. One nucleotide contains an adenine nucleobase and the other, nicotinamide. NAD exists in two forms: an oxidized and reduced form, abbreviated as NAD+ and NADH (H for hydrogen), respectively.

[0054] In cellular metabolism, NAD is involved in redox reactions, carrying electrons from one reaction to another, so it is found in two forms: NAD+ is an oxidizing agent, accepting electrons from other molecules and becoming reduced; with H+, this reaction forms NADH, which can be used as a reducing agent to donate electrons. These electron transfer reactions are the main function of NAD. It is also used in other cellular processes, most notably as a substrate of enzymes in adding or removing chemical groups to or from proteins, in posttranslational modifications. Because of the importance of these functions, the enzymes involved in NAD metabolism are targets for drug discovery. In organisms, NAD can be synthesized from simple building-blocks (de novo) from either tryptophan or aspartic acid, each a case of an amino acid.Alternatively, more complex components of the coenzymes are taken up from nutritive compounds such as niacin; similar compounds are produced by reactions that break down the structure of NAD, providing a salvage pathway that recycles them back into their respective active form. Some NAD is converted into the coenzyme nicotinamide adenine dinucleotide phosphate (NADP), whose chemistry largely parallels that of NAD, though its predominant role is as a coenzyme in anabolic metabolism. Unfortunately, NAD levels decrease with age. Although NAD is not FDA approved and is regulated as a dietary supplement, some studies have suggested that increasing NAD levels via dietary supplementation may have beneficial effects on metabolism and overall health.

[0055] Precursors of NAD include nicotinamide riboside (NR) and nicotinamide mononucleotide (NMN). Taken orally, NMN can be absorbed and converted to NAD+. Studies have suggested that supplementation with NMN has increased NAD+ biosynthesis, suppressed age-related adipose tissue inflammation, enhanced insulin secretion and insulin action, improved mitochondrial function, improved neuronal function in the brain. Proponents of NAD therapy have proposed that NMN may be an effective nutraceutical anti-aging intervention, with beneficial effects on a wide array of physiological functions.

[0056] The term “urolithin A” refers to a metabolite compound resulting from the transformation of ellagitannins by the gut bacteria. It belongs to the class of organic compounds known as benzo-coumarins or dibenzo-a-pyrones. Its precursors - ellagic acids and ellagitannins - are ubiquitous in nature, including edible plants, such as pomegranates, strawberries, raspberries, walnuts, and others. Urolithin A is not known to be found in any food source. Its bioavailability mostly depends on individual microbiota composition, as only some bacteria are able to convert ellagitannins into urolithins. A dietary supplement composed of urolithin A (Uro-A) that may be used to modulate mitochondrial activity, and with potential antioxidant and anti-inflammatory activities. Upon oral administration, urolithin A supplement activates mitophagy, which may improve mitochondrial and cellular health. Urolithin A, one of the natural polyphenolic metabolites of ellagic acid and ellagitannins produced by the gut microbiota, also acts as a direct free radical scavenger, which may protect against inflammation- and reactive oxygen species (ROS)-induced cellular damage.

[0057] The terms “docosahexaenoic acid,” “DHA,” “17-DHA” and “17-HDoHE” refer to an omega-3 fatty acid that is an important component of the human brain, cerebral cortex, skin, and retina. It is given the fatty acid notation 22:6(n-3). It can be synthesized from alpha-linolenic acid or obtained directly from maternal milk (breast milk), fatty fish, fish oil, or algae oil. The consumption of DHA (e.g., from fatty fish such as salmon, herring, mackerel and sardines) contributes to numerous physiological benefits, including cognition. As a component of neuronal membranes, the function of DHA is to support neuronal conduction and to allow the optimal functioning of neuronalmembrane proteins (such as receptors and enzymes). Structurally, DHA is a carboxylic acid (-oic acid) with a 22-carbon chain (docosa- derives from the Ancient Greek for 22) and six (hexa-) cis double bonds (-en-); with the first double bond located at the third carbon from the omega end.

[0058] The term “trimethylglycine” or “TMG” also known as betaine or betaine anhydrous, refers to anti-inflammatory and antioxidative compound. It is an amino acid derivative that is found in plants. It is involved in a chemical process (i.e. , methylation) which is essential for DNA production. Structurally, TMG includes glycine with three attached methyl groups.

[0059] The term “resveratrol” or “3,5,4'-trihydroxy-trans-stilbene” refers to a stilbenoid, a type of natural phenol or polyphenol and a phytoalexin produced by several plants in response to injury or when the plant is under attack by pathogens, such as bacteria or fungi. Sources of resveratrol in food include the skin of grapes, blueberries, raspberries, mulberries, and peanuts. It has been studied for potential therapeutic benefits.

[0060] The term “spermidine” refers to a polyamine compound (C7H19N3) found in ribosomes and living tissues and having various metabolic functions within organisms. Spermidine is an aliphatic polyamine. Spermidine synthase (SPDS) catalyzes its formation from putrescine. It is a precursor to other polyamines, such as spermine and its structural isomer thermospermine. Spermidine synchronizes an array of biological processes, (such as Ca2+, Na+, K+-ATPase) thus maintaining membrane potential and controlling intracellular pH and volume. Spermidine regulates biological processes, such as Ca2+influx by glutamatergic N-methyl-D-aspartate receptor (NMDA receptor), which has been associated with nitric oxide synthase (NOS) and cGMP / PKG pathway activation and a decrease of Na+,K+-ATPase activity in cerebral cortex synaptosomes.

[0061] The term “quercetin” refers to a plant flavonol from the flavonoid group of polyphenols. It is found in many fruits, vegetables, leaves, seeds, and grains; capers, red onions, and kale are common foods containing appreciable amounts of it. It has a bitter flavor and is used as an ingredient in dietary supplements, beverages and foods.

[0062] The term “fisetin” or “7,3',4'-flavon-3-ol” refers to a plant flavonol from the flavonoid group of polyphenols. It can be found in many plants, where it serves as a yellow / ochre coloring agent. It is also found in many fruits and vegetables, such as strawberries, apples, persimmons, onions and cucumbers. The biological activity of fisetin has been studied in many laboratory assays; like other polyphenols it has many activities which can have therapeutic benefit.

[0063] The term “naringenin” refers to a flavanone from the flavonoid group of polyphenols. It is commonly found in citrus fruits, especially as the predominant flavonone in grapefruit. Naringenin controls body lipids by possessing hypocholesterolemic and hypolipidemic effects, as well as anti-estrogenic activities. Generally, flavonoids have been investigated and found to be a major regulator of tumor cell growth, EC migration, and angiogenesis. Additionally, naringenin regulates blood pressure and has antagonistic activities against inflammation.

[0064] The term “transient receptor potential cation channel subfamily M (melastatin) member 8” or “TRPM8,” also known as the cold and menthol receptor 1 or “CMR1” refers to a protein that in humans is encoded by the TRPM8 gene. The TRPM8 channel is the primary molecular transducer of cold somatosensation in humans. In addition, mints can desensitize a region through the activation of TRPM8 receptors (the 'cold' / menthol receptor). TRPM8 is an ion channel: upon activation, it allows the entry of Na+and Ca2+ions into the cell, which leads to depolarization and the generation of an action potential. The signal is conducted from primary afferents (type C- and A-delta) eventually leading to the sensation of cold and cold pain. The TRPM8 protein is expressed in sensory neurons, and it is activated by cold temperatures and cooling agents, such as menthol and icilin whereas WS-12 and CPS-369 are the most selective agonists of TRPM8. As IL-11 over expression is associated with a number of cancers, inhibition of its signaling pathway may have utility in treating cancer, and has also been researched for its role in treating the pathology of aging.

[0065] The term “Interleukin 11 ,” “IL-11” or “adipogenesis inhibitory factor” refers to a protein that in humans is encoded by the IL11 gene. The human IL-11 gene,consisting of 5 exons and 4 introns, is located on chromosome 19, and encodes a 23 kDa protein. IL-11 is a member of the IL-6-type cytokine family, distinguished based on their use of the common co-receptor gp130. Signal specificity is provided by the IL- 11 Ra subunit which is expressed at high levels in fibroblasts and other stromal cells but not immune cells, unlike IL6 receptors that are expressed at highest levels in immune cells and lowly expressed in stromal cells. IL-11 through its binding to its transmembrane IL-11 Ra receptor and resultant activation of downstream signaling pathways has been thought to regulate adipogenesis, osteoclastogenesis, neurogenesis and platelet maturation. More recently it has been discovered that overexpression of IL-11 is associated with a variety of cancers and may provide a link between inflammation and cancer.

[0066] The term “miRNA” or “micro RNA,” “miRNA biomarkers,” or “MicroRNAs” refers to small, single-stranded, non-coding RNA molecules generally about 16 - 27 nucleotides in length. miRNAs base-pair to complementary sequences in mRNA molecules, then silence said mRNA molecules by one or more of the following processes: (a) cleavage of the mRNA strand into two pieces, (b) destabilization of the mRNA by shortening its poly(A) tail, or (c) reducing translation of the mRNA into proteins. In cells of humans and other animals, miRNAs primarily act by destabilizing the mRNA. miRNAs resemble the small interfering RNAs (siRNAs) of the RNA interference (RNAi) pathway, except miRNAs derive from regions of RNA transcripts that fold back on themselves to form short hairpins, whereas siRNAs derive from longer regions of double-stranded RNA. The human genome may encode over 1900 miRNAs. However, only about 500 human miRNAs represent bona fide miRNAs in the manually curated miRNA gene database MirGeneDB. Mature miRNA originates from the 5' arm or the 3' arm of the precursor product and is denoted with a -5p or -3p suffix, respectively.

[0067] miRNA sequences are publicly available. For example, miRBase (mirbase.org) includes a searchable database of annotated miRNA sequences. miRNA sequences are also available through other databases known to those in the art, including the National Center for Biotechnology Information (ncbi.nlm nih.gov). One canalso identify targets for specific miRNAs utilizing public databases and algorithms, for example at MicroCosm Targets (ebi.ac.uk / enright-srv / microcosm / htdocs / targets / ), TargetScan (targetscan.org), and PicTar (pictar.mdc-berlin.de). Based on miRNA sequences from one organism (e.g., a mouse), one in the art can utilize the available databases to determine a corresponding miRNA from another organism (e.g., a human).

[0068] The nomenclature used herein will be understood by those in the art. MicroRNAs are named using the “mir” prefix and a unique identifying number (e.g., miR- 1 , miR-2, . . . miR-89, etc.). The genes that encode the miRNA are also named using the same three-letter prefix, with capitalization, hyphenation, and italics according to the conventions of the organism (for example, mir-1 in C. elegans and Drosophila, MIR156 in Arabidopsis and rice). The identifying numbers are assigned sequentially, with identical miRNAs having the same number, regardless of organism.

[0069] The term “miRNA mimic” or “mimetic” refers to miRNA that has the same sequence as the native or wild type miRNA, but has a modified backbone, a modified base, and / or a 5' or 3' end modification. In some examples an miRNA mimetic is may less susceptible to degradation or nuclease activity. In aspects, an miRNA mimic is an miRNA with at least one sequence modification and having 75% or higher sequence identity to a native or wild type miRNA and that also binds to the same mRNA(s) with similar affinity as the wild type or native miRNA. The disclosed miRNAs may also be both an miRNA mimetic and an miRNA mimic, for example, an miRNA with at least one sequence modification (e.g., 75% or higher sequence identity) to a wild type miRNA, and also having a modified backbone, base, and / or end modification.

[0070] The term “seed sequence” refers to a 6 - 8 nucleotide (nt) long substring within the first 8 nt at the 5'-end of the miRNA (i.e., seed sequence) that is an important determinant of target specificity.

[0071] The term “agomir” refers to a synthetic oligonucleotide or oligonucleotide mimetic that functionally mimics a miRNA. An agomir can be an oligonucleotide with the same or similar nucleic acid sequence to a miRNA or a portion of a miRNA. Incertain embodiments, the agomir has 1 , 2, 3, 4, 5, 6, 7, 8, 9 or 10 nucleotide differences from the miRNA that it mimics. Further, agomirs can have the same length, a longer length or a shorter length than the miRNA that it mimics.

[0072] The term long non-coding RNA (IncRNA) refers to a class of RNA molecules that are typically longer than 200 nucleotides and are not translated into proteins.IncRNAs have been discovered to substantially impact the biology of tumors, especially cancer growth and development, by functioning as tumor drivers or inhibitors. IncRNA instability has been associated with various cancers, notably triple-negative BCs (TNBCs), ovarian, lung, and pancreatic cancer. Several biological functions can be regulated by IncRNAs, such as epigenetic alterations, cell division, growth, division, movement, and death. Further, IncRNAs can engage in various signaling systems, such as the Hippo signaling system, which is important in cancer-associated circuits.

[0073] The term “messenger RNA” or “mRNA” refers to a single-stranded molecule of RNA that corresponds to the genetic sequence of a gene and is read by a ribosome in the process of synthesizing a protein. As used herein, mRNA can also include “miRNA” and small interfering RNAs (siRNAs).

[0074] The term “chondrocyte” refers to the only cells found in healthy cartilage. They produce and maintain the cartilaginous matrix, which consists mainly of collagen and proteoglycans. Although the word chondroblast is commonly used to describe an immature chondrocyte, the term is imprecise, since the progenitor of chondrocytes (which are mesenchymal stem cells) can differentiate into various cell types, including osteoblasts.

[0075] The term “generally recognized as safe” or “GRAS” refers to a United States Food and Drug Administration (FDA) designation that a chemical or substance added to food is considered safe by experts under the conditions of its intended use. An ingredient with a GRAS designation is exempted from the usual Federal Food, Drug, and Cosmetic Act (FFDCA) food additive tolerance requirements.

[0076] The term “analog” or “chemical analog” refers to a compound having astructure similar to that of another compound, but differing from it in respect to a certain component. It can differ in one or more atoms, functional groups, or substructures, which are replaced with other atoms, groups, or substructures.

[0077] The term “prognosis” refers to the forecast or likely outcome of a disease. As used herein, it refers to the probable outcome of a senescence-associated disease or disorder including whether the disease will respond to treatment or mitigation efforts and / or the likelihood that the disease will progress.

[0078] The therapeutic agents in the pharmaceutical compositions can be formulated in a "therapeutically effective amount" or a "prophylactically effective amount." A "therapeutically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result. A therapeutically effective amount may vary depending on the condition to be treated, the severity and course of the condition, the mode of administration, whether the agent is administered for preventive or therapeutic purposes, the bioavailability of the particular agent(s), the ability of the therapeutic small molecule to elicit a desired response in the individual, previous therapy, the age, weight and sex of the patient, the patient's clinical history and response to the agent, the type of the therapeutic small molecule used, discretion of the attending physician, etc. A therapeutically effective amount is also one in which any toxic or detrimental effects is outweighed by the therapeutically beneficial effects. A "prophylactically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result.

[0079] The term “treating” or “treatment” refers to one or more of (1 ) inhibiting the disease, condition or disorder in an individual who is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., arresting further development of the pathology and / or symptomatology); and (2) ameliorating the disease condition or disorder in an individual who is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., reversing the pathology and / or symptomatology) such as decreasing the severity of disease.

[0080] As used herein, the term "prevention" means all of the actions by which theoccurrence of the disease is restrained or retarded.

[0081] The term “prophylactic treatment” refers to any of: halting the onset, reducing the risk of development, reducing the incidence, delaying the onset, reducing the development, as well as increasing the time to onset of symptoms of a particular disorder.

[0082] The term “erastin” refers to a small molecule capable of initiating ferroptotic cell death. Erastin acts through inhibition of the cystine / glutamate transporter, thus causing decreased intracellular glutathione (GSH) levels. Specifically, erastin binds and activates voltage-dependent anion channels (VDAC) by reversing tubulin's inhibition on VDAC2 and VDAC3, and functionally inhibits the cystine-glutamate antiporter system Xc-. Cells treated with erastin are deprived of cysteine and are unable to synthesize the antioxidant glutathione. Depletion of glutathione eventually leads to excessive lipid peroxidation and cell death.

[0083] The term “senolytic” or “senolytic agent” refers to a therapeutic such as a small molecule that can selectively or preferentially induce death of senescent cells. A senolytic agent may kill senescent cells by inducing (i.e. , activating, stimulating or removing inhibition of) an apoptotic pathway that leads to cell death. Senolytic agents can be useful to treat various ailments including diabetes (particularly type 2 diabetes), metabolic syndrome, or obesity; pulmonary diseases, such as chronic obstructive pulmonary disease or idiopathic pulmonary fibrosis; and inflammatory disorders, such as osteoarthritis. Senolytics are also sought-after for cancer therapy, as chemotherapeutics (e.g., DNA damaging agents) can cause cancer cells to become senescent.

[0084] The term “senomorph” refers to one of a range of agents that can modulate the phenotypes of senescent cells (SCs) to those of young cells through interfering with senoinflammation / inflammaging, senescence-related signal pathways and SASP, without induction of SC apoptosis.

[0085] The term “SASP inhibitor” refers to an agent that can inhibit development ofa senescence-associated secretory phenotype (SASP). Suppressing the SASP without eliminating senescent cells is an alternative therapeutic approach for alleviating cellular senescence-related phenotypes or diseases. SASP inhibitors (i.e., senomorphics) can directly or indirectly attenuate the SASP of senescent cells by inhibiting transcription factor nuclear factor (NF)-KB, the JAK-STAT signal transduction pathway, the serine / threonine protein kinase mTOR, mitochondrial complex-1 -related or 4-related targets, or other pathways involved in the induction and maintenance of the SASP

[0086] The term “apoptosis” refers to a caspase-mediated programmed cell death characterized by formation of membrane-enveloped apoptotic bodies that are rapidly phagocytosed by macrophages or neighboring cells. There is evidence of apoptotic mechanisms in animal models of several neurodegenerative diseases, but evidence in human tissues is limited. Caspase-1 , -3, -8, and -9 activation and cytochrome c release seen in models of Huntington’s disease (HD) were also demonstrated in human striatal brain tissue. Similarly, caspase activation and neuronal apoptosis have been demonstrated in ALS and HIV-associated neurodegeneration.

[0087] The term “necroptosis” refers to a form of programmed cell death that is independent from the caspase activation and involves loss of plasma membrane integrity. Two main effector proteins of necroptosis are receptor-interacting serine / threonine-protein kinase 1 (RIPK1 ) and mixed-lineage kinase domain-like (MLKL). Astrocytes release TNF-a, FasL, and TRAIL, which can trigger necroptosis through RIPK1 and MLKL activation, and this mechanism has been demonstrated in murine models of ALS. RIPK1 -mediated axonal pathology was observed in pathological specimens from ALS patients. Necroptotic mechanisms were also observed in MS pathological samples.

[0088] The term "administration" refers to the introduction of an amount of a predetermined substance into a patient by a certain suitable method. The compositions disclosed herein may be administered via any of the common routes, as long as it is able to reach a desired tissue, for example, inhaling, intraperitoneal, intravenous, intramuscular, subcutaneous, intradermal, oral, topical, intranasal, intrapulmonary, orintrarectal administration.

[0089] The term "subject" refers to those who are susceptible to an ailment (e.g., a disease related to senescence) or who are suspected of having or diagnosed with the ailment. However, any subject to be treated with the therapeutic methods described herein is included without limitation.

[0090] The term “gene ontology term enrichment” refers to a technique for interpreting sets of genes making use of the Gene Ontology (GO) system of classification in which genes are assigned to a set of predefined bins depending on their functional characteristics.

[0091] The term “nociceptive stimulus” refers to is the process of communicating noxious stimuli through the central and peripheral nervous system. Nociceptors are sensory receptors that detect potentially damaging stimuli, such as thermal, mechanical, electrical, chemical and ischemic. Cold-water immersion can be used to instigate a nociceptive stimulus. Recent studies indicate benefits of including improved circulation, decreased stress levels, a boost in collagen production, and others that can contribute to one’s health and a reduction in signs of aging.

[0092] The term “exercise” refers to a physical activity that can enhance or maintain fitness and overall health. It is performed for various reasons, including weight loss or maintenance, to aid growth and improve strength, develop muscles and the cardiovascular system, hone athletic skills, improve health, or simply for enjoyment. Types of exercise can be classified as aerobic, anaerobic and flexibility. Types of exercise can also be classified as dynamic or static. Dynamic exercises (e.g., steady running) tend to produce a lowering of the diastolic blood pressure during exercise, due to the improved blood flow. Conversely, static exercise (e.g., weight-lifting) can cause the systolic pressure to rise significantly, albeit transiently, during the performance of the exercise.

[0093] The term “DNA repair-pathway supplements” refers to compounds (e.g., micronutrients) that can promote longevity. Aging adults are prone to deficiencies dueto age-associated functional decline and often to a diet poor in nutrients. Moreover, lack of micronutrients has an indirect impact on the genome. Their low levels reduce the activity of antioxidant enzymes, and therefore inhibit the efficiency of defense against free radicals which can lead to the formation of DNA lesions. The more DNA damage in the genetic material, the faster aging at the cellular level and a higher risk of pathological processes (e.g., carcinogenesis). Supplementation of crucial antioxidative micronutrients (e.g., selenium, zinc, vitamin C, and vitamin E) can positively influence the condition of an aging organism, including minimizing inflammation, enhancing antioxidative defense, and limiting the formation of DNA lesions. In consequence, it may lead to lowering the risk and incidence of age-related diseases such as cardiovascular diseases, neurodegenerative diseases, and malnutrition.

[0094] All numerical designations, e.g., pH, temperature, time, concentration, and molecular weight, including ranges, are to be understood as approximations in accordance with common practice in the art. When used herein, the term “about” may connote variation (+) or (-) 1 %, 5%, 10%, 15% or 20% of the stated amount, as appropriate given the context. It is to be understood, although not always explicitly stated, that the reagents described herein are merely exemplary and that equivalents of such are known in the art.

[0095] Many known and useful compounds and the like can be found in Remington’s Pharmaceutical Sciences (13thEd), Mack Publishing Company, Easton, PA — a standard reference for various types of administration. As used herein, the term “formulation(s)” refers to a combination of at least one active ingredient with one or more other ingredient, also commonly referred to as excipients, which may be independently active or inactive. The term “formulation” may or may not refer to a pharmaceutically acceptable composition for administration to humans or animals and may include compositions that are useful intermediates for storage or research purposes.

[0096] Other technical terms used herein have their ordinary meaning in the art that they are used, as exemplified by a variety of technical dictionaries. The particularvalues and configurations discussed in these non-limiting examples can be varied and are cited merely to illustrate at least one embodiment and are not intended to limit the scope thereof.DETAILED DESCRIPTION

[0097] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the subject technology as claimed. Additional features and advantages of the subject technology are set forth in the description below, and in part will be apparent from the description, or may be learned by practice of the subject technology. The advantages of the subject technology will be realized and attained by the structure particularly pointed out in the written description and claims hereof.

[0098] Applicants identified three distinct target classes and multiple associated therapeutic molecules which either alone or in combinations with the other molecules, can promote longevity (i.e., slowing, stop and / or reverse aging). The three longevity target classes and therapeutic molecules include: (a) transcription regulators, (b) lipid Intermediates and (c) microRNA.Transcription Regulation

[0099] Transcriptional regulation is the means by which a cell regulates the conversion of DNA to RNA (transcription), thereby orchestrating gene activity. A single gene can be regulated in a range of ways, from altering the number of copies of RNA that are transcribed, to the temporal control of when the gene is transcribed. This control allows the cell or organism to respond to a variety of intra- and extracellular signals and thus mount a response. Some examples of this include producing the mRNA that encode enzymes to adapt to a change in a food source, producing the gene products involved in cell cycle specific activities and producing the gene products responsible for cellular differentiation in multicellular eukaryotes.

[0100] Transcriptional regulation is a critical biological process that allows the cell or an organism to respond to a variety of intra- and extra-cellular signals, to define cellidentity during development, to maintain it throughout its lifetime, and to coordinate cellular activity. This highly dynamic mechanism includes a series of biophysical events orchestrated by a huge number of molecules establishing larger networks and occurring through multiple temporal and functional steps that range from specific DNA-protein interactions to the recruitment and assembly of nucleoprotein complexes. Essentially, the key transcription levels include the recruitment and assembly of the entire transcription machinery, the initiation step, pause release and elongation phases, as well as termination of transcription. Additionally, these steps are interconnected with governing chromatin accessibility (such as the unwrapping process, which is controlled by histone modification and chromatin remodeling proteins), and other epigenetic mechanisms (such as enhancer-promoter looping, which is necessary for a successful gene transcription). Finally, various RNA maturation events, such as the splicing that occurs with transcription, constitute an additional level of complexity. Numerous molecules and molecular factors, including transcription factors, cofactors (both coactivators and corepressors), and chromatin regulators, are known to participate to this process. Essential components of the basal transcription machinery include the RNA polymerase II holoenzyme, the general initiation transcription factors (TFIIA, -IIB, - IID, -HE, -HF, and -HH) and the Mediator complex, a multi-subunit compound that joins transcription factors bound at the upstream regulatory elements — such as nuclear receptors — and all the remaining apparatus at the promoter region. It is noteworthy that it also works in close interplay between the basal machinery and factors responsible for the epigenetic modifications; for instance, together with cohesin, it facilitates DNA looping. More recently, a novel multi-subunit complex named Integrator was added as one of the components of the RNA Polymerase Il-mediated transcription apparatus. It is also involved in many stages of eukaryotic transcription for most regulated genes.

[0101] Additionally, the high complexity of transcriptional regulation is also derived from the involvement of non-coding RNAs (ncRNAs). Indeed, research over the last two decades has revealed new classes of ncRNAs, including microRNAs (miRNAs), small nucleolar RNAs (snoRNAs), long ncRNAs (IncRNAs), circular RNAs (circRNAs), and enhancer RNAs (eRNAs), each with different regulatory functions and altogether belonging to a larger RNA communication network ultimately controlling the productionof the final protein.

[0102] Recent advances in “omics” and computational biology have provided novel tools that allow one to integrate different layers of information from biophysical, biochemical, and molecular cell biology studies. In turn, these novel strategies provided a fuller understanding of how DNA sequence information, epigenetic modifications, and transcription machinery cooperate to regulate gene expression. Of note, most of the new molecular biomarkers and therapeutic targets for several human pathologies derive from transcriptome profiling studies, and their number is continuously increasing. Next Generation Sequencing (NGS), mainly RNA-Sequencing (RNA-Seq), has completely revolutionized transcriptome analysis, allowing the quantification of gene expression levels and allele-specific expression in a single experiment, as well as the identification of novel genes, splice isoforms, fusion transcripts, and the entire world of ncRNAs at an unprecedented level.

[0103] It is well known that many human disorders are characterized by global transcriptional dysregulation because most of the signaling pathways ultimately target transcription machinery. Indeed, many syndromes and genetic and complex diseases — cancer, autoimmunity, neurological and developmental disorders, metabolic and cardiovascular diseases — can be caused by mutations / alterations in regulatory sequences, transcription factors, cofactors, chromatin regulators, ncRNAs, and other components of transcription apparatus. Thus, advances in our understanding of molecules and mechanisms involved in the transcriptional circuitry and apparatus lead to new insights into the pathogenetic mechanisms of various human diseases and disorders.

[0104] Applicants have discovered that transcriptional regulation of four particular genes can have particular therapeutic effect related to longevity. These genes include MEG3, IRISIN, MOTS-C and HUMANIN.MEG3

[0105] MEG3 (Maternally Expressed Gene 3) is a long non-coding RNA (IncRNA) essential in many biological activities. The MEG3 gene encodes MEG3, part of theimprinting DLK-MEG3 loci on the human chromosomal region 14q32.3. MEG3 is 35 kb in length and has 10 exons. It is an imprinted gene that comprises ten exons and stores a 1 .6 Kb noncoding RNA (ncRNA). MEG3 uses other substances, such as miRNAs and proteins, to control various biological events. It participates in diverse cellular operations, including epigenetic modification of targeted gene transcription, differentiation of osteogenic tissue, and progression of bone-related conditions involving osteosarcoma, osteoarthritis, and osteoporosis.

[0106] MEG3 may influence targeted gene activity via translation, transcription, post-translational changes, and epigenetic influence. Improper functioning of MEG3 has been associated with poor outcomes and drug resistance (DR) in investigations. MEG3 may influence its target genes via transcriptional and post-translational mechanisms. MEG3, for example, may enhance the levels of p53 by increasing transcriptional activities and post-translational alterations. MEG3 inhibition was related to the nickel-induced promotion of hypermethylation via increased DNMT3b levels, whilst PHLPP1 transcriptional suppression was linked to MEG3's decreased interaction with its suppressive transcription partner c-Jun.Irisin

[0107] Irisin is a novel myokine and adipokine that has gained much attention recently due to its mechanisms of action. Irisin is secreted following proteolytic cleavage of its precursor fibronectin type III domain containing 5 (FNDC5). Following its release, irisin exerts its major action by increasing the expression of mitochondrial uncoupling protein 1 (UCP 1 ), which facilitates the conversion of white adipose tissue (WAT) into beige adipose tissue. Irisin is distributed in various body tissues and several actions have been attributed to its presence in those tissues. It has been suggested that it plays a role in metabolic diseases, ageing, inflammation and neurogenesis. However, the circulating levels of irisin are modulated by several factors such as diet, obesity, exercise, pharmacological agents and different pathological conditions. Modification of circulating irisin level may help in the management of a variety of endocrine and metabolic disorders.

[0108] Irisin is the product of type I membrane protein cleavage encoded by the fibronectin type III domain containing 5 (FNDC5) gene. Irisin can change subcutaneous and visceral adipose tissue into brown adipose tissue, with a consequential increase in thermogenesis. Irisin has also been described as a hormone that may have a key role in glucose homeostasis.MOTS-C

[0109] MOTS-c (mitochondrial open reading frame of the 12S rRNA-c) is a peptide encoded in mitochondrial DNA. It is believed to be involved in skeletal muscle and glucose metabolism. It is upregulated in response to exercise, and is considered an exercise mimetic

[0110] MOTS-C is a nuclear regulatory peptide which plays an important role in aging and age-related disorders, including mechanisms of action and therapeutic potential.Humanin

[0111] Humanin is a micropeptide encoded in the mitochondrial genome by the 16S ribosomal RNA gene, MT-RNR2. Its structure contains a three-turn a-helix, and no symmetry. In in vitro and animal models, it appears to have cytoprotective effects.

[0112] Humanin protects cells from oxidative stress, serum starvation, hypoxia, and other insults in vitro and also improves cardiovascular disease as well as Alzheimer's disease in vivo. Humanin has been shown to extend the lifespan of that nematode by increasing autophagy.Lipid Intermediates

[0113] Lipids are key macromolecules that perform many biological functions ranging from maintaining structural integrity of membranes, energy storage, to signaling molecules. Variations in lipid composition and its levels can influence the functional and physiological state of the cell and its milieu. Recent studies have shown that the regulation of specific lipid species play a critical role in senescence. Moreover, some lipid species even contribute to the low-grade inflammation associated with SASP.Many protein regulators of senescence have been well characterized and are associated with lipid metabolism.

[0114] Applicants have discovered that four particular lipid intermediates can have therapeutic effects related to longevity. These include lipoxin A4 (LXA4), maresins (MaR1 / MaR2), neuroprotection D1.Lipoxin A4 (LXA4)

[0115] Lipoxin A4 (LXA4) and 15-epi-lipoxin A4 belong to the lipoxin family of eicosanoids, which are derived from arachidonic acid metabolism. LXA4 is a potent lipid mediator that plays a crucial role in resolving inflammation and promoting tissue repair.

[0116] LXA4 is an endogenous eicosanoid mediator that has anti-inflammatory and pro-resolving effects on various cell types. It is produced from arachidonic acid, and is expressed by inflammatory cells, especially monocytes and macrophages. LXA4 has been shown to: a) Suppress leukocyte-mediated injury b) Promote chemotaxis of monocytes c) Promote phagocytosis of apoptotic neutrophils d) Inhibit the production of proinflam matory cytokines and cell proliferation e) Inhibit TNF-a-stimulated neutrophil adherence to epithelial monolayers f) Function as putative braking signals for PMN neutrophilmediated tissue injury g) Reduce the degree of lung injury in ARDS rats h) Inhibit the release of pro-inflammatory factors TNF-a and IL-1 [3 in lung tissue homogenate i) Inhibit the production of reactive oxygen species (ROS) and neutrophil extracellular traps (NETs) in peripheral blood neutrophils of ARDS rats.Maresins

[0117] Maresins (MaR1 / MaR2 are specialized pro-resolving mediators that are synthesized from docosahexaenoic acid (DHA) by macrophages and have antiinflammatory and pro-resolving capacities as well as tissue regenerating and pain- relieving properties.

[0118] Maresins are a family of lipid mediators that are biosynthesized by macrophages and are important for: restoring tissue homeostasis, wound repair, reducing nerve sensitivity to painful stimuli, and resolving inflammation.

[0119] Maresins are generated from n-3 polyunsaturated fatty acids, such as: Eicosapentaenoic acid (EPA), Docosapentaenoic acid (DPA), and Docosahexaenoic acid (DHA).

[0120] Maresins are also known as specialized pro-resolving lipid mediators (SPMs). They are lipid mediators that interact with G-protein-coupled receptors and resolve the pro-inflammatory activities of other fatty acyls, such as prostaglandins. Maresins have anti-inflammatory and pro-resolving capacities, as well as tissue regenerating and pain-relieving properties. They play a critical role in: a) Initiating the pro-resolving functions of phagocytes b) Decreasing the magnitude of the overall inflammatory response c) Protecting against inflammation-related disorders d) Maresins may play pivotal roles in: e) Inflammatory diseases, Neurological disorders, Respiratory diseases, Diabetes and obesity, Kidney disease, Liver, Arthritis, Colitis, and Infectious diseases.Neuroprotectin D1

[0121] Protectins are signaling molecules that are produced enzymatically from unsaturated fatty acids. Their molecular structure is characterized by the presence of a conjugated system of double bonds.

[0122] Protectin D1 , also known as Neuroprotectin D1 (NPD1 ) or PD1 , is a lipid messenger that is produced from the omega-3 fatty acid docosahexaenoic acid (DHA). It is found in many tissues, including the retina, the lungs, and the nervous system. Protectin D1 has many effects, including: a) Reducing the occurrence of apoptosis induced by oxidative stress b) Conferring cell survival c) Inhibiting the activation of COX-2 and NF-K[3, which are genes that have pro- inflammatory characteristicsd) Reducing the infarct area in brain ischemia e) Increasing phagocytosis of apoptotic polymorphonuclear leukocytes (PMNs) by macrophages f) Decreasing PMN infiltration in a zymosan-induced mouse model of inflammation g) Decreasing pulmonary edemaProtectin D1 is also a specialized proresolving mediator that: Improves fin fold regeneration, Accelerates the resolution of inflammation, and Promotes macrophage polarization switch towards non-inflammatory states.

[0123] PD1 also influences various signaling pathways that regulate inflammation, cell survival, and apoptosis. For example, it activates the phosphatidylinositol 3-kinase (PI3K) / Akt pathway, which promotes cell survival and growth, and the extracellular signal-regulated kinase (ERK) pathway, which is involved in cell differentiation and survival. NPD1 regulates the expression of genes involved in inflammation, oxidative stress, and apoptosis. It can modulate transcription factors such as nuclear factor-kappa B (NF-KB) and activator protein-1 (AP-1 ), which play key roles in these processes.MicroRNA

[0124] MicroRNAs (miRNAs) belong to a class of small non-coding RNAs that are involved in development and diseases. miRNAs control gene expression by targeting mRNAs based on sequence complementarity. In addition to the dissection of their relevance for cellular processes, miRNAs also show great potential for diagnostic and therapeutic applications.

[0125] To identify miRNAs of potential regulatory, biologic, and / or therapeutic importance, Applicant employed an integrated approach that combined structural and functional genomic analyses. The Applicant compared analysis of expression of miRNAs and inversely correlated mRNAs from a validation data set. Integration of data and a functional screen of an miRNA library uncovered five miRNAs of particular relevance: MIR-145-3p, Let-7c-3p, MIR-548aj-3p, MIR-548aj-4p and hsa-MIR-548x-3p.

[0126] Embodiments include methods for prevention and treatment of senescence- associated diseases and disorders using pharmaceutical compositions that includespecific miRNAs. In aspects, the miRNAs include one or more of (a) MIR-145-3p, (b) let-7 c-3p, (c) MIR-383-5p, (d) MIR-548aj-3p, (e)MIR-548aj-4p and (e) MIR-548x-3p. In aspects, the miRNAs are administered using a lipid-based delivery (e.g., liposomes). In embodiments, the miRNAs are non-coding and act as repressors on one or more sites.

[0127] In contrast to small interfering RNA (siRNA), miRNA-targeted therapy can influence not just a single gene, but entire cellular pathways or processes. It is possible to supplement down regulated or non-functional miRNAs by synthetic oligonucleotides, as well as alleviating effects caused by overexpression of malignant miRNAs through artificial antagonists, either oligonucleotides or small molecules. Thus, in aspects, the miRNAs are non-coding and act as repressors on one or more sites.

[0128] In contrast to small interfering RNA (siRNA), miRNA-targeted therapy can influence not just a single gene, but entire cellular pathways or processes. It is possible to supplement down regulated or non-functional miRNAs by synthetic oligonucleotides, as well as alleviating effects caused by overexpression of malignant miRNAs through artificial antagonists, either oligonucleotides or small molecules. Thus, in aspects, the miRNAs are non-coding and act as repressors on one or more sites. hsa-mir-145-3p

[0129] MIR145 is a microRNA that has been found to be involved in various biological processes and diseases. Recent studies have shown that (human) hsa-miR- 145-3p is involved in two processes which are pivotal for the treatment of Alzheimer’s including (a) the epigenetic pattern age regression in neurons and astrocytes and (b) the reduction in both speed and volume of production of beta amyloid protein. hsa-let-7c-3p

[0130] Two major biological roles have been elucidated for the let-7 miRNA: as an essential regulator of terminal differentiation, and as a fundamental tumor suppressor. This non-coding RNA which reduces expression of Interleukin 8 (IL-8) via the CDH11- TGFp pathway. In Alzheimer’s IL-8 is involved upstream of Tau phosphorylation and neurofilament tangle (NFT) formation. Increased cerebrospinal fluid (CSF) levels of IL-8 are correlated with the amyloid A[3-42 / Ap-40 ratio, MMSE[WL1 ], Tau and pTau181. Inaspects, one or more of hsa-let-7c-3p5, hsa-let-7c-3p6 and hsa-let-7c-3p7 is administered. hsa-mir-548aj-3p and 4p

[0131] These microRNAs and differentially and have roles signaling pathways and tumor suppression. hsa-mir-548x-3p

[0132] Recent studies demonstrate that mir-548x-3p Improves overall functional survivability of Neurons. miR-548x can function as tumor suppressor genes in glioblastoma by controlling the PI3K / AKT signaling pathway and may act as gene therapy for clinical treatment of glioblastoma multiforme.

[0133] MicroRNAs (miRNAs) are small, regulatory RNAs, that regulate gene expression by repressing translation and / or break down mRNAs of their downstream target genes. miRNAs typically modulate gene expression (e.g., increase or decrease translation) by promoting cleavage of target mRNAs or by blocking translation of the cellular transcript. miRNAs are processed from primary transcripts known as pri-miRNA to short stem-loop structures called precursor (pre)-miRNA and finally to functional, mature miRNA. Mature miRNA molecules are partially complementary to one or more messenger RNA molecules, and their primary function is to down-regulate gene expression. Mis-regulation of miRNAs in the central nervous system can contribute to neurodegenerative disorders. Applicants have discovered differential expression levels of various miRNAs in senescent cells and aging adults. These miRNAs can serve as therapeutic targets for promoting longevity and preventing / treating senescence- associated diseases and disorders. miRNA Delivery Vehicles

[0134] Therapeutics that utilize miRNA can require efforts to promote stability and / or delivery of miRNA to the target site. To arrive at a target site they must be resistant to nuclease degradation in the extracellular space. To resolve this problem, chemical modifications of oligonucleotides can be designed to enhance delivery efficiency. Presently, there are ten FDA-approved oligonucleotide drugs, most of whichare chemically modified. These drugs are mostly delivered locally or to the liver. The nucleic acid backbone, ribose sugar moiety and nucleobase itself can all be chemically modified to enhance delivery using locked nucleic acids (LNAs), 5' -(E)- vinylphosphonate modification and pyrimidine methylation. Cobomarsen is an oligonucleotide with chemical modification (partially LNA, full PS backbone) that does not require an additional delivery system. However, chemical modification of miRNA inhibitors cannot target specific tumour sites in vivo. Hence, covalent conjugation of specific moieties to miRNAs, mimetics or inhibitors promotes the uptake of oligonucleotide drugs to specific tissues. The moieties vary, including peptides, antibodies, aptamers and sugars. For example, N-acetylgalactosamine (GalNAc)- conjugated miR-122 was developed to treat hepatitis C virus infection of the liver (Miravirsen, Roche, Switzerland). GalNAc specifically binds to the Asialoglycoprotein receptor on the cell surface of hepatocytes. The interaction between GalNAc and the Asialoglycoprotein receptor leads to the endocytosis of GalNAc-conjugated oligonucleotide drugs. Asialoglycoprotein receptor is highly expressed on hepatocytes, so GalNAc-conjugated oligonucleotides can be specifically taken up by hepatocytes. It should be noted that chemical modification might prevent miRNA mimetic recognition and loading into Argonaute and RISC.

[0135] In addition to the conjugation of various moieties, miRNAs, miRNA mimetics or inhibitors can also be packaged into nanoparticles to increase uptake efficiency. Advances in nanotechnology and material science present versatile solutions for oligonucleotide drug delivery. The most commonly used nanotechnology for nucleic acid drugs is lipid formulations. MRX34 is a double-stranded miR-34 mimetic in liposome nanoparticles. Pharmacodynamic data indicated that MRX34 suppressed miR-34 targets in the white blood cells of enrolled patients.

[0136] Among those, lipid-based delivery is a popular approach. The classical approach consists of a mixture of lipids with cationic head groups and helper lipids, including some with polyethylene glycol chains for masking of the surface charge. Polyanionic nucleic acids are electrostatically complexed to the cationic lipid, yielding lipoplexes. A high degree of optimization of those formulations, both in terms ofstructures and multi-component compositions, has been achieved, and loading capacity and delivery efficiency have been increased to considerably lower the dose necessary for functional effects. However, they still suffer from inherent toxicity, which is, like uptake and re-release efficiency, closely linked to the cationic surface charge.Examples of the in vivo use of lipoplexes include the delivery of miR-133b and miR-29b with a mixture of DOTMA, cholesterol and a PEG lipid, pre-miR-107 with DDAB, cholesterol and PEG lipids, and the use of solid lipid nanoparticles consisting of DDAB, cholesterol and other components for delivery of miR-34a.Polymers

[0137] The cationic polymer polyethylene imine (PEI) is the most widely used polymeric delivery system for plasmid DNA and siRNA. Efficient packaging and a net cationic charge ensure adequate shielding and sufficient interaction with anionic polysaccharides on cell membranes. PEI is thought to increase endosomal escape by the proton sponge effect, an influx of hydrogen ions into acidic endosomes, resulting in swelling and disruption of the intracellular vesicles. Thus, PEI has quickly been adopted for delivery of miRNA mimics.

[0138] PEI can also be used as carrier system for targeted delivery by attaching specific ligands to the polymer. For an miRNA application, the rabies peptide RVG was attached to PEI for transport to and across the blood-brain barrier. In mice, fluorescently tagged miR-124a was found in higher accumulation in the brain compared to underivatized PEI. However, no functional effects were reported, and mannitol was necessary for sufficient blood-brain barrier permeabilization, which limits the therapeutic utility.

[0139] Poly(lactic-co-glycolic acid) (PLGA) is a polymer that has been utilized for antisense and siRNA delivery. PLGA needs to be coated or functionalized for efficient oligonucleotide delivery, but affords long-term dissociation from the carrier for a prolonged effect. While no studies with miRNA mimics delivered with PLGA have been reported, oligonucleotides for antagonizing miRNAs have been successfully delivered to tumours. Coating with cationic peptides nona-arginine or penetratin afforded passivetumour accumulation of an antisense peptide nucleic acid targeted at miR-155 in lymphoma in vivo.

[0140] The use of targeted silica nanoparticles for miRNA delivery resulted in reduction of neuroblastoma growth. The oligonucleotide cargo is noncovalently entrapped in the silica matrix, and dissociates upon hydrolysis of the matrix. Since this anorganic carrier will not be taken up into cells on its own, a receptor-targeting ligand is necessary to achieve intracellular delivery. Grafting an antibody against the cell surface antigen disialoganglioside GD2 afforded successful delivery of miR-34a into neuroblastoma cells in a murine xenograft model.Conjugates

[0141] Conjugation of lipids or receptor-binding molecules directly to the nucleic acid is a promising way to increase cellular uptake of siRNA, and has also been explored for miRNA applications. By attaching cholesterol to the 3'-end of the passenger strand, an accumulation in liver tissue can be achieved. The restriction to liver targeting and the high doses needed for sufficient delivery limit the use of cholesterol- and similar conjugates to scientific rather than therapeutic applications.

[0142] The asialoglycoprotein receptor ligand N-acetylgalactosamine (GalNAc) has been intensively used for targeting siRNA and antisense oligonucleotides to hepatocytes

[0012] , with several agents already advanced to clinical evaluation. For achieving hepatocyte delivery of an anti-miR-122, a covalent conjugate with GalNAc has been developed. The respective compound RG-101 , developed for treatment of HCV infections, has recently begun testing in human volunteers

[0156] , In preclinical animal models, RG-101 showed efficient reduction of viral titers, and a good safety profile.Exosomes and Bacteriophages

[0143] Circulating miRNAs are found in body fluids (plasma, saliva, etc.) and are exchanged between cells despite the abundance of nucleases throughout the body. Natural shielding of endogenous miRNAs is afforded through extracellular vesicles, called exosomes. They are small membrane vesicles (up to 100 nm), and are producedby many cell types, including epithelial, dendritic, and immune cells. Lately, exosomes have been used to encapsule and deliver synthetic or endogenously expressed siRNAs and miRNAs in vivo. The oligonucleotide cargo can be introduced by transfection of corresponding plasmid into exosome-producing cells, or synthetic oligonucleotides can be inferred through electroporation of the mature exosomes. For miRNA delivery, transfection of exosome-producing HEK293 cells with synthetic let-7 was employed to produce miRNA containing exosomes. A peptide binding to the EGF-receptor was introduced by means of a peptide-encoding plasmid. Exosomes distribute preferentially to the reticuloendothelial system (RES), and thus the spleen and liver, but are also found in other organs. Targeting can be achieved through specific ligands, which can be expressed through genetic engineering methodology. Using the RVG peptide, siRNA-containing exosomes were even shown to cross the blood-brain barrier. Targeting and biodistribution of exosomes is also dependent on the cell type they are isolated from. Exosomes are similar to liposomes in terms of consisting of bilayered phospholipids, but the biogenesis of exosomes ensures their biocompatibility and low toxicity. It also significantly complicates pharmaceutical development, production and safety profiling (immunogenicity, and potential biological impurities).

[0144] Bacteriophages have been used to develop virus-like particles for oligonucleotide and drug delivery. The MS2 bacteriophage was modified to produce particles with covalent linkage to pre-m iR-146a. After grafting the HIV-TAT peptide to the particles, the system induced a two-fold higher expression of m iR-146a in vivo. A similar approach used RNA from bacteriophage Phi29 for packaging miRNAs targeted at coxsackievirus B3. Conjugation of folic acid was used for folate-specific cellular uptake to result in reduction of viral replication in an in vitro model. Similar to exosomes, concerns of immune responses triggered by virus-like particles need to be addressed before further development of bacteriophage-derived particles into clinical evaluation. lnterlueken-11

[0145] lnterleukin-11 (IL-11 ), a cytokine produced by bone marrow mesenchymal cells, has an effect on hematopoietic, adipocytic, epithelial, osseous, and cartilaginoustissues. IL-11 stimulates cell migration via a PI3K / AKT and nuclear factor (NF)-KP- dependent pathway, ultimately leading to increased expression of the cell adhesion molecule, ICAM-1.

[0146] Recent studies have demonstrated a link between age and expression of IL- 11 . After about the age of 55 more IL-11 is produced in humans. The increased levels have been linked to chronic inflammation, fibrosis in organs, disorders of metabolism, muscle wasting (sarcopaenia), frailty and cardiac fibrosis. Studies have shown that blocking IL-11 can reduce the occurrence of senescent related ailments and / or promote longevity.

[0147] Accordingly, embodiments include the use of an antibody or antagonist against IL-11 . Commercially available inhibitors (i.e. , blockers) include anakinra, canakinumab, and rilonacept. In aspects, the inhibitor binds to IL-11 and / or blocks its activity or its activity pathway. In aspects, the inhibitor reduces expression of IL-11 .EXAMPLES

[0148] The following non-limiting examples are provided for illustrative purposes only in order to facilitate a more complete understanding of representative embodiments now contemplated. These examples are intended to be a mere subset of all possible contexts in which the components of the formulation may be combined. Thus, these examples should not be construed to limit any of the embodiments described in the present specification, including those pertaining to the type and amounts of components of the formulation and / or methods and uses thereof.Example 1Treatment for Longevity and Senescence-related Ailments

[0149] In this example, a patient (58-year-old male) visits a physician’s clinic and presents signs and symptoms of atherosclerosis and hypertension. The patient also appears overweight with high blood pressure (i.e., 130 / 90). A healthcare provider suspects that a therapy aimed at longevity and removal of senescent cells will improve the patient's cardiovascular ailments.In support of this theory we have tested GRAS and prescription compounds with similar activities or that directly interact with our target pathways. In order to execute on this we have developed a protocol whereby NAD+ and repair pathway based supplements are taken in the morning, senomorphics are taken post workout and senolytics are taken in the evening. This is combined with exercise and nociceptive stimulus including cold plunge.

[0150] As shown in FIG. 2, the patient is provided a treatment plan that includes compounds for consumption in the morning and evening. In this example, the morning routine includes supplements that can promote DNA repair pathways. The evening routine includes supplements that act as senolytic compounds.

[0151] Specifically, the following compounds that are consumed within one hour after awakening: NAD+, urolithin A, DHA and TMG. The following compounds are consumed in the evening (i.e. , within one year of going to sleep): resveratrol, spermidine, quercetin, fisetin.

[0152] The patient is also advised to improve his lifestyle with attention toward exercise, diet and stress. Resveratrol is also consumed after exercise and / or nociceptive stimulus (e.g., ice-bath immersion). The patient also practices intermittent fasting. After two months, the patient reports improvements in muscle recovery, neurocognitive functions and athletic performance. The patient also presents a healthier BMI (with loss of abdominal fat) and a healthy blood pressure (i.e., 120 / 80).Methods of Use

[0153] Embodiments include methods of improving longevity and / or treating an ailment (i.e., a senescence-associated disease or disorder). The methods can include therapies for treating a senescence-associated disease or disorder. In one embodiment, a method includes administering to a pharmaceutical formulation containing a therapeutic agent that selectively kills senescent cells (i.e., selectively kills senescent cells over non-senescent cells). A treatment regimen can include administering a pharmaceutical formulation for a time sufficient and in an amount sufficient to selectively kill senescent cells.

[0154] In aspects, one or more compounds disclosed herein are incorporated into a formulation. For example, a first (e.g., AM) formulation can include NAD (or NAD precursors), urolithin A, DHA and TMG. A second (e.g., PM) formulation can include resveratrol, spermidine, quercetin and fisetin.

[0155] Without being bound be theory, Applicants propose that Naringenin and analogs are also involved in these cascades, in combination with NAD+ which can repair or perform spliceosome products. TRPM8, defines a small and discrete population of sensory afferents that innervate tissues known to be highly sensitive to cold and nociceptive stimuli. TRPM8 can also affect the various transcription regulators, in conjunction with NAD+ and other sirtuin effecting small molecule precursors and pro drugs including nicotinamide mononucleotide and nicotinamide rhiboside. Melastatin 3 (TRPM3) channels play roles in heat hypersensitivity and spontaneous pain after nerve injury. Control of insulin release by transient receptor potential Melastatin 3 (TRPM3) Ion Channels. The TRPM3 ion channel mediates nociception but is not evoked by endogenous pruritogenic mediators. PPARy agonism by naringenin and its analogs is involved by browning of adipose tissue thereby altering the metabolic state of these and surrounding cells.

[0156] Each of the compounds described herein can be present in an amount from about 0.0001 mg, about 0.0005 mg, about 0.0006 mg, about 0.001 mg, about 0.003 mg, about 0.006 mg, about 0.01 mg, about 0.03 mg, about 0.06 mg, about 0.1 mg, about 0.3 mg, about 0.6 mg, about 1 mg, about 2 mg, about 3 mg, about 5 mg, about 6 mg, about 10 mg, about 30 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 300 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000 mg, about 2000 mg, about 3000 mg, about 5000 mg, or about 10,000 mg. As expected, the dosage will be dependent on the condition, size and age of the patient or subject.

[0157] In aspects, an effective amount of the proposed compounds of the present invention is administered to a subject or a patient in the range of from about 1 ng / kg body weight to about 10 mg / kg body weight, about 1 ng / kg body weight to about 1mg / kg body weight, about 1 ng / kg body weight to about 100 g / kg body weight, about 1 ng / kg body weight to about 10 g / kg body weight, about 1 ng / kg body weight / day to about 1 g / kg body weight, about 1 ng / kg body weight to about 100 ng / kg body weight, about 1 ng / kg body weight to about 10 ng / kg body weight, about 10 ng / kg body weight to about 100 mg / kg body weight, about 10 ng / kg body weight to about 10 mg / kg body weight, about 10 ng / kg body weight to about 1 mg / kg body weight, about 10 ng / kg body weight / to about 100 g / kg body weight, about 10 ng / kg body weight to about 10 mg / kg body weight, about 10 ng / kg body weight to about 1 mg / kg body weight, 10 ng / kg body weight to about 100 ng / kg body weight, about 100 ng / kg body weight to about 100 mg / kg body weight, about 100 ng / kg body weight to about 10 mg / kg body weight, about 100 ng / kg body weight to about 1 mg / kg body weight, about 100 ng / kg body weight to about 100 mg / kg body weight, about 100 ng / kg body weight to about 10 mg / kg body weight, about 100 ng / kg body weight to about 1 mg / kg body weight, about 1 mg / kg body weight to about 10 mg / kg body weight, about 1 mg / kg body weight to about 100 mg / kg body weight, about 1 mg / kg body weight / day to about 5 mg / kg body weight / day, about 1 mg / kg body weight to about 10 mg / kg body weight, about 1 mg / kg body weight / day to about 100 mg / kg body weight / day, about 5 mg / kg body weight / day to about 10 mg / kg body weight / day, about 10 mg / kg body weight / day to about 50 mg / kg body weight / day, about 10 mg / kg body weight / day to about 100 mg / kg body weight / day, about 10 mg / kg body weight / day to about 1 g / kg body weight / day, about 1 mg / kg body weight to about 1 g / kg body weight / day, about 10 mg / kg body weight / day to about 10 g / kg body weight / day, about 1 mg / kg body weight / day to about 10 mg / kg body weight / day, about 0.1 mg / kg body weight to about 10 mg / kg body weight, about 0.1 mg / kg body weight / day to about 10 mg / kg body weight / day, about 1 mg / kg body weight to about 1 g / kg body weight, about 0.01 mg / kg body weight to about 100 mg / kg body weight.

[0158] In some embodiments, the formulations and methods described herein promote longevity by changing the microbiome in an aged subject, decreasing fibrosis, decreasing inflammation, decreasing inflammatory response in an aged subject, and decreasing production of reactive oxidation species (ROS). In some embodiments, the formulations and methods decrease fibrosis. In some embodiments, the formulations and methods increase muscle mass. In some embodiments, the formulations andmethods increase stem cell self-renewal. In some embodiments, the stem cells are neuronal stem cells. In some embodiments, the stem cells are satellite cells. In some embodiments, the stem cells are muscle stem cells. In some embodiments, the formulations and methods improve glucose homeostasis. In some embodiments, the formulations and methods increase cognitive function. In some embodiments, the formulations and methods improve memory. In some embodiments, the formulations and methods decrease progerin levels, SRSF1 levels or both. In some embodiments, the formulations and methods increase chondrocyte survival. In some embodiments, the formulations and methods treat cancer. In some embodiments, the formulations and methods reduce the risk of developing cancer.

[0159] In embodiments, the formulations and methods described herein increase life expectancy. In embodiments, the formulations and methods decrease the senescent cell burden in a subject (e.g., by about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80% or about 90% or more). In aspects, the formulations and methods reduce the incidence of one or more ailments associated with aging. In aspects, the formulations and methods increase telomere length and / or impede the shortening of telomeres.

[0160] In one embodiment, a therapeutic composition or treatment disclosed herein is capable of reducing the signs / symptoms of a senescence-associated disease or disorder by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% as compared to a patient not receiving the same treatment. In other aspects of this embodiment, a therapeutic is capable of reducing the signs / symptoms of a senescence-associated disease or disorder in an individual by, e.g., about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 90%, about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% toabout 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70% as compared to a patient not receiving the same treatment.

[0161] In one embodiment, a therapeutic composition or treatment disclosed herein is capable of reducing signs / symptoms of aging in an individual by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% as compared to a patient not receiving the same treatment. In other aspects of this embodiment, a therapeutic is capable of reducing signs / symptoms of aging in a subject by, e g., about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 90%, about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% to about 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70% as compared to a patient not receiving the same treatment.

[0162] In one embodiment, a therapeutic composition or treatment disclosed herein is capable of extending the lifespan of an individual by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% as compared to a patient not receiving the same treatment. In other aspects of this embodiment, a therapeutic is capable of extending the lifespan of an individual by, e.g., about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 90%, about 20% to about 90%, about 30% to about 90%,about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% to about 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70% as compared to a patient not receiving the same treatment.

[0163] Certain embodiments of the present invention are described herein, including the best mode known to the inventors for carrying out the invention. Of course, variations on these described embodiments will become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventor expects skilled artisans to employ such variations as appropriate, and the inventors intend for the present invention to be practiced otherwise than specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described embodiments in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

[0164] Groupings of alternative embodiments, elements, or steps of the present invention are not to be construed as limitations. Each group member may be referred to and claimed individually or in any combination with other group members disclosed herein. It is anticipated that one or more members of a group may be included in, or deleted from, a group for reasons of convenience and / or patentability. When any such inclusion or deletion occurs, the specification is deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.

[0165] Unless otherwise indicated, all numbers expressing a characteristic, item, quantity, parameter, property, term, and so forth used in the present specification and claims are to be understood as being modified in all instances by the term “about.” As used herein, the term “about” means that the characteristic, item, quantity, parameter,property, or term so qualified encompasses a range of plus or minus ten percent above and below the value of the stated characteristic, item, quantity, parameter, property, or term. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and attached claims are approximations that may vary. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical indication should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and values setting forth the broad scope of the invention are approximations, the numerical ranges and values set forth in the specific examples are reported as precisely as possible. Any numerical range or value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements. Recitation of numerical ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate numerical value falling within the range. Unless otherwise indicated herein, each individual value of a numerical range is incorporated into the present specification as if it were individually recited herein.

[0166] The terms “a,” “an,” “the” and similar referents used in the context of describing the present invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein is intended merely to better illuminate the present invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the present specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[0167] Specific embodiments disclosed herein may be further limited in the claims using consisting of or consisting essentially of language. When used in the claims, whether as filed or added per amendment, the transition term “consisting of” excludes any element, step, or ingredient not specified in the claims. The transition term“consisting essentially of” limits the scope of a claim to the specified materials or steps and those that do not materially affect the basic and novel characteristic(s). Embodiments of the present invention so claimed are inherently or expressly described and enabled herein.

[0168] Groupings of alternative embodiments, elements, or steps of the present invention are not to be construed as limitations. Each group member may be referred to and claimed individually or in any combination with other group members disclosed herein. It is anticipated that one or more members of a group may be included in, or deleted from, a group for reasons of convenience and / or patentability. When any such inclusion or deletion occurs, the specification is deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.

[0169] All patents, patent publications, and other publications referenced and identified in the present specification are individually and expressly incorporated herein by reference in their entirety for the purpose of describing and disclosing, for example, the compositions and methodologies described in such publications that might be used in connection with the present invention. These publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representation as to the contents of these documents is based on the information available to the applicants and does not constitute any admission as to the correctness of the dates or contents of these documents.

[0170] In closing, it is to be understood that although aspects of the present specification are highlighted by referring to specific embodiments, one skilled in the art will readily appreciate that these disclosed embodiments are only illustrative of the principles of the subject matter disclosed herein. Therefore, it should be understood that the disclosed subject matter is in no way limited to a particular methodology, protocol, and / or reagent, etc., described herein. As such, various modifications or changes to oralternative configurations of the disclosed subject matter can be made in accordance with the teachings herein without departing from the spirit of the present specification. Lastly, the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention, which is defined solely by the claims. Accordingly, the present invention is not limited to that precisely as shown and described.

Claims

1. CLAIMSWhat is claimed is:1 . A formulation comprised of: a) one or more transcription regulators, b) one or more lipid intermediates, and c) one or more microRNAs.

2. The formulation of claim 1 , wherein the one or more transcription regulators target one or more genes are selected from MEG3, IRISIN, MOTS-C and HUMANIN.

3. The formulation of claim 1 , wherein the one or more lipid intermediates are selected from lipoxin A4 (LXA4), maresins (MaR1 / MaR2), neuroprotection D1.

4. The formulation of claim 1 , wherein the one or more microRNAs are selected from MIR-145 3p, Let-7c-3p, MIR-548aj-3p, MIR-548aj-4p and hsa-MIR-548x-3p.

5. The formulation of claim 1 , further comprising a senolytic agent.

6. The formulation of claim 7, wherein the senolytic agent is selected from one or more of Navitoclax, Venetoclax, Dasatinib / Quercetin, Fisetin, 17-DMAG / IPI504, UBX0101 / FGX04-DRI peptide, cardiotonic steroids, GMD / SSK1 , erastin, vacuolin-1 and apilimod.

7. The formulation of claim 1 , further comprising a senomorphic agent.

8. The formulation of claim 5, wherein the senomorphic agent is selected from one or more of Fisetin, Metformin, Rapamycin, Resveratrol, Quercetin, Dasatinib, Kinase inhibitors, an inhibitor of IkB kinase (IKK), an inhibitor of nuclear factor (NF)-kB5, a free radical scavenger and a Janus kinase (JAK) pathway inhibitor.

9. A method of slowing the aging process or reducing signs of aging in a subject, the method comprising administering a therapeutic amount of the formulation of claim 1 to the subject.

10. A method of treating an ailment in a subject, the method comprised of administering a therapeutic amount of the formulation of claim 1 to the subject.11 . The method of claim 10, wherein the ailment is a senescence-associated disease or disorder.

12. The method of claim 11 , wherein the senescence-associated disease or disorder is one or more of atherosclerosis, osteoarthritis, osteoporosis, hypertension, arthritis, cataracts, cancer, Alzheimer’s disease, chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis, hair graying, sarcopenia, adiposity, neurogenesis, fibrosis and glaucoma.

13. A method of slowing the aging process or reducing signs of aging, the method comprising administering a therapeutic amount of the formulation of claim 1 .

14. A method of treating an ailment in a subject, the method comprised of administering a formulation comprised of: a) one or more transcription regulators, b) one or more lipid intermediates, and c) one or more microRNAs.

15. The method of claim 14, wherein the ailment is a senescence-associated disease or disorder.

16. The method of claim 15, wherein the senescence-associated disease or disorder is one or more of atherosclerosis, osteoarthritis, osteoporosis, hypertension, arthritis, cataracts, cancer, Alzheimer’s disease, chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis, hair graying, sarcopenia, adiposity, neurogenesis, fibrosis and glaucoma.

17. The method of claim 14, wherein the one or more transcription regulators are selected from MEG3, IRISIN, MOTS-C and HUMANIN.

18. The method of claim 14, wherein the one or more lipid intermediates are selected from lipoxin A4 (LXA4), maresins (MaR1 / MaR2), neuroprotection D1 and protecting D1 / NPD1 / PD1.

19. The method of claim 1 , wherein the one or more microRNAs are selected from MIR-145 3p, Let-7 c-3p, MIR-548aj-3p and hsa-MIR-548x-3p.

20. The method of claim 14, further comprising administering a senolytic agent.21 . The method of claim 20, wherein the senolytic agent is selected from one or more of Navitoclax, Venetoclax, Dasatinib / Quercetin, Fisetin, 17-DMAG / IPI504, UBX0101 / FOX04-DRI peptide, cardiotonic steroids, GMD / SSK1 , erastin, vacuolin-1 and apilimod.

22. The method of claim 14, further comprising administering a senomorphic agent.

23. The method of claim 22, wherein the senomorphic agent is selected from one or more of Fisetin, Metformin, Rapamycin, Resveratrol, Quercetin, Dasatinib, Kinase inhibitors, an inhibitor of IkB kinase (IKK), an inhibitor of nuclear factor (NF)-kB5, a free radical scavenger and a Janus kinase (JAK) pathway inhibitor.

24. A formulation comprised of: a) nicotinamide adenine dinucleotide (NAD), b) urolithin A, c) docosahexaenoic acid (DHA), and d) trimethylglycine (TMG).

25. The formulation of claim 24, wherein the formulation is further comprised of: e) resveratrol, f) spermidine, g) quercetin and h) fisetin.

26. The formulation of claim 24, further comprising a senolytic agent.

27. The formulation of claim 24, wherein the senolytic agent is selected from one or more of Navitoclax, Venetoclax, Dasatinib / Quercetin, Fisetin, 17-DMAG / IPI504, UBX0101 / FQX04-DRI peptide, cardiotonic steroids, GMD / SSK1 , erastin, vacuolin-1 and apilimod.

28. The formulation of claim 27, further comprising a senomorphic agent.

29. The formulation of claim 28, wherein the senomorphic agent is selected from one or more of Fisetin, Metformin, Rapamycin, Resveratrol, Quercetin, Dasatinib, Kinase inhibitors, an inhibitor of IkB kinase (IKK), an inhibitor of nuclear factor (NF)-kB5, a free radical scavenger and a Janus kinase (JAK) pathway inhibitor.

30. A method of slowing the aging process or reducing signs of aging in a subject, the method comprising administering a therapeutic amount of the formulation of claim 24 or 25 to the subject.31 . A method of treating an ailment in a subject, the method comprised of administering a therapeutic amount of the formulation of claim 24 or 25 to the subject.

32. The method of claim 31 , wherein the ailment is a senescence-associated disease or disorder.

33. The method of claim 32, wherein the senescence-associated disease or disorder is one or more of atherosclerosis, osteoarthritis, osteoporosis, hypertension, arthritis, cataracts, cancer, Alzheimer’s disease, chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis, hair graying, sarcopenia, adiposity, neurogenesis, fibrosis and glaucoma.

34. A method of slowing the aging process or reducing signs of aging, the method comprising administering a therapeutic amount of the formulation of claim 24 or 25.

35. A method of slowing the aging process or reducing signs of aging in a subject, the method comprising administering a therapeutic amount of a formulation comprised of:a) nicotinamide adenine dinucleotide (NAD), b) urolithin A, c) docosahexaenoic acid (DHA), and d) trimethylglycine (TMG), wherein the formulation is administered in the morning.

36. The method of claim 35, further comprising administering a second formulation comprised of: e) resveratrol, f) spermidine, g) quercetin and h) fisetin, wherein the formulation is administered in the evening.

37. The method of claim 35, further comprising administering a therapeutic amount of resveratrol following exercise.

38. A method of treating an ailment in a subject, the method comprising administering a therapeutic amount of a formulation comprised of: a) nicotinamide adenine dinucleotide (NAD), b) urolithin A, c) docosahexaenoic acid (DHA), and d) trimethylglycine (TMG), wherein the formulation is administered in the morning.

39. The method of claim 38, further comprising administering a second formulation comprised of: e) resveratrol, f) spermidine, g) quercetin and h) fisetin, wherein the formulation is administered in the evening.

40. The method of claim 38, wherein the ailment is a senescence-associated disease or disorder.41 . The method of claim 40, wherein the senescence-associated disease or disorder is one or more of atherosclerosis, osteoarthritis, osteoporosis, hypertension, arthritis, cataracts, cancer, Alzheimer’s disease, chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis, hair graying, sarcopenia, adiposity, neurogenesis, fibrosis and glaucoma.

42. The method of claim 38, further comprising administering a therapeutic amount of resveratrol following exercise.

43. A method of slowing the aging process or reducing signs of aging in a subject, the method comprising administering a therapeutic amount of a formulation comprised of: a) nicotinamide adenine dinucleotide (NAD), b) one or more DNA repair-pathway based supplements, and c) one or more senolytic agents.

44. The method of claim 43, wherein the NAD and the one or more DNA repairpathway based supplements administered in the morning.

45. The method of claim 43, wherein the one or more senolytic agents are administered in the evening.

46. The method of claim 43, wherein the one or more DNA repair-pathway based supplements are selected from Urolithin A, DHA and TMG.

47. The method of claim 43, wherein the one or more senolytic agents are selected from resveratrol, spermidine, quercetin, fisetin.

48. The method of claim 43, further comprising a step of administering a senomorphic agent after exercise.

49. The method of claim 43, wherein the senomorphic agent is resveratrol.

50. The method of claim 43, further comprising a step of administering a nociceptive stimulus.51 . The method of claim 50, wherein the nociceptive stimulus is immersion in an icebath or cold water.

52. The method of claim 43, further comprising a step of intermittent fasting.

53. The method of claim 10 or claim 35, further comprising administering an IL-11 antagonist or an antibody against IL-11 .

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