Dendrimer compositions for targeted delivery of psychedelic therapeutics

Abstract

Dendrimer formulations containing one or more psychedelic or hallucinogenic agents and methods of use thereof are described. Preferably, the dendrimer-conjugated agents selectively bind to one or more receptors on or within target cells. The formulations are suitable for enteral and / or parenteral delivery to treat one or more receptor-mediated disorders, including psychological, cognitive, behavioral, and / or mood disorders.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention is generally in the field of hallucinogenic drug formulations, and more particularly in the field of dendrimer hallucinogen-conjugates that improve receptor binding, cell selectivity, and reduce side effects.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 401,470, filed August 26, 2022, entitled "DENDRIMER COMPOSITIONS FOR TARGETED DELIVERY OF PSYCHEDELIC THERAPEUTICS," by The Johns Hopkins University, listed inventors Kannan Rangaramanujam, Kunal Parikh, Sujatha Kannan, and Anjali Sharma, which is incorporated by reference in its entirety.

[0003] STATEMENT OF FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT none. [Background technology]

[0004] Background of the Invention The word "psychedelic" (psyche, i.e., mind or soul, and delos, i.e., to show) was first coined in 1956 by psychiatrist Humphry Osmond, who was conducting research on lysergic acid diethylamide (LSD) at the time. Psychedelic drugs such as N,N-DMT / DMT (N,N-dimethyltryptamine), 5-MeO-DMT (5-methoxy-N,N-dimethyltryptamine), LSD (lysergic acid diethylamide), MDMA (3,4-methylenedioxymethamphetamine), and psilocybin have held significant value as entheogens in spiritual, religious (shamanic), and sociocultural rituals in Central and South American cultures for millennia.

[0005] Until recently, the scientific environment has not allowed exploration of the spiritual, religious, and medical value of these drugs. More recently, a second wave of psychedelic research is currently focusing on psychedelics as psychopharmaceuticals for treating alcohol and tobacco addiction, general mood and anxiety disorders, and cancer-related depression. Psychedelics and other classes of compounds related to them (e.g., entactogens, known to induce a temporary alteration of consciousness accompanied by acute changes in physical, perceptual, and cognitive properties) are increasingly gaining attention. This is due, in part, to their widespread use, novel synthesis and purification methods, improved mechanistic understanding, and ongoing clinical trials. Their puzzling potential to affect thought and consciousness also makes it important to understand their mechanisms of action and to target their effects to desired cells and tissue regions to avoid side effects.

[0006] plant-derived indoleamines (e.g., N,N-dimethyltryptamine (DMT), 5-methoxy-DMT (5-MeO-DMT), psilocybin, 4-hydroxy-DMT (psilocyn, the active metabolite of psilocybin)), phenylalkylamines (e.g., mescaline and synthetic "amphetamines" such as 2,5-dimethoxy-4-iodoamphetamine (DOI) and 2,5-dimethoxy-4-bromoamphetamine (DOB)), Psychedelics and hallucinogenic drugs, such as mianserin and semi-synthetic ergolines (e.g., LSD), have shown great therapeutic potential for the treatment of several mental health and neurological disorders (e.g., depression, treatment-resistant depression, suicidal ideation, autism, bipolar disorder, anxiety, drug dependence, substance abuse disorders, post-traumatic stress disorder, obesity, headache, pain, fibromyalgia, obsessive-compulsive disorder, anorexia nervosa, inflammation, Alzheimer's disease, attention deficit / hyperactivity disorder, narcolepsy).

[0007] Psychedelic molecules generally achieve their therapeutic effects through activation of 5-HT2A receptors in the brain cortex. 5-HT2A is a type of serotonin receptor thought to mediate brain plasticity and is found in high concentrations in areas of the cortex involved in high-level cortical processing (frontal cortical regions: cingulate cortex and posterior cingulate cortex). However, the practical implementation of these potential treatments faces significant challenges, including solubility, bioavailability, absorption, side effects / toxicity, time to effect, duration of effect, and hallucinogenic activity. Psychedelics have limited access to these critical brain regions, specifically brain and immune cells involved in disease processes. Collectively, these issues limit the applicability, efficacy, and translatability of this class of drugs, preventing the realization of their broad therapeutic potential.

[0008] It is therefore an object of the present invention to provide formulations that allow for more selective delivery of psychedelic and hallucinogenic drugs. Summary of the Invention [Means for solving the problem]

[0009] Summary of the Invention The group of hallucinogens and dendrimer compositions formulated herein includes (i) psychedelics, i.e., a group of serotonergic, typically Schedule I, agonists (e.g., psilocybin, lysergic acid, mescaline, etc.); (ii) entactogens, Schedule I monoamine releasers and reuptake inhibitors (e.g., 3,4-methylenedioxy-methamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA) known to evoke feelings of emotional tolerance and emotional connection); (iii) dissociative agents, i.e., glutamatergic NMDA antagonists (e.g., ketamine, dextromethorphan (DXM), and nitrous oxide, etc.); (iv) atypical hallucinogens with diverse mechanisms (e.g., Δ 9Dendrimer conjugate compositions have been developed that can deliver drugs to receptors on specific cells (neuronal cells, glial cells, macrophages), including targets on their surface and within their interiors. These formulations, through superior binding to target receptors, can enhance the efficacy of these drugs, allow for lower doses, provide new mechanistic insights, reduce side effects, improve solubility, formulation, PK, and other aspects of the use of these drugs, and open up new clinical uses.

[0010] The formulations are based on conjugation of drugs to dendrimers, particularly PAMAM (such as G3, G4, G5, and G6 hydroxyl-terminated PAMAM dendrimers) and glucose dendrimers (such as G1, G2, and G3 glucose dendrimers), which provide improved solubility, uptake into brain and other specific cell types, and selectivity of uptake and receptor binding. [Brief explanation of the drawings]

[0011] [Figure 1A-B] 1A and 1B are schematic diagrams showing an exemplary synthetic route to dendrimer-psilocin bearing a cleavable ester linker using click chemistry. Psilocin is first conjugated via the ester linker to a linker bearing an azide moiety (FIG. 1A), and then conjugated to a dendrimer modified with surface alkyne groups via an azide-alkyne click reaction (FIG. 1B).

[0012] [Figure 2A-B]2A and 2B are schematic diagrams showing an exemplary synthetic route to a dendrimer-psilocin analog with a non-cleavable amide linker using click chemistry. The psilocin analog is first conjugated via the amide linker to a linker containing an azide moiety (FIG. 2A), and then conjugated to a dendrimer modified with surface alkyne groups via an azide-alkyne click reaction (FIG. 2B).

[0013] [Figure 3A-B] 3A and 3B are schematic diagrams showing an exemplary synthetic route to dendrimer-ketamine bearing a non-cleavable amino-alkyl linker using copper-catalyzed alkyne-azide click chemistry. Ketamine hydrochloride (1) is first modified with an alkyne (FIG. 3A) and then conjugated to a dendrimer modified with surface azide groups via an azide-alkyne click reaction (FIG. 3B).

[0014] [Figure 4A-B] 4A and 4B are schematic diagrams showing an exemplary synthetic route to a dendrimer-DMT analog with a non-cleavable amide linker using copper-catalyzed alkyne-azide click chemistry. An N,N-dimethyltryptamine analog (DMT analog) is first conjugated to a linker bearing an azide moiety via the amide linker (FIG. 4A), and then conjugated to a dendrimer modified with surface alkyne groups via an azide-alkyne click reaction (FIG. 4B).

[0015] [Figure 5A-B] Figures 5A and 5B are schematic diagrams of the synthesis of dendrimer-DMT with non-cleavable amino-alkyl linkers. Figure 5A shows a DMT drug modified with an alkyne group. Figure 5B shows conjugation to a dendrimer modified with surface azide groups via an azide-alkyne click reaction.

[0016] [Figure 6A-B]Figures 6A and 6B are schematic diagrams of dendrimer-lysergic acid diethylamide (dendrimer-LSD) with a non-cleavable amino-alkyl linker. Figure 6A shows an LSD modified with an alkyne group. Figure 6B shows conjugation to a dendrimer modified with surface azide groups via an azide-alkyne click reaction.

[0017] [Figure 7] FIG. 7 is a schematic diagram of a stepwise synthetic route for the synthesis of a glucose dendrimer-psilocin conjugate with a cleavable ester linker.

[0018] [Figure 8] FIG. 8 is a schematic diagram of a stepwise synthetic route for the synthesis of a glucose dendrimer-psilocin analog conjugate with a non-cleavable amide linker.

[0019] [Figure 9] FIG. 9 is a schematic diagram of a stepwise synthetic route for the synthesis of glucose dendrimer-ketamine conjugates bearing non-cleavable amino-alkyl linkers.

[0020] [Figure 10] FIG. 10 is a schematic diagram of a stepwise synthetic route for the synthesis of glucose dendrimer N,N dimethyltryptamine analog (DMT analog) conjugates with non-cleavable amide linkers.

[0021] [Figure 11] FIG. 11 is a schematic diagram of a stepwise synthetic route for the synthesis of glucose dendrimer-DMT analog conjugates bearing non-cleavable amino-alkyl linkers.

[0022] [Figure 12]FIG. 12 is a schematic diagram of a stepwise synthetic route for the synthesis of a glucose dendrimer-lysergic acid diethylamide (LSD) conjugate with a non-cleavable amino-alkyl linker.

[0023] [Figure 13] FIG. 13 is a schematic overview of the major pharmacological targets of LSD, psilocybin, DMT, MDMA, and ketamine, the signaling cascades involved, hormonal modulation, and the main behavioral outcomes following their administration in both animals and humans.

[0024] [Figure 14A] Figure 14A is a schematic representation of the synthesis of a PAMAM dendrimer-norketamine conjugate. Figure 14B is a schematic representation of the synthesis of a glucose dendrimer-norketamine conjugate. [Figure 14B] Same as above.

[0025] [Figure 15A] Figure 15A shows a graph of the NMDAR1A / 2B antagonist assay for glucose dendrimer-ketamine (IC50 = 4.54 μM), hydroxyl dendrimer-ketamine (IC50 > 100), and norketamine (IC50 = 6.96 μM). Figure 15B shows the % binding efficacy of logarithmic concentrations (micromolar) of compounds in an assay for the D2L human dopamine GPCR cell-based agonist cAMP. Norketamine (filled circles), glucose dendrimer-ketamine EC50 = 13.08 μmolar (open circles), and hydroxyl dendrimer-ketamine EC50 = 4.263 μmolar (triangles). Figure 15C shows the % efficacy of logarithmic concentrations (micromolar) of ketamine in an assay for the TA1 human trace amine GPCR cell-based agonist cAMP. Norketamine (closed circles), glucose dendrimer-ketamine EC50 = 13.08 micromolar (open circles) and hydroxyl dendrimer-ketamine EC50 = 4.263 micromolar (triangles). [Figure 15B-C] Same as above.

[0026] [Figure 16A-B] Figures 16A and 16B are graphs of the composite neurobehavioral score (Figure 16A) and survival probability after birth (Figure 16B) of wild-type, knockout saline (control) versus knockout mice treated with dendrimer-ketamine conjugate. Figure 16C is a graph of the distance traveled (m). Figure 16D is a graph of the speed at which the mice traveled. Figure 16E is a graph of the time spent in the corner. [Figure 16C-D] Same as above. [Figure 16E] Same as above. DETAILED DESCRIPTION OF THE INVENTION

[0027] Detailed Description of the Invention I. Definition The term "hallucinogen" refers to a group of chemically heterogeneous compounds, all of which have the ability to induce altered states of consciousness (ASCs), characterized by profound changes in mood, thought processes, perceptions, and experiences of self and the environment rarely experienced outside of dreams, meditative, and religious highs, as well as acute psychosis. Not all hallucinogenic compounds reliably produce visual and auditory hallucinations. Therefore, hallucinogens are sometimes referred to as psychotomimetic, psycholytic, or psychedelic agents, reflecting the wide variety of attitudes and intentions with which these substances have been utilized.

[0028] The term "psychedelics" refers to a subset of hallucinogenic drugs whose primary action is to induce non-ordinary states of consciousness. This results in specific psychological visual and auditory changes and often substantial alterations in states of consciousness. Psychedelic states are often compared to meditative, psychodynamic, or transcendental types of altered mind. The "classic" psychedelics—those with the greatest scientific and cultural influence—are mescaline, LSD, psilocybin, and DMT. The majority of psychedelics fall into one of three families of chemical compounds: tryptamines, phenethylamines, or lysergamides, and many tend to act via serotonin 2A receptor agonism. When compounds bind to serotonin 5-HT2A receptors, they modulate the activity of key brain circuits involved in sensory perception and cognition. Although the exact nature of how psychedelics induce changes in sensory perception and cognition via 5-HT2A receptors remains unknown, the resulting reduction in default mode network activity and increased functional connectivity between brain regions may be one of the most important pharmacological mechanisms underlying psychedelic experiences, particularly ego death.

[0029] The terms "active agent" or "biologically active agent" are used interchangeably to refer to a chemical or biological compound that induces a desired pharmacological and / or physiological effect, which may be prophylactic, therapeutic, or diagnostic. These may be nucleic acids, nucleic acid analogs, small molecules with a molecular weight of less than 2 kD, more typically less than 1 kD, peptidomimetics, proteins or peptides, carbohydrates or sugars, lipids, or combinations thereof. The term also encompasses pharmaceutically acceptable, pharmacologically active derivatives of drugs, including, but not limited to, salts, esters, amides, prodrugs, active metabolites, and analogs. The term "therapeutic agent" refers to an agent that can be administered to treat one or more symptoms of a disease or disorder. The term "diagnostic agent" generally refers to an agent that can be administered to locate, precisely target, and confirm a pathological process. Diagnostic agents can label target cells, allowing for subsequent detection or imaging of these labeled target cells.

[0030] An "analog" in relation to a given compound refers to another compound that is structurally similar, functionally similar, or both to the specified compound. Structural similarity can be determined using any criterion known in the art, such as the Tanimoto coefficient, which provides a quantitative measure of similarity between two compounds based on their molecular descriptors. Preferably, molecular descriptors are 2D properties such as fingerprints, topological indices, and maximum common substructures, or 3D properties such as overall shape and molecular field. The Tanimoto coefficient ranges between 0 and 1 (for pairs of dissimilar molecules and pairs of identical molecules, respectively). A compound is considered an analog of the specified compound if it has a Tanimoto coefficient between 0.5 and 1.0 (inclusive), preferably between 0.7 and 1.0 (inclusive), and most preferably between 0.85 and 1.0 (inclusive) of the specified compound. A compound is functionally similar to the specified compound if it induces the same pharmacological or physiological effect, or both, as the specified compound. "Analog" can also refer to modifications, including, but not limited to, hydrolysis, reduction, or oxidation products of a compound. Hydrolysis, reduction, and oxidation reactions are known in the art.

[0031] The term "therapeutically effective amount" refers to the amount of a therapeutic agent that, when incorporated into and / or onto a dendrimer, produces a desired effect at a reasonable benefit / risk ratio applicable to any medical treatment. The effective amount may vary depending on factors such as the disease or condition being treated, the particular targeting construct being administered, the size of the subject, or the severity of the disease or condition. One of ordinary skill in the art can empirically determine the effective amount of a particular compound without necessitating undue experimentation. In some embodiments, the term "effective amount" refers to the amount of a therapeutic or prophylactic agent that relieves or reduces one or more disease symptoms.

[0032] The terms "inhibit" or "reducing," in the context of inhibition, refer to a decrease or reduction in activity and amount. This can be complete inhibition, or a reduction in activity or amount, or partial inhibition or reduction. Inhibition or reduction can be relative to a control or standard level. Inhibition can be 5, 10, 25, 50, 75, 80, 85, 90, 95, 99, or 100%. For example, a dendrimer composition comprising one or more inhibitors can inhibit or reduce the activity and / or amount of affected neurons by about 10%, 20%, 30%, 40%, 50%, 75%, 85%, 90%, 95%, or 99% from the activity and / or amount of the same cells in an equivalent tissue of a subject not administered or treated with the dendrimer composition. In some embodiments, inhibition and reduction are compared at the mRNA, protein, cell, tissue, and organ levels. For example, inhibition and reduction of the rate of neuronal loss, the rate of brain weight loss, or the rate of hippocampal volume loss compared to untreated control subjects.

[0033] The terms "treating" or "preventing" refer to improving, reducing, or otherwise halting the development or progression of a disease, disorder, or condition in an animal that may be susceptible to, but has not yet been diagnosed with, a disease, disorder, and / or condition; inhibiting, e.g., slowing the progression of, a disease, disorder, or condition; and alleviating, e.g., causing regression of, a disease, disorder, and / or condition. Treating a disease or condition includes improving at least one symptom of a particular disease or condition even when the underlying pathophysiology is unaffected, such as treating a subject's pain by administering an analgesic drug even when such a drug does not treat the cause of the pain. Desired effects of treatment include slowing the rate of disease progression, improving or alleviating the disease state, and remission or improved prognosis. For example, an individual is successfully "treated" when one or more symptoms associated with depression are reduced or eliminated, including, but not limited to, reduced levels of anxiety, agitation, or restlessness; amelioration of feelings of sadness, fear, emptiness, or hopelessness; an increase in the quality of life of an individual affected by the disease; a reduction in the dosage of other medications required to treat the disease; and a slowing of the progression of the disease.

[0034] The phrases "pharmaceutically acceptable" or "biocompatible" refer to compositions, polymers, and other materials and / or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. The phrase "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable substance, composition, or vehicle, such as a liquid or solid filler, diluent, solvent, or encapsulating substance, that is involved in carrying or transporting a subject composition from one organ or body part to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the subject composition and not injurious to the patient.

[0035] The term "biodegradable" generally refers to a material that, under physiological conditions, breaks down or erodes into smaller units or chemical species that can be metabolized in vivo, excreted, or excreted. The degradation time is a function of composition and geometry.

[0036] The term "dendrimer" includes, but is not limited to, a molecular architecture having an inner core, an inner layer or "generation" of repeating units regularly attached to this initial core, and an outer surface of terminal groups attached to the outermost generation.

[0037] The term "functionalize" means to modify a compound or molecule in a manner that results in the attachment of a functional group or moiety. For example, a molecule can be functionalized by introducing a molecule that makes it a strong nucleophile or a strong electrophile.

[0038] The term "targeting moiety" refers to a moiety that is localized to or distant from a specific location. The moiety can be, for example, a protein, a nucleic acid, a nucleic acid analog, a carbohydrate, or a small molecule. The location can be a tissue, a specific cell type, an intracellular compartment, or a molecule such as a receptor.

[0039] The term "prolonged residence time" refers to an increase in the time required for a drug to clear from a patient's body or from an organ or tissue of the patient. In certain embodiments, "prolonged residence time" refers to a drug that is cleared with a half-life that is 10%, 20%, 50%, or 75% longer than a comparative standard, such as a comparable drug that is not conjugated to a delivery vehicle, such as a dendrimer. In certain embodiments, "prolonged residence time" refers to a drug that is cleared with a half-life that is 2, 5, 10, 20, 50, 100, 200, 500, 1000, 2000, 5000, or 10,000 times longer than a comparative standard, such as a comparable drug that does not carry a dendrimer that specifically targets a particular cell type.

[0040] The terms "incorporated" and "encapsulated" refer to incorporating, formulating, or otherwise including an agent within and / or on the surface of a composition that allows for release, such as sustained release, of the agent in a desired application. Drugs or other substances can be incorporated into dendrimers by binding (by covalent, ionic, or other bonding interactions) to one or more surface functional groups of such dendrimers, by physical admixture, by enveloping the agent within the dendrimer structure, and / or by encapsulating the agent within the dendrimer structure.

[0041] As used herein, the central nervous system ("CNS") includes the brain and spinal cord. As used herein, the peripheral nervous system ("PNS") refers to nerves other than the brain and spinal cord.

[0042] "Hydroxyl-terminated" in reference to a dendrimer refers to a dendrimer having hydroxyl groups on its surface that are not attached to the termini of the dendrimer via a sugar moiety (such as a saccharide moiety).

[0043] "Sugar-terminated" in reference to a dendrimer refers to a dendrimer that contains sugar moieties (such as saccharide moieties) on its surface but not within its core.

[0044] "Sugar-based" in reference to dendrimers refers to dendrimers that contain sugar moieties (such as saccharide moieties) in their core, or in their core and on their surface. II. Composition

[0045] Compositions of dendrimers conjugated or complexed with one or more hallucinogens and / or dissociative compounds have been developed for preventing and / or treating symptoms associated with one or more psychological, cognitive, behavioral, mood, and / or non-neurological disorders in subjects in need thereof. The compositions are particularly suitable for treating and / or ameliorating one or more symptoms of mental health and neurological disorders (e.g., depression (major depressive disorder, treatment-resistant depression, postpartum depression), suicidal ideation, autism, bipolar disorder, anxiety, drug addiction, substance abuse disorders, post-traumatic stress disorder, obesity, headache, cluster headache, migraine, epilepsy, pain, fibromyalgia, obsessive-compulsive disorder, anorexia nervosa, inflammation, Alzheimer's disease, attention deficit / hyperactivity disorder, narcolepsy, Tourette's syndrome). In preferred embodiments, the dendrimer is a glucose dendrimer or a hydroxyl-terminated dendrimer, such as a hydroxyl-terminated PAMAM or sugar-modified dendrimer.

[0046] Exemplary psychedelics include psilocin, ketamine (R-ketamine, S-ketamine, (R / S)-ketamine), norketamine, ketamine analogs, ketamine metabolites, N,N-dimethyltryptamine (DMT), 4-acetoxy-N,N-dimethyltryptamine, 5-methoxyDMT, 5-chloroDMT, lysergide (LSD), 3,4-methylenedioxymethamphetamine (MDMA), psilocybin, ibogaine, mescaline, mianserin, and norbeocystine.

[0047] Generally, hallucinogens and / or their derivatives bind to receptors on the surface of target cells and / or receptors inside the target cells. Exemplary target cells include, but are not limited to, brain cells such as microglia, astrocytes, and / or neurons, e.g., cells within the site of pathology in the brain or CNS; peripheral neurons, glia, and / or their supporting cells, e.g., cells in the peripheral nervous system, such as intestinal cells, cardiovascular cells, and immune system cells. The microglia and / or astrocytes to which hallucinogens and / or their derivatives are delivered may be activated or inactive microglia and / or astrocytes. Classical / serotonergic psychedelic compounds also exhibit immunomodulatory properties and therefore have applications in autoimmune disorders.

[0048] The hallucinogen and / or derivative thereof of the dendrimer-active agent conjugate binds to a target receptor on the surface of or inside a target cell. In some embodiments, when the hallucinogen and / or derivative thereof binds to the target receptor, the agent remains conjugated to the dendrimer. In these embodiments, after binding, the agent can be released from the dendrimer or can remain conjugated to the dendrimer as an intact dendrimer-active agent conjugate. In some embodiments, the hallucinogen and / or derivative thereof is released from the dendrimer in close proximity to the target receptor and then binds to the target receptor on a target nervous system cell and / or glial cell. A. Dendrimer

[0049] Dendrimers are three-dimensional, hyperbranched, monodisperse, spherical, polyvalent macromolecules containing surface end groups (Tomalia, D.A., et al., Biochemical Society Transactions, 35, 61 (2007); and Sharma, A., et al., ACS Macro Letters, 3, 1079 (2014)).

[0050] The term "dendrimer" includes, but is not limited to, a molecular architecture having an inner core ("GO") and layers (or "generations") of repeating units attached to and extending from the inner core, each layer having one or more branch points and an outer surface with terminal groups attached to the outermost generation. In some embodiments, dendrimers have a regular, dendritic or "star-shaped" molecular structure.

[0051] Typically, dendrimers have a diameter between about 1 nm and about 60 nm, more preferably between about 1 nm and about 50 nm, about 1 nm and about 40 nm, about 1 nm and about 30 nm, about 1 nm and about 20 nm, about 1 nm and about 10 nm, or about 1 nm and about 5 nm. In some embodiments, the diameter is between about 1 nm and about 2 nm. The preferred size of dendrimers for crossing the blood-brain barrier ("BBB") is less than 5 nm, while those that do not cross the BBB and remain in the peripheral circulation are greater than 5 nm. In some embodiments, the dendrimers have a diameter effective to penetrate the BBB and remain in proximity to or within target nervous system cells and / or glial cells for delivery of a drug conjugated to the dendrimer. In some embodiments, the dendrimer has a diameter effective to penetrate the BBB and internalize into target nervous system cells and / or glial cells for delivery of an agent conjugated to the dendrimer, e.g., neurons, oligodendrocytes, astrocytes, microglial cells, and neuroglial supporting cells. In some embodiments, the dendrimer has a diameter effective to penetrate a barrier interface, such as the blood-nerve barrier ("BNB"), and internalize into nervous system cells and / or glial cells of the peripheral nervous system for delivery of an agent conjugated to the dendrimer, e.g., neurons, Schwann cells, satellite cells, and neuroglial supporting cells. In some embodiments, the dendrimer has a diameter effective to remain in the peripheral circulation for delivery of an agent conjugated to the dendrimer to target cells of the peripheral nervous system, e.g., enteric neurons and glia. A major advantage of using dendrimer conjugates is the ability of dendrimers to enhance the binding of psychedelic drugs to their target receptors on target cells, for example, binding of the compound to serotonergic receptors on neurons in affected regions of the brain.

[0052] In some embodiments, the dendrimer has a molecular weight of between about 500 Daltons and about 100,000 Daltons, inclusive, between about 500 Daltons and about 50,000 Daltons, inclusive, or between about 1,000 Daltons and about 20,000 Daltons, inclusive. Dendrimer sizes less than 30,000 Da are preferred for transport across the BBB, and sizes greater than 50,000 Da are preferred for peripheral containment.

[0053] In some embodiments, the dendrimer has a hypercore (e.g., dipentaerythritol) and one or more monosaccharide branching units. In some embodiments, the monosaccharide branching units are conjugated to the core or previous layer of monomers via a linker, such as a polyethylene glycol chain. In a preferred embodiment, the hypercore is dipentaerythritol and the monosaccharide branching units are glucose-based branching units, such as those shown in Structures II-IV.

[0054] In the most preferred embodiment, the dendrimer is made entirely from glucose building blocks. While sugar-modified PAMAM dendrimers can also function, dendrimers made from sugars, especially glucose, are most preferred. Particularly preferred glucose dendrimers are G1-G3 glucose dendrimers, such as G1, G2, and / or G3 glucose dendrimers.

[0055] Dendrimer scaffolds suitable for use in the conjugates include, but are not limited to, poly(amidoamine) or STARBURST™ dendrimers, also known as PAMAM; polypropylamine (POPAM), polyethyleneimine, polylysine, polyesters, iptycene, aliphatic poly(ether), aromatic polyether dendrimers, and dendrimers of sugars (e.g., glucose, galactose, mannose, fructose, etc.) and copolymers thereof (e.g., copolymers of sugars and alkylene glycols) (e.g., dendrimers formed with glucose and ethylene glycol building blocks). Dendrimers can have multiple surface functional groups, such as carboxylic acid, amine, hydroxyl, and / or acetamide. The terms "surface functional group" and "end group" are used interchangeably herein. In some embodiments, dendrimers have surface hydroxyl groups. In some embodiments, one or more of these surface functional groups are further modified with other molecules, such as sugars (e.g., glucose, galactose, mannose, fructose, etc.) and / or polyalkylene glycols, e.g., polyethylene glycol, thus having sugar molecules and / or polyalkylene glycols as terminal moieties / molecules. The dendrimers can be of any generation, including, but not limited to, first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, or tenth generation. In some embodiments, the dendrimers used as platforms are PAMAM dendrimers modified with functional groups having an increased number of surface hydroxyl groups. Preferred PAMAM dendrimers include hydroxylated PAMAM dendrimers, particularly G3-G6 hydroxyl-terminated PAMAM dendrimers, such as G3, G4, G5, and G6 hydroxyl-terminated PAMAM dendrimers.

[0056] In some embodiments, dendrimer-active agent conjugates can be restricted to the peripheral circulation, specifically targeting particular tissue regions and / or cell types, such as peripheral nervous system cells, glial cells, and / or their supporting cells, e.g., enteric neurons and enteric glia, by using high-generation dendrimers (such as fourth-, fifth-, or sixth-generation PAMAM dendrimers, second-, third-, or higher-generation glucose-based dendrimers). Additionally or alternatively, dendrimer-active agent conjugates can be restricted to the peripheral circulation by appropriate functionalization (such as PEGylation) of the dendrimer.

[0057] In some embodiments, dendrimers can be specifically targeted to particular tissue regions and / or cell types in the central nervous system (CNS), PNS and / or periphery, such as neurons and glia in the CNS and / or peripheral nervous system (PNS), by using certain generations of dendrimers, such as second generation (G2), G3, G4 and G5 PAMAM dendrimers and / or glucose dendrimers. Monosaccharide-based dendrimers

[0058] In a preferred embodiment, the branching unit comprises a monosaccharide. In some embodiments, the monosaccharide branching unit is conjugated to the core or to the previous layer of monomers via a linker such as a polyethylene glycol chain. In a preferred embodiment, the monosaccharide branching unit is a glucose-based branching unit. In some embodiments, the branching unit can include PEG and / or alkyl chain linkers between different dendrimer generations. For example, glucose layers are connected by a PEG linker and a triazole ring. In some embodiments, the branching unit is the same for each generation of dendrimers generated from the core. Thus, for example, the branching unit is a glucose-based branching unit for generating a first-generation dendrimer, a second-generation dendrimer, and a third-generation dendrimer.

[0059] In some embodiments, the dendrimer has a hypercore such as dipentaerythritol and one or more monosaccharide branching units. In some embodiments, the hypercore is dipentaerythritol and the monosaccharide branching units are glucose-based branching units. In further embodiments, the spacer molecule may be an alkyl(CH)-hydrocarbon-like unit.

[0060] In some embodiments, dendrimers synthesized using glucose building blocks and having surfaces made primarily of glucose moieties specifically target cells, including injured neurons, ganglion cells, and other nervous system cells, in the brain, eye, and / or peripheral nervous system. In some embodiments, glucose-based dendrimers selectively target or are concentrated within target nervous system cells and / or glial cells. In some embodiments, glucose-based dendrimers selectively target or are concentrated on the surface of target nervous system cells and / or glial cells. In some embodiments, glucose-based dendrimers selectively target or are concentrated within and on the surface of target nervous system cells and / or glial cells. In some embodiments, glucose-based dendrimers selectively target or are concentrated within and / or on the surface of damaged, diseased, and / or overactive neurons and / or glial cells.

[0061] In some cases, the dendrimer comprises a number of sugar molecules and terminal groups, such as glucose and / or hydroxyl groups, effective for targeting one or more neurons and / or glia in the CNS, PNS, and / or eye. The terminal hydroxyl groups of these dendrimers may be terminal glucose molecules, or extra hydroxyl groups that are not part of glucose molecules, or a combination thereof. In some embodiments, all of the terminal hydroxyl groups are part of terminal glucose molecules. In some embodiments, the number of sugar molecules on the end of the dendrimer is determined by the generation number.

[0062] In some embodiments, the dendrimers are made from glucose and oligoethylene glycol building blocks. Exemplary glucose dendrimers are shown in structures V and VII.

[0063] Some exemplary glucose dendrimers include a first-generation glucose dendrimer with 24 hydroxyl (-OH) end groups, a second-generation glucose dendrimer with 96 hydroxyl (-OH) end groups, a third-generation glucose dendrimer with 396 hydroxyl (-OH) end groups, and a fourth-generation glucose dendrimer with 1584 hydroxyl (-OH) end groups. For example, the glucose dendrimer is a second-generation glucose-based dendrimer with 24 glucose molecules on the periphery and 6 embedded glucose molecules in the backbone held together by PEG segments.

[0064] Dendrimer compositions called "glucose dendrimers," which can selectively accumulate within neurons, particularly in the nuclei of damaged and / or overactive neurons, also accumulate at high levels within activated microglia. However, these dendrimers are primarily targeted to neurons, as opposed to hydroxyl dendrimers, which primarily accumulate within microglia. Glucose dendrimers are described in U.S. Patent No. 63 / 327,610, "Dendrimer Compositions for Targeted Delivery of Therapeutics to Neurons," filed April 5, 2022, by The Johns Hopkins University, inventors Kannan Rangaramanujam, Rishi Sharma, Anjali Sharma, Sujatha Kannan, Nirnath Sah, Mira Sachdeva, and Siva P. Kambhampati.

[0065] Glucose dendrimers comprise (a) a central core, (b) one or more branching units based on the monosaccharide glucose, optionally conjugated with linkers, and, optionally, (c) one or more therapeutic, prophylactic, and / or diagnostic agents. Generally, the one or more branching units are conjugated to the central core, and the surface groups of the dendrimer are monosaccharide glucose molecules. In some embodiments, the central core is dipentaerythritol or a hexa-propargylated derivative thereof. In some embodiments, the branching units are conjugated to the central core via a linker, such as a hydrocarbon or oligoethylene glycol chain. In a preferred embodiment, the branching unit is β-D-glucopyranoside tetraethylene glycol azide, having the following structure: [ka] or a peracetylated derivative thereof.

[0066] In some embodiments, the glucose dendrimer is a first, second, third, fourth, fifth or sixth generation dendrimer. In one embodiment, the dendrimer has the following structure: [ka] It is a first generation dendrimer having the following structure: In a preferred embodiment, the dendrimer has the following structure: [ka] It is a second generation dendrimer having the following structure:

[0067] In some embodiments, one or more therapeutic, prophylactic, and / or diagnostic agents are encapsulated, attached, and / or conjugated within the dendrimer at a concentration of between about 0.01% and about 30% by weight, preferably between about 1% and about 20% by weight, and more preferably between about 5% and about 20% by weight. The dendrimer may also be conjugated to one or more diagnostic agents, such as fluorescent dyes, near-infrared dyes, SPECT imaging agents, PET imaging agents, and radioisotopes.

[0068] In some embodiments, the dendrimer and the drug are conjugated via one or more linkers or coupling agents, such as one or more hydrocarbon or oligoethylene glycol chains. Exemplary linking groups are disulfide, ester, ether, thioester, and amide linking groups. 1. Core

[0069] In some embodiments, dendrimers are prepared using methods in which dendrimers are assembled from a multifunctional core that is extended outward through a series of reactions, allowing for the stepwise addition of branching units (i.e., generations) around the core.

[0070] Exemplary chemical structures suitable as core moieties include dipentaerythritol, pentaerythritol, 2-(aminomethyl)-2-(hydroxymethyl)propane-1,3-diol, 2-ethyl-2-(hydroxymethyl)propane-1,3-diol, 3,3',3",3"'-silanetetrayltetrakis(propane-1-thiol), 3,3-divinylpenta-1,4-diene, 3,3',3"-nitrilotripropionic acid, 3,3',3"-nitrilotris(N-(2-aminoethyl)propanamide), 3,3',3"-nitrilotripropionic acid, ... ",3'''-(ethane-1,2-diylbis(azanetriyl))tetrapropanamide, 3-(carboxymethyl)-3-hydroxypentanedioic acid, 2,2'-((2,2-bis((2-hydroxyethoxy)methyl)propane-1,3-diyl)bis(oxy))bis(ethan-1-ol), tetrakis(3-(trichlorosilyl)propyl)silane, 1-thioglycerol, 2,2,4,4,6,6-hexachloro-1,3,5,2l5,4l5,6l5-triazatriphosphinine, 3-(hydroxymethyl)-5,5 -Dimethylhexane-2,4-diol, 4,4',4"-(ethane-1,1,1-triyl)triphenol, 2,4,6-trichloro-1,3,5-triazine, 5-(hydroxymethyl)benzene-1,2,3-triol, 5-(hydroxymethyl)benzene-1,3-diol, 1,3,5-tris(dimethyl(vinyl)silyl)benzene, carbosiloxane core, nitrilotrimethanol, ethylenediamine, propane-1,3-diamine, butane-1,4-diamine, 2,2',2"-nitrilotris(ethane-1- Examples of suitable core moieties include cyclodextrins, alpha cyclodextrins, beta cyclodextrins, gamma cyclodextrins, cucurbituril, benzene-1,2,3,4,5,6-hexathiol, monosaccharides, disaccharides, trisaccharides, oligosaccharides, or azide- or alkyne-modified portions thereof. In some embodiments, the core moiety is chitosan. Thus, azide- or alkyne-modified chitosans are suitable for conjugation to branching units using click chemistry.In a preferred embodiment, the central core is dipentaerythritol or its hexapropargylated derivative.

[0071] In some embodiments, the core moiety is ethylenediamine or tetra(ethylene oxide). In some embodiments, the core moiety is dipentaerythritol. Exemplary chemical structures suitable for use as the core moiety are shown in Table 1 below. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] 2. Branching Unit

[0072] Exemplary chemical structures suitable as branching units include monosaccharides. In some embodiments, the monosaccharide branching units are conjugated to the core or previous layer of monomers via a linker, such as a polyethylene glycol chain. In a preferred embodiment, the monosaccharide branching units are glucose-based branching units. Exemplary glucose-based branching units are shown in Structures II-IV. These are spacer molecules and, therefore, can also be alkyl(CH)n-hydrocarbon-like units.

[0073] The branching units are PEG or alkyl chain linkers between different dendrimer generations, e.g., glucose layers are attached via PEG linkers and triazole rings.

[0074] In a preferred embodiment, the branching units are the same for each generation of dendrimer produced from the core. Thus, in one embodiment, the branching units are glucose-based branching units to produce the first generation dendrimers shown in structures V-VII.

[0075] In some embodiments, the branching unit is a hypermonomer, i.e., an ABn building block. Exemplary hypermonomers include AB4, AB5, AB6, AB7, and AB8 building blocks. The hypermonomer strategy dramatically increases the number of available end groups. An exemplary AB4 hypermonomer is peracetylated β-D-glucopyranoside tetraethylene glycol azide, shown in Structure III.

[0076] The chemical structures listed in Table 1 are also suitable as building blocks for forming branching units of dendrimers. For example, branching units of dendrimers include dipentaerythritol, pentaerythritol, 2-(aminomethyl)-2-(hydroxymethyl)propane-1,3-diol, 2-ethyl-2-(hydroxymethyl)propane-1,3-diol, 3,3',3",3'"-silanetetrayltetrakis(propane-1-thiol), 3,3-divinylpenta-1,4-diene, 3,3',3"-nitrilotripropionic acid, 3,3',3"-nitrilotris(N-(2-aminoethyl)propane)), 3,3',3"-nitrilotriisopropyl ether, ... )propanamide), 3,3',3",3'''-(ethane-1,2-diylbis(azanetriyl))tetrapropanamide, 3-(carboxymethyl)-3-hydroxypentanedioic acid, 2,2'-((2,2-bis((2-hydroxyethoxy)methyl)propane-1,3-diyl)bis(oxy))bis(ethan-1-ol), tetrakis(3-(trichlorosilyl)propyl)silane, 1-thioglycerol, 2,2,4,4,6,6-hexachloro-1,3,5,2l5, 4l5,6l5-triazatriphosphinine, 3-(hydroxymethyl)-5,5-dimethylhexane-2,4-diol, 4,4',4"-(ethane-1,1,1-triyl)triphenol, 2,4,6-trichloro-1,3,5-triazine, 5-(hydroxymethyl)benzene-1,2,3-triol, 5-(hydroxymethyl)benzene-1,3-diol, 1,3,5-tris(dimethyl(vinyl)silyl)benzene, carbosiloxane core, nitrilotrimethanol, Formed by ethylenediamine, propane-1,3-diamine, butane-1,4-diamine, 2,2',2"-nitrilotris(ethan-1-ol), alpha cyclodextrin, beta cyclodextrin, gamma cyclodextrin, cucurbituril, benzene-1,2,3,4,5,6-hexathiol, monosaccharides, disaccharides, trisaccharides, oligosaccharides, or their azide-modified moieties, alkyne-modified moieties, or combinations thereof.Other examples of chemical structures suitable for forming the branching units of dendrimers include, but are not limited to, sugar moieties such as glucose, galactose, mannose, and fructose, and alkylene glycols such as ethylene glycol, and combinations thereof. In some embodiments, the branching unit is chitosan. Thus, azide-modified chitosan or alkyne-modified chitosan is suitable for conjugation to a core moiety or additional, same or different, branching units using click chemistry. In some embodiments, the branching unit is methyl acrylate or ethylenediamine, or a combination thereof. In some embodiments, the branching unit is linear or branched polyethyleneglycerol. In some embodiments, the branching unit is a copolymer of an alkylene glycol (such as ethylene glycol) and a sugar moiety such as glucose, galactose, mannose, and / or fructose. 3.Surface functional groups

[0077] The surface functional groups / molecules of the dendrimer include, but are not limited to, primary amine end groups, hydroxyl end groups, carboxylic acid end groups, acetamide end groups, sugar molecules, oligo- or poly-alkylene glycols, and / or thiol end groups. In some embodiments, the desired terminal functional groups can be added by one of the conjugation methods to the core and branching units.

[0078] In some embodiments, the surface functional groups are hydroxyl groups, such as the hydroxyl groups of the PAMAM dendrimer of the second generation OEG dendrimer shown in Structure I, or the terminal glucose hydroxyl groups of dendrimers prepared with glucose-based branching units shown in Structures V and VII. In some embodiments, the desired surface functional groups can be modified or added by one of the conjugation methods to the core and branching units. Exemplary surface functional groups include hydroxyl, amine, carboxylic acid, acetamide, and thiol end groups, and combinations thereof.

[0079] In some embodiments, dendrimers may specifically target particular tissue regions and / or cell types, such as cells and tissues of the central nervous system (CNS), peripheral nervous system (PNS), and / or eye. In some embodiments, dendrimers specifically target neurons and / or glia of the CNS. In some embodiments, dendrimers specifically target neurons and / or glia of the PNS. In some embodiments, dendrimers specifically target non-neural and / or non-glial cells, such as digestive system cells, cardiovascular cells, and / or immune system cells. In some embodiments, the glucose dendrimers are of generation 1 (G1), G2, G3, G4, and G5, preferably G1, G2, and / or G3.

[0080] In some embodiments, the dendrimer comprises a number of terminal glucose and / or hydroxyl groups effective to target one or more neurons and / or glia of the CNS, PNS, and / or eye, hi some embodiments, the dendrimer comprises a number of terminal glucose and / or hydroxyl groups effective to target one or more non-neuronal and / or non-glial cells, such as cells of the digestive system, cardiovascular cells, and / or immune system cells.

[0081] In some embodiments, the dendrimers are made from glucose and oligoethylene glycol building blocks. An exemplary first generation glucose dendrimer is shown in Structure VI, and a second generation glucose dendrimer is shown in Structure VIII.

[0082] In some embodiments, dendrimers have a plurality of surface functional groups (also referred to herein as surface or peripheral functional groups), such as hydroxyl (-OH), amine, acetamide, and / or carboxyl groups, at the periphery of the dendrimer. In some embodiments, the surface density of such peripheral functional groups is at least 1 group / nm 2 (number of surface functional groups / surface area (nm 2For example, in some embodiments, the surface density of surface functional groups, such as hydroxyl groups, is at least 10, 15, 20, 25, 30, 35, 40, 45, 50, or more than 50 OH groups / nm 2 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more OH groups / nm 2 In some embodiments, the volume density of surface functional groups, such as hydroxyl groups, is from about 1 to about 50 groups / nm 3 Between about 5 and about 30 groups / nm 3 Between about 10 and about 20 groups / nm 3 In a further embodiment, the surface density of surface functional groups, such as hydroxyl groups, is between about 1 and about 50, preferably 5 to 20 groups / nm 2 (number of surface functional groups / surface area (nm 2 )), while each of the surface functional group moieties has a molecular weight between about 100 Da and about 10 kDa, preferably between about 100 Da and 1000 Da.

[0083] In some embodiments, the amount of surface functional groups of the dendrimer, such as any one of the above, e.g., hydroxyl groups, is at least 30%, at least 40%, at least 50%, greater than 40%, greater than 50%, or in the range of greater than 30% to 100%. In preferred embodiments, the amount of surface hydroxyl groups of the dendrimer is preferably greater than 35%.

[0084] In some embodiments, one or more of the surface functional groups, such as any one of those described above, on the outer periphery of the dendrimer are further modified by conjugation with one or more carbohydrate molecules and / or multiple or multiple polyalkylene glycols (e.g., polyethylene glycol). In these embodiments, the surface density of the terminal carbohydrate moieties / molecules and / or polyalkylene glycols can have any of the ranges described above for hydroxyl groups. Hydroxyl-terminated PAMAM dendrimers, PAMAM dendrimers modified on the surface with sugar moieties (>10% of the surface groups are modified with sugars, particularly glucose), and glucose dendrimers (where the dendrimer is made from glucose building blocks) are preferred. For delivery to the brain, constructs with a total molecular weight of <30,000 Da are preferred. For primary confinement in the peripheral circulation, constructs with a total molecular weight of >50,000 Da are preferred. When dendrimers are formed from or include terminal sugar moieties / sugar molecules (e.g., glucose), the terminal hydroxyl groups of these dendrimers may be part of the terminal sugar moiety / sugar molecule, or extra hydroxyl groups that are not part of the sugar moiety / sugar molecule, or a combination thereof. In some embodiments, all of the terminal hydroxyl groups are part of the terminal sugar moiety / sugar molecule. a. Hydroxyl-terminated dendrimers

[0085] In some embodiments, the dendrimer contains multiple hydroxyl groups. Some exemplary densely hydroxyl-containing dendrimers include commercially available polyester dendritic polymers, such as hyperbranched 2,2-bis(hydroxyl-methyl)propionic acid polyester polymers (e.g., hyperbranched bis-MPA polyester-64-hydroxyl, 4th generation), and dendritic polyglycerols. In some embodiments, the hydroxyl-terminated dendrimers include hydroxyl-terminated PAMAM dendrimers, particularly G3-G6 hydroxyl-terminated PAMAM dendrimers, such as G3, G4, G5, and G6 hydroxyl-terminated PAMAM dendrimers.

[0086] In some embodiments, the dense hydroxyl-containing dendrimer is an oligoethylene glycol (OEG)-like dendrimer. For example, the second-generation OEG dendrimer (D2-OH-60) shown in Structure I can be synthesized using highly efficient, robust, and atom-economical chemical reactions, such as Cu(I)-catalyzed alkyne-azide click and photocatalytic thiol-ene click chemistry. Highly dense polyol dendrimers of very low generation with minimal reaction steps can be achieved by using an orthogonal hypermonomer and hypercore strategy, as described, for example, in WO2019094952. In some embodiments, the dendrimer backbone has non-cleavable polyether bonds throughout the structure (non-biodegradable), avoiding dendrimer collapse in vivo and enabling the excretion of such dendrimers from the body as a single entity. [ka]

[0087] In some embodiments, the dendrimer has a plurality of hydroxyl (-OH) groups at the periphery of the dendrimer. In some embodiments, the surface density of the hydroxyl (-OH) groups is at least 1 OH group / nm 2 (number of surface hydroxyl groups / surface area (nm 2 For example, in some embodiments, 1 nm 2 The surface density of hydroxyl groups per nanometer is at least 10, 15, 20, 25, 30, 35, 40, 45, 50, or more than 50 OH groups / nm 2 More than 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 OH groups / nm 2 In some embodiments, the volume density of hydroxyl groups is from about 1 to about 50 groups / nm 3 Between about 5 and about 30 groups / nm 3 Between about 10 and about 20 groups / nm 3 In a further embodiment, the surface density of hydroxyl (—OH) groups is between about 1 and about 50 groups / nm2 or 5-20 OH groups / nm 2 (number of surface hydroxyl groups / surface area (nm 2 )), while having a molecular weight between about 100 Da and about 1000 Da. In some embodiments, the amount of surface hydroxyl groups on the dendrimer is preferably greater than 35%, at least 40%, at least 50%, greater than 40%, greater than 50%, or in the range of greater than 40% to 100%. In some embodiments, the dendrimer has a percentage of hydroxyl groups exposed on the outer surface, with other hydroxyl groups present in the interior core of the dendrimer.

[0088] In some embodiments, the dendrimer specifically targets a particular tissue region and / or cell type after administration to the body. In some embodiments, the dendrimer specifically targets a particular tissue region and / or cell type without a targeting moiety. In some embodiments, the dendrimer contains a number of hydroxyl groups effective to target CNS cells and / or PNS cells, such as microglia, astrocytes, and / or neurons, associated with a disease, disorder, or injury of the central or peripheral nervous system. In some embodiments, the dendrimer specifically targets a particular tissue region and / or cell type without a targeting moiety, and the active agent conjugated to the dendrimer binds directly to receptors on the surface and / or within the target nervous system cells and / or glial cells. Unmodified PAMAM dendrimers with hydroxyl end groups do not concentrate in neurons in the brain and / or retinal ganglion cells (RGCs) in the eye to the same extent as glucose dendrimers. Glucose dendrimers with terminal glucose monosaccharides and a high density of hydroxyl functional groups effectively target neurons in a generation-dependent manner. Generation 2 (G2), G3, and G4 should be effective. G5 and above are more difficult to use.

[0089] In a preferred embodiment, the dendrimer comprises an effective number of terminal glucose and / or hydroxyl groups for targeting one or more neurons in the CNS or eye.The hydroxyl groups on the dendrimer surface are part of glucose molecules.Other than the glucose molecules on the surface, there are no extra hydroxyls.The number of sugar molecules on the surface is determined by the number of generations.It is expected that all generations will target neurons.

[0090] Some exemplary glucose dendrimers include a first-generation glucose dendrimer with 24 hydroxyl (-OH) end groups, a second-generation glucose dendrimer with 96 hydroxyl (-OH) end groups, a third-generation glucose dendrimer with 396 hydroxyl (-OH) end groups, and a fourth-generation glucose dendrimer with 1584 hydroxyl (-OH) end groups. In a preferred embodiment, the glucose dendrimer is a second-generation glucose-based dendrimer with 24 glucose molecules on the periphery and 6 glucose molecules incorporated into the backbone held together by PEG segments. B. Carbohydrate-modified dendrimers

[0091] In some embodiments, the dendrimer contains one or more carbohydrate molecules at its terminal end. These terminal carbohydrate molecules can be prepared by conjugating one or more surface functional groups of the dendrimer, such as amine, carboxyl, or hydroxyl groups, with one or more carbohydrate molecules. In a preferred embodiment, the dendrimer prior to carbohydrate conjugation is a hydroxyl-terminated dendrimer, such as a hydroxyl-terminated PAMAM dendrimer, and one or more of the hydroxyl groups are conjugated to one or more carbohydrate molecules.

[0092] In some embodiments, hydroxyl-terminated dendrimers modified with surface glucose molecules selectively target central and / or peripheral nervous system and / or glial cells in vitro and in vivo; and / or selectively accumulate on the surface and / or within these targets, such that active agents conjugated to the dendrimers bind to one or more receptors on / within the target nervous system cells and / or glial cells. In some embodiments, hydroxyl-terminated dendrimers modified with surface glucose molecules selectively target digestive system cells, cardiovascular cells, and / or immune system cells in vitro and in vivo; and / or selectively accumulate on the surface and / or within these targets, such that active agents conjugated to the dendrimers bind to one or more receptors on / within the target digestive system cells, cardiovascular cells, and / or immune system cells.

[0093] In some embodiments, the carbohydrate moiety used to modify one or more surface functional groups of a dendrimer is a monosaccharide. Exemplary monosaccharides suitable for modifying a dendrimer include glucose, glucosamine, galactose, mannose, fructose, dehydroascorbic acid, urate, and myo-inositol. In some embodiments, the dendrimer is conjugated to glucose and thus contains glucose as a terminal moiety / molecule. In some embodiments, hydroxyl-terminated dendrimers are modified with one or more glucose moieties ("D-Glu"). In some embodiments, the dendrimer is conjugated to galactose. In some embodiments, the dendrimer is conjugated to mannose. In some embodiments, the dendrimer is conjugated to fructose. In some embodiments, the dendrimer is conjugated to one or more monosaccharides other than glucose, such as galactose, mannose, and / or fructose. For example, the carbohydrate moiety may be an oligosaccharide terminated with one or more monosaccharides including glucose, glucosamine, mannose, fructose, thus exposing these sugar moieties on the surface for attachment.

[0094] In preferred embodiments, the glucose- or hydroxyl-terminated PAMAM dendrimer or carbohydrate-functionalized dendrimer has affinity for, and is conjugated to, one or more active agents suitable for binding directly or indirectly to, one or more serotonin (5HT) receptors (e.g., 5HT-1A, 5HT-2B, 5HT-2A, 5HT-2B, 5HT-2C, 5HT-3, 5HT-4, 5HT-6, and 5HT-7 receptors). In some embodiments, the dendrimer binds to one or more norepinephrine (NE) receptors, e.g., α 2A -adrenergic receptor, α 2B -adrenergic receptor, α 2CIn some embodiments, the dendrimer is conjugated to one or more carbohydrate moieties that have affinity for and are suitable for binding to β-adrenergic receptors and / or β-adrenergic receptors. In some embodiments, the dendrimer is conjugated to one or more carbohydrate moieties that have affinity for and are suitable for binding to dopamine D1 and D2 receptors, directly or indirectly. In some embodiments, the dendrimer is conjugated to one or more carbohydrate moieties that have affinity for and are suitable for binding to one or more monoamine transporters (e.g., vesicular monoamine transporter 2 (VMAT2), serotonin reuptake transporter (SERT), noradrenaline transporter (NAT), dopamine transporter (DAT)). In some embodiments, the dendrimer is conjugated to one or more carbohydrate moieties that have affinity for and are suitable for binding to one or more of AMPA receptors, NMDA receptors, EGFR1 receptors, EGFR2 receptors, histamine (H1) receptors, GABA receptors, and trace amine-associated receptor 1 (TAAR1). In some embodiments, the dendrimer is conjugated to one or more active agents that have affinity for and are suitable for transport by one or more of GLUT1, GLUT2, GLUT3, GLUT4, GLUT5, GLUT6, GLUT7, GLUT8, GLUT9, GLUT10, GLUT11, GLUT12, GLUT13, and GLUT14, with or without the carbohydrate moiety. In further embodiments, the dendrimer is conjugated to one or more glucose and / or glucosamine moieties. In some embodiments, the dendrimer contains a carbohydrate moiety capable of transporting the active agent to a target cell / receptor, where activity at the target cell or receptor is driven by the active agent. In these embodiments, the carbohydrate and glucose moieties allow for better targeting of the drug to the cells and / or receptors of interest. For example, in some embodiments, the dendrimer is conjugated to one or more glucose and / or glucosamine moieties.In other embodiments, the dendrimer is conjugated to one or more oligosaccharides terminated with glucose and / or glucosamine moieties, i.e., the glucose and / or glucosamine moieties are exposed on the surface of the dendrimer conjugate suitable for binding to one or more of GLUT, 5HT receptor, NE receptor, DA receptor and / or transporter.

[0095] FIG. 13 is a schematic overview of the major pharmacological targets of LSD, psilocybin, DMT, MDMA, and ketamine, the signaling cascades involved, hormonal modulation, and the main behavioral outcomes following their administration in both animals and humans.

[0096] In some embodiments, the dendrimer has a plurality of monosaccharide (e.g., glucose, etc.) carbohydrate moieties / molecules at the periphery of the dendrimer. In some embodiments, the surface density of monosaccharide (e.g., glucose, etc.) carbohydrate molecules is at least 1 carbohydrate molecule / nm 2 (number of surface carbohydrate groups / surface area (nm 2 In some embodiments, 1 nm 2 The surface density of carbohydrate molecules per nanometer is at least 10, 15, 20, 25, 30, 35, 40, 45, 50, or more than 50 OH groups / nm 2 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more OH groups / nm 2 For example, 1 nm 2 In some embodiments, the surface density of carbohydrate molecules per nanometer is greater than 10. In some embodiments, the volume density of surface carbohydrate molecules is from about 1 to about 50 groups / nm. 3 Between about 5 and about 30 groups / nm 3 Between about 10 and about 20 groups / nm 3 In a further embodiment, the surface density of carbohydrate molecules is between 1 nm 2 Between about 1 and about 50, and between about 5 and about 20 per 1000 (number of surface carbohydrate molecules / surface area (nm 2)) while each carbohydrate moiety has a molecular weight between about 100 Da and about 1000 Da. That is, in these embodiments, one or more surface functional groups of the dendrimer are modified at the terminus to introduce one or more sugar moieties / molecule(s), and the terminal hydroxyl groups may be part of a terminal sugar moiety / molecule, or may be extra hydroxyl groups that are not modified by a sugar moiety / molecule and therefore not part of a sugar moiety / molecule, or a combination thereof.

[0097] In some embodiments, the carbohydrate molecule, such as a monosaccharide, e.g., glucose, is present in an amount by weight that is between about 1% and 40% of the total weight of the glycosylated dendrimer, e.g., between about 2% and 20%, between about 5% and 15%, or between 9% and 12% of the total weight of the glycosylated dendrimer. For example, in some embodiments, the carbohydrate moiety is present in an amount that is about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% of the total weight of the glycosylated dendrimer after conjugation. In some embodiments, conjugation of carbohydrate molecules to one or more surface functional groups occurs with about 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, or 25% of the total number of surface functional groups, preferably hydroxyl groups, available prior to conjugation of the dendrimer. In other embodiments, conjugation of carbohydrate molecules occurs with less than 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40% of the total number of available surface functional groups on the dendrimer prior to conjugation. c. Dendrimers modified with polyalkylene glycol

[0098] In some embodiments, the dendrimer contains one or more terminal polyalkylene glycols. These terminal polyalkylene glycols can be prepared by conjugating one or more of the dendrimer's surface functional groups, such as hydroxyl groups, with a polyalkylene glycol, such as PEG. In some embodiments, the dendrimer prior to conjugation is a hydroxyl-terminated dendrimer, such as a hydroxyl-terminated PAMAM dendrimer, and at least a portion of the surface hydroxyl groups are conjugated with PEG.

[0099] In some embodiments, the dendrimer has a plurality of polyalkylene glycols, such as PEG, at the periphery of the dendrimer. In some embodiments, the surface density of the polyalkylene glycol, such as PEG, is at least 1 polyalkylene glycol / nm 2 (number of surface polyalkylene glycols / surface area (nm 2 In some embodiments, 1 nm 2 The surface density of polyalkylene glycols per nanometer is at least 10, 15, 20, 25, 30, 35, 40, 45, 50, or more than 50 polyalkylene glycols per nanometer. 2 Polyalkylene glycols of 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, and more than 10 units / nm 2 For example, 1 nm 2 In some embodiments, the surface polyalkylene glycol volume density is from about 1 to about 50 groups / nm 3 Between about 5 and about 30 groups / nm 3 Between about 10 and about 20 groups / nm 3 In a further embodiment, the surface density of the polyalkylene glycol, such as PEG, is between 1 nm 2 The number of surface polyalkylene glycols per surface area (nm) is between about 1 and about 50, and between about 5 and about 20 per surface area (nm 2 )) On the other hand, it has a molecular weight between about 100 Da and about 1000 Da.

[0100] In some embodiments, the polyalkylene glycol molecule, such as PEG, may be present in an amount by weight that is between about 1% and 40% of the total weight of the PEGylated dendrimer, e.g., between about 2% and 20%, between about 5% and 15%, or between 9% and 12% of the total weight of the PEGylated dendrimer. For example, in some embodiments, the polyalkylene glycol molecule, such as PEG, is present in an amount that is about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% of the total weight of the PEGylated dendrimer after conjugation.

[0101] In some embodiments, conjugation of polyalkylene glycol molecules such as PEG via one or more surface functional groups of the dendrimer is achieved with about 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, or 25% of the total number of surface functional groups, preferably hydroxyl groups, available prior to conjugation of the dendrimer. In other embodiments, conjugation of polyalkylene glycol molecules such as PEG is achieved with less than 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40% of the total number of available surface functional groups of the dendrimer prior to conjugation. B. Activators

[0102] The dendrimer is conjugated to or complexed with one or more hallucinogens. Hallucinogens are a diverse group of drugs that alter a subject's awareness of their surroundings as well as their own thoughts and emotions. Collectively referred to as "psychedelics," hallucinogens can be divided into two categories: classical hallucinogens (such as LSD) and dissociative drugs (such as PCP). Both types of hallucinogens can cause hallucinations or agitation, and images that appear real but are not. Furthermore, dissociative drugs can cause a subject to feel a loss of control or a sense of detachment from their body and environment. Exemplary hallucinogens that can be conjugated to dendrimer compositions include, but are not limited to, a range of drug classes, including classical psychedelics, dissociative inducers, and delirium inducers. Hallucinogens and their derivatives typically bind to one or more receptors, thereby modulating neurotransmitter signaling in the central and peripheral nervous systems. Hallucinogens typically inhibit the reuptake of neurotransmitters, particularly serotonin, dopamine, and noradrenaline, via selective receptors, thereby increasing the concentrations of these particular neurotransmitters in the synaptic cleft. Thus, partial antagonism, functional selectivity, and inverse agonism all play important roles in determining cellular responses to specific neurotransmitter receptor ligands.

[0103] In a preferred embodiment, the dendrimer comprises 5-HT 2A / C and 5-HT 1APsilocin, ketamine (R-ketamine, S-ketamine, (R / S)-ketamine), norketamine, ketamine analogs, ketamine metabolites, N,N-dimethyltryptamine (DMT), 4-acetoxy-N,N-dimethyltryptamine, 5-methoxy-DMT, 5-chloro-DMT, LSD, 3,4-methylenedioxymethamphetamine (MDMA), psilocybin, ibogaine, mescaline, norbeocystine, 2C compounds (designer agents structurally similar to ecstasy and MDMA), which exhibit high binding affinity to serotonin receptors, 3,4-methylenedioxyethylamphetamine (MDE), d-lysergic acid diethylamide (LSD), or their analogs. The psychedelic hallucinogens are conjugated to one or more psychedelic hallucinogens, such as 25I-NBOMe (4-iodo-2,5-dimethoxy-N-(2-methoxybenzyl)phenethylamine (25I-NBOMe), an N-benzylmethoxy derivative of a 2C family hallucinogen), a synthetic psychedelic drug belonging to the group of psychedelic hallucinogens (25I-NBOMe), and NBOMe (4-iodo-2,5-dimethoxy-N-(2-methoxybenzyl)phenethylamine (25I-NBOMe), an N-benzylmethoxy derivative of a 2C family hallucinogen). In some embodiments, the psychedelic hallucinogen is functionalized with, for example, ester, disulfide, phosphodiester, triglycyl peptide, hydrazine, amide, ether, and aminoalkyl linking groups, optionally with one or more spacers / linkers, for ease of conjugation to the dendrimer and / or desired release rate.

[0104] The majority of psychedelics have poor aqueous solubility in the microgram / mL range. Thus, in some embodiments, dendrimer conjugation of these psychedelics results in an improvement in aqueous solubility of between about 2-fold and about 200-fold, inclusive; between about 5-fold and about 150-fold, inclusive; or between about 10-fold and about 100-fold, inclusive, compared to the free drug, i.e., not conjugated to a dendrimer. 1. Classic Psychedelics

[0105] The composition can include a dendrimer conjugated to one or more classical psychedelics. Classical or serotonergic psychedelic compounds are so named because they primarily interact with the serotonergic system and because most of them are derived from plants or are semi-synthetic compounds. In some cases, classical psychedelics share part of the chemical structure of the endogenous neurotransmitter serotonin (5-HT), particularly those with an indole scaffold. However, some of them, such as mescaline, do not have an indole but are still considered serotonergic psychedelics. Typically, classical psychedelics exert their effects through serotonin 2A receptor (5HT-2A) agonism.

[0106] Classical psychedelics conjugated to the dendrimer compositions may be one or more of the semi-synthetic ergoline LSD, plant-derived tryptamines and / or phenethylamines. Exemplary serotonergic hallucinogens include indoleamines such as psilocybin and LSD, and phenylethylamines such as mescaline and 2,5-dimethoxy-4-iodoamphetamine (DOI). a tryptamine

[0107] Classic psychedelics conjugated to dendrimer compositions may be one or more tryptamines. Tryptamine is an indoleamine metabolite of the essential amino acid tryptophan. Its chemical structure is defined by an indole, i.e., a fused benzene and pyrrole ring, and a 2-aminoethyl group at the second carbon (representing the third aromatic atom; the first aromatic atom is a heterocyclic nitrogen). Tryptamine activates trace amine-associated receptors expressed in the brain and regulates the activity of dopaminergic, serotonergic, and glutamatergic systems. In the gastrointestinal tract, commensal bacteria convert dietary tryptophan to tryptamine, which activates 5-HT4 receptors and regulates gastrointestinal motility.

[0108] Exemplary psychedelic tryptamines that can be conjugated to dendrimers include DMT, etryptamine, N,N-diethyltryptamine (DET), psilocin and psilocybin, and derivatives thereof. i. Psilocybin

[0109] Psilocybin inhibits serotonin 5-HT, which is particularly abundant in the prefrontal cortex. 2A Psilocybin is a high affinity agonist at the 5-HT receptor. It enhances cortical activity secondary to downstream postsynaptic glutamatergic activity. Psilocybin also inhibits 5-HT 1A , 5-HT 1D and 5-HT 2C Although it is active at receptors, these are thought to play a minor role in its action. Psilocybin has been well tolerated and safe in human studies at oral doses of 8-25 mg and intravenous doses of 1-2 mg. [ka]

[0110] N,N-Dimethyltryptamine (DMT or N,N-DMT) is a substituted tryptamine, both a derivative and a structural analog of tryptamine. DMT has a rapid onset, potent action, and a relatively short duration of action. DMT binds to the following serotonin receptors: 5-HT1A, 5-HT1B, 5-HT1D, 5-HT2A, 5-HT2B, 5-HT2C, 5-HT6, and 5-HT7. DMT acts as an agonist at 5-HT1A, 5-HT2A, and 5-HT2C receptors, with strong binding affinity for the 5-HT2B receptor. DMT also has affinity for dopamine D1, α1-adrenergic, α2-adrenergic, imidazoline-1, and σ1 receptors. DMT has also been shown in vitro to be a substrate for the cell-surface serotonin transporter (SERT) expressed in platelets and vesicular monoamine transporter 2 (VMAT2). The psychedelic effects of DMT can be attributed in large part to functionally selective activation of the 5-HT2A receptor. [ka] b. Lysergic acid diethylamide (LSD)

[0111] The dendrimer composition may be conjugated to one or more lysergamides. Amides of lysergic acid, collectively known as lysergamides, include several compounds with potent agonist and / or antagonist activity at various serotonin and dopamine receptors. Exemplary lysergamides include, but are not limited to, d-lysergic acid amide (or d-lysergamide; LSA or LAA), lysergic acid diethylamide (LSD), ergometrine, DAM-57, elagotamine, methergine, methysergide, amesergide, 2-bromo-LSD, LSD-Pip, 12-methoxy-LSD, 1P-LSD, 1B-LSD, 1V-LSD, 1cP-LSD, 13-fluoro-LSD, and 14-hydroxy-LSD. Preferably, the lysergamid conjugated to the dendrimer composition is lysergic acid diethylamide (LSD) or an LSD derivative.

[0112] LSD is a semisynthetic ergosterol that can be derived from the natural ergot alkaloid lysergic acid, which is found in the rye parasite, Claviceps purpurea. The mechanism by which LSD works is primarily mediated by activation of serotonin receptors (i.e., 5-HT2A receptors or 5-hydroxytryptamine 2A receptors, or 5-HT2AR) with modulation of 5-HT2C and 5-HT1A receptors. The interplay between receptor activation and the resulting cognitive impairment and hallucination induction remains poorly understood. LSD-induced 5-HT2AR activation leads to disruption of inhibitory processes in the prefrontal cortex of the hippocampus. Specifically, LSD reduces brain activity in the right middle temporal gyrus, superior anterior / middle prefrontal / inferior frontal gyrus, anterior cingulate cortex, left superior frontal gyrus and postcentral gyrus, and cerebellum. Right hemisphere activation alters thalamic function and increases activity in limbic structures and the frontal cortex, resulting in the formation of induced visual imagery. [ka] C. Phenethylamine

[0113] The dendrimer composition may be conjugated to one or more phenethylamines. Phenethylamine (PEA) is an organic compound, a natural monoamine alkaloid and trace amine, which acts as a central nervous system stimulant in humans. In the brain, phenethylamine binds to trace amine-associated receptor 1 (TAAR1) and regulates monoamine neurotransmission by inhibiting vesicular monoamine transporter 2 (VMAT2) in monoamine neurons. To a lesser extent, phenethylamine also acts as a neurotransmitter in the human central nervous system. Phenethylamine is produced from the amino acid L-phenylalanine by enzymatic decarboxylation via the enzyme aromatic L-amino acid decarboxylase.

[0114] Phenethylamines bind to the trace amine associated receptor 1 (TAAR1) as agonists, releasing norepinephrine and dopamine and inducing acetylcholine release via a glutamate-mediated mechanism. Exemplary phenethylamines that can be conjugated to the present dendrimer compositions include mescaline and MDMA. i. Mescaline

[0115] Mescaline or mescalin (3,4,5-trimethoxyphenethylamine) is a naturally occurring hallucinogenic incomplete alkaloid of the substituted phenethylamine class and is the active ingredient of hallucinogenic cacti such as peyote (Lophophora williamsii) and huachuma (Echinopsis pachanoi, also known as San Pedro). Mescaline is biosynthesized from tyrosine, which in turn is derived from phenylalanine by the enzyme phenylalanine hydroxylase.

[0116] Like other classic psychedelics, mescaline is a 5HT 2A / 2C Mescaline is a serotonergic agonist and one of the most selective psychedelics. It also binds to and modulates the activity of noradrenergic receptors α1 and α2A, as well as the TAAR1 receptor. In some forms, the dendrimer compositions can be complexed with mescaline in an amount effective to deliver between about 300 and about 500 mg. In some forms, the dendrimer compositions can be complexed with mescaline in an amount effective to produce a hallucinogenic effect for about 6 to about 8 hours. [ka] 2. Enteractogen

[0117] The dendrimer composition can include one or more entactogens. Entactogens are Schedule I monoamine releasers and reuptake inhibitors known to evoke feelings of emotional tolerance and emotional connection, such as 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA). Exemplary entactogens that can be conjugated to dendrimers include, but are not limited to, 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxy-N-ethyl-amphetamine (MDEA), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxy-N-hydroxyamphetamine (MDOH), 1,3-benzodioxolyl-N-methylbutanamine (MBDB), 6-APB, methylone, mephedrone, GBL, αMT, MDAI, and related compounds. a.MDMA

[0118] The dendrimer composition may be complexed with MDMA. MDMA increases the amount of serotonin in the synaptic space of serotonergic neurons by inhibiting the uptake of serotonin into neurons and directly releasing serotonin from neurons. The released serotonin binds to various serotonin receptors, overactivating them, which is the primary mechanism by which MDMA causes addiction. MDMA also causes significant norepinephrine release.

[0119] Extracellular MDMA binds to presynaptic serotonin (SERT), norepinephrine (NET), and dopamine transporters (DAT) as a reuptake inhibitor, resulting in reduced uptake of the monoamine neurotransmitters. The potency of MDMA inhibition is highest for NET and SERT, but significantly lower for DAT. As a result, more norepinephrine and serotonin remain in the synaptic space. These monoamine transporters (SERT, NET, and DAT) reuptake their respective neurotransmitters according to ion gradients. Typically, extracellular sodium (Na+) and chloride (Cl-) ion concentrations are high and potassium (K+) ion concentrations are low relative to intracellular concentrations. During each reuptake cycle, one monoamine, Na+ and Cl-, is simultaneously taken into the cell. One intracellular K+ is then transported out of the cell. In experimental situations, high extracellular K+ concentrations can trigger reverse transport. MDMA can substitute for K+ in this transport, and therefore high extracellular MDMA can reverse the transport.

[0120] Intracellular MDMA binds to VMAT2 proteins in synaptic vesicles as an inhibitor. Each VMAT2 transports cytosolic monoamines into the vesicle by dissolving the proton gradient across the vesicle membrane. Therefore, VMAT2 inhibition leads to more free cytosolic monoamines, such as serotonin, in serotonergic neurons. These monoamines can then be released by the monoamine transporter reversed by MDMA. Intracellular MDMA can also bind to monoamine oxidase A (MAO-A) as an inhibitor, thus preventing monoamine oxidase A from destroying cytosolic serotonin.

[0121] As an agonist, MDMA binds to the following receptors: 5-HT1A-, 5-HT2A-, 5-HT2B-, and 5-HT2C-serotonin receptors; α1-, α2A-, and β-adrenergic receptors, D1 and D2 dopamine receptors, M1 and M2 muscarinic receptors, H1 histamine receptors, and TAT (TAAR1) receptors. [ka] 3. Dissociative anesthetics

[0122] Another class of drugs with hallucinogenic properties that can be conjugated to the present dendrimer compositions are psychedelic or dissociative anesthetics, including arylcyclohexamines (also known as arylcyclohexylamines). Arylcyclohexamines are a group of compounds containing a cyclohexamine unit in which the aryl moiety (usually a phenyl ring) is attached to the same atom as the amine group. They all exhibit dissociative activity due to antagonism of N-methyl-d-aspartate (NMDA) receptors. Exemplary arylcyclohexamines that can be conjugated to the present dendrimer compositions include phencyclidine (1-(1-phencyclohexyl)piperidine; PCP), dextromethorphan (DXM), nitrous oxide, ketamine, and methoxetamine (a ketamine analog). Ketamine

[0123] The dendrimer compositions may be conjugated to ketamine (R-ketamine, S-ketamine, (R / S)-ketamine), ketamine analogs, ketamine metabolites, 2-(2-chlorophenyl)-2-(methylamino)-cyclohexanone or analogs thereof, such as KEA-1010, methoxetamine, norketamine, and 2-fluorodeschloroketamine.

[0124] Ketamine (ketamine) is a non-barbiturate dissociative anesthetic. Its beneficial effect is the "dissociation" of brainstem function from higher brain regions, thereby altering the sensation of pain and other stimuli and producing amnesia during medical procedures. It is a cyclohexanone derivative that acts rapidly and produces deep anesthesia and analgesia. Its chemical name is ±)-2-(o-chlorophenyl)-2-(methylamino)cyclohexanone hydrochloride, and its structural formula is CHClNO. Ketamine is a noncompetitive N-methyl-D-aspartate (NMDA) and glutamate receptor antagonist. Ketamine blocks HCN1 receptors. Its unique dissociative and partial agonist effects at the opiate mu receptor allow for painful procedures to be performed with sedation and patient comfort.

[0125] Ketamine's effects in chronic pain, comparable to actual drug levels, and as an antidepressant may be mediated by a secondary increase in structural synaptic connectivity mediated by neuronal responses to the ketamine-induced hyperglutamatergic state.

[0126] Ketamine can interact with sigma receptors. Ketamine tends to act by reducing central sensitization, wind-up (the development of persistent, worsening, or chronic pain), and pain memory. The cholinergic, aminergic, and opioid systems appear to play both positive and negative regulatory roles in both sedation and analgesia. Ketamine reverses tolerance to opioids. Ketamine is metabolized via the hepatic system by N-dealkylation, hydroxylation, conjugation, and dehydration. The half-life of ketamine is approximately 45 minutes.

[0127] In some forms, primarily in the context of adjuvant drug use, the recommended dose of ketamine for anesthesia induction is approximately 1 to 4.5 mg / kg IV and approximately 6.5 to 13 mg / kg IM, with alternative off-label recommendations of 0.5 to 2 mg / kg IV and 4 to 10 mg / kg IM. For use in depression, ketamine is most commonly administered at a subanesthetic dose of 0.5 mg / kg IV over 40 minutes.

[0128] Conjugation to dendrimers can further improve ketamine safety and efficacy. For example, dendrimer conjugation can alter specific receptor activity and / or modify biodistribution (e.g., using higher generation dendrimers that restrict ketamine to the peripheral nervous system to prevent its psychoactive effects). [ka] b.PCP

[0129] Phencyclidine (PCP) (also known as penylcyclohexylpiperidine) is a hallucinogen, specifically a dissociative anesthetic, that can produce a wide variety of physical and behavioral effects. PCP can cause hallucinations and distorted perception of sound.

[0130] The most unusual feature of PCP is that oral doses of 5 to 10 mg can induce acute schizophrenia, including agitation, psychosis, auditory hallucinations, paranoid delusions, and catatonia. Doses greater than 10 mg usually result in coma. More than 50% of adult patients exhibit the classic toxic syndrome of PCP poisoning: violent behavior, nystagmus, tachycardia, hypertension, anesthesia, and analgesia.

[0131] As an NMDA receptor antagonist, PCP primarily acts on ionotropic glutamate receptors, which are NMDA receptors. PCP also inhibits nicotinic acetylcholine receptors (nAChRs). In some forms, the dendrimer composition contains PCP analogs with varying potency at nACh and NMDA receptors. PCP-induced presynaptic nAChR and NMDA receptor interactions affect postsynaptic maturation of glutamatergic synapses, thus affecting synaptic development and plasticity in the brain. These effects can result in the inhibition of excitatory glutamate activity in certain brain regions, such as the hippocampus and cerebellum.

[0132] PCP, like ketamine, also acts as a potent dopamine D2 receptor partial agonist, with affinity for the cloned D2High receptor. This activity may be associated with some of the additional psychotic features of PCP addiction, as evidenced by the successful use of D2 receptor antagonists (such as haloperidol) in the treatment of PCP psychosis. [ka] 4. Ibogaine and analogs and their derivatives

[0133] Ibogaine is an indole alkaloid isolated from the root of the West African shrub Tabernanthe iboga. The therapeutic and psychotropic (dream-like) properties of iboga root have been described in ethnobotanical literature for centuries, and ingestion of ibogaine root preparations has ritual and medicinal uses.

[0134] U.S. Patent No. 4,499,096 to Lots of, H.S. describes a method for interrupting opioid addiction syndrome by administering ibogaine. Oral ibogaine was described as an effective treatment for opioid detoxification. Subsequent studies have shown that ibogaine, when administered in the 500-1000 mg range, reduces drug craving and improves depressive symptoms. This dosage range appears to be a safe and effective treatment for interrupting opioid addiction syndrome. However, concerns regarding safety and cardiotoxicity remain. Similar benefits have been observed in recently abstinent cocaine abusers seeking to interrupt a refractory cycle of drug abuse.

[0135] The molecular structures of ibogaine and noribogaine indicate that ibogaine undergoes O-demethylation to form 12-hydroxyibogamine (noribogaine) by the action of cytochrome P4502D6 (CYP2D6). Ibogaine is metabolized to noribogaine in the gastrointestinal wall and liver.

[0136] 18-MC, a synthetic derivative of ibogaine, is an alpha-3-beta-4 nicotinic receptor antagonist with a distinct mechanism of action that modulates excessive dopamine fluctuations in the mesolimbic system of the brain. 18-MC is a synthetic organic molecule designed around the coronaridine chemical backbone, which is common to several plant-based medicinal compounds, including ibogaine. In preclinical efficacy models, 18-MC demonstrated potent activity in reducing both withdrawal symptoms and self-administration of opioids, stimulants, and other substances of abuse. Extensive preclinical characterization demonstrated that 18-MC has a strong safety and tolerability profile. 18-MC has the potential to overcome the safety limitations of ibogaine and has been demonstrated to have no arrhythmic or neurotoxic activity. Other ibogaine derivatives include ME-18-MC, 18-MAC, voacangine, ibogamine, and coronaridine. 5. Atypical hallucinogens

[0137] In some embodiments, the active agent is one or more atypical hallucinogens with diverse mechanisms, such as delta-9-tetrahydrocannabinol or Δ-9-tetrahydrocannabinol (THC) and ibogaine. C. Coupling Agents and Spacers

[0138] The dendrimer-active agent conjugates can be formed from one or more active agents covalently conjugated to the dendrimer or non-covalently bound to the dendrimer. In a preferred embodiment, the one or more active agents are covalently conjugated to the dendrimer.

[0139] Optionally, one or more active agents are conjugated to the dendrimer via one or more spacers. The term "spacer" includes chemical moieties and functional groups used to link the active agent to the dendrimer. A spacer can be either a single chemical entity or two or more chemical entities linked together. A spacer can include any small chemical entity, peptide, or polymer with sulfhydryl, thiopyridine, succinimidyl, maleimide, vinyl sulfone, carbonate, etc.

[0140] In some embodiments, the spacer through which the active agent is conjugated to the dendrimer contains various linking groups, such as disulfide, ester, carbonate, carbamate, thioester, hydrazine, hydrazide, ether, and amide linking groups. The spacer between the dendrimer and the active agent can be designed to provide a releasable or non-releasable form of the dendrimer conjugate in vivo. In some embodiments, conjugation between the active agent and the dendrimer is via an appropriate spacer, including an ester bond between the active agent and the dendrimer. In some embodiments, one or more spacers between the dendrimer and the active agent can provide a desired effective release rate in vivo. These spacers may contain cleavable linking groups (e.g., ester, disulfide, phosphodiester, triglycyl peptide, hydrazine) or non-cleavable linking groups (e.g., amide, ether, and aminoalkyl). Conjugation between the active agent and the dendrimer can be performed using reactions known in the art, such as click chemistry, acid-amine coupling, and Steglich esterification.

[0141] In some embodiments, conjugation between the active agent and the dendrimer is via a spacer containing a disulfide, ester, ether, thioester, carbamate, carbonate, hydrazine, ether, or amide linking group, or a combination thereof. In some embodiments, conjugation between the active agent and the dendrimer is via a suitable spacer, including an ester or amide linking group between the agent and the dendrimer, depending on the desired release rate of the agent.

[0142] The spacer can be selected from the classes of compounds terminated with sulfhydryl, thiopyridine, succinimidyl, maleimide, vinyl sulfone, and carbonate groups. The spacer can include a thiopyridine-terminated compound such as dithiodipyridine, N-succinimidyl 3-(2-pyridyldithio)-propionate (SPDP), succinimidyl 6-(3-[2-pyridyldithio]-propionamido)hexanoate LC-SPDP, or sulfo-LC-SPDP. The spacer can also include peptides, linear or cyclic, inherently containing a sulfhydryl group, such as glutathione, homocysteine, cysteine ​​and its derivatives, arg-gly-asp-cys (RGDC), cyclo(Arg-Gly-Asp-d-Phe-Cys) (c(RGDfC)), cyclo(Arg-Gly-Asp-d-Tyr-Cys), cyclo(Arg-Ala-Asp-d-Tyr-Cys), etc. The spacer can also be a mercapto acid derivative, such as 3-mercaptopropionic acid, mercaptoacetic acid, 4-mercaptobutyric acid, thiolan-2-one, 6-mercaptohexanoic acid, 5-mercaptovaleric acid, and other mercapto derivatives, such as 2-mercaptoethanol and 2-mercaptoethylamine. The spacer may be thiosalicylic acid and its derivatives, (4-succinimidyloxycarbonyl-methyl-alpha-2-pyridylthio)toluene, (3-[2-pyridylthio]propionyl hydrazide). The spacer may have a maleimide terminus, in which case the spacer comprises a polymer or a small chemical entity such as bis-maleimidodiethylene glycol and bis-maleimidotriethylene glycol, bis-maleimidoethane, bismaleimidohexane, etc. The spacer may comprise a vinyl sulfone such as 1,6-hexane-bis-vinyl sulfone. The spacer may comprise a thioglycoside such as thioglucose. The spacer may be a reduced protein such as bovine serum albumin and human serum albumin, or any thiol-terminated compound capable of forming a disulfide bond.The spacer can include maleimide, succinimidyl and / or thiol-terminated polyethylene glycol. D. Dendrimer-Drug Conjugates or Complexes

[0143] Dendrimer-active agent conjugates can be formed from antidepressants and / or antipsychotics covalently or non-covalently conjugated to dendrimers, dendritic polymers, or hyperbranched polymers. Methods for conjugating one or more active agents to dendrimers are known, such as those described in U.S. Patent Application Publication Nos. 2011 / 0034422, 2012 / 0003155, and 2013 / 0136697. In general, conjugation to dendrimers can further improve the safety and efficacy of these agents. For example, dendrimer conjugation can change specific receptor activity and / or modify biodistribution. For example, the use of higher-generation dendrimers and / or dendrimers with molecular weights greater than 24 kDa can restrict these agents to the peripheral nervous system to prevent their psychoactive effects.

[0144] In some embodiments, one or more active agents are covalently conjugated to one or more terminal groups of the dendrimer, such as a terminal hydroxyl group. In some embodiments, the dendrimer conjugate comprises one or more active agents conjugated to the dendrimer via one or more spacers. The spacer between the dendrimer and the active agent can be designed to provide a releasable or non-releasable form of the dendrimer conjugate in vivo. For example, the spacer may be cleavable or may contain a chemical linking group that is cleavable, for example, upon exposure to the intracellular compartment of a target nervous system cell and / or glial cell, or upon binding to a receptor on or within a target nervous system cell and / or glial cell in vivo. Examples of cleavable linking groups that can be used in the spacer of a dendrimer-active agent conjugate include an esterase-sensitive ester bond, a glutathione-sensitive disulfide bond, a phosphatase-sensitive phosphodiester bond, an oligopeptide such as a triglycyl peptide linker capable of lysosomal release, an acid-cleavable hydrazine linking group, and the like. In some embodiments, the spacer between the dendrimer and the active agent can provide a desired effective release rate in vivo, hi some embodiments, the spacer between the dendrimer and the active agent can be non-cleavable or can include non-cleavable chemical linking groups such as amide, ether, and aminoalkyl linking groups.

[0145] Generally, the spacer between the dendrimer and the active agent is long enough to allow the active agent conjugated to the spacer to reach and bind to the target receptor on the surface and / or inside the target cell. For example, the spacer between the dendrimer and the active agent can have a length ranging from 50 Da to 2000 Da, depending on the desired release rate and the desired receptor binding flexibility. The length of the spacer can vary depending on the location of the target receptor (e.g., on the cell surface, in the cytoplasm of the cell, or in the intercellular compartment of the cell) and / or the density of the receptor if located on the cell surface.

[0146] Dendrimers can be second, third, fourth, fifth, sixth and up to tenth generation. In some embodiments, dendrimers are conjugated to one or more active agents via spacers containing cleavable (ester, disulfide, phosphodiester, triglycyl peptide and hydrazine) or non-cleavable (amide, ether and aminoalkyl) linking groups.

[0147] The density of active agents covalently conjugated to the dendrimer or non-covalently bound to the dendrimer can be adjusted based on the particular antidepressant and / or antipsychotic agent being delivered, the target receptor, the target nervous system cells and / or glial cells, the location of the target nervous system cells and / or glial cells, etc. For example, multiple active agents conjugated to the dendrimer may be present on the outer periphery of the dendrimer, with a surface density of active agents of at least 1 active agent / nm 2 (number of conjugated active agents / surface area (nm 2 For example, in some embodiments, 1 nm 2 The surface density of the activator per nanoparticle is at least 10, 15, 20, 25, 30, 35, 40, 45, 50, or more than 50 OH / nm 2 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more OH / nm 2 In some embodiments, the volume density of the active agent is from about 1 to about 50 groups / nm 3 Between about 5 and about 30 groups / nm 3 Between about 10 and about 20 groups / nm 3 It is between.

[0148] Typically, dendrimer-active agent conjugates have hydrodynamic volumes in the nanometer range. For example, in some embodiments, glucose dendrimer-active agent conjugates comprising one or more antidepressants and / or antipsychotics conjugated to a dendrimer have diameters of about 2 nm to about 100 nm or greater than 100 nm, up to 500 nm, depending on the generation of the dendrimer and the chemical composition and amount of active agent conjugated thereto. In some embodiments, dendrimer-active agent conjugates comprising one or more antidepressants and / or antipsychotics conjugated to a dendrimer have diameters effective to penetrate brain tissue and retain the active agents on the surface and / or within target nervous system cells and / or glial cells for a period of time sufficient for the active agents to bind to targeted receptors on the surface and / or within the target nervous system cells and / or glial cells. In some embodiments, dendrimer-active agent conjugates comprising one or more antidepressants and / or antipsychotics conjugated to a dendrimer have a diameter effective to be retained in the peripheral circulation and to be retained on the surface of and / or within target nervous system cells and / or glial cells (such as, for example, nervous system and / or glial cells of the gastrointestinal system) for a period of time sufficient for the active agent to bind to targeted receptors on the surface of and / or within the target nervous system cells and / or glial cells.

[0149] The dendrimer-active agent conjugate may be neutral, positively charged, or negatively charged. In some embodiments, the dendrimer-active agent conjugate is neutral. The presence of an antidepressant and / or antipsychotic derivative can affect the surface charge of the dendrimer-active agent conjugate. In some embodiments, the surface charge of a dendrimer conjugated to an antidepressant and / or antipsychotic is between -100 mV and 100 mV, between -50 mV and 50 mV, between -25 mV and 25 mV, between -20 mV and 20 mV, between -10 mV and 10 mV, between -10 mV and 5 mV, between -5 mV and 5 mV, or between -2 mV and 2 mV. The above ranges include all values ​​between -100 mV and 100 mV. In a preferred embodiment, the surface charge of the dendrimer-active agent conjugate is neutral or near neutral, ie, from about −10 mV to about 10 mV, inclusive.

[0150] An exemplary dendrimer-active agent conjugate is represented by formula (I): The dendrimer of the exemplary conjugate contains surface hydroxyl groups, one or more of which are conjugated to one or more active agents via one or more spacers, as shown in formula (I) below. [ka]

[0151] wherein D can be a 1st to 10th generation or a 2nd to 10th generation dendrimer such as any one of those described above, for example, PAMAM (such as a hydroxyl-terminated PAMAM dendrimer) or a glucose-based dendrimer; L, at each occurrence, can be any suitable chemical moiety, preferably containing a triazole moiety; Y can be a bond or a secondary amide (-CONH-), a tertiary amide (-CONR-), a sulfonamide (-S(O)-NR-), a secondary carbamate (-OCONH-; -NHCOO-), a tertiary carbamate (-OCONR-; -NRCOO-), a carbonate (-OC(O)-O-), a urea (-NHCONH-; -NRCONH-; -NHCONR-, -NRCONR-), a carbinol (-CHOH-, -CROH-), a disulfide group, a phosphodiester group ( [ka] R may be a linking group selected from a hydrazino group, a hydrazone, a hydrazide, an ester (-C(O)-O-), an ether (-O-), and an oligopeptide (e.g., a triglycyl peptide); R is an alkyl group, an aryl group, or a heterocyclic group; X, at each occurrence, may be an antidepressant and / or antipsychotic; the functional group of X (e.g., an amino group, including a primary amino, secondary amino, or tertiary amino group, a carboxyl group; or a hydroxyl group) forms part of the linking group Y; n may be an integer between 1 and 100; and m may be an integer between 16 and 4096. The dendrimer may be a PAMAM (e.g., hydroxyl-terminated PAMAM) or a glucose dendrimer, which is 100% hydroxyl. m and n depend on the size of dendrimer D, and n should be such that the weight percent of the drug in the total conjugate is 5-20%. This range is also appropriate for binding and internalization.

[0152] The oxygen atoms shown in formula (I) are derived from surface functional groups of the dendrimer, such as surface hydroxyl groups, which may or may not be part of a terminal sugar moiety / molecule (e.g., glucose). Although not depicted in formula (I), one or more hydroxyl groups of the dendrimer that are not conjugated to an active agent may be modified with one or more carbohydrates and / or polyalkylene glycols, such as PEG.

[0153] When administered to a subject in need thereof, the antidepressant and / or antipsychotic agent X of Formula (I) can bind to a target receptor on the surface of or inside a target cell. In some embodiments, when the antidepressant and / or antipsychotic agent X binds to the target receptor, the agent X remains conjugated to the dendrimer. In these embodiments, after binding, the agent X can be released from the dendrimer or can remain conjugated to the dendrimer as an intact dendrimer-active agent conjugate. In some embodiments, the antidepressant and / or antipsychotic agent X is released from the dendrimer in close proximity to the target receptor and then binds to the target receptor.

[0154] In some embodiments, L, at each occurrence, can be represented by -A'-L1-B'-L2-, where A' can be carbonyl (-C(O)-) or a bond (including single, double, and triple bonds, e.g., a single bond), B' can be a bond (including single, double, and triple bonds, e.g., a single bond), an amide, an ester, an ether, a thiol, a dithiol, an aryl, a heteroaryl, a polyaryl, a heteropolyaryl, or a heterocyclic, and L1 and L2 can be independently a bond, an alkylene, a heteroalkylene, an aryl, an aralkyl, an ether, a polyether, L1 and L2 may be a thiol, dithiol, thiol ether, polythioether, oligopeptide, polypeptide, oligo(alkylene glycol) or polyalkylene glycol, or L1 and L2 may independently be composed of combinations of these groups, such as a combination of alkylene and polyether, a combination of alkylene and thiol or dithiol, a combination of alkylene and oligopeptide, a combination of alkylene, polyether and thiol or dithiol, or a combination of polyether and thiol or dithiol. In some forms, L1-B'-L2- together form a chemical moiety selected from -alkylene-triazole-di(alkylene glycol)-, -di(alkylene glycol)-triazole-alkylene-, -alkylene-triazole-oligo(alkylene glycol)-, -oligo(alkylene glycol)-triazole-alkylene-, -alkylene-triazole-poly(alkylene glycol)-, -poly(alkylene glycol)-triazole-alkylene-, -alkylene-triazole-ether-, -alkylene-triazole-alkylene-, -alkylene-amide-alkylene-, and combinations thereof.

[0155] In some embodiments, B' can be a bond (including single, double and triple bonds, eg, single bonds), an amide group, or a heterocyclic group such as a triazole group.

[0156] In some embodiments, L1 is a bond, C1-C 10alkylene, such as C1-C8 alkylene, C1-C6 alkylene, C1-C5 alkylene, C1-C4 alkylene or C1-C3 alkylene, or [ka]

[0157] wherein p is an integer of 1 to 20, 1 to 18, 1 to 16, 1 to 14, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 or 2.

[0158] In some embodiments, L2 is a bond, C1-C 10 alkylene such as alkylene, C1-C8 alkylene, C1-C6 alkylene, C1-C5 alkylene, C1-C4 alkylene or C1-C3 alkylene; [ka] wherein p is an integer from 1 to 20, 1 to 18, 1 to 16, 1 to 14, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 or 2), an oligo- or poly-peptide such as triglycyl peptide, a thiol, or a dithiol, or L2 is comprised of a combination of two or more of alkylene, oligo- or poly-(alkylene glycol), oligo- or poly-peptide, thiol, and dithiol. For example, L2 can be [ka] wherein p, q, r, s, t, and u are independently integers from 0 to 10, 0 to 8, 0 to 6, 0 to 5, 0 to 4, 0 to 3, or 0 to 2, such as 0, 1, or 2, and G' is a thiol, dithiol, oligo-peptide (such as triglycyl peptide), or poly-peptide.

[0159] In some embodiments, Y is a linking group that is minimally cleavable in vivo. In some embodiments, Y is a linking group that is cleavable in vivo. In some embodiments, Y is an amide (-CONH-), ester (-C(O)-O-), ether (-O-), phosphodiester, or disulfide group.

[0160] In some embodiments, both L and Y are single bonds and D is conjugated directly to X (the active agent or analog thereof) through an ether linking group.

[0161] In some embodiments, D is a second generation PAMAM dendrimer, a third generation PAMAM dendrimer, a fourth generation PAMAM dendrimer, a fifth generation PAMAM dendrimer, a sixth generation PAMAM dendrimer, a first generation glucose dendrimer, a second generation glucose dendrimer, a third generation glucose dendrimer, a fourth generation glucose dendrimer, a fifth generation glucose dendrimer, or a sixth generation glucose dendrimer.

[0162] More specific exemplary dendrimer-active agent conjugates are shown in the Examples below. E. Exemplary Dendrimer-Drug Conjugates

[0163] In a preferred embodiment, the dendrimer is conjugated to psilocin or a psilocin analog, shown in structures A-D below. [ka]

[0164] n is an integer from 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0165] In a preferred embodiment, the dendrimer is conjugated to ketamine or a ketamine analog, shown in structures E and F below. [ka]

[0166] In other preferred embodiments, the dendrimer is conjugated to DMT or a DMT analog, as shown in structures GJ below. [ka] [ka]

[0167] In a preferred embodiment, the dendrimer is conjugated to an LSD or an LSD analog, shown in structures K and L below. [ka] [ka] III. Methods of Making Dendrimer Conjugates

[0168] Methods for synthesizing dendrimers and preparing dendrimer nanoparticles are also described. A. Methods for Making Dendrimers

[0169] Dendrimers can be prepared by various chemical reaction steps. Dendrimers are usually synthesized according to methods that allow control of their structure at each stage of construction. Dendritic structures are mostly synthesized by two main different methods: divergent or convergent.

[0170] In some embodiments, dendrimers are prepared using a divergent method, in which dendrimers are assembled from a multifunctional core, which is then extended outward through a series of reactions, typically the Michael reaction. This strategy involves coupling monomer molecules bearing reactive and protecting groups with the multifunctional core moiety, resulting in the stepwise addition of generations around the core, followed by removal of the protecting groups. For example, PAMAM-NH2 dendrimers are first synthesized by coupling N-(2-aminoethyl)acrylamide monomers to an ammonia core.

[0171] In other embodiments, dendrimers are prepared using convergent methods, where dendrimers are built from small molecules that terminate on the surface of a sphere, and the reaction proceeds inward, building inward and finally attaching to the core.

[0172] Many other synthetic routes exist for the preparation of dendrimers, such as the orthogonal, accelerated, two-step convergent or hypercore approach, the hypermonomer or branched monomer approach, the double exponential, the orthogonal coupling or two-step approach, the two-monomer approach, the AB2-CD2 approach, etc.

[0173] In some embodiments, the dendrimer core, one or more branching units, one or more spacers, and / or one or more surface functional groups can be modified to allow conjugation to additional functional groups (such as branching units, spacers, surface functional groups), monomers, and / or drugs via click chemistry using one or more of copper-assisted azide-alkyne cycloaddition (CuAAC), Diels-Alder reaction, thiol-ene reaction, thiol-yne reaction, and azide-alkyne reaction (Arseneault M et al., Molecules 2015 May 20;20(5):9263-94). In some embodiments, prefabricated dendrons are clicked onto high-density hydroxyl polymers. "Click chemistry" involves coupling two different moieties (e.g., a core group and a branching unit, or a branching unit and a surface functional group) via a 1,3-dipolar cycloaddition reaction between, for example, an alkyne moiety (or its equivalent) on the surface of a first moiety and an azide moiety (e.g., present in a triazine composition or its equivalent) or any reactive end group (e.g., a primary amine end group, a hydroxyl end group, a carboxylic acid end group, a thiol end group, etc.) on a second moiety. In some embodiments, one or more hydroxyl groups on the surface of a dendrimer (a hydroxyl-terminated PAMAM dendrimer or a glucose dendrimer) are modified to contain an alkyl group, and the drug is modified to contain an azide group. Alternatively, one or more hydroxyl groups on the surface of a dendrimer (a hydroxyl-terminated PAMAM dendrimer or a glucose dendrimer) are modified to contain an azide group, and the drug is modified to contain an alkyne group. The azide and alkyne then react via a 1,3-dipolar cycloaddition reaction to form the triazole moiety.

[0174] In some embodiments, dendrimer synthesis relies on one or more reactions such as thiol-ene click reaction, thiol-yne click reaction, CuAAC, Diels-Alder click reaction, azide-alkyne click reaction, Michael addition, epoxy ring-opening, esterification, silane chemistry, and combinations thereof.

[0175] In some embodiments, the method includes one or more protection and deprotection steps of functional groups (e.g., hydroxyl groups) on the central core, branching units, and / or therapeutic, prophylactic, or diagnostic agents to facilitate the addition of branching units to generate the desired dendrimer molecule, or the addition of the therapeutic, prophylactic, or diagnostic agent to generate the desired dendrimer conjugate. In the case of hydroxyl groups, the hydroxyl group may be protected by the formation of an ether, ester, or acetal. Other exemplary protecting groups include Boc and Fmoc.

[0176] Any existing dendritic platform can be used to create dendrimers with the desired functionality, i.e., a high density of surface hydroxyl groups, by conjugating higher hydroxyl-containing moieties such as 1-thio-glycerol or pentaerythritol. Exemplary dendritic platforms such as polyamidoamine (PAMAM), poly(propyleneimine) (PPI), poly-L-lysine, melamine, poly(etherhydroxylamine) (PEHAM), poly(esteramine) (PEA), and polyglycerol can be synthesized and explored.

[0177] Dendrimers can also be prepared by combining two or more dendrons. A dendron is a wedge-shaped section of a dendrimer that carries a reactive focal group. Numerous dendron scaffolds are commercially available. These dendrons have appeared in the first, second, third, fourth, fifth, and sixth generations, with 2, 4, 8, 16, 32, and 64 reactive groups, respectively. In certain embodiments, one type of drug is linked to one type of dendron, and a different type of drug is linked to another type of dendron. The two dendrons are then linked to form a dendrimer. The two dendrons can be linked via click chemistry, i.e., a 1,3-dipolar cycloaddition reaction, between an azide moiety on one dendron and an alkyne moiety on another dendron to form a triazole linker.

[0178] Exemplary methods of making dendrimers are described in detail in International Patent Publication Nos. WO2009 / 046446, WO2015168347, WO2016025745, WO2016025741, WO2019094952, and U.S. Pat. No. 8,889,101. 1. Method for preparing glucose dendrimers

[0179] Glucose-based dendrimers are assembled from a multifunctional core, which is extended outward through a series of reactions. The strategy involves coupling monomer molecules bearing reactive and protecting groups with the multifunctional core moiety, which results in the stepwise addition of generations around the core, followed by removal of the protecting groups.

[0180] In some embodiments, glucose dendrimers are synthesized by coupling AB4 peracetylated β-D glucose-PEG4-azide monomers to a hexapropargylated core. In a preferred embodiment, the hypercore is prepared from dipentaerythritol, for example, by propargylating dipentaerythritol to obtain a hexapropargylated core. An exemplary scheme for preparing such glucose dendrimers is shown in Scheme I. [ka]

[0181] In some embodiments, the branching unit is a hypermonomer, i.e., AB n These are building blocks. Exemplary hypermonomers include the AB3, AB4, AB5, AB6, AB7, and AB8 building blocks. The hypermonomer strategy dramatically increases the number of available end groups. An exemplary hypermonomer is the orthogonal hypermonomer AB4, which contains one azide functional group and four allyl groups and is prepared by reacting dipentaerythritol, which has five allyl groups, with monotosylated triethylene glycol azide.

[0182] In some embodiments, the branching unit is a linear or branched polyethyleneglycerol, for example, as shown by Formula III. Other monomers include disaccharides and oligosaccharides, as well as saccharides such as fructose, lactose, and sucrose. a. Synthesis of AB4 building blocks

[0183] Some exemplary synthesis methods for hypermonomer AB4 are described below. In some embodiments, hypermonomer AB4 is based on a glucose molecule. In a preferred embodiment, hypermonomer AB4 is conjugated to polyethyleneglycerol, e.g., tetraethyleneglycol (PEG4). In one embodiment, hypermonomer AB4 is peracetylated β-D-glucopyranoside tetraethyleneglycol azide.

[0184] In some embodiments, the synthesis of glucose-Oac-TEG-Ots comprises the following steps: a solution of peracetylated β-D-glucopyranoside (10 g, 25.6 mmol) was dissolved in 50 mL of anhydrous dichloromethane (DCM), and then 2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (6.2 g, 17.9 mmol) was added, and the reaction mixture was cooled to 0°C. Boron trifluoride diethyl etherate (2.5 equivalents) was added, and the reaction was allowed to reach room temperature. The reaction was monitored by TLC and, after 5 hours, quenched at 0°C by adding saturated sodium bicarbonate solution. After stirring for 10 minutes, DCM (300 mL) was added, and the organic layer was washed three times with saturated sodium bicarbonate solution until foaming disappeared. The reaction mixture was dried over sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The crude product was purified by combiflash® chromatography using a mixture of ethyl acetate / hexane (70:30) as the eluent. The desired compound was obtained in 60% yield. The structure of Glucose-Oac-TEG-Ots is shown below: [ka]

[0185] In some embodiments, the synthesis of Glucose-Oac-TEG-N3 involves the following steps: A solution of Glucose-Oac-TEG-Ots (6 g, 8.8 mmol) is dissolved in 40 mL of anhydrous DMF, then sodium azide (2 equivalents) is added, and the reaction mixture is heated to 50° C. overnight. Upon completion, the reaction mixture is filtered and the DMF is evaporated. Once dry, the crude reaction mixture is passed through a combiflash® column using ethyl acetate:hexane (70:30) as the eluent. The structure of Glucose-Oac-TEG-N3 is shown below: [ka]

[0186] In some embodiments, the synthesis of glucose-OH-TEG-N3 involves the following steps: dissolving peracetylated β-D-glucopyranoside tetraethylene glycol azide in anhydrous methanol and adding sodium methoxide to adjust the pH to about 8.5-9. The reaction is stirred overnight at room temperature, then diluted with methanol and adjusted to about pH 6-7 with Amberlist IR-120+. The reaction mixture is isolated by filtration, and the solvent is removed by rotary evaporation. The structure of glucose-OH-TEG-N3 is shown below. [ka] b. Synthesis of glucose dendrimers

[0187] In some embodiments, glucose dendrimers are synthesized by coupling AB4 peracetylated β-D-glucose-PEG-azide monomers to a hexapropargylated core. In a preferred embodiment, the hexapropargylated core is linked to the AB4 β-D-glucose-PEG-azide building block (2) via a click reaction to yield a first-generation dendrimer.

[0188] In some embodiments, the first-generation dendrimer D1-Glu6-Oac24 is prepared as follows: The hexapropargylated compound (0.5 g, 1 mmol) and the azide derivative (4.1 g, 7.4 mmol, 1.2 equivalents per acetylene) are suspended in a 1:1 mixture of DMF and water in a 20 mL microwave vial equipped with a magnetic stir bar. CuSO4·5H2O (5 mol% / acetylene, 75 mg) and sodium ascorbate (5 mol% / acetylene, 60 mg) dissolved in a minimum amount of water are added. The reaction is microwaved at 50 °C for 6 h. The reaction mixture is dialyzed against DMF followed by water dialysis containing EDTA. The EDTA is further removed by extensive water dialysis. The product is lyophilized to afford D1-Glu6-Oac24. The structure of D1-Glu6-Oac24 is shown below. [ka]

[0189] In some embodiments, the first generation dendrimer D1-Glu6-OH 24 is prepared as follows: peracetylated first-generation glucose dendrimer (1 g, 0.26 mmol) is dissolved in anhydrous methanol and sodium methoxide is added to adjust the pH to approximately 8.5-9. The reaction is stirred overnight at room temperature, then diluted with methanol and the pH adjusted to approximately 6-7 with AMBERLIST® IR-120+. The reaction mixture is separated by filtration, the solvent removed by rotary evaporation, and then subjected to water dialysis. The structure of first-generation glucose dendrimer D1-Glu6-OH24 is shown below. [ka]

[0190] In some embodiments, the first generation glucose dendrimer D1-Glu6-OH 24 is propargylated to give D1-acetylene 24 as follows: D1-Glu6-OH 24D1-acetylene 24 (2 g, 0.721 mmol) was dissolved in anhydrous dimethylformamide (DMF, 50 mL) by sonication. Sodium hydride (60% dispersion in mineral oil) (951 mg, 39.65 mmol) was added slowly in small portions to this solution with stirring at 0°C. The solution was stirred for an additional 15 minutes at 0°C. After this, propargyl bromide (3.85 mL, 34.608 mmol, 80% w / w solution in toluene) was added at 0°C, and stirring was continued for an additional 6 hours at room temperature. The reaction mixture was quenched with ice and water, filtered, and dialyzed against DMF followed by water dialysis to give D1-acetylene 24. The structure of D1-acetylene 24 is shown below. [ka]

[0191] In some embodiments, the first generation dendrimer D1-acetylene 24 is further reacted with AB4β-D-glucose-PEG4-azide to give a second generation dendrimer with 24 glucose molecules, containing 96 surface hydroxyl groups.

[0192] An exemplary second-generation dendrimer, D2-Glu24-Oac96, was prepared as follows: D1-acetylene dendrimer 24 (0.5 g, 0.13 mmol) and glucose-Oac-TEG-azide (2.2 g, 4 mmol) were suspended in a 1:1 mixture of DMF and water in a 20 mL microwave vial equipped with a magnetic stir bar. To this, CuSO4·5H2O (5 mol% / acetylene, 5 mg) and sodium ascorbate (5 mol% / acetylene, 10 mg) dissolved in a minimal amount of water were added. The reaction was microwaved at 50 °C for 8 h. Upon completion, the reaction mixture was dialyzed against DMF, followed by water dialysis containing EDTA. The EDTA was further removed by extensive water dialysis. The product was lyophilized to give D2-Glu 24 -Oac 96 is obtained.

[0193] In some embodiments, second-generation dendrimer D2-Glu24-OH96 is prepared as follows: peracetylated second-generation glucose dendrimer D2-Glu24-OH96 is dissolved in anhydrous methanol and sodium methoxide is added to adjust the pH to about 8.5-9.0. The reaction is stirred overnight at room temperature, then diluted with methanol and the pH adjusted to about 6-7 with AMBERLIST® IR-120+. The reaction mixture is filtered to remove the resin, and the filtrate is rotary evaporated, followed by water dialysis to give the product as an off-white solid.

[0194] Second generation glucose dendrimer D2-Glu 24 -OH 96 The structure of is shown below. [ka]

[0195] In some embodiments, the second generation dendrimer D2-Glu 24 -OH 96 is propargylated at one or more terminal hydroxyl groups suitable for further conjugation to one or more therapeutic, prophylactic, or diagnostic agents. In some embodiments, one or more terminal hydroxyl groups of second-generation dendrimer D2-Glu24-OH96 are propargylated as follows: D2-Glu24-OH96 (5b) (200 mg, 0.016 mmol) is dissolved in anhydrous dimethylformamide (DMF, 10 mL) by sonication. To this stirred solution, sodium hydride (60% dispersion in mineral oil) (22 mg, 0.934 mmol) is added slowly in small portions at 0° C. The solution is further stirred at 0° C. for 15 minutes. After this, propargyl bromide (18.0 μL, 80% w / w solution in toluene) is added at 0° C., and stirring is continued for an additional 6 hours at room temperature. The solvent is evaporated using a V10 evaporator system and the crude product is purified by passing through a PD10 SEPHADEX® G25M column. The aqueous solution is lyophilized to give the product as an off-white solid.

[0196] In some embodiments, one or more fluorescent dyes, such as infrared-fluorescent Cy5 dyes, are conjugated to the second-generation dendrimer D2-Glu24-OH96. In one embodiment, Cy5-D2-Glu24-OH96 (compound 7 in Figure 1B) is prepared as follows: Compound 6 (200 mg, 0.016 mmol) and Cy5 azide (20.7 mg, 0.02 mmol) are suspended in a 1:1 mixture of DMF and water in a 25 mL round-bottom flask equipped with a magnetic stir bar. To this, CuSO4·5H2O (5 mol% / acetylene, 0.3 mg) and sodium ascorbate (10 mol% / acetylene, 0.5 mg) dissolved in a minimum amount of water are added. The reaction is stirred at room temperature for 24 hours. Upon completion, the DMF is evaporated using a V10 column, and purification is performed using a PD10 Sephadex G25 M column. The aqueous solution is lyophilized to give the product as a blue solid.

[0197] In some embodiments, the total hydroxyl groups available for further conjugation to active agents, including therapeutic and / or diagnostic agents, are about 1-30, 2-20, or 5-10 of the total 96 hydroxyl groups available for an exemplary second generation dendrimer having 24 glucose molecules containing 96 surface hydroxyl groups. B. Methods of Making Dendrimer-Drug Conjugates

[0198] Methods for conjugating drugs to dendrimers are generally known in the art, for example, as described in US2011 / 0034422, US2012 / 0003155 and US2013 / 0136697.

[0199] In some embodiments, one or more drugs are covalently attached to the dendrimer. In some embodiments, the drugs are attached to the dendrimer via a spacer designed to be non-cleavable in vivo. In some embodiments, the drugs are attached to the dendrimer via a spacer designed to be cleavable in vivo. For example, the spacer can be designed to be cleaved by hydrolysis, enzymes, or a combination thereof to achieve sustained release of the drug in vivo. In some embodiments, both the chemical structure of the spacer and its point of attachment to the drug can be selected so that cleavage of the spacer releases either the drug or a suitable prodrug thereof. The chemical structure of the spacer can also be selected taking into account the desired release rate of the drug.

[0200] In some embodiments, the conjugation between the drug and the dendrimer is via one or more of a disulfide, ester, ether, phosphodiester, triglycyl peptide, hydrazine, amide, or aminoalkyl linking group. In some embodiments, the conjugation between the drug and the dendrimer is via a suitable spacer that results in an ester or amide bond between the drug and the dendrimer, depending on the desired release rate of the drug. In some cases, an ester or disulfide bond is introduced for a releasable form of the drug. In other cases, an amide or aminoalkyl bond is introduced for a non-releasable form of the drug.

[0201] The spacer generally contains one or more organic functional groups. Examples of suitable organic functional groups contained in the spacer include secondary amide (-CONH-), tertiary amide (-CONR-), sulfonamide (-S(O)2-NR-), secondary carbamate (-OCONH-; -NHCOO-), tertiary carbamate (-OCONR-; -NRCOO-), carbonate (-OC(O)-O-), urea (-NHCONH-; -NRCONH-; -NHCONR-, -NRCONR-), carbinol (-CHOH-, -CROH-), disulfide group, hydrazone, hydrazide, ether (-O-) and ester (-COO-, -CHOC-, CHROC-), where R is an alkyl group, an aryl group or a heterocyclic group. Generally, the one or more organic functional groups in the spacer are selected taking into account the desired release rate of the drug. Additionally, one or more organic functional groups can be selected to facilitate covalent conjugation of a drug to the dendrimer. In some embodiments, conjugation between the drug and the dendrimer is via a suitable spacer that results in a disulfide bridge between the drug and the dendrimer. In some embodiments, the dendrimer-active agent conjugate is capable of rapid release of the drug in vivo by a thiol exchange reaction under reducing conditions found in the body.

[0202] In certain embodiments, the spacer comprises one or more of the organic functional groups described above in combination with a linking group. The linking group may be composed of any collection of atoms, including oligomeric and polymeric chains, for example, where the total number of atoms in the linking group is between 3 and 200 atoms, between 3 and 150 atoms, between 3 and 100 atoms, or between 3 and 50 atoms. Examples of suitable linking groups include alkyl groups, heteroalkyl groups, alkylaryl groups, oligoethylene glycol chains and polyethylene glycol chains, and oligo(amino acid) chains and poly(amino acid) chains. Variations in the linking group provide further control over in vivo drug release. In embodiments in which the spacer comprises a linking group, one or more organic functional groups are typically used to link the linking group to both the anti-inflammatory agent and the dendrimer.

[0203] Reactions and strategies useful for covalently conjugating drugs to dendrimers are known in the art. See, for example, March, "Advanced Organic Chemistry," 5th Edition, 2001, Wiley-Interscience Publication, New York) and Hermanson, "Bioconjugate Techniques," 1996, Elsevier Academic Press, USA. The appropriate method for covalently conjugating a given drug involves functional group compatibility, protecting group strategy, and the presence of labile bonds, and can be selected taking into account the desired linking moiety, as well as the structure of the drug and dendrimer.

[0204] The amount of active agent (drug loading) in the dendrimer-active agent conjugate depends on numerous factors, including the choice of active agent, the dendrimer structure and size, and the tissue to be treated. In some embodiments, the one or more antidepressants and / or antipsychotics are conjugated to the dendrimer at a concentration of between about 0.01% and about 45% by weight (inclusive), between about 0.1% and about 30% by weight (inclusive), between about 0.1% and about 20% by weight (inclusive), between about 0.1% and about 10% by weight (inclusive), between about 1% and about 10% by weight (inclusive), between about 1% and about 5% by weight (inclusive), between about 3% and about 20% by weight (inclusive); or between about 3% and about 10% by weight (inclusive). However, the specific drug loading for any given active agent, dendrimer and targeting site can be determined by routine methods such as those described.

[0205] In some embodiments, drug / spacer conjugation occurs on about 1%, 2%, 3%, 4%, or 5% of the total number of available surface functional groups, such as hydroxyl groups, on the dendrimer prior to conjugation. In other embodiments, drug / spacer conjugation occurs on less than 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of the total number of available surface functional groups on the dendrimer prior to conjugation with the active agent. In some embodiments, the dendrimer-active agent conjugate retains an effective amount of surface functional groups for targeting to target nervous system cells and / or glial cells while simultaneously being conjugated to an effective amount of drug for treating, preventing, and / or imaging a disease or disorder. In some embodiments, the dendrimer-active agent conjugate carries an amount of active agent effective to target the target nervous system cells and / or glial cells and to bind to a target receptor on the surface of or within the target nervous system cells and / or glial cells.

[0206] More specific methods for preparing exemplary dendrimer-active agent conjugates are described in the Examples below. IV. Pharmaceutical Preparations

[0207] Pharmaceutical compositions comprising the dendrimer-active agent conjugates can be formulated in a conventional manner using one or more physiologically acceptable carriers, optionally including additives and adjuvants that facilitate processing into preparations of the active compound that can be pharmaceutically used for oral, mucosal (intranasal, buccal, sublingual, vaginal, rectal or pulmonary), transdermal or injectable (intravenous, subcutaneous, intraperitoneal, intramuscular or intrathecal administration) administration.

[0208] Representative additives include aqueous buffers, wetting agents, viscosity modifiers, tonicity agents, stabilizers, and combinations thereof. Suitable pharmaceutically acceptable additives are preferably selected from generally recognized as safe (GRAS) substances and can be individually administered without causing undesired biological side effects or unwanted interactions.

[0209] Generally, pharmaceutically acceptable salts of active substances can be prepared by reacting a drug in the form of a free acid or free base with a stoichiometric amount of an appropriate base or acid in water, an organic solvent, or a mixture of the two; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Pharmaceutically acceptable salts include drug salts derived from inorganic acids, organic acids, alkali metal salts, alkaline earth metal salts, and salts formed by the reaction of a drug with a suitable organic ligand (e.g., quaternary ammonium salts). A list of suitable salts can be found, for example, in Remington's Pharmaceutical Sciences, 20th ed., Lippincott Williams & Wilkins, Baltimore, MD, 2000, p. 704. Examples of ophthalmic drugs that may be administered in the form of pharmaceutically acceptable salts include timolol maleate, brimonidine tartrate, and diclofenac sodium.

[0210] The compositions are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. The phrase "dosage unit form" refers to a physically discrete unit of the conjugate appropriate for the patient being treated. However, it will be understood that the total number of single doses of the composition will be determined by the attending physician within the scope of sound medical judgment. The therapeutically effective dose can be initially estimated in cell culture assays or animal models, usually mice, rabbits, dogs, or pigs, or extrapolated from human data. Animal models are also used to obtain a desirable concentration range and route of administration. Such information will then be useful for determining effective doses and routes for administration in humans. The therapeutic efficacy and toxicity of the conjugates can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, such as ED50 (the dose therapeutically effective in 50% of the population) and LD50 (the dose lethal to 50% of the population). The dose ratio of toxic to therapeutic effects is the therapeutic index, and the LD 50 / ED 50 Pharmaceutical compositions that exhibit large therapeutic indices are preferred. The data obtained from cell culture assays and animal studies can be used in formulating a range of dosage for human use.

[0211] In certain embodiments, the composition is administered locally, for example, by direct injection or implantation at the site to be treated. In some embodiments, the composition is injected, topically applied, or otherwise directly administered to the vasculature at the site of injury, surgery, or implantation or on vascular tissue adjacent to the site. For example, in embodiments, the composition is applied topically to exposed vascular tissue during a surgical procedure, or as a cream, gel, or lotion. Typically, local administration results in an increased local concentration of the composition, which is higher than the concentration that can be achieved by systemic administration.

[0212] Pharmaceutical compositions formulated for administration by parenteral (intramuscular, intraperitoneal, intravenous or subcutaneous injection) and enteral routes of administration are described. A. Parenteral Administration

[0213] The dendrimer-active agent conjugate composition can be administered parenterally. The terms "parenteral administration" and "parenterally administered" are art-recognized terms and include modes of administration other than enteral and topical administration. Dendrimers can be administered orally, intranasally, subcutaneously, intraperitoneally, transdermally, or intramuscularly. For liquid formulations, pharmaceutically acceptable carriers can be, for example, aqueous or non-aqueous solutions, suspensions, emulsions, or oils. Parenteral vehicles (for subcutaneous, intravenous, intraarterial, or intramuscular injection) include, for example, sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's, and fixed oils. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, and injectable organic esters such as ethyl oleate. Aqueous carriers include, for example, water, alcoholic / aqueous solutions, emulsions or suspensions, including cyclodextrins, saline, and buffered media. Dendrimers can also be administered in emulsion, for example, in water in oil.Examples of oils are those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, olive oil, sunflower oil, fish liver oil, sesame oil, cottonseed oil, corn oil, olive, petrolatum and inorganic.The fatty acids suitable for use in parenteral formulations include, for example, oleic acid, stearic acid and isostearic acid.Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters.

[0214] Formulations suitable for parenteral administration can include aqueous and non-aqueous sterile suspensions, which can contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, as well as suspending agents, solubilizers, thickeners, stabilizers, and preservatives. Intravenous vehicles can include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer's dextrose). In general, water, saline, aqueous dextrose and related sugar solutions, and glycols such as propylene glycol or polyethylene glycol are preferred liquid carriers, particularly for injectable solutions.

[0215] Injectable pharmaceutical carriers for injectable compositions are well known to those skilled in the art (see, e.g., Pharmaceutics and Pharmacy Practice, J.B. Lippincott Company, Philadelphia, PA, Banker and Chalmers, eds., pages 238-250 (1982), and ASHP Handbook on Injectable Drugs, Trissel, 15th ed., pages 622-630 (2009)). B. Enteral Administration

[0216] The dendrimer-active agent conjugate compositions may be administered enterally (oral, sublingual, vaginal, rectal, buccal, nasal, pulmonary, or transdermally). The carrier or diluent may be a solid carrier such as a capsule or tablet or diluent for solid formulations, a liquid carrier or diluent for liquid formulations, or a mixture thereof.

[0217] For liquid preparations, pharmaceutically acceptable carriers can be, for example, aqueous or non-aqueous solutions, suspensions, emulsions or oils.Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, and injectable organic esters such as ethyl oleate.Aqueous carriers include, for example, water, alcoholic / aqueous solutions, cyclodextrins, emulsions or suspensions including saline and buffered media.

[0218] Examples of oils include those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, olive oil, sunflower oil, fish-liver oil, sesame oil, cottonseed oil, corn oil, olive, petrolatum, and inorganic. Fatty acids suitable for use in parenteral formulations include, for example, oleic acid, stearic acid, and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters.

[0219] Vehicles include, for example, sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's, and fixed oils. Formulations include, for example, aqueous and non-aqueous isotonic sterile injection solutions (which may contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient), and aqueous and non-aqueous sterile suspensions (which may contain suspending agents, solubilizers, thickeners, stabilizers, and preservatives). Vehicles may include, for example, fluid and nutrient supplements, electrolyte supplements (such as those based on Ringer's dextrose). In general, water, saline, aqueous dextrose, and related sugar solutions are preferred liquid carriers. They may also be formulated with proteins, fats, saccharides, and other ingredients of infant formula.

[0220] Oral preparations may be in the form of chewing gum, gel strips, tablets, capsules or lozenges. Encapsulating materials for preparing enteric-coated oral preparations include cellulose acetate phthalate, polyvinyl phthalate acetate, hydroxypropylmethylcellulose phthalate and methacrylic acid ester copolymers. Solid oral preparations such as capsules or tablets are preferred. Elixirs and syrups are also well-known oral preparations.

[0221] Formulations for administration to mucosal surfaces such as the nose, oral surfaces or lungs will usually contain pharmaceutically acceptable excipients such as those used for parenteral administration, alone or in combination with various surfactants, penetration enhancers, etc.

[0222] The present compositions can also be made into aerosol formulations (i.e., they can be "nebulized") to be administered by inhalation. Aerosol formulations can be placed into pressurized acceptable propellants, such as dichlorodifluoromethane, propane, nitrogen, and air. For administration by inhalation, the compounds are delivered in the form of an aerosol spray supply from a pressurized pack or nebulizer, using a suitable propellant. V. How to use

[0223] Methods of using dendrimer-psychedelic drug compositions are described. The methods can treat or alleviate one or more symptoms of psychological, cognitive, behavioral, and / or mood disorders. The compositions are administered in an amount effective to produce a therapeutic effect but insufficient to produce adverse effects.

[0224] In preferred embodiments, the dendrimer compositions cross the central and peripheral nervous system barrier interface and selectively target specific cells and specific receptors on cells to address various diseases, disorders, injuries, and conditions. The method comprises administering to a subject in need thereof the composition in an amount effective to increase the permeability of hallucinogenic compounds across the central and peripheral nervous system barrier interface and / or to increase binding of hallucinogenic compounds at specific receptors in specific cells, particularly serotonergic receptors, dopaminergic receptors, adrenergic transporters, and / or monoamine transporters in cells in the central nervous system, peripheral nervous system, and / or peripheral circulation, e.g., nervous system cells, glial cells, gastrointestinal cells, and / or immune cells. A. Treatment Method

[0225] The composition can be administered to prevent, treat, and / or manage the symptoms of various disorders, diseases, and conditions, including, but not limited to, anxiety disorders, mood disorders, eating disorders, personality disorders, stress disorders, and / or psychiatric disorders.In some embodiments, when the dendrimer is conjugated to one or more hallucinogens, the composition can be administered to treat one or more neurological disorders, such as mental health disorders, for example, mood disorders, anxiety disorders, eating disorders, substance-related disorders, and stress disorders, for example, post-traumatic stress disorder, learning disorders, for example, autism, and pain disorders, for example, neuropathic pain.In other embodiments, the dendrimer-hallucinogen conjugate can be administered to subjects who need, for example, to stabilize mood in bipolar disorder, to reduce anxiety in anxiety disorders, and to reduce cancer-related neuropathological distress.In still other embodiments, the dendrimer is conjugated to one or more hallucinogens to treat non-neuronal diseases, such as autoimmune disorders.

[0226] Typically, an effective amount of a dendrimer complex comprising a combination of a dendrimer and one or more therapeutic, prophylactic and / or diagnostic active agents is administered to an individual in need thereof. The dendrimer also comprises a targeting agent, although, as demonstrated by the examples, these are not required for delivery to damaged tissue in the spinal cord, brain and related regions.

[0227] In some embodiments, the dendrimer complex comprises a drug bound to or conjugated to a dendrimer, which is capable of preferentially releasing the drug at the target receptor. The drug can be covalently bound or dispersed or encapsulated within the molecule. The amount of dendrimer complex administered to a subject is selected to deliver an amount effective to reduce, prevent, or otherwise alleviate one or more clinical or molecular symptoms of the disease or disorder being treated, compared to a control, for example, a subject treated with an active agent without a dendrimer. B. Conditions and Diseases to be Treated 1. Site-specific targeting

[0228] The present composition and method are designed to avoid the existing difficulties in selective drug delivery to the central nervous system and peripheral nervous system.The present composition and method can improve the bioavailability of drugs in the central nervous system and peripheral nervous system by one or more of the following: (i) increasing the drug density that crosses the brain barrier, especially the blood-brain and blood-spinal fluid barrier; (ii) improving drug solubility; (iii) promoting target engagement, i.e., increasing site-specific binding; (iv) improving the pharmacokinetics of drugs; and (v) increasing intracerebral distribution.For example, in some forms, the present composition and method can selectively deliver compounds to the peripheral nervous system, thereby increasing the possibility that the present composition can be used to selectively treat peripheral-specific diseases and disorders, including but not limited to neuropathic pain, traumatic nerve injury and inflammatory disorders. A. Improving drug permeability across barrier interfaces

[0229] The present dendrimer compositions and methods can improve the pathway of hallucinogenic compounds across one or more barrier interfaces in the brain and nervous system, particularly the blood-brain barrier (BBB), CSF-blood barrier, and blood-nerve barrier. These barrier interfaces normally protect neurons from blood-borne infectious agents and help maintain water homeostasis and an appropriate environment for neuronal function in the blood. Due to the clinical importance of hallucinogens for the treatment of mental health disorders, the present dendrimer compositions can be used to deliver hallucinogenic compounds with improved permeability across these barrier interfaces for site-specific targeting. i. Blood-brain barrier

[0230] The "blood-brain barrier" (BBB) ​​is a continuous endothelial membrane that, together with pericytes and other components of the neurovascular unit, restricts the entry of toxins, pathogens, and blood cells into the brain. However, the BBB also poses an obstacle to drug delivery to the central nervous system (CNS), in part because (1) delivery of drugs targeted to the brain via systemic routes can result in unacceptably high drug levels in the periphery, and (2) the complex interactions of cells and molecules that contribute to the structure and function of the BBB make it difficult to determine drug permeability across the BBB, drug distribution in the brain, and target engagement in the brain.

[0231] Brain microvascular endothelial cells, pericytes, astrocytes, tight junctions, neurons, and basement membrane structures physically stiffen brain capillaries at the BBB. Brain capillary endothelial cells lack fenestrae, limiting the diffusion of small molecules and proteins. Interendothelial junctions connect endothelial cells to form a continuous barrier, severely restricting the penetration of water-soluble substances. Pericytes, astrocytes, and basement membranes surround endothelial cells, ultimately forming the impermeable BBB. Furthermore, efflux transporters are located in brain capillary endothelial cells, further obstructing the entry of substances into the brain. BBB permeability is primarily controlled by interendothelial junctions, which are protein complexes including adherens junctions, tight junctions, and gap junctions. Adherens junctions primarily regulate the permeability of the endothelial barrier. Tight junctions play a key role in maintaining the permeability barrier between epithelial and endothelial cells, thereby controlling tissue homeostasis. Gap junctions, composed of six connexin molecules, direct electrical and chemical communication between endothelial cells. Finally, instead of having a static structure, the components of the BBB continuously adapt in response to various physiological changes in the brain. The dendrimer compositions and methods overcome the aforementioned challenges and are suitable for delivering hallucinogenic compounds across the blood-brain barrier by one or more of the above-mentioned transport mechanisms.

[0232] Molecules cross the BBB via the paracellular route (between adjacent cells) or the transcellular route (through cells). In the paracellular route, ions and solutes cross the BBB by passive diffusion, taking advantage of their concentration gradient. The transcellular route involves various mechanisms, such as passive diffusion, receptor-mediated transport, and transcytosis.

[0233] Physicochemical factors affecting BBB permeability include molecular weight, charge, lipid solubility, surface activity, and relative size of molecules. BBB permeability can also be affected by physiological factors such as efflux transporters, e.g., P-glycoprotein (P-gp), enzyme activity, plasma protein binding, and cerebral blood flow. Hydrophilic molecules, such as proteins and peptides, enter the brain via specific receptor- and saturable receptor-mediated transport mechanisms, such as glucose transporter-1 (GLUT-1), insulin transporter, and transferrin transporter. These endogenous transporters are expressed in the luminal and abluminal endothelial cell membranes. Among these transport mechanisms, receptor-mediated transcytosis has been extensively studied for delivering drugs to the brain. The dendrimer compositions and methods of the present application are suitable for delivering hallucinogenic compounds across the blood-brain barrier via one or more of the above-mentioned mechanistic pathways. ii. Blood-nerve barrier (BNB)

[0234] The blood-nerve barrier (BNB) defines a physiological space within which axons, Schwann cells, and other associated cells of peripheral nerves function, ensuring proper peripheral nerve function and maintaining homeostasis of the endoneurial environment. The BNB consists of endoneurial microvessels within nerve bundles and the surrounding perineurium. Tight junctions between endothelial cells and pericytes in the endoneurial vasculature isolate the endoneurial membrane from the blood, preventing uncontrolled leakage of molecules and ions from the circulatory system into peripheral nerves. Additionally, a diffusion barrier exists within the perineurium, formed by tight junctions between adjacent perineurial cells and the basement membrane surrounding each perineurial cell layer. Endoneurial capillaries and perineurial passages are strict barriers that isolate the endoneurial extracellular environment of peripheral nerves from both the epineurial perifascicular space and the systemic circulation, thus protecting the endoneurial microenvironment from dramatic concentration changes in the vascular space and other extracellular spaces.

[0235] For drug targets located in peripheral nerves, BNBs can be difficult to resolve, since they can restrict or prevent drugs from reaching their sites of action, thus negatively affecting the efficacy of drugs.In addition, the expression profile of transporters in peripheral nerves can be very different from that in the central nervous system.The dendrimer composition of the present application can be used to improve the permeability of hallucinogenic compounds through BNBs, thereby improving the delivery of hallucinogenic compounds to peripheral nerve targets. iii. Blood-CSF barrier

[0236] The composition may be used to improve the delivery of hallucinogenic compounds to target sites through the blood-cerebrospinal fluid barrier (blood-CSF barrier) and cerebral ventricles. The choroid plexus is a vascular tissue found in all ventricles. The functional unit of the choroid plexus is composed of capillaries surrounded by a layer of differentiated ependymal epithelium. Unlike the capillaries that form the blood-brain barrier, choroid plexus capillaries are fenestrated and do not have tight junctions. Therefore, the endothelium does not form a barrier to the movement of small molecules. Instead, the blood-CSF barrier in the choroid plexus is formed by epithelial cells and the tight junctions that connect them. The other part of the blood-CSF barrier is the arachnoid membrane that surrounds the brain. The cells of this membrane are also connected by tight junctions.

[0237] The CSF space and brain structures adjacent to the CSF compartment are pathological targets of interest in CNS diseases. For example, the subarachnoid, perivascular, or periventricular space are areas of pathogenic lymphocyte, monocyte, and neutrophil accumulation in autoimmune disorders, eosinophilic inflammation, and / or neuroinflammatory disorders such as asthma. The B cell nests that produce potentially harmful antibodies found in various CNS autoimmune diseases are thought to be primarily located in the pia mater. Therefore, in some forms, the dendrimer compositions can be used to deliver antidepressants and / or antipsychotics to targeted areas via the blood-CSF space connected to deep cervical lymph nodes to ameliorate or treat symptoms associated with altered cytokine responses following neuroinflammatory disorders, such as stroke and allergic attacks.

[0238] In some embodiments, dendrimer compositions can be used to deliver hallucinogenic agents to target sites to ameliorate or treat neuropathological and inflammatory symptoms associated with tumor development. Thus, in some embodiments, dendrimer compositions can be used to harness pharmacological pressure from the CSF to achieve therapeutic concentrations of hallucinogenic compounds within the tumor microenvironment and reduce inflammatory responses and tumor development. b. Improved target-specific binding

[0239] The dendrimer compositions are suitable for delivering hallucinogenic compounds that modulate serotonin (5HT) receptors, e.g., 5HT-1A, 5HT-2B, 5HT-2A, 5HT-2B, 5HT-2C, 5HT-3, 5HT-4, 5HT-6, and 5HT-7 receptors, and / or one or more dopamine receptors, e.g., dopamine D1 and D2 receptors, and therefore have improved binding affinity and specificity to one or more receptors. The dendrimer compositions also modulate one or more norepinephrine (NE) receptors, e.g., α 2A -adrenergic receptor, α 2B -adrenergic receptor, α 2C The dendrimer compositions described herein may be used to deliver hallucinogenic compounds for directly or indirectly modulating adrenergic receptors, beta-adrenergic receptors, monoamine transporters, such as the serotonin reuptake transporter (SERT), dopamine transporter (DAT), and / or vesicular monoamine transporter (VMAT2). Additionally, hallucinogenic compounds that have affinity for and are suitable for binding to one or more AMPA receptors, NMDA receptors, EGFR1 receptors, EGFR2 receptors, histamine (H1) receptors, GABA receptors, and trace amine associated receptor 1 (TAAR1) can also be delivered using the dendrimer compositions described herein. Finally, the dendrimer compositions can be used to deliver hallucinogenic drugs with improved binding affinity and specificity to one or more receptors to modulate one or more of the GLUT1, GLUT2, GLUT3, GLUT4, GLUT5, GLUT6, GLUT7, GLUT8, GLUT9, GLUT10, GLUT11, GLUT12, GLUT13 and GLUT14 transporters. i. Improved serotonin receptor binding

[0240] The present dendrimer compositions can improve binding to one or both serotonin receptors and modulate signal transduction in a cell- and tissue-specific manner.

[0241] In the central nervous system (CNS), serotonin is produced almost exclusively in neurons originating from the raphe nuclei located in the midline of the brainstem. These serotonin-producing neurons form the largest and most complex efferent system in the human brain. The most caudal raphe innervates the spinal cord, while the more rostral raphes, the dorsal and medial raphe nuclei, innervate most of the rest of the CNS through generalized projections. The vast majority of every cell in the brain is close to serotonergic fibers, and nearly all behavior and numerous other brain functions are regulated by serotonin.

[0242] Serotonin exerts its effects through interaction with 13 serotonin G protein-coupled receptors (GPCRs). Serotonin receptors are widespread throughout the body and regulate a range of diverse processes, including but not limited to, learning and memory, control of the sleep / wake cycle, thermoregulation, appetite, male and female sexual behavior, pain, motor activity, and aspects of autonomic function such as arterial blood pressure and heart rate.

[0243] 5-HT2A receptors are widely expressed in the cerebral cortex, particularly layers I and IV-V, the piriform and entorhinal cortices, the claustrum, the endopiriform nucleus, and the olfactory bulb / anterior olfactory nucleus, the brainstem, and the limbic system and basal ganglia, particularly the nucleus accumbens and caudate nucleus. Therefore, dysfunction of the serotonergic system is associated with several diseases and disorders, such as anxiety and depressive disorders, migraine, personality disorders, obsessive-compulsive disorders, drug addiction, and neurodegenerative disorders. Activation of the 5-HT2A receptor is required for the action of "classic" psychedelics such as LSD, psilocybin, and mescaline, which act as full or partial agonists at this receptor and represent the three major classes of 5-HT2A agonists: ergolines, tryptamines, and phenethylamines, respectively. Therefore, in some embodiments, dendrimer-hallucinogen compositions can be used to activate the 5-HT2A receptor and ameliorate symptoms associated with one of the aforementioned disorders. For example, dendrimer-R-DOI conjugates can be administered to subjects in need of alleviating anti-inflammatory responses associated with neurodegenerative or anxiety disorders through functional selectivity. Functional selectivity refers to the process by which different ligands induce slightly different receptor conformations to mobilize different sets of effector pathways. In a second example, the composition may activate 5-HT2A receptors located in the apical dendrites of pyramidal cells in the prefrontal cortex, inducing hallucinogenic effects. In a third example, the composition can be used to deliver classical psychedelics to subjects in need of modulating the activity of the 5-HT2A-mGlu2 receptor heterodimer and dopamine receptors to enhance PFC activity and improve learning, memory, and attention deficits.

[0244] 5-HT2A receptor is also widely distributed in peripheral tissues, and serotonergic dysregulation is also involved in diseases in peripheral tissues, such as pulmonary hypertension, bile duct cancer, chronic renal failure and inflammatory bowel disease.In some embodiments, the present composition can be used to modulate the 5-HT2A receptor activity in immune-related tissues, such as spleen, thymus and circulating lymphocytes.In some embodiments, the present composition is used to modulate the 5-HT2A receptor protein level in peripheral blood mononuclear cells (PBMCs), eosinophils and T cells, in order to regulate innate and / or adaptive immune response.For example, the present composition can be administered to a subject who needs to reduce inflammation and eosinophil infiltration due to allergy-induced asthma. ii. Improved dopamine receptor binding

[0245] There are five types of dopamine receptors, including D1, D2, D3, D4, and D5, each with a different function. The dendrimer composition can improve binding to one or more dopamine receptors, preferably D1 and / or D2 receptors, and modulate signal transduction in a cell- and tissue-specific manner. D1 receptors are involved in memory, attention, impulse control, regulation of renal function, and movement. They bind to G stimulatory sites and activate adenylyl cyclase. Activation of adenylyl cyclase leads to the production of the second messenger cAMP, which in turn leads to the production of protein kinase A (PKA), which leads to further transcription in the nucleus. Meanwhile, D2 receptors are associated with movement, attention, sleep, memory, and learning. They bind to G inhibitory sites, inhibit adenylyl cyclase, and activate K+ channels.

[0246] Dopamine receptors are expressed in the central nervous system, specifically in the hippocampal dentate gyrus and subventricular zone. D1 receptors are highly concentrated in the striatum, nucleus accumbens, olfactory bulb, and substantia nigra. These receptors regulate reward systems, motor activity, memory, and learning. D1 receptors, along with stimulatory adenyl cyclase, also activate phospholipase C, which induces intracellular calcium release and activates protein kinase C. Protein kinase C is a calcium-dependent protein kinase. Calcium also modulates neurotransmitter release via exocytosis. D2 receptors are primarily expressed in the striatum, the external segment of the globus pallidus, the core of the nucleus accumbens, the hippocampus, the amygdala, and the cerebral cortex, where they modulate postsynaptic receptor-mediated extrapyramidal activity. D2 receptors are important in signal transduction for the survival and development of human dopamine neurons. Dopamine receptors are also expressed peripherally, more prominently in the kidney and vasculature. For example, in the kidney, D1 receptors inhibit Na / K ATPase via the PKA and PKC pathways, thereby increasing electrolyte excretion and renal vasodilation.

[0247] Thus, in some forms, the compositions are used to deliver LSD, alone or in combination with L-dopa, to modulate D1 and D2 receptor activity and ameliorate symptoms of Parkinson's-induced psychosis, such as depression, anxiety, limb pain, fatigue, sleep disturbances and cognitive impairment. iii. Improved noradrenergic receptor binding

[0248] The present dendrimer compositions can improve binding to one or both noradrenergic receptors and modulate signal transduction in a cell- and tissue-specific manner.

[0249] Norepinephrine, also known as noradrenaline, is a brain neurotransmitter that plays an essential role in regulating arousal, attention, cognitive function, and stress response. Norepinephrine also functions peripherally as a hormone as part of the sympathetic nervous system in the "fight-or-flight" response. During states of stress or anxiety, norepinephrine and epinephrine are released and bind to adrenergic receptors throughout the body, exerting effects such as dilating the pupils and bronchioles, increasing heart rate and constricting blood vessels, increasing renin secretion from the kidneys, and inhibiting peristalsis. The noradrenergic system plays a role in the pathogenesis of some serious neuropsychiatric disorders and has become an important pharmacological target in a variety of neuropathological, neurological, and cardiopulmonary disorders.

[0250] The central noradrenergic system consists of two major ascending projections originating in the brainstem: the dorsal noradrenergic bundle (DNB) and the ventral noradrenergic bundle (VNB). The DNB originates from the A6 locus coeruleus, located in the dorsal pons, and is composed primarily of noradrenergic neurons. The DNB serves as the predominant site of norepinephrine production in the central nervous system. The DNB sends projections that exclusively innervate the cerebral cortex, hippocampus, and cerebellum, and has overlapping projections with those from the VNB to innervate regions of the amygdala, hypothalamus, and spinal cord. The VNB originates from nuclei in the pons and medulla and sends projections that innervate the amygdala, hypothalamus, and regions of the midbrain and medulla.

[0251] The sympathetic nervous system and neuroendocrine chromaffin cells (located in the adrenal medulla) are primarily responsible for the synthesis and exocytosis of norepinephrine and other catecholamines into the circulation. Hormones act on alpha- and beta-adrenergic receptors in smooth muscle cells and adipose tissue, located throughout the body.

[0252] After an action potential at the presynaptic terminal, voltage-gated calcium channels are stimulated, resulting in calcium influx from the extracellular space to the intracellular space. This influx causes norepinephrine (stored in vesicles) to bind to the cell membrane and be released into the synaptic cavity by exocytosis. Norepinephrine can then target and bind to three major receptors: alpha-1 (alpha-1), alpha-2, and beta receptors. These receptors are classified as G-protein-coupled receptors, which have either inhibitory or excitatory effects and have different binding affinities for norepinephrine.

[0253] Alpha-1 receptors are further subdivided into alpha-1a, alpha-1b, and alpha-1d receptors. These receptors are located postsynaptically in brain regions, including the locus coeruleus, olfactory bulb, cerebral cortex, dentate gyrus, amygdala, and thalamus. Alpha-1 receptors have a moderate binding affinity for norepinephrine and are coupled to the Gq protein signaling pathway. In this pathway, phospholipase C (PLC) is activated on the cell membrane to convert phosphatidylinositol 4,5-bisphosphate (PIP2) to inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG). IP3 is released into the cytosol and binds to transmembrane IP3 receptors located in the endoplasmic reticulum (ER), which function as calcium channels. Upon binding, the receptor undergoes a conformational change, resulting in calcium release from the ER into the cytosol. DAG persists in the cell membrane and functions to upregulate protein kinase C (PKC) and phosphorylate other proteins, the combined effects of which result in excitatory cellular effects.

[0254] Alpha-2 receptors are subdivided into alpha-2a, alpha-2b, and alpha-2c receptors. These receptors are located both presynaptically and postsynaptically in brain regions, including the locus coeruleus, amygdala, and hypothalamus. These receptors have the highest binding affinity for norepinephrine and are linked to the Gi / o protein signaling pathway. This pathway reduces cAMP levels, which in turn reduces adenylyl cyclase activity and produces inhibitory cellular effects. Presynaptic noradrenergic terminals contain alpha-2 autoreceptors that block further release of norepinephrine.

[0255] Beta receptors are subdivided into beta-1, beta-2, and beta-3 receptors. These receptors are present in various regions of the brain, with beta-1 and beta-2 receptors being most prevalent in cerebral lipids. These receptors have the lowest binding affinity for norepinephrine and are coupled to the G protein signaling pathway. In this pathway, elevated cAMP levels lead to the activation of protein kinase A (PKA), which then proceeds to phosphorylate other proteins within the cell, resulting in excitatory cellular effects. Beta-2 receptors are also coupled to the Gi protein signaling pathway. Beta-3 receptors are present in adipose tissue.

[0256] In the adrenal medulla, acetylcholine stimulates the release of adrenaline and noradrenaline. Acetylcholine binds to nicotinic acid receptors located on adrenal chromaffin cells, generating an action potential sustained by voltage-gated sodium and potassium channels. This action potential triggers calcium influx into the cytosol, leading to the binding of norepinephrine vesicles to the cell membrane and its release into the circulation, where it travels and binds to alpha and beta receptors on smooth muscle and adipocytes.

[0257] Norepinephrine can be broken down intracellularly or in the synaptic cleft by the enzymes monoamine oxidase (MAO) or catechol-O-methyltransferase (COMT). MAO oxidizes norepinephrine, while COMT metabolizes the deaminated norepinephrine by O-methylation. MAO and COMT are found in adrenal chromaffin cells, while sympathetic nerves contain only MAO. COMT is found in all organs. The liver is responsible for the complete breakdown of norepinephrine to vanillylmandelic acid (VMA). C. Condition to be treated

[0258] This composition is suitable for treating one or more diseases, conditions and injuries in central nervous system and peripheral nervous system.This composition can be used for treating various diseases, disorders and injuries, including mental health disorders, gastrointestinal disorders, and / or treating other tissues in which nerves play a role in disease or injury.This composition and method are also suitable for preventive use. For example, the composition may be administered to a patient in need thereof to improve, treat, or prevent symptoms associated with various disorders, diseases, and conditions, including, but not limited to, mental health disorders and neurological disorders such as depression (including major depressive disorder, treatment-resistant depression, and postpartum depression), posttraumatic stress disorder, panic disorder, social anxiety disorder, anorexia nervosa, suicidal ideation, obsessive-compulsive disorder, premenstrual dysphoric disorder, anorexia, substance abuse disorders, epilepsy, autism spectrum disorder, attention-deficit hyperactivity disorder, schizophrenia, cluster headache, migraine, seizures, fibromyalgia, narcolepsy, obesity, Alzheimer's disease, Tourette's syndrome, etc.; pain disorders, such as neuropathic pain and chronic pain, phobias, and cardiovascular diseases; pain disorders, such as neuropathic pain and / or gastrointestinal disorders. In other forms, the composition may be administered to a subject in need of, for example, stabilizing mood in bipolar disorder, reducing anxiety in anxiety disorders, and reducing inflammation in Parkinson's disease.

[0259] deliver therapeutic, prophylactic or diagnostic agents, preferably less than 20 nm in diameter and at least 0.8 OH groups / nm2 hydroxyl group surface density, preferably less than 10 nm in diameter and at least 1 OH group / nm 2 hydroxyl group surface density of 0.01, more preferably less than 6 nm in diameter and at least 1 OH group / nm 2 and most preferably a diameter of between 5 nm and at least 1.5 OH groups / nm 2 Dendrimer complex compositions having a hydroxyl group surface density of 0.05% selectively target microglia and astrocytes, which play an important role in the pathogenesis of numerous disorders and conditions, including neurodevelopmental disorders, neurodegenerative disorders, neuropsychiatric disorders, and chronic pain. Accordingly, the dendrimer complexes are administered in dosage units effective to treat or alleviate conditions associated with pathologies in the central and peripheral nervous systems. For example, the dendrimer complexes are administered in dosage units effective to treat or alleviate conditions associated with pathologies affecting neuronal microglia and astrocytes. Generally, by targeting these cells, the dendrimers specifically deliver agents to treat neuroinflammation.

[0260] In preferred embodiments, the compositions may comprise glucose or hydroxyl dendrimers having a diameter of 5 nm or less and may be conjugated to antidepressants, antipsychotics, or other agents that act by modulating monoaminergic neurotransmission to treat mental health and CNS disorders. In other preferred embodiments, the compositions may comprise glucose or hydroxyl dendrimers having a diameter of greater than 5 nm and may be conjugated to antidepressants, antipsychotics, or other agents that act by modulating monoaminergic neurotransmission to treat peripheral nervous system disorders. a. Mental health disorders and conditions

[0261] The compositions and methods are suitable for treating a variety of mental health disorders and conditions, including, but not limited to, affective or mood disorders, anxiety disorders, childhood disorders, eating disorders, personality disorders, and substance-related disorders. i. Affective or mood disorders

[0262] Affective or mood disorders are described by marked disruptions of emotion (severe low spirits called depression or severe elevated mood called hypomania or mania). These include bipolar disorder, cyclothymia, hypomania, major depressive disorder, severe mood dysregulation disorder, persistent depressive disorder, premenstrual dysphoric disorder, seasonal affective disorder, depression associated with a medical illness, and depression induced by substance use or medication.

[0263] In some embodiments, this composition and method are suitable for treating depression, treatment-resistant depression and the symptoms associated with suicidal ideation.Up to 30% of people with depression do not respond effectively to antidepressant treatment, and adherence to treatment regimen is low, partly due to the biological differences between patients and the long time it takes to respond to drugs.Therefore, there is an urgent need to expand the repertoire of drugs available to people with depression.

[0264] In exemplary embodiments, the compositions are used to deliver psilocin, psilocybin and / or DMT to subjects suffering from treatment-resistant depression to establish long-term behavioral outcomes, such as improved coping strategies and enhanced cognitive function, e.g., associative learning, a cognitive function commonly impaired by major depressive disorder (MDD). ii. Anxiety disorders

[0265] Anxiety disorders involve excessive fear or apprehension as opposed to normal feelings of tension or apprehension, and include generalized anxiety disorder, panic disorder, social anxiety disorder, and various phobia-related disorders.

[0266] Generalized anxiety disorder (GAD) usually involves persistent feelings of anxiety or fear, which can interfere with daily life. It is not the same as occasionally worrying about things or feeling anxious due to stressful life events. People living with GAD experience frequent anxiety for months, sometimes even years. The composition and method are suitable for treating one or more symptoms of GAD, including but not limited to restlessness, fatigue, difficulty concentrating, irritability, headache, muscle pain, stomach pain, or pain of unknown origin, excessive worry, and sleep problems, such as difficulty falling asleep or staying asleep.

[0267] Panic disorder is an anxiety disorder characterized by unexpected and repeated episodes of intense fear accompanied by physical symptoms, which can include chest pain, palpitations, shortness of breath, dizziness or abdominal distress, or a sense of losing control even when there is no obvious danger or trigger.Persons with panic disorder often worry about when the next attack will occur, and actively try to prevent future attacks by avoiding the places, situations or actions that are associated with panic attacks.Panic attacks can occur frequently, such as several times a day, or infrequently, such as several times a year.The present composition and method are suitable for treating one or more symptoms of panic attack, including but not limited to palpitations, excessive sweating, trembling or tingling, chest pain, and difficulty in controlling emotions, such as feelings of impending doom and uncontrollable emotions.

[0268] Social anxiety disorder is an intense and persistent fear of being observed and judged by others.For people with social anxiety disorder, the fear of social situations can be so intense that it seems uncontrollable.For some people, this fear can prevent them from going to work, going to school or doing everyday things.The composition and method are suitable for treating one or more symptoms of social anxiety disorder, including but not limited to excessive blushing, sweating or trembling, palpitations, stomach pain, rigid body posture or speaking in an excessively low voice, and self-consciousness or fear of negative judgment.

[0269] Phobia is the strong fear or aversion to certain objects or situations.In some situations, feeling anxiety may be realistic, but the fear that phobia people feel is disproportionate to the actual danger caused by that situation or object.The composition and method are suitable for treating one or more symptoms of phobia, including but not limited to the irrational or excessive worry about encountering the object or situation of fear, the strong immediate anxiety when encountering the object or situation of fear, and the endurance of unavoidable objects and situations with strong anxiety.

[0270] In an exemplary embodiment, the composition can be used to deliver MDMA to a subject in need of ameliorating symptoms associated with trauma-related disorders, such as PTSD, such as reducing pathological fear responses, social isolation and emotional numbing, and increasing trust and prosocial behavior. In another exemplary embodiment, the composition is suitable for delivering LSD to a subject in need of treatment for obsessive-compulsive disorder, cancer-related anxiety, and / or alcohol use disorder. iii. Eating disorders

[0271] Eating disorders are serious, often fatal, illnesses that involve severe disturbances in a person's eating behavior and related thoughts and emotions. Eating disorders involve preoccupations with food, weight, and body shape. Common eating disorders include anorexia nervosa, bulimia nervosa, and binge eating disorder.

[0272] Anorexia nervosa is a condition in which a person avoids food, severely restricts food, or eats only small amounts of certain foods. They may repeatedly weigh themselves. They may view themselves as overweight even when dangerously thin. There are two subtypes of anorexia nervosa: the restrictive subtype and the bulimic subtype. People with the restrictive subtype of anorexia nervosa severely restrict the amount and type of food they consume. People with the bulimic subtype of anorexia nervosa also severely restrict the amount and type of food they consume. In addition, they may have episodes of binge eating and vomiting, eating a large amount of food in a short period of time followed by vomiting or using laxatives or diuretics to expel the food. Symptoms of anorexia nervosa include, but are not limited to, thinning of bones (osteopenia or osteoporosis), mild anemia, and muscle wasting and weakness, brittle hair and nails, dry, yellowish skin, thin hair growth (vellus hair) all over the body, severe constipation, low blood pressure, slowed breathing and pulse rate, damage to the structure and function of the heart, brain damage, multiple organ failure, a drop in internal body temperature, feeling constantly cold, lethargy, sluggishness or constant fatigue, and infertility.

[0273] Bulimia nervosa is a condition in which a person has repeated and frequent episodes of eating abnormally large amounts of food and feels unable to control these episodes. This binge eating is followed by behaviors to counteract the binge eating, such as compulsive vomiting, excessive use of laxatives or diuretics, fasting, excessive exercise, or a combination of these behaviors. People with bulimia nervosa may be slightly underweight, normal weight, or overweight. Symptoms of bulimia nervosa include chronic inflammation and pain in the throat, swelling of the salivary glands in the neck and jaw area, worn tooth enamel and increased tooth sensitivity and tooth decay as a result of exposure to stomach acid, acid reflux and other gastrointestinal problems, intestinal discomfort and irritation due to laxative abuse, severe dehydration due to fluid expulsion, and electrolyte imbalances (too low or too high levels of sodium, calcium, potassium, and other minerals) that can lead to stroke or heart attack.

[0274] In exemplary embodiments, the compositions may be used to deliver psilocybin, LSD, and / or ayahuasca to a subject in need of: (1) increasing connectivity between neuronal networks in the subject, creating the potential for action beyond debilitating self-imposed limitations; and / or (2) promoting desirable brain states that may accelerate the healing process, e.g., increasing neuroplasticity and neurogenesis, improving mood, reducing fear responses, and promoting acceptance and empathy for self and others. b. Neurological and neurodegenerative diseases

[0275] The compositions and methods are suitable for treating symptoms associated with neurological and neurodegenerative diseases.

[0276] Neurodegenerative diseases are chronic, progressive disorders of the nervous system that affect neurological and behavioral functions and involve biochemical changes that result in distinct histopathological and clinical syndromes (Hardy H, et al., Science. 1998;282:1075-9). Abnormal proteins that resist cellular degradation accumulate within cells. The pattern of neuronal loss is selective, in the sense that one group is affected while others remain intact. In many cases, there is no clear precipitating event for the disease. Diseases traditionally described as neurodegenerative include Alzheimer's disease, Huntington's disease, and Parkinson's disease.

[0277] Neuroinflammation, mediated by activated microglia and astrocytes, is a major hallmark of various neurological disorders and represents a potential therapeutic target (Hagberg, H et al., Annals of Neurology 2012, 71, 444; Vargas, DL et al., Annals of Neurology 2005, 57, 67; and Pardo, CA et al., International Review of Psychiatry 2005, 17, 485). Several scientific reports suggest that targeting these cells early in the disease process can reduce neuroinflammation, potentially delaying disease onset and providing a longer therapeutic window for treatment (Dommergues, MA et al., Neuroscience 2003, 121, 619; Perry, VH et al., Nat Rev Neurol 2010, 6, 193; Kannan, S et al., Sci. Transl. Med. 2012, 4, 130ra46; and Block, ML et al., Nat Rev Neurosci 2007, 8, 57). Delivering therapeutic agents across the blood-brain barrier (BBB) ​​is challenging. Neuroinflammation causes disruption of the BBB. BBB damage in neuroinflammatory disorders can be exploited to transport drug-loaded nanoparticles through the brain (Stolp, HB et al., Cardiovascular Psychiatry and Neurology 2011, 2011, 10; and Ahishali, B et al., International Journal of Neuroscience 2005, 115, 151).

[0278] The present compositions and methods can also be used to deliver active agents for treating neurological or neurodegenerative diseases, or neurological or neurodegenerative disorders, or central nervous system disorders.In a preferred embodiment, the present compositions and methods are effective for treating and / or improving neuroinflammation associated with neurological or neurodegenerative diseases, or neurological or neurodegenerative disorders, or central nervous system disorders, or peripheral symptoms caused by neurological or neurodegenerative diseases, or neurological or neurodegenerative disorders.The present methods generally include administering an effective amount of the composition to a subject to improve cognition or reduce cognitive decline, improve cognitive function or reduce cognitive decline, improve memory or reduce memory decline, improve learning skills or learning ability, or reduce decline in learning skills or learning ability, or a combination thereof.

[0279] Neurodegeneration refers to the progressive loss of neuron structure or function, including neuron death.For example, the present compositions and methods can be used to treat the subject with diseases or disorders such as Parkinson's disease (PD) and PD-related disorders, Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD) and other dementia, multiple sclerosis (MS), post-encephalitic dementia, cancer and chemotherapy-related cognitive impairment and dementia, and depression-induced dementia and pseudodementia.

[0280] In exemplary embodiments, the compositions may be used to deliver one or more psychedelic compounds to a subject in need of ameliorating symptoms associated with neurodegenerative diseases, such as stimulating neurogenesis, neuroprotection and neuroplasticity, reducing neural and oligodendrocyte cell loss, reducing oxidative stress and BBB disruption, and reducing inflammation and endoplasmic reticulum damage. c.Pain

[0281] The present compositions and methods are suitable for treating neuropathic pain and / or non-neuropathic pain associated with various disorders, such as complex regional pain syndrome, peripheral neuropathy, multiple sclerosis, and cancer-induced pain.For example, psilocybin and LSD can be effective in treating neuropathic (chronic) nerve pain.Chronic nerve pain, also known as neuropathic pain, is caused by nerve damage or other problems involving nerves, and often does not respond to standard painkillers such as paracetamol.In some forms, the present compositions can be used to deliver psilocybin and LSD to subjects who need to reduce the severity of headaches and migraines and extend the remission period between headaches and migraines.

[0282] Although the mechanism of chronic pain development is not fully understood, it is likely to involve the complex interaction between somatic and visceral afferent input, peripheral and central sensitization, emotional state, and behavior and cognition.Distraction and mood changes can have a powerful effect on pain perception.Therefore, in some embodiments, this composition can be used to deliver psilocybin to the subject who needs to alleviate the pain caused by cancer-related anxiety and depression.

[0283] In some embodiments, the composition can be used to treat chronic pain cases that do not involve nerves (non-neuropathic pain).For example, the dendrimer composition can be complexed with LSD or MDMA and used to treat chronic non-neuropathic pain.Conditions that cause non-neuropathic pain that can benefit from treatment with the composition include, but are not limited to, fibromyalgia, chronic lower back pain, and chronic neck pain.For example, the composition can be used to deliver psilocybin or LSD to subjects who need to relieve symptoms associated with fibromyalgia and / or chronic pelvic pain, including, but not limited to, muscle, abdominal, neck, and / or back pain, as well as stiffness, fatigue, sleep disorders, headaches, and migraines. The most important indications are major depressive disorder, treatment-resistant depression, post-traumatic stress disorder, panic disorder, social anxiety disorder, anorexia nervosa, suicidal ideation, obsessive-compulsive disorder, anorexia, substance abuse disorders, epilepsy, bipolar disorder, autism spectrum disorder, attention deficit hyperactivity disorder, schizophrenia, headache, seizures, fibromyalgia, narcolepsy, obesity, Alzheimer's disease and Tourette's syndrome. D. Dosage and Effective Amount

[0284] The dosage and administration regimen will depend on the severity and location of the disorder or injury and / or the method of administration and the particular agent being delivered, and can be determined by one skilled in the art.

[0285] In some embodiments, the dosage is expressed in mg / kg, particularly when expressed as an in vivo dosage of the dendrimer gene-editing composition.

[0286] Typically, dosages range from micrograms per kg of body weight to about 100 mg per kg. Dosages may be, for example, 0.01 mg / kg to about 1,000 mg / kg per dose, or 0.5 mg / kg to about 1,000 mg / kg, or 1 mg / kg to about 1,000 mg / kg, or about 10 mg / kg to about 500 mg / kg, or about 20 mg / kg to about 500 mg / kg, or 20 mg / kg to about 100 mg / kg per dose, or 25 mg / kg to about 75 mg / kg per dose, or about 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, or 75 mg / kg per dose.

[0287] Preferably, the dendrimer-gene editing agent composition does not target or otherwise genetically modify non-target cells or healthy cells that are not within or associated with the diseased tissue, or targets or genetically modifies them at a lower level than cells associated with the disease or disorder, such as cancer and / or a proliferative disorder. In this way, by-products and other side effects associated with the composition are reduced. Thus, in preferred embodiments, the dendrimer composition is administered in an amount that results in improvement, or enhancement, or functioning in an individual with a disease or disorder, such as cancer and / or a proliferative disorder.

[0288] The actual effective amount of the composition may vary depending on factors including the specific drug administered, the specific composition formulated, the administration form, and the age, weight, condition, and route of administration and the disease or disorder of the subject being treated. Generally, in the case of intravenous injection or infusion, the dosage is lower than that in oral administration.

[0289] Dosages can vary and can be administered daily, for one or several days, in one or multiple doses. In some embodiments, dendrimer conjugation can increase the efficacy and duration of treatment, thereby reducing the need for repeated administration for weekly, monthly, 6-monthly, yearly, or other longer-term administration regimens. Some embodiments can be incorporated into drug delivery systems (e.g., implants, pumps, patches, creams, etc.) to provide controlled, sustained delivery in a manner that reduces compliance requirements and the potential for abuse. Guidance can be found in the literature regarding appropriate dosages for a given class of pharmaceutical product. Optimal administration schedules can be calculated from measurements of drug accumulation in the subject's or patient's body. Those skilled in the art can easily determine the optimal dosage, administration method, and repetition rate. Optimal dosages can vary depending on the relative potency of individual pharmaceutical compositions and can generally be estimated based on effective dosages in in vitro and in vivo animal models. In particular, in some cases it may be desirable to minimize or prevent the hallucinations associated with psychedelic compounds, which can be achieved by peripheral confinement or by the controlled and / or sustained activity of the dendrimer composition.

[0290] Dosage forms of pharmaceutical compositions comprising the dendrimer compositions are also provided. A "dosage form" refers to the physical form of a dose of a therapeutic compound, such as a capsule or vial, intended for administration to a patient. The term "dosage unit" refers to the amount of a therapeutic compound administered to a patient in a single dose.

[0291] Generally, the timing and frequency of administration will be adjusted to balance the efficacy of a given treatment or diagnostic schedule with the side effects of a given delivery system.

[0292] In some embodiments, the dosage is administered daily, biweekly, weekly, every two weeks, or less frequently in an amount that provides a therapeutically effective increase in blood levels of the therapeutic agent. When administration is by a route other than oral, the composition may be delivered over a period of more than one hour, e.g., 3 to 10 hours, to produce a therapeutically effective dose within 24 hours. Alternatively, the composition may be formulated for controlled release, in which case the composition is administered as a single dose repeated on a weekly or less frequent regimen.

[0293] It is understood by those skilled in the art that the administration regimen can be any length of time sufficient to treat the disorder in a subject.In some embodiments, the regimen comprises one or more cycles of treatment followed by one or more rest periods (e.g., drug-free).The rest period can be 1, 2, 3, 4, 5, 6 or 7 days; or 1, 2, 3, 4 weeks, or 1, 2, 3, 4, 5 or 6 months. E. Control

[0294] The treatment results of a composition comprising one or more gene-editing compositions bound to or conjugated to a dendrimer can be compared with a control. Suitable controls are known in the art and include, for example, untreated subjects or placebo-treated subjects. A typical control is a comparison of the condition or symptom of a subject before and after administration of a glucose dendrimer composition. The condition or symptom may be a biochemical, molecular, physiological, or pathological readout. For example, the effect of the composition on a particular symptom, pharmacological indicator, or physiological indicator can be compared with the condition of an untreated subject or a subject before treatment. In some embodiments, the symptom, pharmacological indicator, or physiological indicator is measured in the subject before treatment and then initiated again one or more times after treatment. In some embodiments, the control is a reference level or an average value determined based on measurements of the symptom, pharmacological, or physiological indicator in one or more subjects (e.g., healthy subjects) without the disease or condition being treated. In some embodiments, the effect of the treatment is compared with conventional treatments known in the art. In some embodiments, the untreated control subject has the same disease or condition as the treated subject.

[0295] In some embodiments, the control includes an equal amount of the gene editing composition delivered alone or bound to a dendrimer that does not have glucose-based branching units, such as a dendrimer of similar generation, molecular weight and / or surface group density (e.g., hydroxyl groups). VI. Kit

[0296] The composition can be packaged in a kit. The kit can include a single or multiple dose composition comprising one or more psychedelic drugs associated with or conjugated to a dendrimer (e.g., one or more hydroxyl-terminated PAMAM dendrimers or glucose dendrimers described in the Examples), and instructions for administering the composition. Specifically, the instructions instruct the administration of an effective amount of the dendrimer composition to an individual with a specific disease / disorder for which it is indicated. The composition can be formulated as described above with reference to a specific treatment method, and can be packaged in any convenient manner.

[0297] The invention will be further understood by reference to the following non-limiting examples. [Example]

[0298] Example 1 Synthesis of Hydroxyl-Polyamidoamine (PAMAM-OH) Dendrimer Psychedelic Conjugates The synthesis of PAMAM-OH-psychedelic conjugates was achieved using a variety of linking chemistries and linkers (both cleavable and non-cleavable). Briefly, the surface hydroxyl groups of PAMAM-OH are modified with a linker to provide complementary groups on the surface that can further react with complementary groups on the drug-linker. The drug compound is modified with a linker that provides a complementary functional group for reaction with the dendrimer-linker. The linker on the drug is attached by a cleavable or non-cleavable linking group. Examples of cleavable linking groups include esterase-sensitive ester bonds, glutathione-sensitive disulfide bonds, phosphatase-sensitive phosphodiester bonds, lysosomal-releasable triglycyl peptide linkers (CX), and acid-cleavable hydrazine linking groups. Examples of non-cleavable linking groups include ether, aminoalkyl, or amide bonds. The linker can be an amino acid, peptide, polyethylene glycol (n = 2-15), or a hydrocarbon chain. 1. Synthesis of dendrimer-psilocin conjugates with cleavable ester linkers

[0299] Dendrimer-psilocin conjugates bearing cleavable esters were prepared according to the reactions illustrated in Figures 1A and 1B, which are schematic diagrams showing a stepwise synthetic route for conjugating psilocin to dendrimers using click chemistry. 2. Synthesis of dendrimer-psilocin analogs with non-cleavable amide linkers

[0300] Dendrimer-psilocin conjugates bearing non-cleavable amide linkers were synthesized according to the reactions illustrated in Figures 2A and 2B, which are schematic diagrams showing a stepwise synthetic route for conjugating psilocin analogs to dendrimers using click chemistry. 3. Synthesis of dendrimer-ketamine with a non-cleavable amino-alkyl linker

[0301] Dendrimer-ketamine conjugates bearing non-cleavable amino-alkyl linkers were synthesized according to the reactions illustrated in Figures 3A and 3B, which are schematic diagrams showing an exemplary synthetic route for conjugating ketamine to dendrimers using copper-catalyzed alkyne-azide click chemistry (Figure 3B). 4. Synthesis of dendrimer-DMT analogs with non-cleavable amide linkers

[0302] Dendrimer-N,N-dimethyltryptamine (dendrimer-DMT) conjugates bearing non-cleavable amide linkers were synthesized according to the reactions illustrated in Figures 4A and 4B, which are schematic diagrams showing an exemplary stepwise synthetic route for conjugating DMT to dendrimers using copper-catalyzed alkyne-azide click chemistry. 5. Synthesis of dendrimer-DMT analogs bearing non-cleavable amino-alkyl linkers

[0303] Dendrimer-DMT conjugates bearing non-cleavable amino-alkyl linkers were also synthesized according to the reactions illustrated in Figures 5A and 5B, which are schematic diagrams showing an exemplary stepwise synthetic route for conjugating DMT to dendrimers using copper-catalyzed alkyne azide click chemistry. 6. Synthesis of dendrimer-LSD with non-cleavable amino-alkyl linkers

[0304] Dendrimer-lysergic acid diethylamide (dendrimer-LSD) conjugates bearing non-cleavable amino-alkyl linkers were synthesized according to the reactions illustrated in Figures 6A and 6B, which are schematic diagrams showing an exemplary stepwise synthetic route for conjugating LSD to dendrimers using copper-catalyzed alkyne azide click chemistry. conclusion

[0305] Dendrimer conjugation provides site-specific targeting by directing drugs to the target site, reducing dose, enhancing efficacy, and mitigating side effects associated with the free drug. The PAMAM-OH platform has extremely high aqueous solubility (>300 mg / mL). The majority of psychedelics have poor aqueous solubility in the μg / mL range. Dendrimer conjugation improves aqueous solubility by 10-100 times compared to the free drug. Sustained intracellular release at the target avoids the systemic and dose-related side effects of free psychedelics. Dendrimer conjugation can significantly reduce the time required for onset of drug activity. Example 2 Synthesis of glucose dendrimer (GD) psychedelic conjugates

[0306] The synthesis of glucose dendrimer-psychedelic conjugates is achieved using a combination of linking chemistries and linkers (both cleavable and non-cleavable). Briefly, the surface hydroxyl groups of the glucose dendrimer are modified with a linker to facilitate reaction between complementary groups on the surface and those on the drug linker. The drug compound is also modified with a linker to facilitate reaction between the complementary functional groups and the dendrimer-linker. The linker on the drug compound is attached by a cleavable or non-cleavable linking group. Examples of cleavable linking groups include esterase-sensitive ester bonds, glutathione-sensitive disulfide bonds, phosphatase-sensitive phosphodiester bonds, lysosomal-releasable triglycyl peptide linkers (CX), and acid-cleavable hydrazine linking groups. Examples of non-cleavable linking groups include ether or amide bonds. The linker may be an amino acid, peptide, polyethylene glycol (n = 2-15), or a hydrocarbon chain. 1. Synthesis of dendrimer-psilocin with cleavable ester linkers

[0307] Glucose dendrimer-psilocin conjugates with cleavable ester linking groups were prepared according to the reaction illustrated in Figure 7. The procedure for the synthesis of the psilocin-azide compound is shown in Figure 1A. The synthesis of glucose dendrimer and psilocin conjugates is achieved by partially modifying the OH groups of the glucose dendrimer with complementary groups on the surface of the glucose dendrimer, which are then reacted with an azide-containing complementary linker linked to psilocin to generate the glucose dendrimer-psilocin conjugate (Figure 7). 2. Synthesis of dendrimer-psilocin analogue conjugates bearing non-cleavable amide linkers

[0308] Glucose dendrimer-psilocin conjugates with non-cleavable amide linkers were prepared according to the reaction illustrated in Figure 8. The procedure for the synthesis of psilocin analog-azide compounds is shown in Figure 2A. The synthesis of glucose dendrimer and psilocin analog conjugates is achieved by partially modifying the OH groups of the glucose dendrimer with complementary groups on the surface of the glucose dendrimer, which are then reacted with an azide-containing complementary linker linked to the psilocin analog to generate the glucose dendrimer-psilocin conjugate (Figure 8). 3. Synthesis of dendrimer-ketamine with a non-cleavable amino-alkyl linker

[0309] Glucose dendrimer-ketamine conjugates bearing non-cleavable amino-alkyl linking groups were prepared according to the reactions illustrated in Figure 9. Ketamine hydrochloride (1) was first modified with the alkyne shown in Figure 3A. An exemplary synthetic route for glucose dendrimer and ketamine conjugates is shown in Figure 9. 4. Synthesis of dendrimer-DMT analogs with non-cleavable amide linkers

[0310] Glucose dendrimer-DMT conjugates with non-cleavable amide linkers were prepared according to the reaction illustrated in Figure 10. An N,N dimethyltryptamine analog (DMT analog) was first conjugated to a linker bearing an azide moiety via the amide linker (Figure 4A). An exemplary synthetic route for glucose dendrimer and DMT conjugates is shown in Figure 10. 5. Synthesis of dendrimer-DMT with non-cleavable amino-alkyl linkers

[0311] Glucose dendrimer-DMT conjugates bearing non-cleavable amino-alkyl linking groups were prepared according to the reactions illustrated in Figure 11. An N,N dimethyltryptamine analog (DMT analog) was first modified with an alkyne group as shown in Figure 5A. An exemplary synthetic route for glucose dendrimer and DMT conjugates is shown in Figure 11. 6. Synthesis of glucose dendrimer-LSD with non-cleavable amino-alkyl linkers

[0312] Glucose dendrimer-lysergic acid diethylamide (LSD) conjugates bearing non-cleavable amino-alkyl linking groups were prepared according to the reactions illustrated in Figure 12. LSD was first modified with an alkyne group as shown in Figure 6A. An exemplary synthetic route for glucose dendrimer and LSD conjugates is shown in Figure 12. conclusion

[0313] Dendrimer conjugation provides site-specific targeting by directing the drug to the target site, reducing dose, enhancing efficacy, and mitigating side effects associated with the free drug. The glucose dendrimer platform has extremely high aqueous solubility (>500 mg / mL). The majority of psychedelics have poor aqueous solubility in the μg / mL range. Dendrimer conjugation improves aqueous solubility by 10-100 times compared to the unconjugated free drug. Sustained intracellular release at the target avoids the systemic and dose-related side effects of free psychedelics. Dendrimer conjugation can significantly reduce the time required for onset of drug activity.

[0314] The compositions can deliver drugs to receptors on specific cells (neural cells, glial cells, macrophages), including targets on their surface and within them. These formulations can increase the efficacy of these drugs, allow for lower doses, provide new mechanistic insights, reduce side effects, and improve solubility, formulation, and pharmacokinetics.

[0315] Taken together, dendrimer-based psychedelics significantly improve the safety, efficacy, reproducibility and ease of implementation of these molecules. Example 3 Synthesis and binding properties of dendrimer conjugates

[0316] Novel chemical entities for psychedelics and norketamine containing hydroxyl-terminated PAMAM and glucose dendrimers were synthesized and investigated to determine in vivo potency and targeting data in the case of norketamine / ketamine and binding affinity data in the case of D-tryptamine. Materials and Methods

[0317] Figure 14A is a schematic representation of the synthesis of a PAMAM dendrimer-norketamine conjugate. Figure 14B is a schematic representation of the synthesis of a glucose dendrimer-norketamine conjugate.

[0318] Unless otherwise noted, reactions were performed in flame-dried glassware under a positive nitrogen pressure using dry solvents. Commercial-grade reagents and anhydrous solvents were purchased from chemical suppliers and used without further purification. 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC.HCl), N,N-diisopropylethylamine (DIPEA), 4-(dimethylamino)pyridine (DMAP), trifluoroacetic acid (TFA), anhydrous dichloromethane (DCM), and N,N'-dimethylformamide (DMF) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cyanine 5 (Cy5)-mono-NHS ester was purchased from Amersham Bioscience-GE Healthcare. Deuterated solvents dimethyl sulfoxide (DMSO-d6), water (DO), and chloroform (CDCl3) were purchased from Cambridge Isotope Laboratories Inc. (Andover, MA). Ethylenediamine-core polyamidoamine (PAMAM) dendrimer, generation 4.0, hydroxy surface (G4-OH; diagnostic grade; consisting of 64 hydroxyl end groups), methanol solution (13.75% w / w) was purchased from Dendritech Inc. (Midland, MI, USA). Dialysis membranes were purchased from Spectrum Laboratories Inc. (Rancho Dominguez, CA, USA). Hu308, tryptamine, 1-(2-amino-1-(4-methoxyphenyl)ethyl)cyclohexanol, nor-ketamine, 5-hydroxytryptamine, psilocybin analogs, psilocyn analogs, and cannabidiol drugs were purchased from Cayman Chemicals. Synthesis of hydroxyl-PAMAM dendrimer-drug conjugates

[0319] The PAMAM-G4-OH (D4-OH) dendrimer, consisting of approximately 64 terminal hydroxyl groups, was used in the synthesis. After each synthesis step, the product was purified by dialysis in DMF for 24 hours to remove small molecular weight impurities, followed by water dialysis to remove the DMF.1 H NMR (in DMSO-d6 and DO) and HPLC analysis were used to confirm the formation and purity of the intermediates and final products. Monofunctional D4-OH was functionalized with alkyne groups by treatment with 5-hexynoic acid under standard esterification conditions using EDC.HCl and 4-DMAP in DMF at room temperature for 36 hours to yield the D-hexyne bifunctional dendrimer. The number of alkyne groups on the dendrimer surface was selected to maintain approximately 10-15 to maintain the overall aqueous solubility of the conjugate. The crude product was dialyzed against ultrapure water for 24 hours using a 1 kDa membrane to remove low molecular weight impurities by selective diffusion through the semipermeable dialysis membrane. 1 H NMR and HPLC analyses were used to confirm product formation and purity of intermediates and final products, which are shown in Figures 12A-12C. Synthesis of D-hexyne

[0320] A solution of PAMAM G4-OH 1 (10.00 g, 0.7 mmol) in DMF (50 mL) was treated with 5-hexynoic acid (1.40 g, 12.6 mmol) and DMAP (2.41 g, 12.6 mmol) and stirred for 5 minutes at room temperature. EDC.HCl (1.54 g, 12.6 mmol) was then added portionwise to the reaction mixture over 5 minutes. The reaction mixture was stirred at room temperature for 36 hours. The crude product was transferred to cellulose dialysis tubing with a 1 kD MW cutoff and dialyzed against DMF for 12 hours and then against water for 24 hours. The aqueous layer was frozen and lyophilized to afford D-hexyne as a hygroscopic white solid (75% yield). 1 H NMR (500 MHz, DMSO-d6) 8.21-7.57 (m, internal amide H), 4.71 (s, GABA amide H, 50H), 4.01 (t, 22-24 CH2), 3.5-2.1 (m, dendrimer CH2) 1.71-1.59 (m, 22-24 CH2). HPLC C18 retention time 4 minutes: purity approximately 99%. Synthesis of dendrimer-drug conjugates

[0321] A solution of D-hexyne and drug-azide in DMF (5 mL) was treated with copper sulfate pentahydrate (CuSO4.5H2O) and sodium ascorbate in water. The reaction mixture was stirred and heated at 50 °C for 10 h in a microwave synthesizer. Upon completion, the reaction mixture was dialyzed against DMF in 1 KDa cutoff cellulose dialysis tubing. EDTA (50 μL, 0.5 M) solution was added to the solution to remove copper by chelation. DMF dialysis was followed by water dialysis overnight. Drug loading was calculated by proton integration comparing the peaks corresponding to the dendrimer and drug. G2-glucose dendrimer (GD2)-drug conjugate

[0322] The second-generation glucose dendrimer (GD2) consists of 24 glucose molecules (96 surface hydroxyl groups) used for conjugation. Glucose dendrimers are primarily made up of a central core of di-pentaerythritol and glucose moieties consisting of one or more branching units of monosaccharide glucose molecules. Unlike hydroxyl-terminated PAMAM dendrimers, glucose dendrimers are primarily taken up by injured neurons and have been found to specifically target hyperexcitable neurons in both culture and in vivo mouse models. Synthesis and characterization of glucose dendrimer (GD2)

[0323] GD synthesis began by reacting the hexapropargylated core with AB4, a β-D-glucose-PEG-azide building block, via a click reaction to yield the first-generation glucose dendrimer (GD1). Propargylation of the OH groups on GD1 yielded GD1-acetylene 24, which was reacted with β-D-glucose-PEG-azide to yield the second-generation GD2 with 24 glucose moieties, resulting in 96 surface hydroxyl groups. Furthermore, a Cy5 fluorescent tag was attached onto GD2 via propargylation of approximately 2–3 hydroxyl groups to yield an alkyne-containing GD2 dendrimer. GD intermediates and final products were purified using dialysis.1 The physicochemical properties of the GD2 dendrimer were also evaluated (Table 2). [Table 2] Synthesis and characterization of GD2-drug conjugates

[0324] The drugs norketamine, tryptamine, venlafaxine, and Hu308 were conjugated to GD2-hexynoic acid dendrimers using a click chemistry strategy. The linker-attached drug moieties were conjugated to glucose dendrimers using a Cu(I)-catalyzed click (CuAAC) reaction in the presence of catalytic amounts of CuSO4.5HO and sodium ascorbate to yield the GD2-drug conjugates. Traces of copper were removed by dialysis against ethylenediaminetetraacetic acid (EDTA). The final GD2-drug conjugates were characterized by NMR and HPLC. [Table 3] [Table 4] Example 4 Binding assay of dendrimer-ketamine conjugates Materials and Methods Human serotonin 5-HT2A receptor (agonist radioligand):

[0325] Objective: To evaluate the affinity of compounds for the human 5-HT2A receptor in transfected HEK-293 cells as determined in a radioligand binding assay.

[0326] Experimental protocol: Cell membrane homogenates (30 μg of protein) are incubated with 0.1 nM [125I]DOI in the absence or presence of test compounds in a buffer containing 50 mM Tris-HCl (pH 7.4), 5 mM MgCl2, 10 μM pargyline, and 0.1% ascorbic acid at 22°C for 60 min.

[0327] Nonspecific binding is determined in the presence of 1 μM DOI. After incubation, samples are rapidly filtered under vacuum through glass fiber filters (GF / B, Packard) presoaked in 0.3% PEI using a 96-sample cell harvester (Unifilter, Packard) and rinsed several times with ice-cold 50 mM Tris-HCl. Filters are then dried and radioactivity is counted in a scintillation counter (Topcount, Packard) using scintillation cocktail (Microscint 0, Packard).

[0328] Results are expressed as percent inhibition of control radioligand specific binding.

[0329] The standard reference compound is DOI, which is tested in each experiment at several concentrations to obtain a competition curve from which the IC50 of DOI is calculated.

[0330] See Bryant, et al. (1996), A novel class of 5-HT2A receptor antagonist: aryl aminoguanidines, Life Sci., 15: 1259. Assay for the human opioid GPCR cell-based antagonist cAMP delta (DOP):

[0331] Objective: To evaluate the efficacy (IC50) and potency (maximal response) of compounds against the human delta (DOP) receptor in stably transfected CHO-K1 cells. The assay principle is a cAMP cell-based assay.

[0332] Experimental Protocol Cells were seeded into white-walled 384-well microplates in a total volume of 20 μL and incubated overnight at 37° C. Before testing the cells, the plate culture medium was replaced with 10 μL of assay buffer (HBSS + 10 mM HEPES).

[0333] Briefly, intermediate dilutions of sample stock solutions were made to generate 4X samples in assay buffer. 5 μL of 4X sample was added to cells and incubated at 37°C for 30 minutes. 5 μL of 4X EC80 in 4X forskolin reagent was added and cells were incubated at 37°C for 30 minutes. The final assay vehicle concentration was 1%. Results are expressed as percent inhibition of control ligand.

[0334] General information [Table 6]

[0335] TA1 Human Trace Amine GPCR Cell-Based Agonist cAMP Assay

[0336] Objective: To assess compound efficacy (EC50) and potency (maximal response) against the human TA1 receptor in stably transfected CHO-K1 cells as determined in a GPCR cell-based cAMP assay.

[0337] Experimental protocol: Prior to testing, cells were seeded into white-walled 384-well microplates in a total volume of 20 μL and incubated overnight at 37°C.

[0338] Prior to testing the cells, the plating medium was replaced with 15 μL of assay buffer (HBSS + 10 mM HEPES). Briefly, intermediate dilutions of sample stock solutions were made to generate 4X samples in assay buffer. 5 μL of 4X was added to the cells and incubated at 37°C for 30 minutes. The final assay vehicle concentration was 1%.

[0339] Results are expressed as percent efficacy relative to the maximal response of the control ligand. General information [Table 7]

[0340] result [Table 5]

[0341] Figure 15A is a graph of the NMDAR1A / 2B antagonist assay for glucose dendrimer-ketamine (IC50 = 4.54 μM), hydroxyl dendrimer-ketamine (IC50 > 100), and norketamine (IC50 = 6.96 μM). Figure 15B is the % binding efficacy of log concentration (micromolar) of compound in a D2L human dopamine GPCR cell-based agonist cAMP assay. Norketamine (filled circles), glucose dendrimer-ketamine EC50 = 13.08 μmolar (open circles), and hydroxyl dendrimer-ketamine EC50 = 4.263 μmolar (triangles). Figure 15C is the % efficacy of log concentration (micromolar) of ketamine in a TA1 human trace amine GPCR cell-based agonist cAMP assay. Norketamine (closed circles), glucose dendrimer-ketamine EC50 = 13.08 micromolar (open circles) and hydroxyl dendrimer-ketamine EC50 = 4.263 micromolar (triangles).

[0342] Significant efficacy data have been obtained with a clinically important drug, a derivative of ketamine (norketamine), which has been tested for depression and addiction. Hydroxyl dendrimer-drugs target these receptors in microglia / macrophages, while glucose dendrimer-drugs target these receptors in both neurons and microglia.

[0343] The conjugate may be active with or without drug release. Binding affinity measures the activity of the intact conjugate. Typically, psychedelic drug conjugates are prepared in a non-release form (e.g., tryptamine, psilocin, psilocybin, ketamine) and are intended to be active in the intact form. This allows the intact conjugate to be released through the kidney without toxicity due to drug release. Alternatively, the intact conjugate can be designed to be released via an analog.

[0344] Both GD-ketamine and HD-ketamine exhibit unique effects that may be beneficial for neuropsychiatric conditions. Both dendrimer-ketamine conjugates exhibit stronger binding to opiate μ, opiate k, and sigma 2 receptors than nor-ketamine, indicating that dendrimer conjugation provides a positive benefit for binding to these receptors. Norketamine does not bind to opioid μ receptors (Ki of 53 μM for GD-ket and 40 μM for HD-ket). The Ki for norketamine against opioid k is 381 μM, which is approximately four-fold better than GD-ket (Ki = 82 μM) and approximately seven-fold better than HD-ket (Ki = 556 μM). The antidepressant effects of ketamine are thought to be mediated by opioid k receptors, and dendrimer conjugation increases its affinity for these receptors.

[0345] Improved binding to sigma 2 receptors based on dendrimer conjugation has also been observed. The greater the binding ability of HD ketamine to sigma 2 receptors, the greater the neuroprotective effect. The mechanism of neuroprotection may be related to increased NGF and BDNF production. Activation of sigma 2 receptors may also be beneficial for neuropsychiatric conditions such as schizophrenia and psychosis. Reduced anxiety and increased antidepressant effects may be observed with sigma 2 receptor targeting. Ketamine binds to sigma 2 receptors at mM concentrations (Pergolizi, 2023; Bonaventura, 2021).

[0346] Binding to the dopamine 2 receptor is only observed with dendrimer-conjugated ketamine. Free norketamine does not appear to bind to the dopamine 2 receptor. Similarly, ketamine also shows no affinity for the dopamine 2 receptor. The dopaminergic effects of ketamine are thought to be indirect. However, glucose dendrimer-ketamine has an RC50 of 13.07 μM at the D2 receptor, while hydroxyl dendrimer-ketamine has an RC50 of 4.3 μM, demonstrating its greater potency. Ketamine's antidepressant and neuroprotective effects are largely mediated by these receptors and dopamine receptors. Ketamine, norketamine, and ketamine metabolites do not appear to have a direct effect on dopamine receptors and do not bind to them. The enhanced activity observed at the D2 receptor in functional assays indicates that dendrimer-ketamine is more potent than an antidepressant.

[0347] GD-ketamine and HD-ketamine exhibit distinct characteristics. Conjugation with hydroxyl dendrimers (HD) compared to glucose dendrimers (GD) appears to alter ketamine's function, which is highly expected. While HD-ketamine does not demonstrate blockage of NMDAR1A / 2B ion channels, GD-ketamine demonstrates that the type of dendrimer conjugated to ketamine is important for the functional differences between them. GD-ketamine also exhibits agonist activity at the 5HT1A receptor, which is not observed in the case of HD-ketamine.

[0348] These results indicate that dendrimer conjugation produces unexpected advantages over free drug for receptor binding and that they depend on dendrimer structure. The microglial targeting of hydroxyl dendrimers and the additional neuronal targeting of glucose dendrimers yields unexpected results with clinical significance.

[0349] The conjugates are targeted to specific receptors on whatever cells they are present on. This includes neurons, microglia, macrophages, and other cells. These receptors can be found anywhere in the brain. Examples include serotonin receptors such as 5HT1A, 5HT2A, and NMDA. Serotonin receptors can also be found on microglia. The compounds bind to and act on these receptors. The "intrinsic cellular targeting" of these dendrimers (hydroxyl dendrimers—microglia / macrophages and glucose dendrimers—to neurons) is somewhat secondary to the action on the specific receptors targeted by the compositions. Important receptors include serotonin receptors [5HT1A, 5HT2A (agonist, antagonist, reverse, or inverse agonist)] and NMDA receptors. In many cases, binding to specific receptors and not binding to other receptors using dendrimers can increase binding potency and reduce the side effects of these drugs.

[0350] Combined with the cell targeting capabilities of glucose dendrimers (e.g., against injured neurons (primary), microglia / macrophages (secondary), or hydroxyl dendrimers (microglia / macrophages)), dendrimer conjugation has demonstrated improved aqueous solubility of >5-200 fold, offering the distinct advantages of modifiable linkages, improved targeting selectivity, ease of formulation and delivery, and reduced side effects.

[0351] Using tryptamine as an example, the binding affinity of drugs conjugated to these dendrimers was shown to have unexpected properties.

[0352] Both hydroxyl and glucose dendrimers have OH surface groups. When tryptamine was conjugated to these dendrimers using the same linking chemistry, very different affinities were observed for serotonin and other receptors. The conjugates exhibited lower affinities than the free drug in cell-based binding assays (an indicator of in vivo efficacy). Lower affinity may allow for less strong binding and may modulate the drug's undesirably strong effects on the receptor. Second, the hydroxyl dendrimer conjugate was not active, whereas the glucose dendrimer conjugate was. This was unexpected and may be due to differences in the internal structure of the glucose and hydroxyl dendrimers. The drug may fold within the hydrophobic core of the hydroxyl dendrimer but open outward in the hydrophilic interior of the glucose dendrimer. Example 5 Treatment of Animal Models of Rett Syndrome with Ketamine and Ketamine-Dendrimer Conjugates

[0353] Rett syndrome (RTT) is a genetic neurodevelopmental disorder affecting females, occurring in 1 in 10,000–15,000 live births. Affected females develop normally for 6–18 months but then lose voluntary movement, including speech and manual dexterity. Most RTT patients are heterozygous for mutations in the X-linked gene MECP2, which encodes a protein that binds to methylation sites in genomic DNA, resulting in gene silencing. The symptoms, progression, and severity of Rett syndrome can vary dramatically from person to person. A wide range of disorders can potentially be associated with Rett syndrome. Symptoms generally appear gradual. RTT is usually characterized by a period of normal postnatal growth followed by regression of speech and hand movements, gait abnormalities, irregular hand movements, and slowed head growth. Other diagnostic criteria for RTT include irregular breathing, gastrointestinal and musculoskeletal disorders, seizures, poor sleep, decreased response to physical pain, and behavioral problems.

[0354] Ketamine is a well-established anesthetic that produces "dissociative anesthesia," exerting both central and peripheral effects, including hypnosis, analgesia, and sympathetic activity, leading to hypertension and tachycardia. The primary mechanism is thought to be its role as an antagonist at N-methyl-d-aspartate (NMDA) receptors. Unlike SSRIs and SNRIs, which only have a delayed effect on depression control, this can result in rapid action and response, as observed in the treatment of treatment-resistant depression, MDD, and suicidal ideation. Ketamine is also effective as a treatment for chronic pain, again due to its effect on NMDAR inhibition. However, its increased dopamine release and its role in ketamine also exert neuroprotective effects through non-NMDAR-mediated mechanisms, such as increased BDNF and mTOR. Ketamine also exerts effects on opioid receptors, increasing dopamine. These effects are beneficial for chronic pain, treatment-resistant depression, MDD, and treatment-resistant epilepsy / seizures.

[0355] However, ketamine has several short-term and long-term side effects. At high doses, ketamine can cause respiratory control and systemic side effects such as increased heart rate, high blood pressure, hyperthermia, loss of coordination, dizziness, nausea, vomiting, sensory-perceptual alternation disorders, and a high incidence of auditory and visual hallucinations. More than half of patients treated with ketamine develop and manifest themselves after the effects of ketamine wear off, a phenomenon characterized by euphoria, vivid dreams, hallucinations, illusions, body image and object distortions, and delirium, which can be extremely upsetting and lead to self-harm. Some of these symptoms correlate with those of schizophrenia. Long-term use of ketamine can result in memory impairment and impaired executive function. Ketamine is also addictive, resulting in tolerance and withdrawal and dependence. Due to these significant side effects, ketamine can only be administered in a controlled environment.

[0356] Ketamine has been shown to be effective in a mouse model of Rett syndrome when delivered IP at a large dose of 8 mg / kg daily for 40 days (total dose of 320 mg / kg), improving survival by 50% at postnatal day 80 (Patrizi et al., 2016). Materials and Methods

[0357] The dendrimer conjugates were prepared as described above.

[0358] As reported by Guy et al., Mecp2 null mutant mice develop neurological symptoms mimicking Rett syndrome. Nat Genet. 2001 Mar;27(3):322-6. doi: 10.1038 / 85899. PMID: 11242117. Both Mecp2 null mutant mice and mice lacking Mecp2 in the brain develop severe neurological symptoms at approximately 6 weeks of age.

[0359] Patrizi A, et al. Chronic Administration of the N-Methyl-D-Aspartate Receptor Antagonist Ketamine Improves Rett Syndrome Phenotype. Biol Psychiatry. 2016 May 1;79(9):755-764. doi: 10.1016 / j.biopsych.2015.08.018. Epub 2015 Aug 24. PMID: 26410354; PMCID: PMC7410367 reported a systematic randomized preclinical study of chronic administration of low-dose (8 mg / kg, i.p.) NMDAR antagonist ketamine in Mecp2 null mice, initiated either early at disease onset or after the development of the RTT phenotype. Mice were treated with 8 mg ketamine / kg / day ip from days 15 to 55 or from days 30 to 55 (total dose of 320 mg / kg for 40 days or 200 mg / kg for 25 days). Treatment from day 30 was ineffective, while treatment from day 15 showed some efficacy at a total dose of 320 mg / kg. Experimental paradigm for MECP2 knockout ("KO") mice:

[0360] Treated from 28 days of age (4 weeks old, symptomatic). Untreated animals die around 55-60 days.

[0361] Mice were treated with ketamine or dendrimer-conjugated ketamine, and controls included wild-type, saline-treated, and saline-treated knockout.

[0362] Animals were treated ip twice weekly at 2.5 mg / kg / dose, with a total dose at the time of evaluation (60 days of age) of approximately 22.5 mg / kg.

[0363] Dendrimer ketamine was tested in a Rett syndrome mouse model at a dose of 2.5 mg / kg (ketamine) IP every other week for 8 weeks (total dose of ketamine 40 mg / kg).

[0364] Four-week-old Mecp2 knockout (KO) mice were randomly treated with saline or 2.5 mg / kg i.p. ketamine or D-ketamine (WT-saline, KO-saline, KO-ketamine, KO-D-ketamine groups) every other week for 8 weeks. Neurobehavioral assessments were performed weekly by recording a composite neurobehavioral score (NBS), which includes assessments of mobility, gait, paw grip, tremor, and respiration, each on a 0-3 scale; higher scores indicate worse phenotypes. The D-ketamine-treated group showed a slowing of disease phenotype progression, accompanied by better neurobehavioral scores after treatment, whereas untreated KO mice did not. [Table 8]

[0365] Open field test: Long-term behavioral changes in saline (WT), ketamine, and D-ketamine groups compared to KO-saline at 8.5 weeks of treatment. Motor function was assessed by recording mice in an open field arena (10.5 inches × 19 inches × 8 inches). Mice were recorded in the same room where they were housed to avoid stress and variability in the testing procedure. Each mouse was placed in a clean open field arena and allowed to explore for 10 minutes, and activity was recorded. Animals were placed in the same open field, facing the long wall and 5 cm apart. result

[0366] Animal survival, neurobehavioral score and activity (total distance traveled, speed and time spent in the corner) were assessed.

[0367] Figures 16A and 16B are graphs of the composite neurobehavioral score (Figure 16A) and postnatal survival probability (Figure 16B) of wild-type, knockout saline (control) versus knockout mice treated with dendrimer-ketamine conjugates. Figure 16C is a graph of the distance traveled (m). Figure 16D is a graph of the speed at which the mice traveled. Figure 16E is a graph of the time spent in the corner.

[0368] The dendrimer conjugated with ketamine provides increased efficacy and increased binding to NMDAR without associated side effects.The dose used is significantly lower than that of free ketamine, which reduces side effects.This is tested in a mouse model of Rett syndrome as proof of concept, because Rett syndrome is a disease that increases glutamate production and increases NMDAR expression / activation.

[0369] A significant improvement in survival was observed with D-ketamine compared to untreated and free ketamine-treated animals, with 100% survival up to 90 days of age. Previously published data by others in a similar model showed only 50% survival after treatment with 320 mg / kg ketamine (an 8-fold higher dose).

[0370] A significant improvement in motor function was observed with D-ketamine, with behavior similar to that of normal healthy controls.

[0371] During a 10-minute period in the open field, a significant difference was observed in the total distance traveled by WT compared to KO mice. Significant improvements were also observed in motor function, expressed as an increase in total distance traveled, by D-ketamine-treated KO in the open field test. Significant improvements were observed in maximum speed and time spent in the corner with D-ketamine compared to the KO saline group.

[0372] The video shows a dramatic improvement in phenotype, with D-ketamine-treated KO mice approaching healthy mice.

Claims

1. A composition comprising a hydroxyl-terminated dendrimer, a sugar-terminated dendrimer, and / or a sugar-based dendrimer, which are covalently conjugated to at least one or more psychedelic agents or hallucinogens via spacers as needed, wherein the psychedelic agent or hallucinogen is not a cannabinoid.

2. The composition according to claim 1, wherein the dendrimer is a first-generation, second-generation, third-generation, fourth-generation, fifth-generation, sixth-generation, seventh-generation, or eighth-generation dendrimer.

3. The composition according to claim 1, wherein the dendrimer is a glucose dendrimer, preferably a first-generation, second-generation, or third-generation glucose dendrimer.

4. The composition according to claim 1, wherein the dendrimer is preferably a hydroxyl-terminated poly(amideamine) (PAMAM) dendrimer having more than 40% or 50% of hydroxyl surface groups.

5. The composition according to claim 1, wherein the dendrimer is a third-generation, fourth-generation, fifth-generation, sixth-generation, or seventh-generation dendrimer.

6. The composition according to claim 1, wherein the composition comprises sugar-terminated dendrimers, and the sugar-terminated dendrimers comprises one or more monosaccharides selected from the group consisting of glucose, galactose, glucosamine, galactose, mannose, and fructose, preferably more than 10 surface monosaccharide portions on their surface.

7. The composition according to claim 1, wherein the dendrimer is prepared from a monosaccharide and optionally from an ethylene glycol building block, preferably from galactose or glucose and optionally from ethylene glycol, and has more than 10 surface sugar moieties.

8. The composition according to claim 1, wherein the monosaccharide building block is one or more selected from the group consisting of glucose, galactose, glucosamine, galactose, mannose, and fructose, and the dendrimer is preferably a glucose dendrimer.

9. The composition according to claim 1, wherein the dendrimer is conjugated to one or more psychedelic agents via spacers having cleavable or incleavable linking groups.

10. The composition according to claim 9, wherein the cleavable bond is selected from the group consisting of esters, disulfides, phosphodiesters, triglycyl peptides, and hydrazine linking groups.

11. The composition according to claim 9, wherein the inclementable bond is selected from the group consisting of amides, ethers, and aminoalkyl linking groups.

12. The composition according to claim 1, wherein the spacer connecting the dendrimer and the psychedelic agent comprises a hydrocarbon such as alkylene, a diethylene glycol portion and / or an oligoethylene glycol chain.

13. The composition according to claim 1, wherein the spacer includes a triazole portion.

14. The composition according to claim 1, wherein the psychedelic agent is selected from the group consisting of classical serotonergic hallucinogens, for example, the drug is an enteractogen, a dissociative anesthetic, or an atypical hallucinogen.

15. The composition according to claim 1, wherein the at least one psychedelic agent or hallucinogen comprises psilocin, ketamine (R-ketamine, S-ketamine, (R / S)-ketamine), norketamine, ketamine metabolites, N,N-dimethyltryptamine (DMT), 4-acetoxy-N,N-dimethyltryptamine, 5-methoxyDMT, 5-chloroDMT, lysergicide (LSD), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), psilocybin, ibogaine, mescaline, mianserin, 2,5-dimethoxy-4-iodoamphetamine (DOI), ayahuasca, 1-(1-phencyclohexyl)piperidine (PCP), norbeocystine, or an analog of a derivative thereof.

16. The composition according to claim 14, wherein the classic serotonergic hallucinogen is selected from the group consisting of psilocybin, psilocin, lysergic acid diethylamide (LSD), mescaline, mianserin, and 2,5-dimethoxy-4-iodoamphetamine (DOI).

17. The composition according to claim 14, wherein the enterogen is selected from the group consisting of 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA).

18. The composition according to claim 14, wherein the dissociative anesthetic agent is selected from the group consisting of ketamine (R-ketamine, S-ketamine, (R / S)-ketamine), norketamine, ketamine metabolites, 1-(1-phencyclohexyl)piperidine (PCP), or derivatives thereof.

19. The composition according to claim 1, wherein the composition provides a sustained release such that an effective amount is obtained over a period of 24 hours.

20. The composition according to claim 1, wherein the one or more psychedelic agents conjugated to the dendrimer are present in a concentration of about 0.01% to about 30% by weight, preferably about 1% to about 20% by weight, and more preferably about 5% to about 20% by weight.

21. The composition according to claim 1, wherein the composition further comprises one or more diagnostic agents, preferably the diagnostic agent being selected from the group consisting of fluorescent dyes, near-infrared dyes, SPECT imaging agents, PET imaging agents, and radioisotopes.

22. The composition according to claim 1, which provides sustained delivery of a drug or sustained receptor activity at a therapeutically effective level, preferably for 24 hours, at a certain level to achieve clinical benefit without hallucinations, or at a relatively constant dose to maintain a mental state.

23. A pharmaceutical formulation comprising a dendrimer according to any one of claims 1 to 22 and a pharmaceutically acceptable carrier or additive.

24. The pharmaceutical formulation according to claim 23, wherein the formulation is formulated for systemic administration.

25. The pharmaceutical formulation according to claim 23, wherein the formulation is formulated for enteral or parenteral administration.

26. The pharmaceutical formulation according to claim 23, wherein the formulation is formulated for oral, mucosal (intranasal cavity, oral cavity, rectum, vagina, sublingual, lung), intramuscular, intravenous, subcutaneous, transdermal, or intrathecal administration.

27. The pharmaceutical preparation according to claim 23, which is in a form selected from the group consisting of hydrogels, nanoparticles or microparticles, suspensions, powders, tablets, capsules, creams, and solutions.

28. A composition according to any one of claims 1 to 22 for treating one or more psychological disorders, cognitive disorders, behavioral disorders and / or mood disorders in a subject requiring treatment for one or more psychological disorders, cognitive disorders, behavioral disorders and / or mood disorders, characterized in that an effective amount of the composition is administered to the subject to treat, alleviate and / or prevent one or more symptoms associated with one or more of the psychological disorders, cognitive disorders, behavioral disorders and / or mood disorders.

29. The composition according to claim 28, wherein one or more of the psychological disorders, cognitive disorders, behavioral disorders and / or mood disorders are selected from apathy, low motivation, attention deficit, executive function and / or cognitive engagement impairment, obsessive-compulsive disorder, and neurocognitive disorder.

30. The composition according to claim 28, characterized in that the composition is administered in an amount effective to produce improvements in motivation, attention, accuracy, response speed, perseveration and / or cognitive engagement without causing adverse side effects.

31. The composition according to claim 28, characterized in that the composition is administered in an amount effective to cause one or more psychedelic agents, covalently conjugated to a dendrimer, to bind to one or more receptors on or inside the surface of a target cell.

32. The composition according to claim 31, wherein the target cells are nervous system cells, glial cells, and macrophages of the peripheral and / or central nervous system.

33. The composition according to claim 31, wherein the receptor is selected from the group consisting of 5-HT receptor subtypes, adrenergic receptors, dopaminergic receptors, and histaminergic receptors.

34. The aforementioned receptor is 5-HT ２ , 5-HT ６ , 5-HT ７ The composition according to claim 33, selected from the group consisting of adrenergic α2, D1, and D2 receptors.

35. The aforementioned receptor is 5-HT ２Ａ The composition according to claim 31.

36. The composition according to claim 28 for treating psychiatric disorders such as obsessive-compulsive disorder, eating disorders, attention deficit and hyperactivity disorder, and schizophrenia.

37. The composition according to claim 28 for treating patients with depression, such as treatment-resistant depression, anxiety disorders, and post-traumatic stress disorder (PTSD).

38. The composition according to claim 28 for treating patients with drug and / or alcohol dependence.

39. The composition according to claim 28 for treating patients having Alzheimer's disease, dementia, multiple sclerosis or other neurodegenerative disorders.

40. The composition according to claim 28 for treating patients with chronic inflammation and autoimmune disorders, such as rheumatoid arthritis, atherosclerosis, Parkinson's disease, Alzheimer's disease, and multiple sclerosis.

41. The composition according to claim 28 for treating pain, including cluster headaches, migraines, chronic pain, postoperative pain, and cancer-related pain.

42. The composition according to claim 28, wherein the composition exerts its effect for at least 24 hours after administration.

43. The composition according to claim 28, wherein the composition comprises a glucose dendrimer-ketamine conjugate.

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