Treatment of polysubstance use disorder

Abstract

A method of treating a substance use disorder (SUD) in a subject, comprising administering a therapeutically effective amount of an oral enhanced NAD+ pharmaceutical formulation to the subject.

Description

TREATMENT OF POLYSUBSTANCE USE DISORDER

[0001] All patents, patent applications and publications cited herein are hereby incorporated by reference in their entirety. Such patent applications include but are not limited to (i) PCT / US23 / 35557, entitled “Enhanced NAD+ Compositions and Methods of Making and Using the Same,” (ii) PCT / US25 / 43556, entitled “IMPROVING IMMUNE FUNCTION WITH ENHANCED NAD+ COMPOSITIONS,” and (lii) PCT / US2025 / 035967, entitled “ENHANCED NAD+ COMPOSITIONS AND METHODS OF MAKING AND USING THE SAME.”

[0002] The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art as know n to those skilled therein as of the date of the invention described and claimed herein.

[0003] This patent disclosure contains material that is subject to copyright protection. The copyright owner has no objection to the facsimile reproduction by anyone of the patent document or the patent disclosure as it appears in the U.S. Patent and Trademark Office patent file or records, but otherwise reserves any and all copyright rights.CROSS-REFERENCE TO RELATED APPLICATIONS

[0004] This application claims priority to U.S. Provisional Application No. 63 / 759,977, entitled “TREATMENT OF POLYSUBSTANCE USE DISORDER,” filed Febniary 18, 2025, the contents of which is incorporated herein by reference in its entirety.FIELD OF THE INVENTION

[0005] The present invention is in the field of medicine, and more particularly addiction- and detoxification-related disorders including PSUD and SUD.BACKGROUND OF THE INVENTION

[0006] It is estimated that between 35% and 75% of individuals suffering from a substance use disorder (“SUD”) take and are addicted to more than one substance, and thus are affected with “polysubstance use disorder “(PSUD). PSUD can refer to a disorder in which an individual uses or takes and are addicted to at least tw o different drugs, including prescription and / or illegal drugs. The affected person may take the drugs together or separately within a short timeperiod and suffers ill effects from the combination. The multiple drugs may be taken intentionally or unintentionally. Intentional polysubstance use occurs when a person takes a drug to increase or decrease the effects of a different drug or wants to experience the effects of the combination. Unintentional polysubstance use occurs when a person takes drugs that have been mixed or cut with other substances, like fentanyl, without their knowledge. Polysubstance use may be more dangerous than monosubstance use because the effects from combining drugs may be stronger and more unpredictable than one drug alone. In certain instances, such a combination can be deadly. Indeed, PSUD is associated with an increased risk of concurrent mental disorders, overdose, and unintentional death. Among patients whose use disorder is at least moderately severe or who are receiving treatment, the PSUD vs. SUD frequency estimates stand at 3-to-l ratio. For example, up to 80% of opioid-related deaths in the US occur in individuals who have also taken another substance. Additionally, up to 80% of illicit drug supply in the US may be contaminated with adulterants (e.g., fentanyl, xylazine), suggesting that, whether intentional or not, PSUD is likely clinically present in many cases of SUD requiring treatment.

[0007] Drug metabolism is one of the pathways altered in predisposition for PSUD and, thus, represents one pharmaceutical target for treatment of PSUD.

[0008] Withdrawal is a step for initiating behavioral drug-free treatment and is an endpoint of long-term substitution treatment paradigms. The acute effects of withdrawal can be severe and life-threatening, necessitating careful management of the detoxification process. The heterogeneity of PSUD significantly complicates the detoxification process due to the overlapping effects of multiple substances. Considerations not only include the pharmacodynamics of the substances involved, but also the biological repercussions of such use, such as, but not limited to. excitotoxicity, neuroadaptation, endothelial injury, liver damage, oxidative and nitrosative stress, inflammation, and metabolic dysregulation. These systems are altered in SUDs and PSUDs and exacerbate the challenges of both acute withdrawal and chronic disease management.

[0009] Unfortunately, clinical studies of PSUD have been limited to date, and no pharmaceutical approaches have been approved for treatment of PSUD in the U.S. This can be attributed, in part, to the treatment of PSUD being particularly challenging due to the complexity conferred by genetic predispositions to addiction, comorbid psychiatric conditions, acute effects of the substance(s). chronic and neuroadaptive changes that follow repeated use, and acute effects of use cessation, withdrawal, and relapse. The mechanisms of action ofdifferent drug substances on the brain and body vary' depending on the substance used but are primarily mediated by alterations in neurotransmitter levels and changes in the corresponding brain receptors’ activity, ultimately leading to profound and lasting changes in receptor density, long-term potentiation and long-term depression, and overall balance of excitatory vs. inhibitory' signaling.

[0010] Thus. PSUD and SUD present unique problems in acute management settings, and methods of treatment are urgently needed.SUMMARY OF THE INVENTION

[0011] It has been discovered that LNAD+, an oral formulation of enhanced NAD+, when administered, significantly increases intracellular levels of NAD+ and primary' NAD+ metabolites, as well as causes changes in several multi-omic biomarkers. These findings demonstrate that administered LNAD is bioavailable. and its safety and identified multi-omic biomarkers of relevance support LNAD as a treatment of SUD and PSUD. These findings have been exploited to develop the present disclosure, which in part is directed to methods of reducing and relieving the symptoms of detoxification after poly substance and monosubstance abuse. This discovery has been exploited to provide the instant disclosure, which includes a method of treating PSUD and SUD.

[0012] In one aspect, a method of treating a substance use disorder (SUD) in a subject is disclosed herein. In examples, the method comprising administering a therapeutically effective amount of an enhanced NAD+ pharmaceutical formulation to the subject.

[0013] In another aspect, the present disclosure relates to a method of treating poly substance use disorder (PSUD) in a subject, the method comprising administering a therapeutically effective amount of an oral enhanced NAD+ formulation to the subject.

[0014] In yet another aspect, the present disclosure provides a method of treating one or more withdrawal symptoms in a subject recovering from substance use disorder (SUD) or polysubstance use disorder (PSUD), the method comprising administering a therapeutically effective amount of an oral enhanced NAD+ formulation to the subject. In particular examples, the enhanced NAD+ is LNAD.

[0015] The various methods described herein can comprise administering a maintenance dose of the enhanced NAD+ pharmaceutical formulation to the subject for a maintenance period. Incertain examples, the maintenance dose is less than or equal to the therapeutically effective amount. The maintenance period can be least about one week.

[0016] In certain examples, the enhanced NAD+ pharmaceutical formulation or the oral enhanced NAD+ formulation is a mouthwash, and the step of administering the therapeutically effective amount of the enhanced NAD+ pharmaceutical formulation comprises instructing the subject to intake a quantity of the mouthwash into the subject’s mouth and perform a swishing motion for less than about one minute.

[0017] In various examples, the SUD is a polysubstance use disorder (PSUD). The PSUD can be an opioid use disorder. In various embodiments, the administration of therapeutically effective amount of an enhanced NAD+ pharmaceutical formulation or the oral enhanced NAD+ formulation changes the level of gene expression of a protein involved in SUD.DESCRIPTION OF THE DRAWINGS

[0018] The foregoing and other objects of the present disclosure, the various features thereof, as well as the disclosure itself may be more fully understood from the following description, when read together with the accompanying drawings in which:

[0019] FIG. 1 provides a tabular representation of certain molecular findings related to the potential pathophysiology7of SUD / PSUD and affected biological systems;

[0020] FIG. 2A is a graphic representation of the averaged intracellular concentration of NAD+ (icNAD+) in blood draws taken from patients administered LNAD (orange) or placebo (black) on 2 baseline days (days 1 and 4), and then on days 11 and 13; and

[0021] FIG. 2B is a graphic representation of the fold change in relative baseline of intracellular NAD+ concentration in the blood draws taken on baseline and days 11 and 13 from each of the patients administered either the LNAD formulation (left) or placebo (right).DESCRIPTION

[0022] The disclosures of these patents, patent applications, and publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art as known to those skilled therein as of the date of the invention described and claimed herein. The instant disclosure will govern in the instance that there is any inconsistency between the patents, patent applications, and publications and this disclosure.

[0023] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. The initial definition provided for a group or term herein applies to that group or term throughout the present specification individually or as part of another group, unless otherwise indicated.

[0024] For the purposes of explaining the invention certain features of medical and pharmacological technologies known to those skilled in the art of clinical testing and therapeutic treatments have been omitted or simplified in order not to obscure the basic principles of the invention. Parts of the following description will be presented using terminology commonly employed by those skilled in the art of optical design. It should also be noted that in the following description of the invention repeated usage of the phrase “in one embodiment” does not necessarily refer to the same embodiment.

[0025] As used herein, the articles “a,” “an,” and “the” refer to one or to more than one (z.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element. The use of the word “a” or “an” when used in conjunction with the term “comprising” in the claims and / or the specification may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.” Furthermore, use of the term “including” as well as other forms, such as “include,” “includes,” and “included,” is not limiting.

[0026] As used herein, the term “about” will be understood by persons of ordinary skill in the art and will vary to some extent on the context in which it is used. As used herein when referring to a measurable value such as an amount, a temporal duration, and the like, the term “about” is meant to encompass variations of ±20% or ±10%, including ±5%, ±1%, and ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods.

[0027] The terms “substance” and “drug” are used interchangeably throughout.

[0028] Wherever any of the phrases “for example,” “such as,” “including” and the like are used herein, the phrase “and without limitation” is understood to follow' unless explicitly stated otherwise. Similarly, “an example,” “exemplary” and the like are understood to be nonlimiting.

[0029] The term “substantially” allows for deviations from the descriptor that do not negatively impact the intended purpose. Descriptive terms are understood to be modified by the term “substantially” even if the w ord “substantially” is not explicitly recited.

[0030] The terms “comprising” and “including” and “having” and “involving” (and similarly “comprises”, “includes,” “has,” and “involves”) and the like are used interchangeably and havethe same meaning. Specifically, each of the terms is defined consistent with the common United States patent law definition of "‘comprising” and is therefore interpreted to be an open term meaning '"at least the following,” and is also interpreted not to exclude additional features, limitations, aspects, etc. Thus, for example, “a process involving steps a, b, and c” means that the process includes at least steps a, b and c. Wherever the terms “a” or “an” are used, “one or more” is understood, unless such interpretation is nonsensical in context.

[0031] Embodiments as described herein can involve isolating, collecting, or obtaining a biological sample from a subject. As used herein, the term “collecting a sample” or “isolating a sample”, for example, can refer to any process for directly or indirectly acquiring a biological sample from a subject. For example, a biological sample may be obtained (e.g., at a point-of-care facility, e.g., a physician's office, a hospital, laboratory facility) by procuring a tissue sample (such as a skin biopsy) from a subject. Alternatively, a biological sample may be obtained by receiving the biological sample (e.g., at a laboratory facility) from one or more persons who procured the sample directly from the subject. The biological sample may be, for example, a tissue (e.g., biopsy), fluid (e.g., cerebrospinal fluid, plasma, blood, serum) or cell (e.g., skin fibroblast cells, peripheral blood cells) of a subject.

[0032] The term “sample” can refer to a biological sample obtained or derived from a source of interest, as described herein. In some embodiments, a source of interest comprises an organism, such as an animal or human. In some embodiments, a biological sample comprises biological tissue or fluid. In some embodiments, a biological sample is or comprises bone marrow; spleen, blood; blood cells; blood mononuclear cells; serum; plasma; ascites; tissue or fine needle biopsy samples; cell -containing body fluids; free floating nucleic acids; sputum; saliva; urine; cerebrospinal fluid, peritoneal fluid; pleural fluid; feces; lymph; gynecological fluids; skin swabs; vaginal swabs; oral swabs; nasal swabs; washings or lavages such as a ductal lavages or bronchoalveolar lavages; aspirates; scrapings; bone marrow specimens; tissue biopsy specimens; surgical specimens; feces, other body fluids, secretions, and / or excretions; and / or cells therefrom, etc. In some embodiments, a biological sample is or comprises cells obtained from an individual. In some embodiments, a sample is a “primary’ sample” obtained directly from a source of interest by any appropriate means. For example, in some embodiments, a primary biological sample is obtained by methods selected from the group consisting of biopsy {e.g., fine needle aspiration or tissue biopsy), surgery, collection of body fluid {e.g., blood, lymph, feces etc.), etc. In some embodiments, the term “sample” refers to a preparation that is obtained by processing (e.g., by removing one or more components of and / orby adding one or more agents to) a primary' sample. For example, filtering using a semi-permeable membrane. Such a "‘processed sample” may comprise, for example nucleic acids or proteins extracted from a sample or obtained by subjecting a primary sample to techniques such as amplification or reverse transcription of rnRNA, isolation and / or purification of certain components, etc.

[0033] The term ‘“subject” or ‘“patient” can refer to any organism to which aspects of the disclosure can be performed, e.g., for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Subjects to which methods as described herein are performed comprise mammals, such as primates, for example humans. For diagnostic or research applications, a wide variety of mammals are suitable subjects, including rodents (e.g.. mice, rats, hamsters), rabbits, primates, and swine such as inbred pigs and the like. The term “living subject” can refer to a subject noted herein or another organism that is alive. The term “‘living subject” can refer to the entire subject or organism and not just a part excised (e.g., a liver or other organ) from the living subject. The term “normal subject” or “control” can refer to a subject that is not afflicted with a disease or condition, such as a subject that is not afflicted with SUD or PSUD.

[0034] As used herein, the phrase “therapeutic agent” can refer to any agent that elicits a desired pharmacological effect when administered to a subject. In some embodiments, an agent is considered to be a therapeutic agent if it demonstrates a statistically significant effect across an appropriate population. In some embodiments, the appropriate population may be a population of model organisms. In some embodiments, an appropriate population may be defined by various criteria, such as a certain age group, gender, genetic background, preexisting clinical conditions, history' of substance abuse, history' of SUD, history of PSUD, etc. In some embodiments, a therapeutic agent is any substance that can be used to alleviate, ameliorate, relieve, inhibit, delay onset of, reduce severity of. and / or reduce incidence of one or more symptoms or features of a disease, disorder, and / or condition. The term “therapeutically effective amount”, as used herein, can refer to an amount of a therapeutic agent whose administration, when viewed in a relevant population, correlates with or is reasonably expected to correlate with achievement of a particular therapeutic effect. The therapeutic effect may be objective (i.e., measurable by some test or marker) or subjective (i.e., subject gives an indication of or feels an effect). “‘Therapeutic effects” can refer to increasing intracellular NAD+, increasing circulating NAD+, or a combination thereof. In certain embodiments, “therapeutic effects” refers to amelioration of one or more symptoms of withdrawal during detoxification.

[0035] In some embodiments, a therapeutically effective amount of a substance is an amount that, when administered to a subject suffering from or susceptible to a disease, disorder, and / or condition, is sufficient to treat, diagnose, prevent, and / or delay and / or alleviate one or more symptoms of the disease, disorder, and / or condition. Disease progression can be monitored by clinical observations, laboratory and imaging investigations apparent to a person skilled in the art. A therapeutically effective amount is administered in a dosing regimen that can comprise multiple unit doses. For a therapeutic agent, a therapeutically effective amount (and / or an appropriate unit dose within an effective dosing regimen) can vary, for example, depending on route of administration, on combination with other pharmaceutical agents. Also, the specific therapeutically effective amount (and / or unit dose) for a patient can depend upon a variety' of factors including the disorder being treated and the severity of the disorder; the activity of the specific pharmaceutical agent employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and / or rate of excretion or metabolism of the therapeutic agent; the duration of the treatment; and like factors known in the art. Furthermore, an effective amount may be administered via a single dose or via multiple doses within a treatment regimen. In some embodiments, individual doses or compositions are considered to contain a “therapeutically effective amount” when they contain an amount effective as a dose in the context of a treatment regimen. Those of ordinary skill in the art will appreciate that a dose or amount may be considered to be effective if it is or has been demonstrated to show statistically significant effectiveness when administered to a population of patients; a particular result need not be achieved in a particular individual patient in order for an amount to be considered to be therapeutically effective as described herein.

[0036] The word “treating” can mean the medical management of a subject, e.g. an animal or human, with the intent that a prevention, cure, stabilization, or amelioration of the symptoms or condition will result. This term includes active treatment, that is, treatment directed specifically toward improvement of the disorder; palliative treatment, that is, treatment designed for the relief of symptoms rather than the curing of the disorder; preventive treatment, that is, treatment directed to prevention of disorder; and supportive treatment, that is, treatment employed to supplement another specific therapy directed toward the improvement of the disorder. The term “treatment” also includes symptomatic treatment, that is, treatment directed toward constitutional symptoms of the disorder. “Treating” a condition with the compounds of the disclosure involves administering such a compound, alone or in combination and by anyappropriate means, to a patient. For example, “treating’' SUD or PSUD can refer to (or be indicated by) raising intracellular NAD+ levels in a patient or subject. Treatment can be administered to a subject who does not exhibit signs of a disease, disorder, and / or condition (e.g., prior to an identifiable disease, disorder, and / or condition), and / or to a subject who exhibits only early signs of a disease, disorder, and / or condition for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and / or condition. In embodiments, treatment is administered humans suffering from SUD or PSUD. In certain embodiments, treatment refers to administration of enhanced NAD+ to a patient who is suffering from SUD or PSUD. Treatment can refer to administration of enhanced NAD+ to a subject who is at risk of developing SUD or PSUD. In certain embodiments, treatment includes administering enhanced NAD to a subjection who is recovering from SUD or PSUD.

[0037] As used herein, the term “NAD” refers to “nicotinamide adenine dinucleotide” and encompasses NAD+ and / or NADH. The term “NAD+” as used in this disclosure describes compositions comprising NAD+.

[0038] The terms “enhanced NAD+ composition.” “enhanced NADU” “enhanced NAD+ formulation,” “enhanced NAD+ pharmaceutical formulation,” as used interchangeably in this disclosure encompass compositions and pharmaceutical formulations comprising NAD+ prepared in accordance with or otherwise disclosed in PCT / US23 / 35557, PCT / US2025 / 035967, or PCT / US25 / 43556. or NAD+ which has substantially similar characteristics as the enhanced NAD+ disclosed in any of the foregoing patent applications, such substantially similar characteristics including, but not limited to any one or more of: an increased shelf life, bioavailability when provided orally, and increasing intracellular NAD+ levels in a subjected following administration to the subject. In certain embodiments, the enhanced NAD+ comprises “LNAD,” “BNAD,” “NADaCell™,” “LathMized NADU" or any combination thereof.

[0039] In various embodiments, the step of administering a pharmaceutical composition or therapeutic compound (such as an enhanced NAD+ composition) can be accomplished by any route of administration known in the art. Exemplary routes of administration include but are not limited to oral, subcutaneous, parenteral (such as, intravenous, intraperitoneal), intramuscular, rectal, epidural, intratracheal, intranasal, dermal, vaginal, buccal, intraperitoneal, intravenous, ocular, or pulmonary. The compositions and preparations can appear in conventional forms, for example, solutions and suspensions for injection, capsulesand tablets, in the form of enteric formulations, e.g. as disclosed in U.S. Pat. No. 5,350,741, and for oral administration.

[0040] Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

[0041] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EM™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, a pharmaceutically acceptable polyol like glycerol, propylene glycol, liquid polyethylene glycol, and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, and thimerosal. In many cases, it can be useful to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

[0042] Sterile injectable solutions can be prepared by incorporating the compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated herein, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basicdispersion medium and the required other ingredients from those enumerated herein. In the case of sterile powders for the preparation of sterile injectable solutions, examples of useful preparation methods are vacuum drying and freeze-drying which yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-fdtered solution thereof.

[0043] Oral compositions can include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed. Oral formula of the drug can be administered once a day, twice a day, three times a day, or four times a day, for example, depending on the half-life of the drug.

[0044] As used herein, the term “unadministered” subject refers to a subject that has not been administered the enhanced NAD+ formulation according to the disclosure and may be also called an “untreated” or subject.

[0045] The present disclosure provides a treatment for PSUD and SUD in which an oral formulation of an enhanced nicotinamide adenine dinucleotide (NAD+) is administered to human subject suffering from PSUD.

[0046] In various embodiments, the present disclosure provides for treatment of PSUD and SUD through administration of an oral enhanced NAD+ formulation, such as, but not limited to, “LN AD.”

[0047] NAD+ is a ubiquitous coenzy me that is involved in diverse biological processes. Such biological processes include, but are not limited to, maintaining mitochondrial redox status and cellular energy homeostasis to participating in post-translational protein modification and DNA repair and generating second messengers. The processes depend not only on the role of NAD+ as the electron donor / acceptor in oxidative / reductive reactions but also its utilization by NAD-consuming enzymes.

[0048] Following administration in accordance with the dosage regimen provided in Example 2, orally administered LNAD significantly increases in intracellular NAD levels (p=4.13xlOr!3) (see, e.g., FIG. 2), primary' NAD+ metabolites, MeNAM and 2PY, as well as changes in several multi-omic biomarkers, relative to placebo (as described in more detail in Example 2,below). Thus, LNAD is bioavavailable, and its safety and identified multi-omic biomarkers of relevance support use of LNAD as a treatment of SUD and PSUD.

[0049] Without being bound by theory, FIG. 1 delineates an overview of certain molecular findings from one study as related to the known or suspected pathophysiology of SUD / PSUD and affected biological systems.

[0050] LNAD Increases Lipid Turnover and Metabolism, and Promotes Tissue Integrity

[0051] There was a significant increase in plasma concentrations of fatty acid binding proteins FABP1 and RBP2 following administration of LNAD. There were no detrimental changes in clinical laboratory biomarkers of liver functioning following administration of LNAD but reduced levels of liver enzymes GGT and ALP, as well as serum bilirubin were observed.

[0052] Pathway analyses further indicated an increase in the activity of the drug metabolism via cytochrome P450 enzymes and xenobiotic metabolism pathways. Without being bound by any particular mechanism, such an increase in drug metabolism may be partially due to observed increases in glutathione transporter GSTA1. Glutathione protects cells from oxidative stress generated during P450-mediated metabolism of toxins and drugs, facilitates the removal of reactive metabolites produced by P450s, and is closely linked with the NAD+ / NADH and NADP+ / NADPH redox balance within the cell.

[0053] Most recreational drugs are lipophilic, enabling them to cross the blood-brain-barrier, and are metabolized by the P450 family of enzymes. Based on the observed effects on lipid and drug metabolism in this study, and without being bound by any particular theory, LNAD may aid the removal of harmful substances accumulated with chronic polysubstance use by enhancing shuttling and clearance of lipids and other hydrophobic molecules, reducing oxidative stress, and aid the removal of harmful substances accumulated with chronic PSUD. Restoration of lipid homeostasis and retinoid availability may further support neuronal membrane integrity and signaling function disrupted by PSUD. Results from the study described in Example 2 show that the blood-brain-barrier maintenance pathway was downregulated following LNAD administration. Peripheral markers of inflammation such as cytokines and chemokines exhibit a heterogeneous pattern of alterations across SUDs.

[0054] TNF-a and IL6 can alter the blood-brain-barrier permeability and can fostering peripheral monocyte activation and migration. These cytokines are overexpressed in blood across SUDs, indicating possible chronic inflammation. Neuroinflammation is a significant element of PSUD that arises at the interplay between endothelial injury, oxidative stress,excitotoxicity. and immune activation. However, after LNAD treatment, there was an overall reduction in peripheral abundance biomarkers of endothelial injury and chemokines such as mitochondria-damaging MMP9, CXCL10 and CXLC12. These results show beneficial changes in peripheral inflammation as well as metabolism, tissue integrity, endothelial functioning, and immune signaling.

[0055] LNAD Increases SIRT1 and NAD-Dependent Deacetylase

[0056] Study results show that LNAD increases circulating sirtuin-1 (SIRT1) and NAD-Dependent Deacetylase which regulates inflammation, metabolism, mitochondrial function, and neuroplasticity. SIRT1 is a nuclear transcriptional regulator that controls a wide variety of cellular functions. As a class III histone deacetylase, SIRT1 directly uses NAD+ during its deacetylation cycle. It plays roles in cellular metabolism and oxidative stress response by regulating multiple transcriptional pathways. It enhances the activity of peroxisome proliferator-activated receptor gamma coactivator 1 -alpha (PGCla), a key regulator of mitochondrial biogenesis and respiration. SIRT1 has also been shown to deacetylate and inhibit sterol regulatory element binding proteins (SREBP1 and SREBP2), transcription factors that control the expression of genes involved in fatty acid and triglyceride synthesis. Without wishing to be bound by theory7, such changes can contribute to the positive lipid-lowering effects observed in this Study that are also consistent with results of a recent meta-analysis of NAD precursor supplementation studies.

[0057] Consistent with the role SIRT1 in regulating mitochondrial biogenesis, the Study also revealed a decrease in mitochondrial apoptosis marker superoxide dismutase 2 (SOD2) and increase in mitochondrial bicarbonate HCO3 transporter carbonic anhydrase 5A (CA5A) in the plasma proteome, suggesting improvements in mitochondrial functioning and reduced oxidative stress and apoptosis. Pathway analyses confirmed a reduction in the mitochondrial apoptosis pathway activity. Mitochondrial dysfunction and oxidative stress are linked to neurotransmitter imbalances, excitotoxicity, and organ damage (in particular, liver, endothelial / cardiac, and neuronal) in the context of chronic substance use and PSUD.

[0058] Many recreational drugs, such as, but not limited to, methamphetamine and opioids, induce oxidative damage. There are significant changes in oxidative metabolites and antioxidant capacity across multiple SUDs.

[0059] LNAD Reduces Circulating Glutamate and Increases Gamma-Glutamyl-Glutamine.

[0060] Together with changes in GGT and GSTA1, results from the Study in Example 2 show that LNAD can reduce peripheral inflammation and affect glutamate release and metabolism. The glutamate-GABA-glutamine cycle is involved in the regulation of the balance of neuronal excitation vs inhibition, and peripheral as well as brain glutamate levels are altered by SUD. In the brain, glutamate released from presynaptic neurons is taken up by astrocytes and converted to glutamine which is then transferred back to neurons and converted back to glutamate by mitochondrial glutaminase. Imbalances in this cycle contribute to excitotoxicity and depressed GABAergic tone associated with PSUD as well as comorbid mental health disorders.

[0061] In PSUD, chronic exposure exacerbates oxidative stress, leading to changes in glutathione homeostasis. This disruption can manifest as glutathione depletion and impaired enzymatic activity, such as in glutathione peroxidase and glutathione S-transferases, crucial for neutralizing reactive oxygen species and detoxifying xenobiotics. Glutathione is a tripeptide of glutamate, cysteine, and glycine, and is crucial for regulating reactive oxygen species and redox balance in the brain. Without being bound by theory, by modulating glutathione-related enzymes like GGT and GSTA1 and glutamate and NAD+ availability for glutathione synthesis and regeneration, LNAD can enhance antioxidant capabilities and reduce inflammation and excitotoxicity. Without wishing to be bound by theory, by normalizing glutamatergic tone, LNAD can rebalance the excitatory / inhibitory signaling disrupted by chronic substance exposure. Without being bound by any particular mechanism, reduced inflammation can also protect against excitotoxic damage to neurons and improve neuroplasticity required for recovery. Without being bound by theory, LNAD's role in modulating glutamate metabolism, evidenced by reduced circulating glutamate and increased gamma-glutamyl-glutamine, can further be complemented by its effects on the body’s antioxidant capacity.

[0062] LNAD Upregulated Amino Group Metabolism and the Urea Cycle.

[0063] Convergently between metabolomic and proteomic data, LNAD administration was associated with an upregulation of the urea cycle and potential reduction in nitrosative stress. The urea cycle is responsible for removing ammonia, a byproduct of protein and amino acid catabolism, in particular catabolism of glutamate. Impairments in urea cycle function can lead to accumulation of ammonia, resulting in neurotoxicity. Without being bound by theory, LNAD administration can enhance clearance of nitrogenous wastes from amino acid metabolism through increased urea production. Such an interpretation is also consistent withthe evidence for improved protein digestion based on reduced pancreatic enzymes in circulation following LNAD administration in the Study.

[0064] LNAD increased peripheral concentrations of serotonin receptor HTR1A, a key neurotransmitter system disrupted in addictive disorders. In the brain, HTR1A modulates glutamate levels and inflammation. Central activation of HTR1A leads to vasodilation in the periphery; whereas peripheral activation of HTR1A has anti-inflammatory effects on immune cells; HTR1A may also help normalize neurotransmission and glutamate levels. Without wishing to be bound by theory, by regulating amino acid breakdown, tryptophan metabolism, and ammonia disposal through the urea cycle, while also modulating serotonin signaling involved in excitotoxicity, LNAD can rebalance disrupted neurotransmitter systems and reduce neurotoxicity in PSUD.

[0065] LNAD Reduces Circulating Cortisone Levels.

[0066] Metabolic pathway analysis indicated that LNAD significantly altered steroid hormone biosynthesis, consistent with its beneficial effects on mitochondrial functioning described above. The HPA axis plays a role of the dysregulation of stress response systems in the development of SUD and PSUD. Elevated salivary, hair, and serum cortisol (as well as cortisone) levels have been documented for various SUDs and are predictive of treatment efficacy. In SUD / PSUD, the overactivation of the HPA axis links together stress, metabolism (via glucose and insulin), and downstream behavioral effects such as disruptions in sleep due to dysregulation of the circadian and cellular clocks. Glucocorticoid receptors participate in opiate withdrawal-induced noradrenergic activity, and changes in the HPA axis affect brain regions that are involved in learning and rew ard processing.

[0067] Circulating and salivary cortisol and cortisone are positively correlated with one another and undergo interconversion. Without being bound by theory, by attenuating cortisol / cortisone levels, LNAD aids in the normalization of HPA axis hyperactivity and downstream metabolic and behavioral sequelae associated with chronic stress in PSUD. Coupled with beneficial effects on mitochondrial energy metabolism, and without being bound by theory, LNAD also counteracts cortisol-induced mitochondrial dysfunction observed in addiction models. A decrease in abundance of other steroid hormone metabolites was also detected following administration of LNAD, which is consistent with altered lipid and steroid hormone biosynthesis and mitochondrial function.

[0068] These results demonstrate LNAD’s ability to address PSUD through its effects on a wide range of biological pathways. In particular, without being bound by theory, LNAD-induced enhancement of lipid metabolism, mitochondrial functioning, and hormone and neurotransmitter balance, alongside the putative reduction in oxidative and nitrosative stress and inflammation, demonstrate that administration LNAD represents an effective treatment option in the comprehensive management of SUD and PSUD.

[0069] In certain embodiments, the present disclosure relates to LNAD administration for treatment of acute PSUD, for withdrawal management, or a combination thereof. Briefly, withdrawal symptoms result, at least in part, due to a complex profile of metabolic, neurotransmitter, inflammatory, cardiovascular and psychological disturbances. Without wishing to be bound by theory, LNAD’s modulation of interconnected systems (including any one or more of glutamatergic signaling, hypothalamic-pituitary-adrenal (HP A) axis regulation, antioxidant capacity, and lipid and cellular metabolism) provides multi-system biological stabilization. For example, without wishing to be bound by theory LNAD’s normalization of glutamatergic tone can mitigate excitotoxicity-related seizure activity. Similarly, and without being bound by theory, LNAD’s influence on the HPA axis can attenuate surges in catecholamines and glucocorticoids that precipitate autonomic dysregulation, hypertension, and arrhythmias. Without wishing to be bound by theory, enhanced antioxidant defenses following LNAD administration can lessen oxidative stress and endothelial injury that can potentiate cognitive deficits and neuronal damage. Furthermore, and without being bound by theory, the increase in serotonin receptor HTR1A, a key neurotransmitter disrupted in addiction, observed following administration of LNAD can alleviate withdrawal -emergent anxiety or provide an anti-inflammatory effect.

[0070] Methods of Treatment and Use

[0071] In various aspects, the present disclosure relates to treatment of a variety of conditions, diseases and disorders with administration of an enhanced NAD+ pharmaceutical formulation. In certain embodiments, the present disclosure relates to the treatment of SUD, PSUD, or both via administration of enhanced NAD+ pharmaceutical formulations to a patient suffering from one or more symptoms of SUD or PSUD. In embodiments, the present disclosure includes methods of treating chronic PSUD, acute PSUD, or a combination thereof.

[0072] The present disclosure includes preventing or ameliorating one or more symptoms associated with SUD. PSUD, or a combination thereof. The disclosure further relates totreatment of withdrawal symptoms in a subject recovering from SUD or PSUD. Methods of disclosed herein relate to treatment of withdrawal symptoms in a subject during detoxification. In certain embodiments, the methods disclosed herein relate to treatment of opioid withdrawal symptoms in a subject suffering from SUD, PSUD, or a combination thereof.

[0073] In various embodiments, administration of an enhanced NAD+ pharmaceutical formulation results in an increase in intracellular NAD+ levels in a subject as compared to an unadministered subject. In some examples, administration of the enhanced NAD+ pharmaceutical formulation increases intracellular levels of NAD+ in blood cells.

[0074] In various embodiments, the enhanced NAD+ pharmaceutical formulations utilized in the methods disclosed herein are stable in a variety of dosage forms. In some embodiments, the NAD+ composition comprises an amorphous component. In addition, the enhanced NAD+ pharmaceutical formulations disclosed herein offer increased biological activity toward increased cellular NAD levels, increased stability, or a combination thereof. Additionally, the inclusion of PEG in certain exemplary formulations provide a “molecular shuttle” for the delivery of NAD+, for example, but not limited to. across the buccal membranes into the blood. This formulation provides certain advantages that are useful to assist in NAD+-dependent cellular activities.

[0075] The enhanced NAD+ pharmaceutical formulations disclosed herein can be administered to a subject (e.g., but not limited to, a human) as a nutrient and / or a therapeutic agent to prevent, ameliorate, and / or treat PSUD or SUD in a subject.

[0076] In some embodiments, the enhanced NAD+ pharmaceutical formulations are administered to increase the NAD+ concentration or NAD+ stability (i.e., half-life) in a subject afflicted with SUD or PSUD.

[0077] In one aspect, the present disclosure relates to a method of treating PSUD or SUD in a subject, comprising administering to the subject a therapeutically effective amount of an NAD+ enhanced pharmaceutical formulation. The present disclosure can relate to a method of treating withdrawal during detoxification in a subject suffering from SUD or PSUD.

[0078] In some embodiments, the administration of the NAD+ enhanced pharmaceutical formulation changes the plasma or serum level of a protein that contributes to the effects or symptoms of SUD, PSUD, or withdrawal during detoxification as compared to that of an unadministered subj ect. In some embodiments, the change includes an increase or decrease in the level of the protein in the administered subject compared to that level in an unadministeredsubject. In some embodiments, the change comprises an increase or decrease in the plasma level of protein in the administered subject compared to that level in an unadministered subject.

[0079] In some embodiments, administration of a therapeutically effective amount of an enhanced NAD+ increases lipid turnover. Administration of a therapeutically effective amount of an enhanced NAD+ can increase lipid metabolism. In certain embodiments, administration of a therapeutically effective amount of an enhanced NAD+ promotes tissues integrity. Administration of a therapeutically effective amount of an enhanced NAD+ can reduce biomarkers of endothelia injury. In embodiments, administration of a therapeutically effective amount of an enhanced NAD+ reduces peripheral chemokines.

[0080] In certain embodiments, administration of a therapeutically effective amount of an enhanced NAD+ increases circulating sirtuin-1 (SIRT1). In embodiments, administration of a therapeutically effective amount of an enhanced NAD+ reduces circulating glutamate in the subject. Administration of a therapeutically effective amount of an enhanced NAD+ can increase gamma-glutamyl-glutamine in a subject.

[0081] In certain embodiments, administration of a therapeutically effective amount of an enhanced NAD+ upregulates the urea cycle. Administration of a therapeutically effective amount of enhanced NAD+ can reduce circulating cortisone levels as compared to an untreated subject.

[0082] In some embodiments, the disclosure provides a method of promoting cellular NAD+ metabolism and NAD+ cellular homeostasis in a subjects suffering from an SUD or PSUD, comprising administering to the subject an amount of the enhanced NAD+ pharmaceutical formulation described herein effective to promote cellular NAD+ metabolism.

[0083] In embodiments, the therapeutically effective amount of enhanced NAD+ is between about 5 mg to about 4000 mg per day, inclusive. The therapeutically effective amount of enhanced NAD+ can be between about 100 mg to about 3500 mg per day. In some embodiments of the present disclosure, the therapeutically effective amount of enhanced NAD+ is between about 20 mg to about 2500 mg per day. In some embodiments the therapeutically effective amount of enhanced NAD+ is between about 50 mg to about 2000 mg per day. The therapeutically effective amount of enhanced NAD+ can be between about 100 mg to about 1500 mg per day. In some embodiments of the present disclosure, the therapeutically effective amount of enhanced NAD+ is between about 200 mg to about 1000 mg per day. In further embodiments, the therapeutically effective amount of enhanced NAD+ is between about 300 mg to about 900 mg per day. The therapeutically effective amount ofenhanced NAD+ can be between about 400 mg to about 800 mg per day. In further embodiments, the therapeutically effective amount of enhanced NAD+ is between about 500 mg to about 700 mg per day. In further embodiments, the therapeutically effective amount of NAD+ is between about 550 mg and about 650 mg per day. In some embodiments, the amount of NAD+ is about 500 mg per day. In some embodiments, the therapeutically effective amount of enhanced NAD+ is about 600 mg per day. In some embodiments, the therapeutically effective amount of enhanced NAD+ is about 700 mg per day. In some embodiments, the therapeutically effective amount of enhanced NAD+ is about 800 mg per day. In some embodiments, the therapeutically effective amount of enhanced NAD+ is about 900 mg per day. In some embodiments, the therapeutically effective amount of enhanced NAD+ is about 1000 mg per day. In some embodiments, the therapeutically effective amount of enhanced NAD+ is about 400 mg per day. In some embodiments, the therapeutically effective amount of enhanced NAD+ is about 300 mg per day. In some embodiments, the therapeutically effective amount of enhanced NAD+ is about 200 mg per day. The therapeutically effective amount of enhanced NAD+ can be more than about 1000 mg per day. In some embodiments, the therapeutically effective amount of enhanced NAD+ is up to about 3000 mg per day. The therapeutically effective amount of enhanced NAD+ can be up to about 2500 mg per day. In certain embodiments, the therapeutically effective amount of enhanced NAD+ is up to about 2000 mg per day. The therapeutically effective amount of enhanced NAD+ can be about 500 mg / day, about 750 mg / day, about 1000 mg / day, about 1250 mg / day, about 1500 mg / day, about 1750 mg / day, about 2000 mg / day, about 2250 mg / day, about 2500 mg / day, about 2750 mg / day, or about 3000 mg / day.

[0084] In various embodiments, the therapeutically effective amount can be administered in a single dose or can be administered over multiple doses in a single day. In embodiments with multiple doses per day, each dose can be provided in equal amounts, or one or more doses can be in an amount that is greater than at least one other dosage amount. In embodiments, the therapeutically effective amount is administered over 1 to 10 doses in a single day. The therapeutically effective amount can be administered in 2 doses per day, 3 doses per day, 4 doses per day, 5 doses per day, 6 doses per day. 7 doses per day, 8 doses per day, 9 doses per day, or 10 doses per day.

[0085] In certain embodiments, the methods disclosed herein comprise administering a therapeutically effective amount of enhanced NAD+ to a subject suffering from SUD or PSUD during an initial treatment period followed by administering a maintenance dose of enhancedNAD+ to the subj ect during a maintenance period. In embodiments, the maintenance does is less than the dose provided during the initial treatment period. The maintenance dose can be equal to or less than the dose administered during the initial treatment period. The maintenance dose can be about 75% of the dose administered during the initial treatment period. In embodiments, the maintenance dose is up to about 50% of the dose administered during the initial treatment period. The maintenance dose can be less than about 50% of the dose administered during the initial treatment period. In certain embodiments, the maintenance dose is up to about 45% of the dose administered during the initial treatment period, up to about 40% of the dose administered during the initial treatment period, up to about 35% of the dose administered during the initial treatment period, up to about 30% of the dose administered during the initial treatment period, up to about 25% of the dose administered during the initial treatment period, up to about 20% of the dose administered during the initial treatment period, up to about 15% of the dose administered during the initial treatment period, up to about 10% of the dose administered during the initial treatment period, or up to about 5% of the dose administered during the initial treatment period. In certain embodiments, the maintenance dose is less than about 5% of the dose administered during the initial treatment period.

[0086] In various embodiments, the maintenance dose can be administered in a single dose or can be administered over multiple doses in a single day. In embodiments with multiple doses per day, each dose can be provided in equal amounts, or one or more doses can be in an amount that is greater than at least one other dosage amount. In embodiments, the maintenance dose is administered over 1 to 10 doses in a single day. The maintenance dose can be administered in 2 doses per day, 3 doses per day, 4 doses per day, 5 doses per day, 6 doses per day, 7 doses per day, 8 doses per day, 9 doses per day, or 10 doses per day.

[0087] In certain embodiments, at least one substance associated with the SUD or PSUD wherein the SUD or PSUD is a substance that is referenced within the Controlled Substances Act (21 U.S.C. § 801 et seq.). The at least one substance can be a narcotic, a stimulant, a depressant, a hallucinogen, or any other drug that has a potential for abuse by a subject. Nonlimiting examples of the at least one substance include opioids, cocaine, methamphetamine, ecstasy (MDMA), prescription stimulants (e.g.. dextroamphetamine), benzodiazepines, cannabis, n LSD, psilocybin mushrooms, DMT, PCP, mescaline, inhalants (e.g., glue, paint thinners, gasoline, nitrous oxide, and other volatile compounds with the potential for abuse), nicotine, sedatives (e.g., barbiturates, zolpidem), anabolic steroids (e.g., testosterone, nandrolone, and other performance-enhancing drugs), over-the-counter medications withpotential for abuse (e.g., cough and cold medications (e.g., dextromethorphan), pain relievers, sleep aids, and the like), or any combination thereof.

[0088] Reference will now be made to specific examples illustrating the disclosure. It is to be understood that the examples are provided to illustrate exemplar}7embodiments and that no limitation to the scope of the disclosure is intended thereby.EXAMPLESEXAMPLE 1Testing of Enhanced NAD+ Formulation Safety and Symptom Mitigation During Polydrug Detoxification Withdrawal

[0089] Five adult subjects were observed after being administered an enhanced NAD+ formulation according to the below dosing schedule to assess: (1) treatment safety; (2) treatment response; treatment effect; and (4) the variance of the effect of treatment. The Clinical Opiate Withdrawal Scale (COWS) was used to quantify the severity of opiate withdrawal symptoms. COWS scores < 6, is the maximum threshold for no detox, and COWS scores < 12, is the maximum threshold for mild detox. Other factors studied were the impact and efficacy of NADaCell™ on the need for PRN '‘comfort” medications typically administered for the treatment of opioid withdrawal symptoms (OPS).

[0090] Dosing Schedule:

[0091] Participants were administered enhanced NAD+ (treatment A) for four (4) days following at least a q.i.d. schedule, with four-five 500 mg daily doses spread approximately 3-4 hours apart within a 10-16-hour window for a total daily dose of 2000 mg to 2500 mg. The enhanced NAD+, as well as the matching placebo, met the FDA criteria for administration in humans (including compliance with GMP regulations).

[0092] Each dose of enhanced NAD+ was delivered by oral administration as a swish and swallow.

[0093] A designated study coordinator actively monitored dose administration and blood draw compliance with the assistance of technology that allows the participant to receive reminders and provide quick responses to questions pertaining to the ongoing study protocol.

[0094] Dose Administration Instructions

[0095] Doses were administered with 3-4 hours between doses. If a dose was missed, it was taken within 2 hours of the scheduled dose; another 2 hours were allowed to pass prior to resuming the dose schedule such that all the doses for that day were completed. Certain subj ects received a 5th dose on at least one day of treatment, which was taken at least about 2 hours after the 4th dose on the same day (see Table 1 for daily dosing schedule).Table 1*Dose may not have been administered to each subject on each day.

[0096] Formulating and administering the enhanced NAD+ for administration in this example:• Added dry powder contents of bottle to about 4 ounces (!4 cup) of water immediately before taking. Stirred well.• After stirring, enhanced NAD+ was administered via small sips of the prepared solution, swished in mouth for approximately 45 seconds, then swallowed.• Repeated swish and swallow until all prepared solution was swallowed (approximately 3 to 5 sips).

[0097] Subject 1• Gender Identity: Male• DOB: 10 / 78• DAST score: 7 / 10 - Substantial Addiction• Occupation: Personal Trainer• Prior Formal Drug Treatment History': None. Self-detox attempted.• Drug of Choice: Percocet, 150+ mg. daily snorted

[0098] Day One - Last reported use 12 hours prior, validated by urine tox screen result of opiates. Subject presented with a slightly flat affect and anxious and a desire to be free of his addiction. Day generally uneventful. Frequent periods of resting free of visible physical distress. Slight restlessness and pacing when awake.• Appetite: Normal• Sleep: Minimally interrupted• Vital Signs: 160 / 94 P71; 160 / 94 P71; 158 / 90 P71; 149 / 83 P80• Clinical Opioid Withdrawal Scale: 6, 6, 4, 0• PRN Medications Administered: None• Reported / Observed Adverse Side Effects to the enhanced NAD+ formulation:None

[0099] Day Two - Subject was up eating breakfast and worked out @ 5:00AM. Reports to nurse no current cravings. Appetite good. Overall, subject states, “I thought I would feel worse” Taking a few naps. Minimal restlessness. Still feeling anxious, however no reports or exhibiting withdrawal symptoms.• Appetite: Normal• Sleep: Minimally interrupted• Vital Signs: 154 / 80 P63; 150 / 78 P69; 153 / 70 P62; 149 / 68 P62• Clinical Opioid Withdrawal Scale: 4, 4, 4, 4• PRN Medications Administered: 650mg. Tylenol for headache due to allergies.• Reported / Observed Adverse Side Effects to NADaCell™: None

[0100] Day Three - Subject was up early working out for over an hour and has a bright affect and outlook. Cravings reported as zero. Appetite good. Complained of headache 5 / 10 and has requested Tylenol. Mild nose running however feels it may be allergy related. His affect is bright. Subject states, “I feel great”. Has been missing family and is tearful at times.• Appetite: Normal• Sleep: Rested peacefully• Vital Signs: 140 / 70 P60; 142 / 70 P58; 140 / 68 P60; 138 / 68 P60• Clinical Opioid Withdrawal Scale: 0, 0, 0, 0• PRN Medications Administered: 650mg. Tylenol for headache due to allergies.

[0101] Day Four- Subject up working out. Bright affect. Thoughts organized. Reports having slept well. Thoughts clearer and more organized. Blood pressure is at an all-time low for him. Subject stated, ‘This is what it would always be, before using”. Subject has no complaints or reports any side effects from drink mix. Spent fourth night at the house.• Appetite: Normal• Sleep: Normal• Vital Signs: 132 / 76 P72; 136 / 68 P58• Clinical Opioid Withdrawal Scale: 0, 0• PRN Medications Administered: None• Reported / Observed Adverse Side Effects to NADaCell™: None

[0102] Follow-up 7-days post DC: Doing well; reports sober. “I can’t believe I have so much energy.” Taking NADaCell™ lOOmg. BID and “back to life”. Checks in regularly with team members.Subject 2• Gender Identity: Female• DOB: 01 / 90• DAST score: 10 / 10 - Severe Addiction• Prior Formal Drug Treatment History: More than 12x• Drug of Choice: Heroin IV

[0103] Day One - Last reported use 2 hours prior, validated by urine tox screen polydrug results of methadone, benzodiazepine. THC, amphetamines, opiates, fentanyl and phenobarbital. Subject admitted showing no signs of withdrawal. Subject states uses every 3-4 hour and thought process appears scattered and reports she is anxious to get her detox underway. Social most of the day with her peers and was eating on and off all day. Reported in PM surprised at not feeling worse.• Appetite: Normal• Sleep: Minimally interrupted• Vital Signs: 90 / 58 P58; 101 / 58 P58; 110 / 62 P60; 105 / 58 P71• Clinical Opioid Withdrawal Scale: 5, 5, 4, 5• PRN Medications Administered: None• Reported / Observed Adverse Side Effects to NADaCell™: None

[0104] Day Two - Subject complained of being anxious. Eating and drinking. Appetite good. Self-reported no cravings. Social with peers when awake.• Appetite: Normal• Sleep: Minimally interrupted• Vital Signs: 115 / 78 P83; 120 / 70 P80; 138 / 84 P84; 116 / 70 P90• Clinical Opioid Withdrawal Scale: 0, 4, 3, 4• PRN Medications Administered: Gabapentin 300mg. HS for complaints of anxiety• Reported / Observed Adverse Side Effects to NADaCell™: None

[0105] Day Three - Subject has bright affect. Slightly somatic at times however can talk through it. Denies cravings. PRN seeking. Self-reports anxiety and loose stools. “I feel better than I thought I would7’. Appetite good. Speaks in detail about plans for her future. Thoughts appear clear and organized. Supportive to peers. Active in conversation.• Appetite: Excellent• Sleep: Normal• Vital Signs: 110 / 68 P82; 114 / 72 P78; 121 / 68 P65; 138 / 78 P88• Clinical Opioid Withdrawal Scale: 5, 5, 5, 6• PRN Medications Administered: Imodium 300mg. HS; Gabapentin 300mg. HS • Reported / Observed Adverse Side Effects to NADaCell™: None

[0106] Day Four - Social with peers. Thoughts organized. Taking active role in supporting peers. Still reporting being anxious on a 4 / 10. Denies cravings. Has conversations with peers about goals and local meetings to attend. Less isolation and has scheduled to attend IOP upon DC. Appears confident. Went out to AA meeting with peer. Spent fourth night at the house.• Appetite: Normal• Sleep: Minimally interrupted• Vital Signs: 124 / 78 P70; 120 / 64 P68• Clinical Opioid Withdrawal Scale: 6. 0PRN Medications Administered: Gabapentin 300mg. HS for complaints of anxietyReported / Observed Adverse Side Effects to NADaCell™: None

[0107] Follow-up 7-days post DC: Subject doing well; reports sober. Enrolled in IOP. Taking NADaCell™ lOOmg. BID. UPDATED 3 / 2022: Subject is currently sober and has a leadership position at a treatment facility.

[0108] Subject s• Gender Identity: Female• DOB: 7 / 1992• DAST score: 10 / 10 - Severe Addiction• Prior Formal Drug Treatment History’: Multiple• Drug of Choice: Heroin IV

[0109] Day One - Last reported use 2 hours prior, validated by urine tox screen poly¬ drug results of opiates, THC. cocaine, benzodiazepines, and amphetamines. No current craving. Social with peers. Some mild yawning. Thoughts disorganized. Appetite is apparent. Calle is optimistic about her detox. As the day progresses, starts to become seclusive, reporting that subject is generally a quiet shy person when not using. Social with peer.• Appetite: Normal• Sleep: Normal• Vital Signs: 118 / 64 P83; 125 / 74 P76; 138 / 81 P86; 127 / 72 P80• Clinical Opioid Withdrawal Scale: 0, 3, 0, 2• PRN Medications Administered: None• Reported / Observed Adverse Side Effects to NADaCell™: None

[0110] Day Two - Subject up eating breakfast. Spent good part of morning in bed watching tv. Subject has taken several showers and is reporting some sweating and restlessness. Has not requested any pms at this time. Less social than the rest of peers. Reports no cravings. PRN for loose stool and nausea given. Calle however is still eating and drinking just not at level of her peers.• Appetite: Diminished but not absent• Sleep: Interrupted• Vital Signs: 124 / 78 P70; 120 / 60 P68; 138 / 89 P93; 127 / 72 P80• Clinical Opioid Withdrawal Scale: 1, 0, 0, 6• PRN Medications Administered: Imodium 4mg. HS; Zofran 4mg. PO HS • Reported / Observed Adverse Side Effects to NADaCell™: None

[0111] Day Three - Subject spending time isolating; eating and drinking but, in room, less social. Thoughts are organized, but more along the lines of self-loathing. States, "I have felt worse. I just get more scared of just the waiting for it to be more unbearable.” PRN administered for loose stool and muscle twitching. Being a medical professional, Calle feels that this is the peak of her detox.• Appetite: Diminished but not absent• Sleep: Interrupted• Vital Signs: 136 / 87 P89; 132 / 89 P93; 156 / 86 P96; 168 / 83 P100• Clinical Opioid Withdrawal Scale: 6, 5, 9, 9• PRN Medications Administered: Zanaflex 8mg. HS twitching; Zofran 4mg. HS • Reported / Observed Adverse Side Effects to NADaCell™: None

[0112] Day Four - Subject ate a small breakfast. Twitching subsided. Reports minimal cravings. Affect brighter; however, appears to struggle. Tearful and slightly somatic, but able to talk through it. By noon is actively conversing with peers and being supportive to them. Ate dinner, showered, put on makeup, and went to local AA meeting with peer from the group. States feeling clearer. Spent 4thnight at the house.• Appetite: Normal• Sleep: Minimally interrupted• Vital Signs: 141 / 88 P86; 128 / 75 P70• Clinical Opioid Withdrawal Scale: 9, 6• PRN Medications Administered: Zanaflex 8mg. PO morning• Reported / Observed Adverse Side Effects to NADaCell™: None

[0113] Follow-up 7-days post DC: Returned to workday following DC. Single lapse after meeting up with former roommate. Reported subject got “right back on track” and moved in with a sober friend. Enrolled in IOP. Taking NADaCell™ lOOmg. BID. Follows up with nurse daily. UPDATED 3 / 2022: Calle moved to Texas, has remained sober, and resumed career.

[0114] Subject 4• Gender Identity: Male• DOB: 2 / 1986• DAST score: 8 / 10 - Substantial Addiction• Prior Formal Drug Treatment History: 26x• Drug of Choice: Heroin / Fentanyl inhaled• Current Medications: Gabapentin 600mg. QID

[0115] Day One - Last reported use 2 hours prior, validated by urine tox screen polydrug results of fentanyl, opiates, methadone, amphetamines, cocaine, THC, benzodiazepines. Subject is very optimistic and reports smoking heroin / fentanyl “all day, every day.” Affect bright. Subject has expressed wanting this more than anything and is tired of the drug ruled life he has been living. Subj ect is eating and socializing the maj ority of the day with staff and peers.• Appetite: Normal• Sleep: Minimally interrupted• Vital Signs: 156 / 80 P89; 151 / 78 P81; 147 / 78 P80; 148 / 82 P84• Clinical Opioid Withdrawal Scale: 3, 4, 3, 4• PRN Medications Administered: None• Reported / Observed Adverse Side Effects to NADaCell™: None

[0116] Day Two - Subject is social. No self-reported cravings. Appetite good. Social with peers. Working at compound. No request for PRN. Had lasagna for dinner and demonstrates a very7healthy appetite to the point of binge eating into the late hours of the night and the early morning hours of day 3. “I feel amazing.”• Appetite: Excessive• Sleep: Minimally interrupted• Vital Signs: 150 / 70 P78; 152 / 74 P78; 143 / 87 P81; 154 / 89 P89• Clinical Opioid Withdrawal Scale: 4, 2, 4, 4• PRN Medications Administered: None• Reported / Observed Adverse Side Effects to NADaCell™: None

[0117] Day Three - Eating excessively from 11:30PM day 2 till 1:30AM day 3.3: 15 AM began complaining of stomach cramping and chills; took shower and bath to mitigate. Dry heaving. Administered PRN for nausea. Denies self-reported cravings. Subject states wanting to get through this and is compliant with regimen. Sleeping on and off and find some solace with a peer and sleeps through most off night although reports being restless.• Appetite: Minimal• Sleep: Interrupted• Vital Signs: 157 / 86 P97; 146 / 70 P74; 148 / 84 P93; 137 / 74 P86• Clinical Opioid Withdrawal Scale: 12, 6. 9, 6• PRN Medications Administered: Zofran 8 mg. 4AM, 8AM; 6PM; Phenergan 5ML HS• Reported / Observed Adverse Side Effects to NADaCell™: None

[0118] Day Four - Subject is labile and is eating and drinking. Intends to leave tonight and not take advantage of the extra night’s stay that is available. Subject denies wanting to use. Received DC instructions. Left at 5:00PM, checking in hours later thanking us and stating will stay clean.• Appetite: Moderate• Sleep: Minimally interrupted• Vital Signs: 146 / 70 P76; 138 / 70 P72• Clinical Opioid Withdrawal Scale: 6, 7• PRN Medications Administered: Phenergan 5ML PO 9:00AM• Reported / Observed Adverse Side Effects to NADaCell™: None

[0119] Follow-up 7-days post DC: Returned to work the day following DC. Reports doing well and is ‘“back to life.” Taking NADaCell™ 25mg. BID and requested NADaCell™ 1 OOmg. tablets be sent to him. Checks in regularly with team.Subject 5• Gender Identity: Male• DOB: 08 / 90• DAST score: 8 / 10 - Substantial Addiction• Prior Formal Drug Treatment History: 8x• Drug of Choice: Heroin IV

[0120] Day One - Last reported use 2 hours prior, validated by urine tox screen polydrug results of amphetamines, opiates, benzodiazepines, THC, and fentanyl. Subject appeared visibly intoxicated with a flat affect, initially minimally social except for peer known prior to arrival. Thoughts disorganized. Ate lunch poolside and became more active with peers. Thoughts continued to appear disorganized throughout the day. Denies any cravings.• Appetite: Normal• Sleep: Restless• Vital Signs: 118 / 70 P68; 116 / 74 P63; 127 / 78 P82; 126 / 80 P84• Clinical Opioid Withdrawal Scale: 4, 4, 4, 4• PRN Medications Administered: None• Reported / Observed Adverse Side Effects to NADaCell™: None

[0121] Day Two - Reports and exhibits sweating, some mild cramping, loose stool and nausea during early morning hours. Refused PRN medication. Advised nurse of prior injection site infection. Warm to touch pain 6 / 10. Vitals within normal limits no fever. Eating and drinking with no issue. No report of elevation in craving scale. At lunchtime, refused both PO and IM antibiotics, stating “I don't want them, they give me a stomachache.” Infection appeared to continue to worsen. Had dinner with peers, then decided to leave AMA.• Appetite: Slightly diminished• Sleep: N / A• Vital Signs: 138 / 87 P76; 132 / 64 P68; 136 / 88 P82• Clinical Opioid Withdrawal Scale: 9, 7, 8• PRN Medications Administered: Refused all PRN’s• Reported / Observed Adverse Side Effects to NADaCell™: NoneSummary' of Results:

[0122] Outcome: Days 1-4 acute detox, 85% of COWS scores were < 6; 100% of COWS scores were < 12.

[0123] Treatment Safety: The formulation / intervention is believed to be safe. There were no known adverse events occurred or were reported. In addition, in a double-blind, placebo-controlled study in a Wellness Cohort (“RCT”), there was a favorable safety profile across 52 biomarkers including liver, kidney and inflammatory as well as self-reported data from participants.

[0124] Treatment Response: Subjects responded to the enhanced NAD+ formulation administered during detox. Detox symptoms are associated with an acute stressor on the body resulting in a reduction in NAD levels. This repletion protocol mitigates the physiological symptoms of withdrawal and the associated psychological components of withdrawal based on a standardized COW Scale.

[0125] Treatment Effect Variance: There was minor (i.e., no non-responders and minimal dropout rate) variance in the effect of treatment with the enhanced NAD+ formulation. The RCT showed every patient in the treatment arm was a responder with no placebo effect exhibited by any participants.EXAMPLE 2LNAD Treatment Feasibility Study

[0126] This study enrolled N=60 healthy aging adults, >45, randomized into the treatment and placebo arms, and examined a wide range of peripheral blood biomarkers, including plasma proteome and metabolome, as well as clinical laboratory7tests, following a 5-day supplementation protocol.

[0127] Methodology

[0128] Study configuration and experimental design was a randomized, doubleblinded, placebo-controlled, parallel group design. The study recruited N=76 participants; of these, N=60 successfully enrolled and were randomized. N=51 received at least one dose of LNAD101 or placebo and provided at least one blood draw on treatment. Analyses reported here are based on the full available dataset from N=51 treated participants (25 women, 26 men). The recruits were adult English-speaking volunteers aged 45 to 75 years (inclusive) in overall self-reported good health (common conditions such as hypertension and hyperlipidemia were permitted). Exclusion criteria included Body Mass Index >35 kg / m2; diagnoses of acute or chronic infectious diseases, autoimmune diseases, diabetes, or cancer; pregnancy; cunent use of NAD+ supplements or precursors.

[0129] Test Product and Placebo Dose and Mode of Administration

[0130] The formulation was an oral, enhanced NAD+ (LN AD) formulation containing 500 mg NAD and 500 mg inactive polyethylene glycol (PEG). For each dose, a measured amount of LNAD powder was dissolved in water and participants took the dose orally using a “swish and swallow” procedure. The formulation meets the FDA criteria for administration in humans (including compliance with GMP regulations) and was provided by BioNADrx (207 N. Boone Street #15. Johnson City, TN 37604. Test Product batch no. 5024-21- 1004-PW1L 3 SYN).

[0131] Placebo contained no active ingredient but otherwise was not discernible from the LNAD formulation and contained inactive PEG as the main filler (50% mass in the active treatment group, and identical mass in the placebo group without the NAD+). Placebo meets the FDA criteria for administration in humans (including compliance with GMP regulations) and was provided by BioNADrx (batch no. 3350-21 -1004-PW1.)

[0132] This w as a 13-day protocol with a 7-day run-in period involving wash-out from all dietary supplements (Days 1-7), followed by a 5-day treatment period (Days 8-12) and 1 day follow-up (Day 13). After the 7-day run-in period, participants were administered the LNAD formulation or placebo for 5 days four times daily (q.i.d.) with one additional dose administered on each of Days 9 and 10. Baseline blood draws and vital signs w ere obtained on Day 4 and Day 6 (prior to treatment) and results were averaged as “baseline” for statistical analysis. Additional blood draws and vital signs were obtained on Day 11 (on-treatment) andDay 13 (post- treatment). The Day 11 blood draw corresponded to ~14 doses; the Day 13 blood draw corresponded to ~22 doses. Inactive placebo was administered using the same mode of administration, schedule, and duration as the formulation.

[0133] Testing Criteria

[0134] Intracellular NAD levels were assessed via a colorimetric assay (Jinfiniti, Augusta, GA) that measured levels of total NAD in red and white blood cells and platelets. Extracellular NAD was assessed in cell-free plasma. Statistically significant changes in the levels of the following peripheral blood biomarkers related to overall health and expected mechanisms of action of NAD were measured to determine:

[0135] Liver and kidney function (serum levels of alkaline phosphatase (ALP), alanine transaminase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), albumin, total bilirubin, blood urea nitrogen (BUN), creatinine);

[0136] Oxidative stress (serum levels of gamma-glutamyl transferase (GGT); cumulative serum levels of reactive oxygen metabolites (ROM);

[0137] Inflammation: (serum levels of cardiac C-reactive protein (CRP) and high-sensitivity CRP (hsCRP), tumor necrosis factor (TNF), interleukin-6 (IL-6));

[0138] Lipid metabolism (serum levels of triglycerides, low-density lipoprotein (LDL), high-density7lipoprotein (HDL), their ratio (HDL / LDL);

[0139] Serum glucose; and

[0140] Bioavailability of NAD+ metabolites (plasma levels of nicotinamide (NAM), quinolinate, nicotinate, trigonelline, 1 -methylnicotinamide (MeNAM) and Nl-methyl-2-pyrudone-5-carboxamyde (2PY).

[0141] The study also examined the levels of a comprehensive array of about 4,000 circulating biomarkers utilizing data from blood draws on Days 6, 11 and 13 (baseline, on-treatment, post-treatment). These included greater than 1,000 metabolites present on Metabolon’s Global Platform used for untargeted metabolomics of plasma samples from the study, including amino, carbohydrates, lipids, nucleotides, peptides, as well as partially characterized molecules. The examined biomarkers also included about 3,000 proteins whose abundance in plasma was assayed using Olink’s Explore 3,072 PEA proteomics platform with NGS readout (Illumina’s HiSeq 2000) including a wide range of interrogated proteins targeted a host of biological pathways, (cellular membrane maintenance, nutrient transport, oxygen species formation, lipid transport and degradation, inflammation, immune signaling, andmitochondrial functioning.). Samples were biobanked and preserved in liquid nitrogen at -80 C for batch processing; assaying of longitudinal samples from the same participant was performed simultaneously.

[0142] Safety

[0143] Individual adverse events were documented and included the self-reported daily Review of Systems (ROS) targeting 14 organ and functional systems. Vital signs, weight and wearable data- were collected at each blood draw visit (systolic and diastolic blood pressure, calculated MAP, resting heart rate, Body Mass Index, waist circumference) as well and as provided by a Fitbit tracker (heart rate, sleep duration by stage). Fifty-two clinical laboratory tests were also conducted to identify out-of-range results after baseline. Subjects reported measures mood and subjective well-being were measured using assessment instruments that had been previously independently validated for daily use. Daily anxiety7and stress were measured by the Clinically Useful Anxiety Outcome Scale - Daily version (CUXOS-D), mood by the Depression, Anxiety, and Stress Scale-21 (DASS-21), and fatigue with the Daily Fatigue Impact Scale (D-FIS).

[0144] Statistical Analyses

[0145] Statistical analyses were performed using a one-stage slope estimand approach under the Generalized Linear Mixed Modeling parametrization. Outcome levels were modeled as a function of a set of predetermined covariates as fixed effects (age, sex, interaction between age, sex for all phenotypes; time-varying Body Mass Index was included as the primary covariate in analyses of blood biomarkers), fixed effect of time (number of full days on treatment in the continuous time analyses, individual timepoint group in the categorical time analyses), treatment Group, interaction between Time and Group. Random effect structure included random individual intercepts and random slopes (primary endpoint) or random intercepts (secondary / exploratory endpoints). Continuous covariates were mean-centered; female sex was chosen as the reference; following unblinding, the placebo group was chosen as the reference. P-values were obtained using the Kenward-Roger approximation for the likelihood ratio test (LRT) statistic. Nonparametric bootstrap-based confidence intervals were obtained for primary' endpoints at Type I error rate of 5% adjusted for multiple-comparisons across study laboratory tests. For NAD+ assays and clinical laboratory tests, both baseline draws were coded as 0, thereby averaging across two baseline blood draws pre-initiation oftreatment. Given the number of blood draws available, change was modeled as a linear parameter in the case of biomarkers.

[0146] The primary analyses were focused on testing the interaction between Time and Group in accordance with a priori expectations for the assayed concentrations of NAD+. P-values were corrected for multiple comparisons separately for each -omic layer where applicable using the minimum effective number of independent tests (Meff) procedure. No endpoint data were imputed. The same statistical methods were used for the Exploratory Biomarkers analysis. Metabolomics data were log2-transformed prior to analyses; proteomic data were expressed on the log2 scale as well using Olink’s relative abundance (NPX) units. In both cases, a change of 1 unit corresponds to approximately a two-fold change in abundance. Descriptive safety analyses were supplemented with statistical analyses of adverse event incidence, vitals, subjective well-being, and other clinical laboratory result.

[0147] Results

[0148] The Study demonstrated a significant increase in intracellular NAD levels (p=4.13xlOrI3). primary NAD+ metabolites, MeNAM and 2PY, as well as changes in several multi-omic biomarkers, relative to placebo. Its short-term use in healthy individuals did not reveal any adverse safety effects.

[0149] The bioavailability of this enhanced form of oral NAD+ is supported by several converging -omic lines of evidence: (1) substantial elevation of plasma levels of primary NAD+ metabolites, MeNAM and 2PY; (2) elevated plasma levels of NAD-dependent and NAD-consuming enzy mes, including SIRT1, following LNAD administration, compared to placebo; (3) NAD+ availability to extra- and intracellular enzy mes evident in the pattern of findings from untargeted metabolomics that identified several NAD-enabled coupled enzymatic reactions t enhanced by LNAD, relative to placebo; and (4) the clinical laboratory data and the joint analyses of plasma proteome and metalome consistent with NAD+ administration leading to a complex cascade of changes that are enabled by its biological relevance to a diverse set of processes.

[0150] Accordingly, administered bioavailability of LNAD demonstrated by this study provided evidence for its use in the treatment of PSUD and SUD.EQUIVALENTS

[0151] Those skilled in the art will recognize, or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific embodiments described specifically herein. Such equivalents are intended to be encompassed in the scope of the following claims.

Claims

CLAIMS1. A method of treating a substance use disorder (SUD) in a subject, the method comprising administering a therapeutically effective amount of an enhanced NAD+ pharmaceutical formulation to the subject.

2. The method of claim 1 , further comprising administering a maintenance dose of the enhanced NAD+ pharmaceutical formulation to the subject for a maintenance period.

3. The method of claim 2. wherein the maintenance dose is less than or equal to the therapeutically effective amount.

4. The method of claim 2. wherein the maintenance period is at least about one week.

5. The method of claim 1, wherein the enhanced NAD+ pharmaceutical formulation is a mouthwash, and the step of administering the therapeutically effective amount of the enhanced NAD+ pharmaceutical formulation comprises instructing the subject to intake a quantity of the mouthwash into the subject’s mouth and perform a swishing motion for less than about one minute.

6. The method of claim 1. wherein the SUD is a polysubstance use disorder (PSUD).

7. The method of claim 1, wherein the enhanced NAD+ pharmaceutical formulation comprises “LNAD,”8. The method of claim 1 or claim 2, wherein the administration of the enhanced NAD+ pharmaceutical formulation changes the level of gene expression of a protein involved in SUD.

9. A method of treating substance use disorder (SUD) or poly substance use disorder (PSUD) in a subject, the method comprising administering a therapeutically effective amount of an oral enhanced NAD+ formulation to the subject.

10. The method of claim 9, further comprising administering a maintenance dose of the oral enhanced NAD+ formulation to the subject for a maintenance period.

11. The method of claim 10, wherein the maintenance dose is less than or equal to the therapeutically effective amount.

12. The method of claim 10, wherein the maintenance period is at least about one week.

13. The method of claim 9. wherein the step of administering the therapeutically effective amount of the oral enhanced NAD+ formulation comprises instructing the subject to intake a quantity of the mouthwash into the subject’s mouth and perform a swishing motion for less than about one minute.

14. The method of claim 9, wherein the oral enhanced NAD+ comprises LNAD.

15. The method of claim 9, wherein the administration of the oral enhanced NAD+ formulation changes the level of gene expression of a protein involved in SUD or PSUD.

16. A method of treating one or more withdrawal symptoms in a subject recovering from substance use disorder (SUD) or polysubstance use disorder (PSUD), the method comprising administering a therapeutically effective amount of an oral enhanced NAD+ formulation to the subject.

17. The method of claim 16, further comprising administering a maintenance dose of the oral enhanced NAD+ formulation to the subject for a maintenance period.

18. The method of claim 17, wherein the maintenance dose is less than or equal to the therapeutically effective amount.

19. The method of claim 17, wherein the maintenance period is at least about one week.

20. The method of claim 16, wherein the step of administering the therapeutically effective amount of the oral enhanced NAD+ formulation comprises instructing the subject to intake a quantity of the mouthwash into the subject’s mouth and perform a swishing motionfor less than about one minute.

21. The method of claim 16, wherein the oral the enhanced NAD+ comprises LNAD.

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