Methods of identifying effective amounts of plastogens

Abstract

Disclosed herein are methods for determining effecting amounts of plastogens. Also described are methods of treating diseases or disorders that are mediated by the loss of synaptic connectivity and / or plasticity, such as neurological diseases and disorders, with therapeutically effective amounts of plastogens.

Description

WSGR Docket No. 55776-731.601METHODS OF IDENTIFYING EFFECTIVE AMOUNTS OF PLASTOGENSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of US Provisional Application No. 63 / 698,518, filed on September 24, 2024, and US Provisional Application No. 63 / 729,840, filed on December 9, 2024, each of which is hereby incorporated by reference in its entirety.FIELD OF THE INVENTION

[0002] Described herein are methods of identifyingplastogens andtherapeutically orbehaviourally relevant doses of plastogens. Plastogens are useful for the treatment of conditions, diseases, or disorders that would benefit from promoting neuronal growth and / or improving neuronal structure.BACKGROUND OF THE INVENTION

[0003] Altered synaptic connectivity and plasticity has been observed in the brains of individuals with neurological diseases and disorders. Psychoplastogens promote neuronal growth and improve neuronal architecture through mechanisms involving the activation of AMPA receptors, the tropomyosin receptor kinase B (TrkB), and the mammalian target of rapamycin (mTOR). Modulators of these biological targets, such as, for example, ketamine, scopolamine, N,N- dimethyltryptamine (DMT), and rapastinel have demonstrated psychoplastogenic properties. For example, ketamine is capable of rectifying deleterious changes in neuronal structure that are associated with neurological diseases and disorders. Such structural alterations include, for example, the loss of dendritic spines and synapses in the prefrontal cortex (PFC) as well as reductions in dendritic arbor complexity. Furthermore, pyramidal neurons in the PFC exhibit top-down control over areas of the brain controlling motivation, fear, and reward. Psychedelic psychoplastogens have demonstrated antidepressant, anxiolytic, and anti-addictive effects of in the clinic.BRIEF SUMMARY OF THE INVENTION

[0004] In one aspect, provided herein is a method of determining an effective amount of a plastogen for treating a brain disorder, comprising: a) administering one or more doses of the plastogen to a mammal; b) measuring1) by electroencephalogram (EEG) or quantitative electroencephalogram (qEEG), a slow wave frequency in the mammal; and, optionally,2) a concentration of the plastogen in the brain of the mammal; and c) determining the effective amount of the plastogen, wherein the effective amount of the plastogen increases power in the slow wave frequency.WSGR Docket No. 55776-731.601

[0005] In one aspect, provided herein is a method of treating a brain disorder in a patient, comprising administering an effective amount of a plastogen, wherein the effective amount of the plastogen is determined by: a) administering one or more doses of the plastogen to a mammal; b) measuring1) by electroencephalogram (EEG) or quantitative electroencephalogram (qEEG), a slow wave frequency in the mammal; and, optionally,2) a concentration of the plastogen in the brain of the mammal; and c) determining the effective amount of the plastogen, wherein the effective amount of the plastogen increases power in the slow wave frequency.

[0006] In another aspect, provided herein is a pharmaceutical composition comprising an effective amount of a plastogen, wherein the effective amount of the plastogen is determined by: a) administering one or more doses of the plastogen to a mammal; b) measuring1) by electroencephalogram (EEG) or quantitative electroencephalogram (qEEG), a slow wave frequency in the mammal; and, optionally,2) a concentration of the lastogen in the brain of the mammal; and c) determining the effective amount of the plastogen, wherein the effective amount of the plastogen increases power in the slow wave frequency.

[0007] Other objects, features, and advantages of the methods and compositions described herein will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating specific embodiments, are given by way of illustration only, since various changes and modifications within the spirit and scope of the instant disclosure will become apparent to those skilled in the art from this detailed description.BRIEF DESCRIPTION OF THE DRAWINGS

[0008] The novel features of the disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawing (also “figure” and “FIG.” herein), of which:

[0009] FIGs. 1A to 1C illustrate Spontaneous EPSC frequency (Hz) in PFC layer V pyramidal neurons for DLX-001 (CMPD-1) (FIG. 1A), DMT (FIG. IB), and ketamine and psilocybin (FIG.WSGR Docket No. 55776-731.601

[0010] FIGs. 2A to 2D illustrate Spontaneous EPSC amplitude (pA) in PFC layer V pyramidal neurons forDLX-001 (CMPD-1) (FIG. 2A), DMT (FIG. 2B), ketamine and psilocybin (FIG. 2C), and DLX-159 (CMPD-2) (FIG. 2D).

[0011] FIGs. 3A to 3D illustrate structural neuroplastic effects of tested compounds. FIGs. 3A to 3C illustrate spinogenesis and spine density of PFC layer V pyramidal neurons treated with DLX- 001 (Comp-2) (FIG. 3A), ketamine and psilocybin (FIG. 3B), and DLX-159 (Comp-2) (FIG. 3C). FIG. 3D illustrates synapse density of PFC layer I neurons following administration of DLX-159 (Comp-2).

[0012] FIGs. 4A to 4C illustrate antidepressive effect of DLX-001 (Comp-1) and ketamine in a forced swim test 24-hours post dose (FIG. 4A), 7-days post dose (FIG. 4B), and 14-days post dose (FIG. 4C)

[0013] FIG. 5 illustrates antidepressive effect of DLX-159 (Comp-2) in an IFN-alpha depression induced forced swim test.

[0014] FIG. 6 illustrates effect of DLX-001 (CMPD-1) and ketamine on delta, theta, alpha, beta, and gamma frequency bands in a qEEG study.

[0015] FIG. 7 illustrates effect of DLX-159 (CMPD-2) and ketamine on delta, theta, alpha, beta, and gamma frequency bands in a qEEG study.

[0016] FIG. 8 illustrates effect of DLX-159 (CMPD-2) and ketamine slow wave amplitude determined in preclinical qEEG.

[0017] FIG. 9 illustrates how preclinical markers of neuroplasticity may be correlatedto determine behavorally or therapeutically effective doses.DETAILED DESCRIPTION OF THE INVENTION

[0018] The present disclosure provides methods of identifying effective amounts of plastogens, methods of treating brain disorders with said effective amount of plastogen, and pharmaceutical compositions comprising the effective amount of plastogen.

[0019] In some embodiments, provided herein is a method of determining an effective amount of a plastogen for treating a brain disorder, comprising: a) administering the plastogen to a mammal; b) measuring1) by electroencephalogram (EEG) or quantitative electroencephalogram (qEEG), a slow wave frequency in the mammal; and, optionally,2) a concentration of the plastogen in the brain of the mammal; and c) determining the effective amount of the plastogen, wherein the effective amount of the plastogen increases power in the slow wave frequency.WSGR Docket No. 55776-731.601

[0020] In some embodiments, provided herein is a method of treating a brain disorder in a patient, comprising administering an effective amount of a plastogen, wherein the effective amount of the plastogen is determined by: a) administering the plastogen to a mammal; b) measuring1) by electroencephalogram (EEG) or quantitative electroencephalogram (qEEG), a slow wave frequency in the mammal; and, optionally,2) a concentration of the plastogen in the brain of the mammal; and c) determining the effective amount of the plastogen, wherein the effective amount of the plastogen increases power in the slow wave frequency.

[0021] In some embodiments, provided herein is a pharmaceutical composition comprising an effective amount of a plastogen, wherein the effective amount of the plastogen is determined by : a) administering the plastogen to a mammal; b) measuring1) by electroencephalogram (EEG) or quantitative electroencephalogram (qEEG), a slow wave frequency in the mammal; and, optionally,2) a concentration of the plastogen in the brain of the mammal; and c) determining the effective amount of the plastogen, wherein the effective amount of the plastogen increases power in the slow wave frequency.

[0022] In some embodiments, determining the effective amount of the plastogenfortreatingthe brain disorder comprises measuring, by electroencephalogram (EEG) or quantitative electroencephalogram (qEEG), a slow wave frequency in the mammal; and, a concentration of the plastogen in the brain of the mammal. In some embodiments, the effective amount of the plastogen increases power in the slow wave frequency and has a measurable concentration of the plastogen in the brain of the mammal. In some embodiments, the effective amount of the plastogen increases power in the slow wave frequency by at least 1 pp, at least 2 pp, at least 3 pp, at least 4 pp, at least 5 pp, at least 6 pp, at least 7 pp, at least 8 pp, at least 9 pp, at least 10 pp, at least 11 pp, at least 12 pp, at least 13 pp, at least 14 pp, at least 15 pp, at least 16 pp, at least 17 pp, at least 18 pp, at least 19 pp, at least 20 pp, at least 21 pp, at least 22 pp, at least 23 pp, at least 24 pp, at least 25 pp, at least 26 pp, at least 27 pp, at least 28 pp, at least 29 pp, or at least 30 pp above baseline. In some embodiments, the measurable concentration of the plastogen in the brain of the mammal is a free fraction of the plastogen in the brain of the mammal. In some embodiments, the free fraction of the plastogen is at least InM, at least 2nM, at least 3nM, at least 4nM, at least 5nM, at least 6nM, at least 7nM, at least 8nM, at least 9nM, at least lOnM, at least 1 InM, at least 12nM, at least 13nM, at least 14nM, at leastWSGR Docket No. 55776-731.60115nM, at least 16nM, at least 17nM, at least 18nM, at least 19nM, at least 20nM, at least21nM, at least 22nM, at least 23nM, at least 24nM, at least 25nM, at least 26nM, at least 27nM, at least 28nM, at least 29nM, at least 30nM, at least 3 InM, at least 32nM, at least 33nM, at least 34nM, at least 35nM, at least 36nM, at least 37nM, at least 38nM, at least 39nM at least 40nM, at least 41nM, at least 42nM, at least 43nM, at least 44nM, at least 45nM at least 46nM, at least 47nM, at least 48nM, at least 49nM, at least 50nM, at least 60nM, at least 70nM, at least 80nM, at least 90nM, or at least lOOnM.

[0023] In some embodiments, measuring the slow wave frequency and the concentration of the plastogen in the brain occurs between 0.5 - 24 hours after administering the plastogen, such as 0.5 - 1 hour, 1 - 1.5 hours, 1.5 - 2 hours, 2 - 2.5 hours, 2.5 - 3 hours, 3 - 3.5 hours, 3.5 - 4 hours, 4 - 4.5 hours, 4.5 - 5 hours, 5 - 6 hours, 6 - 7 hours, 8 - 10 hours, 10 - 12 hours, 12 - 16 hours, 16 - 20 hours, or 20 - 24 hours, after administering the plastogen. In some embodiments, the slow wave frequency is delta wave frequency or theta wave frequency. In some embodiments, the slow wave frequency is measured between 1 to 8 Hz. In some embodiments, the slow wave frequency is delta wave frequency. In some embodiments, the delta wave frequency is measured between 1 to 4 Hz. In some embodiments, the slow wave frequency is theta wave frequency. In some embodiments, the theta wave frequency is measured between 4 to 8 Hz.

[0024] In some embodiments, determining the effective amount of the plastogenfortreatingthe brain disorder further comprises measuring structural neuroplasticity in the mammal, wherein the effective amount of the plastogen increases structural neuroplasticity. In some embodiments, determining the effective amount of the plastogen for treating the brain disorder further comprises measuring functional neuroplasticity in the mammal, wherein the effective amount of the plastogen increases structural neuroplasticity. In some embodiments, determining the effective amount of the plastogen for treating the brain disorder further comprises measuring a behavioral effect, wherein the effective amount of the plastogen reduces pathological symptoms or maladaptive behaviors associated with the behavioral effect.

[0025] In some embodiments, administering the plastogen comprises administering 1 to 8 doses in different amounts to the mammal. In some embodiments, the mammal is a rodent. In some embodiments, the mammal is a human.

[0026] In some embodiments, the effective amount is a behaviorally effective amount or a therapeutically effective amount. In some embodiments, the effective amount is a behaviorally effective amount. In some embodiments, the effective amount is a therapeutically effective amount.

[0027] In some embodiments, the brain disorder is a neurological disease or disorder, a neuropsychiatric disease or disorder, a neurodegenerative disease or disorder, or aWSGR Docket No. 55776-731.601 neuropsychological disease or disorder. In some embodiments, the neuropsychiatric disease is major depressive disorder, treatment resistant depression, generalized anxiety disorder, post -traumatic stress disorder, obsessive compulsive disorder, substance use disorders, or a psychosis. In some embodiments, the psychosis is schizophrenia, bipolar disorder, or psychosis in Alzheimer’s disease (AD-P). In some embodiments, the neurodegenerative disease is a dementia, traumatic brain injury, or Parkinson’s disease. In some embodiments, the dementia is Alzheimer’s disease, vascular dementia, Lewy body dementia, frontotemporal dementia, Huntington’s disease, or mixed dementia. In some embodiments, the brain disorder is behavioral or psychological symptoms in dementia, or a combination thereof. In some embodiments, the brain disorder is motor symptoms, behavioral symptoms, or psychological symptoms in Parkinson’s disease, or a combination thereof.Certain Terminology

[0028] Unless otherwise stated, the following terms used in this application have the definitions given below. It must be noted that, as used in the specification and the appended claims, the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise. In this application, the use of “or” means “and / or” unless stated otherwise. The use of the term “including” as well as other forms, such as “include”, “includes,” and “included,” is not limiting. The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0029] The terms "administer," "administering," "administration," and the like, as used herein, refer to the methods that may be used to enable delivery of compounds or compositions to the desired site of biological action. These methods include, but are not limited to oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular or infusion), topical and rectal administration. Those of skill in the art are familiar with administration techniques that can be employed with the compounds and methods described herein. In some embodiments, the compounds and compositions described herein are administered orally.

[0030] The terms “effective amount,” as used herein, refers to a sufficient amount of an agent or a compound being administered, which will produce a desired biological or physiological effect.

[0031] The term “therapeutically effective amount,” as used herein, refers to a sufficient amount of an agent or a compound being administered, which will relieve to some extent one or more of the symptoms of the disease or condition being treated. The result includes reduction and / or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system.

[0032] The term “subject” or “patient” encompasses mammals. Examples ofmammals include, but are not limited to, any member of the Mammalian class: humans, non-human primates such as chimpanzees, and other apes and monkey species; farm animals such as cattle, horses, sheep, goats,WSGR Docket No. 55776-731.601 swine; domestic animals such as rabbits, dogs, and cats; laboratory animals including rodents, such as rats, mice and guinea pigs, and the like. In one embodiment, the mammal is a human.

[0033] The terms “treat,” “treating,” or “treatment,” as used herein, include alleviating, abating or ameliorating at least one symptom of a disease or condition, preventing additional symptoms, inhibiting the disease or condition, e.g., arresting the development of the disease or condition, relievingthe disease or condition, causing regression of the disease or condition, relieving a condition caused by the disease or condition, or stopping the symptoms of the disease or condition either prophylactically and / or therapeutically.Brain Disorders

[0034] Neuronal plasticity, and changes thereof, have been attributed to many neurological diseases and disorders. For example, during development and in adulthood, changes in dendritic spine number and morphology (e.g., lengths, crossings, density) accompany synapse formation, maintenance and elimination; these changes are thought to establish and remodel connectivity within neuronal circuits. Furthermore, dendritic spine structural plasticity is coordinated with synaptic function and plasticity. For example, spine enlargement is coordinated with long-term potentiation in neuronal circuits, whereas long-term depression is associated with spine shrinkage.

[0035] In addition, dendritic spines undergo experience-dependent morphological changes in live animals, and even subtle changes in dendritic spines can affect synaptic function, synaptic plasticity, and patterns of connectivity in neuronal circuits. For example, disease-specific disruptions in dendritic spine shape, size, and / or number accompany neurological diseases and disorders, such as, for example, neurodegenerative (e.g., Alzheimer’s disease or Parkinson’s disease) and neuropsychiatric (e.g., depression or schizophrenia) diseases and disorders, suggesting that dendritic spines may serve as a common substrate in diseases that involve deficits in information processing.

[0036] In some embodiments, a compound or plastogen of the present invention is used to treat a neuropsychiatric disease. In some embodiments, the neuropsychiatric disease is major depressive disorder, treatment resistant depression, generalized anxiety disorder, post -traumatic stress disorder, obsessive compulsive disorder, substance use disorders, or a psychosis. In some embodiments, the psychosis is schizophrenia, bipolar disorder, or psychosis in Alzheimer’s disease (AD-P).

[0037] In some embodiments, a compound or plastogen of the present invention is used to treat pain. In some embodiments, a compound or plastogen of the present invention is used to treat migraine.

[0038] In some embodiments, a compound or plastogen of the present invention is used to treat a neurodegenerative disease. In some embodiments, the neurodegenerative disease is a dementia, traumatic brain injury, or Parkinson’s disease. In some embodiments, the dementia is Alzheimer’sWSGR Docket No. 55776-731.601 disease, vascular dementia, Lewy body dementia, frontotemporal dementia, Huntington’s disease, or mixed dementia. In some embodiments, the compound or plastogen of the present invention is used to treatbehavioral and / or psychological symptoms in dementia. In some embodiments, the compound or plastogen of the present invention is used to treat motor symptoms, behavioral symptoms, and / or psychological symptoms in Parkinson’s disease.

[0039] In some embodiments, a neurological disease or disorder is a disease or disorder of the central nervous system (CNS) (e.g., brain, spine, and / or nerves) of an individual.

[0040] Types of neurological diseases and disorders include, but are not limited to, neurodegenerative diseases (such as Alzheimer’s disease, Parkinson’s disease, and dementia), headaches (e.g., migraines), brain injury (e.g., stroke or traumatic brain injury), brain cancer, an anxiety disorder (e.g., post-traumatic stress disorder (PTSD) or obsessive-compulsive disorder (OCD)), a mood disorder (e.g., suicidal ideation, depression, or bipolar disorder), apsychotic disorder (e.g., schizophrenia or substance-induced psychotic disorder), a personality disorder, an eating disorder (e.g., binge eating disorder), a sleep disorder, a sexuality disorder, an impulse control disorder (e.g., gambling, compulsive sexuality, or kleptomania), a substance use disorder (e.g., alcohol dependence, opioid addiction, or cocaine addiction), a dissociative disorder (e.g., epilepsy, amnesia, or dissociative identity disorder), a cognitive disorder (e.g., sub stance -induced cognitive impairment), a developmental disorder (e.g., Attention-Deficit / Hyperactivity Disorder (ADHD)), an autoimmune disease (e.g., multiple sclerosis (MS)), pain (e.g., chronic pain), andafactitiousdisorder. In some embodiments, a mammal treated with a compound or plastogen described herein has a disease or disorder that is or is associated with a disease or disorder of the CNS.

[0041] Neurodegenerative diseases or disorders include, but are not limited to, Alzheimer’s disease (AD), Parkinson’s disease (PD), prion disease, frontotemporal dementia, motor neuron disease (MND), Huntington’s disease (HD), Lewy Body dementia (LBD), and the like.

[0042] Substance use disorders include, but are not limited to, substance abuse, addiction and dependence, such as addiction or dependence to alcohol, opioids (e.g., heroin, oxycodone, and hydrocodone), cocaine, amphetamines (e.g., methamphetamine), nicotine, cannabinoids (e.g., tetrahydrocannabinol (THC)), caffeine, phencyclidine, paint thinner, glue, steroids (e.g., anabolic steroids), barbiturates (e.g., phenobarbital), methadone, benzodiazepines (e.g., diazepam), and the like.

[0043] Impulse control disorders include, but are not limited to, gambling, kleptomania, trichotillomania, intermittent explosive disorder, pyromania, skin picking, compulsive buying Tourette syndrome, compulsive sexual behavior, and the like.WSGR Docket No. 55776-731.601

[0044] Neuropsychiatric disorders include, but are not limited to, seizures (e.g., epilepsy), attention deficit disorders (e.g., ADHD and Autism), eating disorders (e.g., bulimia, anorexia, binge eating disorder, and pica), depression (e.g., clinical depression, persistent depressive disorder, bipolar disorder, postpartum depression, suicidal ideation, major depressive disorder, seasonal depression, and the like), anxiety (e.g., panic attacks, social anxiety disorder, panic disorder, and the like), schizophrenia, post-traumatic stress disorder (PTSD), obsessive-compulsive disorder (OCD), substance-induced psychotic disorder, sub stance -induced cognitive impairment, and the like.

[0045] Brain injury includes, but is not limited to, stroke, traumatic brain injury, dementia pugiliistica, chronic traumatic encephalopathy (CTE), or the like.Plastogens

[0046] A plastogen is a therapy or compound, or a pharmaceutically acceptable salt thereof, that increases neuroplasticity. Examples of plastogens include electroconvulsive therapy, ketamine, DMT, certain N-substituted indoles such as DLX-001, and certain TBG analogs such as DLX-159. In some embodiments, a plastogen described herein is a small molecule. A summary of select plastogens and their effect on Total Power, Delta Power, Theta Power, Alpha Power, Beta Power, and Gamma Power is shown in Table 1 .Table 1. Slow Wave changes are Predominant in PlastogensPharmaceutical compositions

[0047] In some embodiments, provided herein is a pharmaceutical composition comprising an effective amount of a plastogen, wherein the effective amount of the plastogen is determined by: a) administering one or more doses of the plastogen to a mammal; b) measuring1) by electroencephalogram (EEG) or quantitative electroencephalogram (qEEG), a slow wave frequency in the mammal; and, optionally,2) a concentration of the lastogen in the brain of the mammal; and c) determining the effective amount of the plastogen, wherein the effective amount of the plastogen increases power in the slow wave frequency.WSGR Docket No. 55776-731.601In some embodiments, the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient.

[0048] Pharmaceutical compositions are formulated in a conventional manner using one or more pharmaceutically acceptable inactive ingredients that facilitate processing of the active compounds into preparations that are used pharmaceutically . Proper formulation i s dependent upon the route of administration chosen. A summary of pharmaceutical compositions described herein is found, for example, in Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y. , 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkinsl999), herein incorporated by reference for such disclosure.

[0049] In some embodiments, the plastogens described herein are administered either alone or in combination with pharmaceutically acceptable carriers, excipients or diluents, in a pharmaceutical composition. Administration of the plastogens and compositions described herein canbe affected by any method that enables delivery of the plastogens to the site of action. These methods include, though are not limited to delivery via enteral routes (including oral, gastric or duodenal feeding tube, rectal suppository and rectal enema), parenteral routes (injection or infusion, including intraarterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural and subcutaneous), inhalational, transdermal, transmucosal, sublingual, buccal and topical (including epicutaneous, dermal, enema, eye drops, ear drops, intranasal, vaginal) administration, although the most suitable route may depend upon for example the condition and disorder of the recipient. By way of example only, plastogens described herein can be administered locally to the area in need of treatment, by for example, local infusion during surgery, topical application such as creams or ointments, injection, catheter, or implant. The administration can also be by direct injection at the site of a diseased tissue or organ.

[0050] In some embodiments, pharmaceutical compositions suitable for oral administration are presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. In some embodiments, the active ingredient is presented as a bolus, electuary or paste.

[0051] Pharmaceutical compositions which can be used orally include tablets, push -fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. Tablets may be made by compression or molding, optionally with one or more accessoryWSGR Docket No. 55776-731.601 ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with binders, inert diluents, or lubricating, surface active or dispersing agents. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. In some embodiments, the tablets are coated or scored and are formulated so as to provide slow or controlled release of the active ingredient therein. All formulations for oral administration should be in dosages suitable for such administration. The push -fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and / or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In some embodiments, stabilizers are added. Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or Dragee coatings for identification or to characterize different combinations of active compound doses.

[0052] In some embodiments, pharmaceutical compositions are formulated for parenteral administration by injection, e.g., by bolus injection or continuousinfusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi -dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and / or dispersing agents. The compositions may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in powder form or in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, saline or sterile pyrogen -free water, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described.

[0053] Pharmaceutical compositions for parenteral administration include aqueous and nonaqueous (oily) sterile injection solutions of the active compounds which may contain antioxidants, buffers, bacteriostats and solutes whichrenderthe formulation isotonicwith theblood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodiumWSGR Docket No. 55776-731.601 carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.

[0054] It should be understood that in addition to the ingredients particularly mentioned above, the plastogens and compositions describedhereinmay include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavoring agents.Methods of Treatment and Dosing

[0055] The plastogens disclosed herein, or pharmaceutically acceptable salts, solvates, or stereoisomers thereof, are useful for promoting neuronal growth and / or improvingneuronal structure.

[0056] Provided herein is a method of treating a brain disorder in a patient, comprising administering an effective amount of a plastogen, wherein the effective amount of the plastogen is determined by: a) administering one or more doses of the plastogen to a mammal; b) measuring1) by electroencephalogram (EEG) or quantitative electroencephalogram (qEEG), a slow wave frequency in the mammal; and, optionally,2) a concentration of the plastogen in the brain of the mammal; and c) determining the effective amount of the plastogen, wherein the effective amount of the plastogen increases power in the slow wave frequency .

[0057] In some embodiments, provided herein is a method of promoting neural plasticity (e.g., cortical structural plasticity) in a patient by administering an effective amount of a plastogen described herein to the patient.

[0058] In some embodiments, provided herein is a method of promoting neuronal growth in a patient, comprising administering to the patient an effective amount of a plastogen described herein.

[0059] In some embodiments, provided herein is a method of improving neuronal structure in a patient, comprising administering to the patient an effective amount of a plastogen described herein.

[0060] In some embodiments, provided herein is a method of treating a disease or disorder in a patient that is mediated by the loss of synaptic connectivity, plasticity, or a combination thereof, comprising administering to the patient an effective amount of a plastogen described herein.

[0061] In some embodiments, the brain disorder disease is a neuropsychiatric disease. In some embodiments, the neuropsychiatric disease is a mood or anxiety disorder. In some embodiments, the neurological disease is a migraine, headaches (e.g., cluster headache), post -traumatic stress disorder (PTSD), anxiety, depression, neurodegenerative disorder, Alzheimer’s disease, Parkinson’s disease,WSGR Docket No. 55776-731.601 psychological disorder, treatment resistant depression, suicidal ideation, major depressive disorder, bipolar disorder, schizophrenia, stroke, traumatic brain injury, and addiction (e.g., substance use disorder). In some embodiments, the neurological disease is a migraine or cluster headache. In some embodiments, the neurological disease is a neurodegenerative disorder, Alzheimer’s disease, or Parkinson’s disease. In some embodiments, the neurological disease is a psychological disorder, treatment resistant depression, suicidal ideation, major depressive disorder, bipolar disorder, schizophrenia, post-traumatic stress disorder (PTSD), addiction (e.g., substance use disorder), depression, or anxiety. In some embodiments, the neuropsychiatric disease is a psychological disorder, treatment resistant depression, suicidal ideation, major depressive disorder, bipolar disorder, schizophrenia, post-traumatic stress disorder (PTSD), addiction (e.g., substance use disorder), depression, or anxiety. In some embodiments, the neuropsychiatric disease or neurological disease is post-traumatic stress disorder (PTSD), addiction (e.g., substance use disorder), schizophrenia, depression, or anxiety. In some embodiments, the neuropsychiatric disease or neurological disease is addiction (e.g., substance use disorder). In some embodiments, the neuropsychiatric disease or neurological disease is depression. In some embodiments, the neuropsychiatric disease or neurological disease is anxiety. In some embodiments, the neuropsychiatric disease or neurological disease is post -traumatic stress disorder (PTSD). In some embodiments, the neurological disease is stroke or traumatic brain injury. In some embodiments, the neuropsychiatric disease or neurological disease is schizophrenia.

[0062] In some embodiments, the brain disorder is a neurological disease or disorder, a neuropsychiatric disease or disorder, a neurodegenerative disease or disorder, or a neuropsychological disease or disorder. In some embodiments, the neuropsychiatric disease is major depressive disorder, treatment resistant depression, generalized anxiety disorder, post -traumatic stress disorder, obsessive compulsive disorder, substance use disorders, or a psychosis. In some embodiments, the psychosis is schizophrenia, bipolar disorder, or psychosis in Alzheimer’s disease (AD-P). In some embodiments, the neurodegenerative disease is a dementia, traumatic brain injury, or Parkinson’s disease. In some embodiments, the dementia is Alzheimer’s disease, vascular dementia, Lewy body dementia, frontotemporal dementia, Huntington’s disease, or mixed dementia. In some embodiments, the brain disorder is behavioral or psychological symptoms in dementia, or a combination thereof. In some embodiments, the brain disorder is motor symptoms, behavioral symptoms, or psychological symptoms in Parkinson’s disease, or a combination thereof.EXAMPLES

[0063] The following examples are provided for illustrative purposes only and not to limit the scope of the claims provided herein.WSGR Docket No. 55776-731.601

[0064] The terms “DLX-001,” Comp-1,” and“CMPD-l” are used interchangeably throughout the examples and figures, and refer to the same compound.

[0065] The terms “DLX-159,” Comp-2,” and “CMPD-2” are used interchangeably throughout the examples and figures, and refer to the same compound.I. BIOLOGICAL EXAMPLESExample 1: Measuring Functional Neuroplasticity

[0066] Spontaneous EPSC frequency, a measure of synaptic connectivity, was determined in PFC layer V pyramidal neurons. The EPSC frequency (Hz) was determined for DLX-001 (lOmg / kg, ip), DMT (lOmg / kg, ip), ketamine (lOmg / kg, ip), and psilocybin (3mg / kg, ip). The Spontaneous EPSC frequency results for DLX-001, DMT, and ketamine and psilocybin, are shown in Figures 1 A, IB, and 1C, respectively.

[0067] Spontaneous EPSC amplitude, a measure of synaptic strength, was determined in PFC layer V pyramidal neurons. The EPSC amplitude (pA) was determined for DLX-001 (lOmg / kg, ip), DMT (lOmg / kg, ip), ketamine (lOmg / kg, ip), psilocybin (3mg / kg, ip), and DLX-159 (lOmg / kg, ip). The Spontaneous EPSC amplitude results for DLX-001, DMT, ketamine and psilocybin, and DLX-159 are shown in Figures 2 A, 2B, 2C, and 2D, respectively.Example 2: Measuring Structural Neuroplasticity

[0068] Dendritogenesis Assays. Phenotypic screening has historically proven more successful than target-based approaches for identifying drugs with novel mechanisms of action. Using a phenotypic assay, the compounds provided herein are tested for their ability to increase dendritic arbor complexity in cultures of cortical neurons. Following treatment, neurons are fixed and visualized using an antibody against MAP2 — a cytoskeletal protein localized to the somatodendritic compartment of neurons. Sholl analysis is then performed, and the maximum number of crossings (Nmax) is used as a quantitative metric of dendritic arbor complexity. For statistical comparisons between specific compounds, the raw Nmaxvalues are compared. Percent efficacies are determined by setting the Nmaxvalues for the vehicle (DMSO) and positive (ketamine) controls equal to 0% and 100%, respectively.

[0069] Animals. For the dendritogenesis experiments, timed pregnant Sprague Dawley rats are obtained from Charles River Laboratories (Wilmington, MA). In some instances, male and female C57BL / 6J mice are obtained from Jackson Laboratory (Sacramento, C. A.). In some instances, mice are housed in a temperature and humidity -controlled room maintained on a 12-h light / dark cycle in groups of 4-5 (same sex).

[0070] Dendritogenesis - Sholl Analysis. Neurons are plated in 96-well format(200 pL of media per well) at a density of approximately 15,000 cells / well in Neurobasal (Life Technologies)WSGR Docket No. 55776-731.601 containing 1% penicillin-streptomycin, 10% heat-inactivated fetal bovine serum, and 0.5 mM glutamine. After 24 h, the medium is replaced with Neurobasal containing lx B27 supplement (Life Technologies), 1% penicillin-streptomycin, 0.5 mM glutamine, and 12.5 pM glutamate. After 3 days in vitro (DIV3), the cells are treated with compounds. Compounds tested in the dendritogenesis assays are treated at 10 uM unless noted otherwise. Stock solutions of the compounds in DMSO are first diluted 100 -fold in Neurobasal before an additional 10 -fold dilution into each well (total dilution = 1 :1000; 0.1% DMSO concentration). Treatments are randomized. After 1 h, the media is removed and replaced with newNeurobasal media containing 1 x B27 supplement, 1 % penicillin -streptomycin, 0.5 mM glutamine, and 12.5 M glutamate. The cells grow for an additional 71 h. At that time, neurons are fixed by removing 80% of the media and replacing it with a volume of 4% aqueous paraformaldehyde (Alfa Aesar) equal to 50% of the working volume of the well. Then, the cells are incubated at room temperature for 20 min before the fixative is aspirated and each well washed twice with DPBS. Cells are permeabilized using 0.2% Triton X-100 (ThermoFisher) in DPBS for 20 minutes atroom temperature without shaking. Plates are blocked with antibody dilutingbuffer(ADB) containing 2% bovine serum albumin (BSA) in DPBS for 1 h at room temperature . Then, plates are incubated overnight at 4°C with gentle shaking in ADB containing a chicken anti-MAP2 antibody (1 : 10,000; EnCor, CPCA-MAP2). The next day, plates are washed three times with DPBS and once with 2% ADB in DPBS. Plates are incubated for 1 h at room temperature in ADB containingan antichicken IgG secondary antibody conjugated to Alexa Fluor 488 (Life Technologies, 1 :500) and washed five times with DPBS. After the final wash, 100 pL of DPBS is added per well and imaged on an ImageXpress Micro XL High-Content Screening System (Molecular Devices, Sunny vale,CA) with a 20x objective.

[0071] Images are analyzed using ImageJ Fiji (version 1.51W). First, images corresponding to each treatment are sorted into individual f olders that are then blinded f or data analysis. Plate controls (both positive and negative) are used to ensure thatthe assay is working properly as well as to visually determine appropriate numerical values for brightness / contrast and thresholding to be applied universally to the remainder of the randomized images. Next, the brightness / contrast settings are applied, and approximately 1-2 individual pyramidal-like neurons per image (i.e., no bipolar neurons) are selected using the rectangular selection tool and saved as separate files. Neurons are selected that did not overlap extensively with other cells or extend far beyond the field of view. The threshold settings are then applied to the individual images. The paintbrush tool is used to eliminate artifacts and dendritic processes originating from adjacent neurons (cleanup phaseNext, the pointtool is used to select the center of the neuron, and the images are saved and processed using the following Sholl analysis batch macro:WSGR Docket No. 55776-731.601 run("Sholl Analysis...", "starting=O ending=NaN radius_step=2 #_samples=l integration=Mean enclosing= l #_primary=4 infer fit linear polynomial=[Best fitting degree] most semi-log normalizer=Area create background=228 save do");Sholl analysis circle radii = 2 pixel increments = 0.67 pm. All images are taken and analyzed by an experimenter blinded to treatment conditions. The number of crossings for each neuron at each distinct radius is averaged to produce an average Sholl plot for each treatment. The Nmaxvalues are simply determined by identifying the maximum of each plot. For each treatment, neurons are selected from at least 6 wells spread across 2 plates (9 sites / well x 3 wells / plate x 2 plates). Each plate is prepared using neurons obtained from independent pregnant dams).

[0072] Spinogenesis Experiments. Spinogenesis experiments were performed on PFC layer V pyramidal neurons. Spine density was determined forDLX-001 , psilocybin, ketamine, and DLX-159 in half -log doses as demonstrated by Golgi staining. The spinogenesis results for DLX-001, psilocybin and ketamine, and DLX-159 are shown in Figures 3 A , 3B, and 3C, respectively. Synapse density for PFC layer I neurons following administration of DLX-159 is shown in FIG. 3D.

[0073] Neurite Outgrowth Assay. Changes in the pattern of neurite outgrowth have been implicated in psychiatric and neuro degenerative disorders as well as traumatic injuries. The discovery of new compounds that can positively affect neuritogenesis are important for developing new therapeutics for neurological diseases. Measurement of neurite outgrowth of rat cortical neurons using an automated image-based assay is used to determine the neuroplastic effects of the compounds of the present invention. The neurite outgrowth assay is performed at Neurofit SAS (France) as described below.

[0074] Pregnant Wistar rats (Janvier; France) are used for the study. They are delivered 6 days before theiruse. Upon arrival atNeurofit animal facility, they are housedone per cage and maintained in a room with controlled temperature (21-22°C) and a reversed light-dark cycle (12h / 12h; lights on: 17:30 - 05 :30; lights off: 05 :30 - 17:30) with food and water available ad libitum.

[0075] Female Wistar rats of 17 days gestation are killed by cervical dislocation and the fetuses are removed from the uterus. Their brains are placed in ice-cold medium of Leibovitz (LI 5, Gibco, Fisher bioblock, France). Cortices are dissected and meninges are carefully removed. The cortical neurons are dissociated by trypsinization for 30 min at 37°C (trypsin -ED TA, Gibco) in presence of 0.1 mg / ml DNAse I (Roche, France). The reaction is stopped by addition of Dulbecco’s Modified Eagle Medium (DMEM; Gibco) with 10% of fetal bovine serum (FBS; Gibco). The suspension is triturated with a 10-ml pipette and using a needle syringe 2 IGand centrifuged at 350 x g for 10 min at room temperature. The pellet of dissociated cells is resuspended in a medium consisting of Neurobasal (Gibco) supplemented with 2% B27 supplement (Gibco), 0.5mML -Glutamine (Gibco),WSGR Docket No. 55776-731.601 an antibiotic-antimicotic mixture. Viable cells are counted in a Neubauer cytometer using the trypan blue exclusion test (Sigma). Cells are seeded at a density of 10000 cells per well in 96 -well plate (Costar) precoated with poly-L-lysine. Test compound at different concentrations is added to the cultures. Donepezil (positive control) is tested at 250 nM.

[0076] After 72h (3 days) of plating, cultures are fixed with paraformaldehyde in PBS (4%, Sigma) for 30 min at 4°C. Then, cells are successively permeabilized with 0.1% Triton X100 for 30 min, saturated with PBS containing 3% of BSA and are incubated Ih with anti -beta III tubulin antibody (Sigma) at 1 / 10 000 in PBS containing 0.5% of BSA. Cells are washed three times with PBS containing 0.5% of BSA, and they are incubated Ih with goat anti-mouse antibody coupled with AF488 (Invitrogen Al 1001) diluted at 1 / 1000 in PBS containing 0.5% of BSA. Finally, nuclei are staining with DAPI 1 mg / ml at 1 / 1000 in PBS containing 0.5% of BSA. After rinsing with PBS, the plate is filmed and neurite networks are examined and analyzed using High-Content Screening (Celllnsight, Thermo Scientific). The average number of neurites per neuron and the average total length of neurites per neuron are the main parameters analyzed. Analysis of data is performed using analysis of variance (ANOVA). The Fisher’s Protected Least Significant Difference test is used for multiple comparisons. A p value < 0.05 is considered significant. The software used is StatView 5.0 from SAS Institut.

[0077] In some embodiments, a compound of the present invention increases the pattern of neurite outgrowth. In some embodiments, a compound of the present invention increases neurite average length compared to a control. In some embodiments, a compound of the present invention increases neurite branch points compared to a control. In some embodiments, a compound of the present invention significantly increases the number of new neurites and / or the average neurite length compared to a control.Example 3: Measuring Behavoral Effects

[0078] Forced Swim Test (FST). Male Sprague Dawley rats from Envigo (Indianapolis, IN) are obtained and housed 3 rats per cage following an IACUC approved protocol. All experiments are carried out at ambient temperatures (20 and 23 °C) under artificial lighting during the light-on part of the light / dark cycle in a Forced Swim chamber constructed of clear acrylic (height = 40 cm; diameter = 20.3 cm). Only one rat is placed in the swim chamber at a time for each swim test. The water is changed and the chamber cleaned between each animal. All rats are exposed to two swim sessions. The water depth is 16 cm in the first swim session and 30 cm in the second swim session, and the water temperature is maintained at 23±1 °C for all swim sessions. During the FST, animals undergo a 15 min swim session (pre-swim), lasting for 15 minutes, dried with paper towels, and returned to the home cage. Rats are injected with either saline, ketamine (positive control), or test compoundWSGR Docket No. 55776-731.601 after the habituation session, returnedto home cage, and then tested in a second FST lasting 5 minutes ~24 hours (second swim test) later. The second swim test is video recorded for scoring. Body weights are measured on both days. Scoring of the second swim test is performed by trained technicians using a time sampling technique in which the animal in the video recorded test is viewed every 5 seconds and the behavior seen is noted. The measures noted are immobility, climbing, and swimming behaviors. The effect of DLX-1 and Ketamine in the FST are shown in FIG. 4 A (24 hr post-dose), FIG. 4B (7 -days post dose), and FIG. 4C (14 days post dose).

[0079] IFN-alpha administered for 4 weeks induces a depression -like phenotype on the FST.

[0080] A single dose of DLX-159 (low dose, ip) or ketamine (5 mg / kg, ip) reversed this effect 24 hours after administration. The effect of ketamine and DLX-159 in the IFN-alpha depression induced FST are shown in FIG. 5.

[0081] Statistical analysis. Treatments are randomized, and data are analyzed by experimenters blinded to treatment conditions. Statistical analyses are performed using GraphPad Prism (version 8.1.2). Comparisons are planned prior to performing each experiment.Example 4: Measuring EEG and qEEG Power

[0082] DLX-001 (30 mg / kg, ip) and ketamine (10 mg / kg, ip) were administered to Wistar Kyoto rats (depressed phenotype) andqEEGwas performed. As shown in FIG. 6, DLX-001 increased power in delta, theta, and sigma bands 4-hours post administration during wake in WK rats at 30 mg / kg IP.

[0083] DLX-159 (half -log dosing, mg / kg, ip) and ketamine (10 mg / kg, ip) were administered to rats and qEEG was performed. Animals are dosed during waking states. After the plastogen has exerted its influence and cleared the animal’s system SWAs can be measured duringNREM sleep episodes. As shown in FIG. 7, DLX-159 increased power in delta and theta bands 4-hours post administration during wake in rats.

[0084] Slow wave amplitude (SWA) was determined in preclinical qEEG. SWA increased during NREM episode 10-12 hours post administration of DLX-159 (half-log dosing, mg / kg, ip) and ketamine (10 mg / kg, ip). As shown in FIG. 8, DLX-159 and ketamine increased SWA power.Example 5: Measuring Plastogen Free Fraction in Brain

[0085] Levels of plastogen in the brain are determined by conventional methods in the art, for example using MS-HPLC.Example 6: Correlating preclinical measurements qEEG values to arrive at an an Efficacious Dose

[0086] Markers of pre-clinical neuroplasticity may be correlated to identify therapeutically or behaviorally efficacious doses, as shown in FIG. 9.WSGR Docket No. 55776-731.601II. PHARMACEUTICAL COMPOSITIONSExample A-l: Parenteral Pharmaceutical Composition

[0087] To prepare a parenteral pharmaceutical composition suitable for administrationby injection (subcutaneous, intravenous), 1 -1000 mg of a water-soluble salt of a plastogen described herein, or a pharmaceutically acceptable salt or solvate thereof, is dissolved in sterile water and then mixed with 10 mL of 0.9% sterile saline. A suitable buffer is optionally added as well as optional acid or base to adjust the pH. The mixture is incorporated into a dosage unit form suitable for administration by injection.Example A-2: Oral Solution

[0088] To prepare a pharmaceutical composition for oral delivery, a sufficient amount of a plastogen described herein, or a pharmaceutically acceptable salt thereof, is added to water (with optional solubilizer(s), optional buffer(s) and taste masking excipients) to provide a 20 mg / mL solution.Example A-3: Oral Tablet

[0089] A tablet is prepared by mixing 20-50% by weight of a plastogen described herein, or a pharmaceutically acceptable saltthereof, 20-50% by weight of microcrystalline cellulose, and 1 -10% by weight of magnesium stearate or other appropriate excipients. Tablets are prepared by direct compression. The total weight of the compressed tablets is maintained at 100 -500 mg.Example A-4: Oral Capsule

[0090] To prepare a pharmaceutical composition for oral delivery, 1 -1000 mg of a plastogen described herein, or a pharmaceutically acceptable saltthereof, is mixed with starch or other suitable powder blend. The mixture is incorporated into an oral dosage unit such as a hard gelatin capsule, which is suitable for oral administration.

[0091] In another embodiment, 1 -1000 mg of a plastogen described herein, or a pharmaceutically acceptable saltthereof, is placed into Size 4 capsule, or size 1 capsule (hypromellose or hard gelatin) and the capsule is closed.

[0092] The examples and embodiments described herein are for illustrative purposes only and various modifications or changes suggested to persons skilled in the art are to be included within the spirit and purview of this application and scope of the appended claims.

Claims

WSGR Docket No. 55776-731.601CLAIMSWHAT IS CLAIMED IS:

1. A method of determining an effective amount of a plastogen for treating a brain disorder, comprising: d) administering one or more doses of the plastogen to a mammal; e) measuring1) by electroencephalogram (EEG) or quantitative electroencephalogram (qEEG), a slow wave frequency in the mammal; and, optionally,2) a concentration of the plastogen in the brain of the mammal; and f) determining the effective amount of the plastogen, wherein the effective amount of the plastogen increases power in the slow wave frequency.

2. The method of claim 1 , wherein determining the effective amount of the of the plastogen for treating a brain disorder comprises measuring1) by electroencephalogram (EEG) or quantitative electroencephalogram (qEEG), a slow wave frequency in the mammal; and,2) a concentration of the plastogen in the brain of the mammal.

3. The method of claim 1 or 2, wherein the effective amount of the plastogen increases power in the slow wave frequency and has a measurable concentration of the plastogen in the brain of the mammal.

4. The method of any one of claims 1 -3 , wherein the effective amount of the plastogen increases power in the slow wave frequency by at least 1 pp, at least 2 pp, at least 3 pp, at least 4 pp, at least 5 pp, at least 6 pp, at least 7 pp, at least 8 pp, at least 9 pp, at least 10 pp, at least 11 pp, at least 12 pp, at least 13 pp, at least 14 pp, at least 15 pp, at least 16 pp, at least 17 pp, at least 18 pp, at least 19 pp, at least 20 pp, at least 21 pp, at least 22 pp, at least 23 pp, at least 24 pp, at least 25 pp, at least 26 pp, at least 27 pp, at least 28 pp, at least 29 pp, or at least 30 pp above baseline.

5. The method of claim 3 or 4, wherein the measurable concentration of the plastogen in the brain of the mammal is a free fraction of the plastogen in the brain of the mammal.

6. The method of claim 5 , wherein the free fraction of the plastogen is at least 1 nM, at least 2nM, at least 3 nM, at least 4nM, at least 5nM, at least 6nM, at least 7nM, at least 8nM, at least 9nM, at least lOnM, at least 1 InM, at least 12nM, at least 13nM, at least 14nM, at least 15nM, at least 16nM, at least 17nM, at least 18nM, at least 19nM, at least 20nM, at least 2 InM, at least 22nM, at least 23nM, at least 24nM, at least 25nM, at least 26nM, at least 27nM, at least 28nM, at least 29nM, at least 30nM, at least 3 InM, at least 32nM, at least 33nM, at leastWSGR Docket No. 55776-731.60134nM, atleast 35nM, atleast36nM, atleast 37nM, atleast38nM, at least 39nMatleast40nM, at least 41nM, at least 42nM, at least 43nM, at least 44nM, at least 45nM at least 46nM, at least 47nM, at least 48nM, at least 49nM, at least 50nM, at least 60nM, at least 70nM, at least 80nM, at least 90nM, or at least lOOnM.

7. The method of any one of claims 1 -6, wherein measuring the slow wave frequency and the concentration of the plastogen in the brain occurs between 0.5 - 6 hours after administering the plastogen, such as 0.5 - 1 hour, 1 - 1.5 hours, 1.5 - 2 hours, 2 - 2.5 hours, 2.5 - 3 hours, 3 - 3.5 hours, 3.5 - 4 hours, 4 - 4.5 hours, 4.5 - 5 hours, 5 - 5.5 hours, or 5.5 - 6 hours, after administering the plastogen.

8. The method of any one of claims 1 -7, wherein the slow wave frequency is delta wave frequency or theta wave frequency.

9. The method of claim 8, wherein the slow wave frequency is measured between 1 to 8 Hz.

10. The method of any one of claims 1 -8, wherein the slow wave frequency is delta wave frequency.

11. The method of claim 10, wherein the delta wave frequency is measured between 1 to 4 Hz.

12. The method of any one of claims 1 -8, wherein the slow wave frequency is theta wave frequency.

13. The method of claim 12, wherein the theta wave frequency is measured between 4 to 8 Hz.

14. The method of any one of claims 1 -13, wherein administering the plastogen comprises administering 1 to 8 doses in different amounts to the mammal.

15. The method of any one of claims 1 -14, wherein the mammal is a rodent.

16. The method of any one of claims 1 -14, wherein the mammal is a human.

17. The method of any one of claims 1 -16, wherein the effective amount is a behaviorally effective amount or a therapeutically effective amount.

18. The method of any one of claims 1 -17, wherein the effective amount is a behaviorally effective amount.

19. The method of any one of claims 1 -17, wherein the effective amount is a therapeutically effective amount.

20. The method of any one of claims 1 -19, wherein the brain disorder is a neurological disease or disorder, a neuropsychiatric disease or disorder, a neurodegenerative disease or disorder, or a neuropsychological disease or disorder.

21. The method of claim 20, wherein the neuropsychiatric disease is major depressive disorder, treatment resistant depression, generalized anxiety disorder, post -traumatic stress disorder, obsessive compulsive disorder, substance use disorders, or a psychosis.WSGR Docket No. 55776-731.60122. The method of claim 21, wherein the psychosis is schizophrenia, bipolar disorder, or psychosis in Alzheimer’s disease (AD-P).

23. The method of claim 21 , wherein the neurodegenerative diseaseis a dementia, traumatic brain injury, or Parkinson’s disease.

24. The method of claim 23, wherein the dementia is Alzheimer’s disease, vascular dementia, Lewy body dementia, frontotemporal dementia, Huntington’s disease, or mixed dementia.

25. The method of any one of claims 1 -19, wherein the brain disorder is behavioral or psychological symptoms in dementia, or a combination thereof.

26. The method of any one of claims 1 -19, wherein the brain disorder is motor symptoms, behavioral symptoms, or psychological symptoms in Parkinson’s disease, or a combination thereof.

27. A method of treating a brain disorder in a patient, comprising administering an effective amount of a plastogen, wherein the effective amount of the plastogen is determined by: d) administering one or more doses of the plastogen to a mammal; e) measuring1) by electroencephalogram (EEG) or quantitative electroencephalogram (qEEG), a slow wave frequency in the mammal; and, optionally,2) a concentration of the plastogen in the brain of the mammal; and f) determining the effective amount of the plastogen, wherein the effective amount of the plastogen increases power in the slow wave frequency.

28. The method of claim 27, wherein determining the effective amount of the of the plastogen for treating a brain disorder comprises measuring1) by electroencephalogram (EEG) or quantitative electroencephalogram (qEEG), a slow wave frequency in the mammal; and,2) a concentration of the plastogen in the brain of the mammal.

29. The method of claim 27 or 28, wherein the effective amount of the plastogen increases power in the slow wave frequency and has a measurable concentration of the plastogen in the brain of the mammal.

30. The method of any one of claims 27-29, wherein the effective amount of the plastogen increases power in the slow wave frequency by at least 1 pp, at least 2 pp, at least 3 pp, at least 4 pp, at least 5 pp, at least 6 pp, at least 7 pp, at least 8 pp, at least 9 pp, at least 10 pp, at least 11 pp, at least 12 pp, at least 13 pp, at least 14 pp, at least 15 pp, at least 16 pp, at least 17 pp, at least 18 pp, at least 19 pp, at least 20 pp, at least 21 pp, at least 22 pp, at least 23 pp,WSGR Docket No. 55776-731.601 at least 24 pp, at least 25 pp, at least 26 pp, at least 27 pp, at least 28 pp, at least 29 pp, or at least 30 pp above baseline.

31. The method of claim 29 or 30, wherein the measurable concentration of the plastogen in the brain of the mammal is a free fraction of the plastogen in the brain of the mammal.

32. The method of claim 31, wherein the free fraction of the plastogen is at least InM, at least2nM, at least 3nM, at least 4nM, at least 5nM, at least 6nM, at least 7nM, at least 8nM, at least 9nM, at least lOnM, at least 1 InM, at least 12nM, at least 13nM, at least 14nM, at least 15nM, at least 16nM, at least 17nM, at least 18nM, at least 19nM, at least 20nM, at least2 InM, at least 22nM, at least 23nM, at least 24nM, at least 25nM, at least 26nM, at least27nM, at least 28nM, at least 29nM, at least 30nM, at least 3 InM, at least 32nM, at least33nM, at least 34nM, at least 35nM, at least 36nM, at least 37nM, at least 38nM, at least39nMatleast40nM, at least 4 InM, atleast 42nM, atleast 43nM, atleast44nM, atleast45nM at least 46nM, at least 47nM, at least 48nM, at least 49nM, at least 50nM, at least 60nM, at least 70nM, at least 80nM, at least 90nM, or at least lOOnM.

33. The method of any one of claims 27-32, wherein measuring the slow wave frequency and the concentration of the plastogen in the brain occurs between 0.5 - 6 hours after administering the plastogen, such as 0.5 - 1 hour, 1 - 1.5 hours, 1.5 - 2 hours, 2 - 2.5 hours, 2.5 - 3 hours, 3 - 3.5 hours, 3.5 - 4 hours, 4 - 4.5 hours, 4.5 - 5 hours, 5 - 5.5 hours, or 5.5 - 6 hours, after administering the plastogen.

34. The method of any one of claims 27-33, wherein the slow wave frequency is delta wave frequency or theta wave frequency.

35. The method of claim 34, wherein the slow wave frequency is measured between 1 to 8 Hz.

36. The method of any one of claims 27-34, wherein the slow wave frequency is delta wave frequency.

37. The method of claim 36, wherein the delta wave frequency is measured between 1 to 4 Hz.

38. The method of any one of claims 27-34, wherein the slow wave frequency is theta wave frequency.

39. The method of claim 38, wherein the theta wave frequency is measured between 4 to 8 Hz.

40. The method of any one of claims 27-39, wherein administering the plastogen comprises administering 1 to 8 doses in different amounts to the mammal.

41. The method of any one of claims 27-40, wherein the mammal is a rodent.

42. The method of any one of claims 27-40, wherein the mammal is a human.

43. The method of any one of claims 27-42, wherein the effective amount is a behaviorally effective amount or a therapeutically effective amount.WSGR Docket No. 55776-731.60144. The method of any one of claims 27-43, wherein the effective amount is a behaviorally effective amount.

45. The method of any one of claims 27-43, wherein the effective amount is a therapeutically effective amount.

46. The method of any one of claims 27-45, wherein the brain disorder is a neurological disease or disorder, a neuropsychiatric disease or disorder, a neurodegenerative disease or disorder, or a neuropsychological disease or disorder.

47. The method of claim 46, wherein the neuropsychiatric disease is major depressive disorder, treatment resistant depression, generalized anxiety disorder, post -traumatic stress disorder, obsessive compulsive disorder, substance use disorders, or a psychosis.

48. The method of claim 47, wherein the psychosis is schizophrenia, bipolar disorder, or psychosis in Alzheimer’s disease (AD-P).

49. The method of claim 46, wherein the neurodegenerative diseaseis a dementia, traumatic brain injury, or Parkinson’s disease.

50. The method of claim 49, wherein the dementia is Alzheimer’s disease, vascular dementia, Lewy body dementia, frontotemporal dementia, Huntington’s disease, or mixed dementia.

51. The method of any one of claims 27-45, wherein the brain disorder is behavioral or psychological symptoms in dementia, or a combination thereof.

52. The method of any one of claims 27-45, wherein the brain disorder is motor symptoms, behavioral symptoms, or psychological symptoms in Parkinson’s disease, or a combination thereof.

53. A pharmaceutical composition comprising an effective amount of a plastogen, wherein the effective amount of the plastogen is determined by: d) administering one or more doses of the plastogen to a mammal; e) measuring1) by electroencephalogram (EEG) or quantitative electroencephalogram (qEEG), a slow wave frequency in the mammal; and, optionally,2) a concentration of the plastogen in the brain of the mammal; and f) determining the effective amount of the plastogen, wherein the effective amount of the plastogen increases power in the slow wave frequency.

54. The method of claim 53, wherein determining the effective amount of the of the plastogen for treating a brain disorder comprises measuring1) by electroencephalogram (EEG) or quantitative electroencephalogram (qEEG), a slow wave frequency in the mammal; and,WSGR Docket No. 55776-731.6012) a concentration of the plastogen in the brain of the mammal.

55. The method of claim 53 or 54, wherein the effective amount of the plastogen increases power in the slow wave frequency and has a measurable concentration of the plastogen in the brain of the mammal.

56. The method of any one of claims 53-55, wherein the effective amount of the plastogen increases power in the slow wave frequency by at least 1 pp, at least 2 pp, at least 3 pp, at least 4 pp, at least 5 pp, at least 6 pp, at least 7 pp, at least 8 pp, at least 9 pp, at least 10 pp, at least 11 pp, at least 12 pp, at least 13 pp, at least 14 pp, at least 15 pp, at least 16 pp, at least 17 pp, at least 18 pp, at least 19 pp, at least 20 pp, at least 21 pp, at least 22 pp, at least 23 pp, at least 24 pp, at least 25 pp, at least 26 pp, at least 27 pp, at least 28 pp, at least 29 pp, or at least 30 pp above baseline.

57. The method of claim 55 or 56, wherein the measurable concentration of the plastogen in the brain of the mammal is a free fraction of the plastogen in the brain of the mammal.

58. The method of claim 57, wherein the free fraction of the plastogen is at least InM, at least2nM, at least 3nM, at least 4nM, at least 5nM, at least 6nM, at least 7nM, at least 8nM, at least 9nM, at least lOnM, at least 1 InM, at least 12nM, at least 13nM, at least 14nM, at least 15nM, at least 16nM, at least 17nM, at least 18nM, at least 19nM, at least 20nM, at least2 InM, at least 22nM, at least 23nM, at least 24nM, at least 25nM, at least 26nM, at least27nM, at least 28nM, at least 29nM, at least 30nM, at least 3 InM, at least 32nM, at least33nM, at least 34nM, at least 35nM, at least 36nM, at least 37nM, at least 38nM, at least39nMatleast40nM, at least 4 InM, atleast 42nM, atleast 43nM, atleast44nM, atleast45nM at least 46nM, at least 47nM, at least 48nM, at least 49nM, at least 50nM, at least 60nM, at least 70nM, at least 80nM, at least 90nM, or at least lOOnM.

59. The method of any one of claims 53-58, wherein measuring the slow wave frequency and the concentration of the plastogen in the brain occurs between 0.5 - 6 hours after administering the plastogen, such as 0.5 - 1 hour, 1 - 1.5 hours, 1.5 - 2 hours, 2 - 2.5 hours, 2.5 - 3 hours, 3 - 3.5 hours, 3.5 - 4 hours, 4 - 4.5 hours, 4.5 - 5 hours, 5 - 5.5 hours, or 5.5 - 6 hours, after administering the plastogen.

60. The method of any one of claims 53-59, wherein the slow wave frequency is delta wave frequency or theta wave frequency.

61. The method of claim 60, wherein the slow wave frequency is measured between 1 to 8 Hz.

62. The method of any one of claims 53-60, wherein the slow wave frequency is delta wave frequency.

63. The method of claim 62, wherein the delta wave frequency is measured between 1 to 4 Hz.WSGR Docket No. 55776-731.60164. The method of any one of claims 53-60, wherein the slow wave frequency is theta wave frequency.

65. The method of claim 64, wherein the theta wave frequency is measured between 4 to 8 Hz.

66. The method of any one of claims 53-65, wherein administering the plastogen comprises administering 1 to 8 doses in different amounts to the mammal.

67. The method of any one of claims 53-66, wherein the mammal is a rodent.

68. The method of any one of claims 53-66, wherein the mammal is a human.

69. The method of any one of claims 53-68, wherein the effective amount is a behaviorally effective amount or a therapeutically effective amount.

70. The method of any one of claims 53-69, wherein the effective amount is a behaviorally effective amount.

71. The method of any one of claims 53-69, wherein the effective amount is a therapeutically effective amount.

72. The method of any one of claims 53-71, wherein the brain disorder is a neurological disease or disorder, a neuropsychiatric disease or disorder, a neurodegenerative disease or disorder, or a neuropsychological disease or disorder.

73. The method of claim 72, wherein the neuropsychiatric disease is major depressive disorder, treatment resistant depression, generalized anxiety disorder, post -traumatic stress disorder, obsessive compulsive disorder, substance use disorders, or a psychosis.

74. The method of claim 73, wherein the psychosis is schizophrenia, bipolar disorder, or psychosis in Alzheimer’s disease (AD-P).

75. The method of claim 73 , wherein the neurodegenerative diseaseis a dementia, traumatic brain injury, or Parkinson’s disease.

76. The method of claim 75, wherein the dementia is Alzheimer’s disease, vascular dementia, Lewy body dementia, frontotemporal dementia, Huntington’s disease, or mixed dementia.

77. The method of any one of claims 53-71, wherein the brain disorder is behavioral or psychological symptoms in dementia, or a combination thereof.

78. The method of any one of claims 53-71, wherein the brain disorder is motor symptoms, behavioral symptoms, or psychological symptoms in Parkinson’s disease, or a combination thereof.

79. The method of any one of claims 1 -78, wherein the plastogen is a small molecule.

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