Dendrimer compositions for targeted delivery of rosemary therapeutic agents

Abstract

Dendrimer formulations comprising one or more fantastic or fantastic agents and methods of using the same are described. Preferably, the dendrimer conjugates selectively bind to one or more receptors on the surface or within the target cell. The formulations are suitable for enteral and / or parenteral administration for the treatment of one or more receptor-mediated diseases, including psychological, cognitive, behavioral and / or mood disorders.

Description

Field of the Invention

[0001] The present invention relates generally to the field of hallucinogenic drug formulations, and more particularly to dendrimer hallucinogenic drug conjugates that improve receptor binding, cell selectivity, and reduce side effects.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Application No. 63 / 401,470, filed on August 26, 2022, entitled “Dendrimer Compositions for Targeted Delivery of Psychedelic Therapeutics,” filed by Johns Hopkins University, with inventors including Kannan Rangaramanujam, Kunal Parikh, Sujatha Kannan, and Anjali Sharma, which is hereby incorporated in its entirety.

[0004] Statement Regarding Federally Funded Research

[0005] none. Background of the Invention

[0007] The word "psychedelic" (psyche, meaning mind or soul, and delos, meaning display) was first coined by psychiatrist Humphry Osmond in 1956 while studying lysergic acid diethylamide (LSD). For thousands of years, 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 played an important role in the hallucinations of Central and South America.

[0008] Until recently, the medicinal value of these drugs has been explored in a scientific context. Recently, a second wave of psychedelic research is focusing on psychedelics as neuropharmaceuticals to treat alcohol and tobacco addiction, general mood and anxiety disorders, and cancer-related depression. Psychedelics and other classes of compounds related to them (e.g., elicitors known to temporarily alter consciousness, involving dramatic changes in somatic, perceptual, and cognitive properties) are receiving increasing attention. This is due in part to their wider range of uses, new methods for synthesis and purification, in-depth mechanistic understanding, and ongoing clinical trials. Their vast potential for affecting mind and consciousness also makes it important to understand their mechanisms of action and to target their effects to cells and tissue regions of interest to avoid side effects.

[0009] Psychedelic drugs and hallucinogens, such as plant-derived indoleamines (e.g., N,N-dimethyltryptamine (DMT), 5-methoxy-DMT (5-MeO-DMT), dimethyl-2-hydroxytryptamine phosphate, 4-hydroxy-DMT (psilocin, the active metabolite of dimethyl-2-hydroxytryptamine phosphate)), phenylalkylamines (e.g., mescaline), and synthetic "amphetamines" such as 2,5-dimethoxy-4-iodoamphetamine (DOI) and 2,5- Dimethoxy-4-bromoamphetamine (DOB), mianserin, and semisynthetic ergolines (e.g., LSD) have shown great therapeutic potential for the treatment of a variety of mental health and neurological disorders (e.g., depression, refractory 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).

[0010] In general, psychedelic molecules achieve their therapeutic effects by activating 5-HT2A receptors in the cerebral cortex. 5-HT2A is a serotonin receptor that is thought to mediate brain plasticity and is found in higher densities in cortical areas involved in higher-order cortical processing (prefrontal cortical areas: cingulate cortex and posterior cingulate cortex). However, the practical implementation of these potential therapies faces significant challenges, including solubility, bioavailability, absorption, side effects / toxicity, time to onset, duration of efficacy, and hallucinogenic effects. Psychedelic drugs have limited access to these critical areas of the brain, particularly into brain cells and immune cells involved in disease processes. Collectively, these issues limit the applicability, efficacy, and translational potential of this class of drugs and prevent them from realizing their widespread therapeutic potential.

[0011] It is therefore an object of the present invention to provide formulations that allow for a more selective delivery of psychedelic and hallucinogenic drugs. SUMMARY OF THE INVENTION

[0013] The group of hallucinogens and dendrimer compositions formulated according to the present invention include the following active ingredients, for example: (i) hallucinogens, a group of serotonergic agonists, generally class I agonists, such as dimethyl-2-hydroxytryptamine phosphate, lysergic acid, mescaline; (ii) elicitors, class I monoamine releasers and reuptake inhibitors, known to evoke feelings of emotional openness and connection, such as 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA); (iii) dissociative agents, glutamatergic NMDA antagonists, such as ketamine, dextromethorphan (DXM) and nitrous oxide; (iv) atypical hallucinogens, which have a variety of mechanisms, such as Δ 9- Tetrahydrocannabinol (THC) and ibogaine. Dendrimer conjugate compositions have been developed that are capable of delivering drugs to receptors on specific cells (neuronal cells, glial cells, macrophages), including targets on their surface and inside. These formulations may enhance the potency of these drugs through superior binding to target receptors, thereby reducing dosage, bringing new mechanistic insights, reducing side effects, improving solubility, formulation, PK and other aspects of the use of these drugs, thereby opening new avenues for clinical use.

[0014] The formulations are based on conjugation of the drug 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 have enhanced solubility, uptake into the brain and other specific cell types, and selectivity of uptake and receptor binding. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1A and 1B Schematic diagram of an exemplary synthetic route for synthesizing a dendrimer psilocin with a cleavable ester linker using click chemistry. First, psilocin is conjugated to a linker with an azide moiety via an ester linkage ( Figure 1A ), and then conjugated to the surface alkyne-modified dendrimers via an azide-alkyne click reaction ( Figure 1B ).

[0017] Figure 2A and 2B Schematic diagram of an exemplary synthetic route for synthesizing a dendrimer-dimethyl-4-hydroxytryptamine analog with a non-cleavable amide linkage using click chemistry. First, the dimethyl-4-hydroxytryptamine analog is conjugated to a linker with an azide moiety via an amide linkage ( Figure 2A ), and then conjugated to the surface alkyne-modified dendrimers via an azide-alkyne click reaction ( Figure 2B ).

[0018] Figure 3A and 3B Schematic diagram of an exemplary synthetic route for synthesizing dendrimer-ketamine with non-cleavable amino-alkyl linkages using copper-catalyzed alkyne azide click chemistry. Ketamine hydrochloride (1) ( Figure 3A ), followed by conjugation via an azide-alkyne click reaction with a dendrimer modified with surface azide groups ( Figure 3B ).

[0019] Figure 4Aand 4B Schematic diagram of an exemplary synthetic route for synthesizing dendrimer-DMT analogs with non-cleavable amide linkages using copper-catalyzed alkyne azide click chemistry. N,N-dimethyltryptamine analogs (DMT analogs) are first conjugated to a linker with an azide moiety via an amide linkage ( Figure 4A ), and then conjugated to the dendrimer modified with surface alkyne groups via an azide-alkyne click reaction ( Figure 4B ).

[0020] Figure 5A and 5B Schematic diagram of the synthesis of dendrimer-DMT with non-cleavable amino-alkyl linkages. Figure 5A A DMT drug modified with an alkyne group is shown. Figure 5B Conjugation to a dendrimer modified with surface azide groups via an azide-alkyne click reaction is shown.

[0021] Fig. 6A and 6B is a schematic diagram of a dendrimer-lysergic acid diethylamide (dendrimer-LSD) having a non-cleavable amino-alkyl linkage. Fig. 6A An LSD modified with an alkyne group is shown. Figure 6B Conjugation to a dendrimer modified with surface azide groups via an azide-alkyne click reaction is shown.

[0022] Figure 7 Schematic diagram of the step-by-step synthetic route for the synthesis of glucose dendrimer-dimethyl-4-hydroxytryptamine conjugates with cleavable ester linkages.

[0023] Figure 8 Schematic diagram of the stepwise synthetic route of glucose dendrimer-dimethyl-4-HT analog conjugates with non-cleavable amide linkages.

[0024] Fig. 9 Schematic diagram of a stepwise synthetic route for the synthesis of glucose dendrimer-ketamine conjugates with non-cleavable amino-alkyl linkages.

[0025] Fig.10 Schematic diagram of a step-by-step synthetic route for the synthesis of glucose dendrimer N,N-dimethyltryptamine analog (DMT analog) conjugates with non-cleavable amide linkages.

[0026] Fig.11 Schematic diagram of the stepwise synthetic route of glucose dendrimer-DMT analog conjugates with non-cleavable amino-alkyl linkages.

[0027] Fig.12Schematic diagram of the step-by-step synthetic route for the synthesis of glucose dendrimer-lysergic acid diethylamide (LSD) conjugates with non-cleavable amino-alkyl linkages.

[0028] Fig.13 Schematic representation of the major pharmacological targets of LSD, dimethylhydroxytryptamine phosphate, DMT, MDMA, and ketamine, the signaling cascades involved, hormonal regulation, and the main behavioral consequences following administration in animals and humans.

[0029] Fig.14A Schematic diagram of the synthesis of PAMAM dendrimer-norketamine conjugate. Fig. 14B Schematic diagram of the synthesis of glucose dendrimer-norketamine conjugate.

[0030] Fig.15A Figure 1 is a graph of the NMDAR 1A / 2B antagonist assays of glucose dendrimer-ketamine (IC50 = 4.54 μM), hydroxy dendrimer-ketamine (IC50>100), and norketamine (IC50 = 6.96 μM). Fig. 15B is the binding efficiency % of the micromolar log concentration of the compound in the D2L human dopamine GPCR cell agonist cAMP assay. Norketamine (closed circles), glucose dendrimer-ketamine EC50 = 13.08 micromolar (open circles), and hydroxy dendrimer-ketamine EC50 = 4.263 micromolar (triangles). Fig. 15C is the binding efficiency in % of micromolar log concentration of ketamine in the TA1 human trace amine GPCR cell agonist cAMP assay. Norketamine (closed circles), glucose dendrimer-ketamine EC50 = 13.08 micromolar (open circles), and hydroxy dendrimer-ketamine EC50 = 4.263 micromolar (triangles).

[0031] Fig.16A and 16B is the composite neurobehavioral score of wild-type, knockout saline (control) and knockout mice treated with dendrimer-ketamine conjugate ( Fig.16A ) and postnatal survival ( Fig. 16B ) in the table below. Fig. 16C is a graph of distance travelled (m); Fig.16D is a graph of the speed at which mice travel; Fig.16E is a graph of the time spent in the corner. DETAILED DESCRIPTION OF THE INVENTION

[0033] I. Definitions

[0034] The term "hallucinogen" refers to a group of chemically distinct compounds that are able to induce altered states of consciousness (ASCs) characterized by profound changes in mood, thought processes, perceptions, and the experience of self and environment, which occur only in dreams, meditation, and acute psychosis. Not all hallucinogenic compounds reliably produce visual and auditory hallucinations. Therefore, hallucinogens may also be called psychotomimetic agents (mimicking psychosis), psycholytic agents (relaxing the mind), or psychedelics (manifesting the mind), reflecting different attitudes and intentions towards these substances.

[0035] The term "psychedelic" refers to a class of hallucinogenic drugs whose primary effect is to induce an abnormal state of consciousness. This results in specific mental, visual, and auditory changes, and often a markedly altered state of consciousness. The psychedelic state is often compared to meditative, psychodynamic, or transcendental types of mental alterations. The "classic" psychedelics, those that have had the greatest impact on science, include mescaline, LSD, dimethylhydroxytryptamine phosphate, and DMT.

[0036] Most psychedelic drugs belong to one of three families of compounds: tryptamines, phenethylamines, or lysergic acid amides, and many tend to act by agonizing serotonin 2A receptors. When compounds bind to serotonin 5-HT2A receptors, they modulate the activity of key brain circuits involved in sensory perception and cognition, however, the exact nature of how psychedelics induce perceptual and cognitive changes through 5-HT2A receptors remains unclear, although a reduction in default mode network activity and an increase in functional connectivity between brain regions may be one of the most relevant pharmacological mechanisms of psychedelic experiences, especially ego death.

[0037] The terms "active agent" or "bioactive agent" are used interchangeably and refer to a chemical or biological compound that induces a desired pharmacological and / or physiological effect, which may be preventive, therapeutic or diagnostic. These may be nucleic acids, nucleic acid analogs, small molecules having a molecular weight of less than 2 kD and more typically less than 1 kD, peptide mimetics, proteins or peptides, carbohydrates or sugars, lipids or combinations thereof. These terms also include pharmaceutically acceptable, pharmacologically active drug derivatives, 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 reveal, precisely locate and define the location of a pathological process. A diagnostic agent can label target cells so that these labeled target cells can be subsequently detected or imaged.

[0038] "Analogs" are related to a given compound and refer to another compound that is structurally similar, functionally similar, or both to a specified compound. Structural similarity can be determined using any standard known in the art, such as the Tanimoto coefficient, which provides a quantitative measure of the similarity between the two compounds based on their molecular descriptors. Preferably, the molecular descriptors are 2D properties, such as fingerprints, topological indices, and maximum common substructures, or 3D properties, such as overall shape and molecular fields. For different and identical molecular pairs, the range of the Tanimoto coefficient is between 0 and 1 (inclusive). If the Tanimoto coefficient of the compound and the specified compound is 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), then the compound can be considered to be an analog of the specified compound. If the pharmacological effect, physiological effect, or both induced by the compound are the same as those of a specific compound, the compound is functionally similar to a specific compound. "Analogs" can also refer to modifications of compounds, including but not limited to hydrolysis, reduction, or oxidation products. Hydrolysis, reduction, and oxidation reactions are known in the art.

[0039] The term "therapeutically effective amount" refers to an amount of a therapeutic agent that, when incorporated into and / or onto a dendrimer, produces some 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 undue experimentation. In some embodiments, the term "effective amount" refers to an amount of a therapeutic or prophylactic agent that reduces or alleviates the symptoms of one or more diseases.

[0040] In the context of inhibition, the terms "inhibit" or "reduce" refer to reducing or decreasing activity and number. This can be a complete inhibition or reduction of activity or number, or a partial inhibition or reduction. The inhibition or reduction can be compared to a control or standard level. The 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 number of diseased neurons by about 10%, 20%, 30%, 40%, 50%, 75%, 85%, 90%, 95% or 99% compared to the activity and / or number of the same cells in an equivalent tissue of a subject that has not received or been 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, rate of brain weight loss or rate of hippocampal volume loss compared to an untreated control subject.

[0041] The terms "treat" or "prevent" refer to ameliorating, alleviating or otherwise preventing a disease, disorder or condition from occurring or developing in an animal that may be susceptible to the disease, disorder and / or condition but has not yet been diagnosed as having the disease, disorder or condition; inhibiting the disease, disorder or condition, e.g., preventing its progression; and relieving the disease, disorder or condition, e.g., causing the disease, disorder and / or condition to regress. Treating the disease or condition includes ameliorating at least one symptom of the particular disease or condition, even if the underlying pathophysiology is not affected, such as treating pain in a subject by administering an analgesic, even if such agent does not treat the cause of the pain. Desired effects of treatment include reducing the rate of disease progression, improving or alleviating the disease state, and alleviating or improving prognosis. For example, an individual is successfully "treated" if one or more symptoms associated with depression are alleviated or eliminated, including but not limited to reducing the degree of anxiety, agitation or restlessness, improving feelings of sadness, tearfulness, emptiness or helplessness, improving the patient's quality of life, reducing the dose of other drugs required to treat the disease, and delaying the progression of the disease.

[0042] The phrases "pharmaceutically acceptable" or "biocompatible" refer to compositions, polymers and other materials and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications, and at a reasonable benefit / risk ratio. The phrase "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, solvent or encapsulating material, used to deliver or transport any subject composition from one organ or part of the body 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.

[0043] The term "biodegradable" generally refers to materials that, under physiological conditions, will degrade or erode into smaller units or chemicals that can be metabolized, eliminated, or excreted from the body. Degradation time depends on composition and form.

[0044] The term "dendrimer" includes, but is not limited to, a molecular architecture having an inner core, inner layers or "generations" of repeating units regularly attached to the initiating core, and an outer surface attached to the terminal groups of the outermost generations.

[0045] The term "functionalization" refers to modifying a compound or molecule in some way to attach 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.

[0046] The term "targeting moiety" is a moiety that is designated to be located at or away from a particular 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 particular cell type, a subcellular compartment, or a molecule such as a receptor.

[0047] The term "extended residence time" refers to an increase in the time required for an agent to be cleared from a patient's body, or an organ or tissue of the patient. In certain embodiments, "extended residence time" refers to an agent whose clearance half-life is 10%, 20%, 50%, or 75% longer than a control standard (e.g., a comparable agent not bound to a delivery vehicle (e.g., a dendrimer). In certain embodiments, "extended residence time" refers to an agent whose clearance half-life is 2, 5, 10, 20, 50, 100, 200, 500, 1000, 2000, 5000, or 10000 times longer than a control standard (e.g., a comparable agent not bound to a delivery vehicle (e.g., a dendrimer)) that is specifically targeted to a particular cell type.

[0048] The terms "binding" and "encapsulation" refer to binding, formulating or otherwise including an agent into and / or onto a composition so as to allow release (e.g., sustained release) of the agent in a desired application. The agent or other material can be bound to the dendrimer by binding to one or more surface functional groups of the dendrimer (through covalent, ionic or other binding interactions), by physical admixture, by encapsulating the agent within the dendritic structure, and / or by encapsulating the agent within the dendritic structure.

[0049] As used herein, the central nervous system ("CNS") includes the brain and spinal cord. As used herein, the peripheral nervous system ("PNS") refers to the nerves outside the brain and spinal cord.

[0050] "Hydroxyl terminated" with respect to dendrimers refers to dendrimers having hydroxyl groups on their surface. These hydroxyl groups are not attached to the termini of the dendrimer via a sugar moiety (eg, a saccharide moiety).

[0051] "Sugar-terminated" with respect to a dendrimer means that the dendrimer contains sugar moieties (eg, carbohydrate moieties) on the surface of the dendrimer rather than in its core.

[0052] "Sugar-based" with respect to dendrimers refers to dendrimers that contain sugar moieties (eg, saccharide moieties) in the core of the dendrimer or in both its core and its surface.

[0053] II. Composition

[0054] Dendrimer compositions conjugated or complexed with one or more hallucinogen and / or dissociative agent compounds have been developed for use in preventing and / or treating symptoms associated with one or more psychological, cognitive, behavioral, mood disorders and / or non-neurological disorders in a subject in need thereof. The compositions are particularly useful for treating and / or ameliorating one or more symptoms of mental health and neurological disorders (e.g., depression (major depressive disorder, refractory depression, postpartum depression), suicidal ideation, autism, bipolar disorder, anxiety, drug dependence, 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 syndrome). In a preferred embodiment, the dendrimer is a glucose dendrimer or a hydroxyl-terminated dendrimer, such as a hydroxyl-terminated PAMAM or a sugar-modified dendrimer.

[0055] Exemplary hallucinogens include dimethyl-4-hydroxytryptamine, 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, lysergic acid monourea (LSD), 3,4-methylenedioxymethamphetamine (MDMA), dimethyl-2-hydroxytryptamine phosphate, ibogaine, mescaline, mianserin, and norbaeocystin.

[0056] Typically, hallucinogens and / or their derivatives bind to receptors on the surface of target cells and / or receptors within target cells. Exemplary target cells include, but are not limited to, brain cells, such as microglia, astrocytes and / or neurons, for example, cells within pathological sites in the brain or CNS; cells in the peripheral nervous system, such as peripheral neurons, glial cells and / or their supporting cells, such as intestinal cells, cardiovascular cells and immune system cells. The microglia and / or astrocytes to which the hallucinogens and / or their derivatives are delivered can be active or inactive microglia and / or astrocytes. Classical / serotonin psychedelic compounds also exhibit immunomodulatory properties and therefore have applications in autoimmune diseases.

[0057] The hallucinogen and / or derivative thereof of the dendrimer-active agent conjugate binds to a target receptor on the surface of a target cell or inside a target cell. In some embodiments, the agent remains conjugated to the dendrimer while the hallucinogen and / or derivative thereof binds to the target receptor. In these embodiments, after binding, the agent may be released from the dendrimer or 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 proximity to the target receptor and then binds to the target receptor on the target neural and / or glial cells.

[0058] A. Dendrimers

[0059] Dendrimers are three-dimensional, hyperbranched, monodisperse, globular and multivalent macromolecules that include surface end groups (Tomalia, DA, et al., Biochemical Society Transactions, 35, 61 (2007); and Sharma, A., et al., ACS Macro Letters, 3, 1079 (2014)).

[0060] 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 attached to the end groups of the outermost generation. In some embodiments, the dendrimer has a regular dendrimer or "starburst" molecular structure.

[0061] Typically, the diameter of the dendritic macromolecule is between about 1 nm and about 60 nm, more preferably between about 1 nm and about 50 nm, between about 1 nm and about 40 nm, between about 1 nm and about 30 nm, between about 1 nm and about 20 nm, between about 1 nm and about 10 nm, or between 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 the dendritic macromolecule to cross the blood-brain barrier ("BBB") is less than 5 nm, while the size of the dendritic macromolecule that cannot cross the BBB and stays in the peripheral circulation is greater than 5 nm. In some embodiments, the diameter of the dendritic macromolecule can effectively penetrate the BBB and remain near or inside the target neural and / or glial cells to deliver the agent bound thereto. In some embodiments, the dendritic macromolecule has a diameter that effectively penetrates the BBB and is internalized into the target neural and / or glial cells (e.g., neurons, oligodendrocytes, astrocytes, microglia, and glial support cells) to deliver the agent bound thereto. In some embodiments, the dendrimer has a diameter effective to penetrate a barrier interface (e.g., the blood-nerve barrier ("BNB")) and be internalized into neural and glial cells (e.g., neurons, Schwann cells, satellite cells, and glial support cells) of the peripheral nervous system to deliver the agent bound thereto. In some embodiments, the dendrimer has a diameter effective to be retained in the peripheral circulation to deliver the agent bound thereto to target cells of the peripheral nervous system, e.g., enteric neurons and glial cells. A major benefit of using dendrimer conjugates is that the dendrimer is able to enhance the ability of the psychedelic drug to bind to a target receptor on a target cell, e.g., binding of the compound to a serotonin receptor on neurons in an affected area of ​​the brain.

[0062] In some embodiments, the molecular weight of the dendrimer is between about 500 Daltons and about 100,000 Daltons, between about 500 Daltons and about 50,000 Daltons, or between about 1,000 Daltons and about 20,000 Daltons, inclusive. Dendrimers with a size less than 30,000 Da are more suitable for transport across the BBB, while those with a size greater than 50,000 Da are more suitable for confinement in the periphery.

[0063] In some embodiments, the dendrimer has a supercore (e.g., dipentaerythritol) and one or more monosaccharide branching units. In some embodiments, the monosaccharide branching units are bound to the core or previous monomer layers by a linker such as a polyethylene glycol chain. In a preferred embodiment, the supercore is dipentaerythritol and the monosaccharide branching units are glucosyl branching units, as shown in structures II-IV. In a most preferred embodiment, the dendrimer is made entirely of glucose building blocks. PAMAM dendrimers modified with sugars can also work, but dendrimers made of sugars, especially glucose, are most preferred. Particularly preferred glucose dendrimers are G1 to G3 glucose dendrimers, such as G1, G2 and / or G3 glucose dendrimers.

[0064] Dendrimer scaffolds suitable for conjugation include, but are not limited to, polyamidoamine (also known as PAMAM) or STARBURST TM Dendritic macromolecules; polypropylamine (POPAM), polyethyleneimine, polylysine, polyester, polystyrene, aliphatic polyethers, aromatic polyether dendritic macromolecules, dendritic macromolecules of sugars (e.g., glucose, galactose, mannose, fructose, etc.) and copolymers thereof, such as copolymers of sugars and alkylene glycols (e.g., dendritic macromolecules formed from glucose and ethylene glycol building blocks). Dendritic macromolecules can have multiple surface functional groups, such as carboxylic acids, amines, hydroxyls, and / or acetamides. The terms "surface functional groups" and "terminal groups" are used interchangeably herein. In some embodiments, the dendritic macromolecule has surface hydroxyls. 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), thereby having sugar molecules and / or polyalkylene glycols as terminal moieties / molecules. The dendrimer can be of any generation, including but not limited to generation 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. In some embodiments, the dendrimer is a PAMAM dendrimer used as a platform and modified with functional groups to increase the number of surface hydroxyl groups. Preferred PAMAM dendrimers include hydroxyl-terminated PAMAM dendrimers, particularly G3 to G6 hydroxyl-terminated PAMAM dendrimers, such as G3, G4, G5 and G6 hydroxyl-terminated PAMAM dendrimers.

[0065] In some embodiments, by using higher generation dendrimers (e.g., 4th, 5th, or 6th generation PAMAM dendrimers, 2nd, 3rd, or higher generation glucosyl dendrimers), the dendrimer-active agent conjugates can be confined to the peripheral circulation and specifically targeted to specific tissue regions and / or cell types, such as peripheral nerve cells and macrophages, glial cells, and / or their supporting cells, such as enteric neurons and glial cells. In addition, alternatively, by appropriate functionalization of the dendrimers (e.g., PEGylation), the dendrimer-active agent conjugates can be confined to the peripheral circulation.

[0066] In some embodiments, dendrimers can be specifically targeted to specific tissue regions and / or cell types of the central nervous system (CNS), peripheral nervous system (PNS), and / or periphery, such as neurons and glial cells of the CNS, and / or neurons and glial cells of the PNS, by using dendrimers of a certain generation, such as PAMAM dendrimers of generation 2 (G2), G3, G4, and G5, and / or glucose dendrimers.

[0067] Monosaccharide-based dendrimers

[0068] In preferred embodiments, the branching units comprise monosaccharides. In some embodiments, the monosaccharide branching units are attached to the core or a previous monomer layer via a linker such as a polyethylene glycol chain. In preferred embodiments, the monosaccharide branching units are glucose-based branching units. In some embodiments, the branching units may include PEG and / or alkyl chain linkers between different dendrimer generations. For example, the glucose layers are connected via a PEG linker and a triazole ring. In some embodiments, the branching units are the same for each generation of dendrimers generated from the core. Thus, for example, for the generation of the 1st generation of dendrimers, for the generation of the 2nd generation of dendrimers, and for the generation of the 3rd generation of dendrimers, the branching units are glucose-based branching units.

[0069] In some embodiments, the dendrimer has a supercore such as dipentaerythritol and one or more monosaccharide branching units. In some embodiments, the supercore is dipentaerythritol and the monosaccharide branching units are glucosyl branching units. In further embodiments, the spacer molecule may also be an alkyl CH2) n – Hydrocarbon-like units.

[0070] In some embodiments, dendrimers synthesized using glucose building blocks, whose surfaces are composed primarily of glucose moieties, specifically target cells, including damaged neurons, ganglion cells, and other neuronal cells in the brain, eye, and / or peripheral nervous system. In some embodiments, glucose-based dendrimers selectively target or are enriched in target neural and / or glial cells. In some embodiments, glucose-based dendrimers selectively target or are enriched in target neural and / or glial cells. In some embodiments, glucose-based dendrimers selectively target or are enriched on the surface of target neural and / or glial cells. In some embodiments, glucose-based dendrimers selectively target or are enriched in and / or on the surface of damaged, diseased, and / or overactive neurons and / or glial cells.

[0071] In some cases, dendrimers include an effective number of sugar molecules and terminal groups, such as glucose and / or hydroxyl groups, for targeting one or more neurons and / or glial cells of the CNS, PNS, and / or eye. The terminal hydroxyl groups of these dendrimers can be part of a terminal glucose molecule or additional hydroxyl groups that are not part of a glucose molecule, or a combination thereof. In some embodiments, all terminal hydroxyl groups are part of a terminal glucose molecule. In some embodiments, the number of sugar molecules at the end of the dendrimer is determined by the number of generations.

[0072] In some embodiments, the dendrimer is composed of glucose and oligoethylene glycol building blocks. Exemplary glucose dendrimers are shown in structures V and VII.

[0073] 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, a glucose dendrimer is a second generation glucose-based dendrimer with 24 glucose molecules on the periphery and 6 glucose molecules embedded in the main chain, held together by PEG segments.

[0074] Dendrimer compositions that can selectively accumulate within neurons, particularly in the nuclei of injured and / or overactive neurons, termed "glucose dendrimers," can also accumulate in large quantities within activated microglia. However, in contrast to hydroxy dendrimers that accumulate primarily in microglia, these dendrimers primarily enter neurons. Johns Hopkins University describes glucose dendrimers in USSN 63 / 327,610, "Dendrimer compositions for targeted delivery of therapeutic agents to neurons," with inventors Kannan Rangaramanujam, Rishi Sharma, Anjali Sharma, Sujatha Kannan, Nirnath Sah, Mira Sachdeva, and Siva P. Kambhampati, filed April 5, 2022.

[0075] Glucose dendrimers include (a) a central core, (b) one or more branching units, wherein the branching units are monosaccharide glucosyl branching units, optionally with linkers attached thereto; and optionally (c) one or more therapeutic, prophylactic and / or diagnostic agents. Typically, one or more branching units are attached 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 hexapropargylated derivative thereof. In some embodiments, the branching units are attached to the central core via linkers such as hydrocarbon or oligoethylene glycol chains. In a preferred embodiment, the branching unit is β-D-glucopyranoside tetraethylene glycol azide having the structure

[0076]

[0077] or its peracetylated derivatives.

[0078] In some embodiments, the glucose dendrimer is a generation 1, 2, 3, 4, 5, or 6 dendrimer. In one embodiment, the dendrimer is a generation 1 dendrimer having the following structure:

[0079]

[0080] In a preferred embodiment, the dendrimer is a second generation dendrimer having the following structure:

[0081]

[0082] In some embodiments, one or more therapeutic, prophylactic and / or diagnostic agents are encapsulated, bound and / or incorporated into the dendrimer at a concentration of about 0.01% to about 30%, preferably about 1% to about 20%, more preferably about 5% to about 20% by weight. The dendrimer is conjugated to one or more diagnostic agents, such as fluorescent dyes, near infrared dyes, SPECT imaging agents, PET imaging agents, and radioisotopes.

[0083] In some embodiments, the dendrimer and one or more pharmaceutical agents are linked via one or more linkers or coupling agents (e.g., one or more hydrocarbon or oligoethylene glycol chains). Exemplary linkages are disulfide bonds, ester bonds, ether bonds, thioester bonds, and amide bonds.

[0084] 1. Core

[0085] In some embodiments, dendrimers are prepared using a process that assembles from a multifunctional core and extends outward through a series of reactions. The multifunctional core moiety allows for the stepwise addition of branching units (ie, generations) around the core.

[0086] 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"'-silanetetrayltetra(propane-1-thiol), 3,3-divinylpenta-1,4-diene, 3,3',3"-nitrilotripropionic acid, 3,3',3"-nitrilotripropionic acid, 3,3',3"- tris(N-(2-aminoethyl)propionamide), 3,3',3",3"-(ethane-1,2-diylbis(azatriyl))tetrapropionamide, 3-(carboxymethyl)-3-hydroxyglutaric acid, 2,2'-((2,2-bis((2-hydroxyethoxy)methyl)propane-1,3-diyl)bis(oxy))bis(ethanol-1-ol), tetrakis(3-(trichlorosilyl)propyl)silane, 1-thioglycerol, 2,2,4, 4,6,6-hexachloro-1,3,5,2l5,4l5,6l5-triazatriphosphine, 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"-nitrilotri(ethane-1-ol), α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, cucurbituril, benzene-1,2,3,4,5,6-hexathiol, monosaccharide, disaccharide, trisaccharide, oligosaccharide or its azide, alkyne modified part. In some embodiments, the core part is chitosan. Therefore, azide-modified chitosan or alkyne-modified chitosan is suitable for binding to branch units using click chemistry. In a preferred embodiment, the central core is dipentaerythritol or its hexapropargylated derivative.

[0087] In some embodiments, the core moiety is ethylenediamine or tetraethylene oxide. In some embodiments, the core moiety is dipentaerythritol. Examples of chemical structures suitable for use as core moieties are shown in Table 1 below.

[0088] Table 1. Structural representation of various building blocks (core, branching units, surface functional groups, monomers) used for dendrimer synthesis.

[0089]

[0090]

[0091]

[0092]

[0093]

[0094]

[0095] 2. Branch unit

[0096] Exemplary chemical structures suitable as branching units include monosaccharides. In some embodiments, the monosaccharide branching units are conjugated to the monomer core or the previous layer through 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 (CH2)n-type hydrocarbon units.

[0097] The branching units are PEG or alkyl chain linkers between different generations of dendrimers, for example, glucose layers are connected via PEG linkers and triazole rings.

[0098] In preferred embodiments, the branching units are identical for each generation of dendrimers generated from the core. Thus, in one embodiment, the branching units are glucose-based branching units used to generate the first generation dendrimers shown in structures V-VII.

[0099] In some embodiments, the branching unit is a supermonomer, i.e., AB n Building blocks. Exemplary supermonomers include AB4, AB5, AB6, AB7, AB8 building blocks. The supermonomer strategy greatly increases the number of available end groups. An exemplary AB4 supermonomer is peracetylated β-D-glucopyranoside tetraethylene glycol azide, as shown in structure III.

[0100] The chemical structures listed in Table 1 are also suitable as building blocks for forming branch units of dendritic macromolecules. For example, branch units of dendritic macromolecules are formed from the following: dipentaerythritol, pentaerythritol, 2-(aminomethyl)-2-(hydroxymethyl)propane-1,3-diol, 2-ethyl-2-(hydroxymethyl)propane-1,3-diol, 3,3',3",3"'-silanetetrayltetra(propane-1-thiol), 3,3-divinylpenta-1,4-diene, 3,3',3"-nitrilotripropionic acid, 3,3',3" -nitrilotris(N-(2-aminoethyl)propionamide), 3,3',3",3"-(ethane-1,2-diylbis(azatriyl))tetrapropionamide, 3-(carboxymethyl)-3-hydroxyglutaric acid, 2,2'-((2,2-bis((2-hydroxyethoxy)methyl)propane-1,3-diyl)bis(oxy))bis(ethanol-1-ol), tetrakis(3-(trichlorosilyl)propyl)silane, 1-thioglycerol, 2 ,2,4,4,6,6-hexachloro-1,3,5,2l5,4l5,6l5-triazatriphosphine, 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 -tri(dimethyl(vinyl)silyl)benzene, carbosiloxane core, nitrilotrimethanol, ethylenediamine, propane-1,3-diamine, butane-1,4-diamine, 2,2',2"-nitrilotri(ethanol-1-ol), α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, cucurbituril, benzene-1,2,3,4,5,6-hexathiol, monosaccharides, disaccharides, trisaccharides, oligosaccharides, or their azide, alkyne-modified parts, or combinations thereof.

[0101] Other examples of chemical structures suitable for forming 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 with a core moiety or other identical 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 a linear or branched polyglycerol. In some embodiments, the branching unit is a copolymer of an alkylene glycol (e.g., ethylene glycol) and a sugar moiety (e.g., glucose, galactose, mannose, and / or fructose).

[0102] 3. Surface functional groups

[0103] The surface functional groups / molecules of the dendrimers are not limited to primary amine end groups, hydroxyl end groups, carboxylic acid end groups, acetamide end groups, sugar molecules, oligo- or polyalkylene glycols and / or thiol end groups. In some embodiments, the desired terminal functional groups can be added by one of the conjugation methods of the core and branch units.

[0104] In some embodiments, the surface functional groups are hydroxyl groups, such as hydroxyl groups of PAMAM dendrimers, hydroxyl groups of 2nd generation OEG dendrimers as shown in structure I, or hydroxyl groups of terminal glucose of dendrimers prepared with glucosyl branching units as 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 of the core and branching units. Exemplary surface functional groups include hydroxyl end groups, amine end groups, carboxylic acid end groups, acetamide end groups, and thiol end groups, and combinations thereof.

[0105] In some embodiments, dendrimers can be specifically targeted to specific 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 glial cells of the CNS. In some embodiments, dendrimers specifically target neurons and / or glial cells of the PNS. In some embodiments, dendrimers specifically target non-neural and / or non-glial cells, such as gastrointestinal 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 of generation G1, G2, and / or G3.

[0106] In some embodiments, the dendrimers contain an effective number of terminal glucose and / or hydroxyl groups for targeting one or more neurons and / or glial cells of the CNS, PNS, and / or eye. In some embodiments, the dendrimers contain an effective number of terminal glucose and / or hydroxyl groups for targeting one or more non-neural and / or non-glial cells, such as gastrointestinal cells, cardiovascular cells, and / or immune system cells.

[0107] In some embodiments, the dendrimer is composed of glucose and oligoethylene glycol building blocks. An exemplary first generation glucose dendrimer is shown in Structure VI, while a second generation glucose dendrimer is shown in Structure VIII.

[0108] In some embodiments, the dendrimer has a plurality of surface functional groups, such as hydroxyl (-OH) groups, amine groups, acetamide groups, and / or carboxyl groups (also referred to herein as surface functional groups or peripheral functional 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 / nm 2 For example, in some embodiments, the surface density of surface functional groups (e.g., hydroxyl groups) is greater than 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 OH groups / nm 2 , for example at least 10, 15, 20, 25, 30, 35, 40, 45, 50 or more 50 OH groups / nm 2 In some embodiments, the volume density of surface functional groups (eg, hydroxyl groups) is from about 1 to about 50 groups / nm 3 Between about 5 and about 30 groups / nm 3 Between or about 10 to about 20 groups / nm 3 In further embodiments, the surface density of surface functional groups (e.g., hydroxyl groups) is from about 1 to about 50 groups / nm 2 Between, preferably 5-20 groups / nm 2 (Number of surface functional groups / nm 2 The molecular weight of each surface functional portion is between about 100 Da and about 10 kDa, preferably between about 100 Da and 1000 Da.

[0109] In some embodiments, the amount of surface functional groups (e.g., any of the above, such as hydroxyl groups) of the dendrimer 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 surface hydroxyl content of the dendrimer is preferably greater than 35%.

[0110] In some embodiments, one or more surface functional groups (e.g., any of the above) at the periphery of the dendrimer are further modified by conjugation with one or more carbohydrate molecules and / or one or more polyalkylene glycols (e.g., polyethylene glycol). In these embodiments, the surface density of the terminal carbohydrate moiety / molecule and / or polyalkylene glycol may have any range of hydroxyl groups as described above. Hydroxyl terminated PAMAM dendrimers, PAMAM dendrimers with surfaces modified with sugar moieties (where >10% of the surface groups are modified with sugars (especially glucose)), and glucose dendrimers (where the dendrimer is made of 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 to the peripheral circulation, constructs with a total molecular weight of >50,000 Da are preferred. When the dendrimers are formed of sugar moieties / molecules (e.g., glucose) or include sugar moieties / molecules at the termini, the terminal hydroxyl groups of these dendrimers may be part of the terminal sugar moiety / molecule or additional hydroxyl groups that are not part of the sugar moiety / molecule, or a combination thereof. In some embodiments, all terminal hydroxyl groups are part of a terminal sugar moiety / molecule.

[0111] a. Hydroxyl-terminated dendrimers

[0112] In some embodiments, the dendrimer comprises a plurality of hydroxyl groups. Some exemplary high density hydroxyl-containing dendrimers include commercially available polyester dendrimers, such as hyperbranched 2,2-bis(hydroxy-methyl)propionic acid polyester polymers (e.g., hyperbranched bis-MPA polyester-64-hydroxy, 4th generation), dendritic polyglycerols. In some embodiments, the hydroxyl-terminated dendrimers include hydroxyl-terminated PAMAM dendrimers, particularly G3 to G6 hydroxyl-terminated PAMAM dendrimers, such as G3, G4, G5, and G6 hydroxyl-terminated PAMAM dendrimers.

[0113] In some embodiments, the high-density hydroxyl-containing dendrimer is an oligoethylene glycol (OEG)-based dendrimer. For example, the second-generation OEG dendrimer (D2-OH-60) shown in Structure I can be synthesized using efficient, robust, and atom-economical chemical reactions, such as Cu(I)-catalyzed alkyne-azide click and photocatalyzed thiol-ene click chemistry. Orthogonal supermonomers and supercore strategies can be used to achieve very low-generation high-density polyol dendrimers with minimal reaction steps, for example as described in WO2019094952. In some embodiments, the dendrimer backbone has non-cleavable polyether bonds throughout the structure to avoid disintegration of the dendrimer in vivo and allow such dendrimers to be eliminated from the body as a single entity (non-biodegradable).

[0114]

[0115] Structure I. Second generation (G2) oligoethylene glycol dendrimers

[0116] 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 hydroxyl (-OH) groups is at least 1 OH group / nm 2 (Number of surface hydroxyl groups / nm 2 For example, in some embodiments, per nm 2 The surface density of hydroxyl groups is greater than 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 OH groups / nm 2 , for example at least 10, 15, 20, 25, 30, 35, 40, 45, 50 or more than 50 OH groups / nm 2 In some embodiments, the volume density of hydroxyl groups is between about 1 and about 50 groups / nm 3 Between about 5 and about 30 groups / nm 3 Between or between about 10 and about 20 groups / nm 3 In further embodiments, the surface density of hydroxyl (-OH) groups is from about 1 to about 50, or from 5 to 20 OH groups / nm 2 (number of surface hydroxyl groups / surface area in nm2), and a molecular weight of about 100 Da to 1000 Da. In some embodiments, the amount of surface hydroxyl groups of 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 may have a small fraction of the hydroxyl groups exposed on the outer surface, while the remaining hydroxyl groups are located in the inner core of the dendrimer.

[0117] In some embodiments, the dendrimers specifically target specific tissue regions and / or cell types after administration into the body. In some embodiments, the dendrimers specifically target specific tissue regions and / or cell types without a targeting moiety. In some embodiments, the dendrimers include an effective number of hydroxyl groups for targeting CNS cells and / or PNS cells, such as microglia, astrocytes, and / or neurons associated with a disease, disorder, or injury of the central nervous system or peripheral nervous system. In some embodiments, the dendrimers specifically target specific tissue regions and / or cell types without a targeting moiety, and the active agent bound thereto binds directly to receptors on the surface and / or inside the target neural and / or glial cells.

[0118] Unmodified PAMAM dendrimers with hydroxyl end groups are not as enriched in neurons of the brain and / or retinal ganglion cells (RGCs) of the eye as the 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.

[0119] In a preferred embodiment, the dendrimer contains an effective number of terminal glucose and / or hydroxyl groups for targeting one or more neurons of the CNS or eye. The hydroxyl groups on the surface of the dendrimer are part of a glucose molecule. There are no additional hydroxyl groups other than the glucose molecules on the surface. The number of sugar molecules on the surface is determined by the generation. All generations are expected to target neurons.

[0120] Some exemplary glucose dendrimers include a 1st generation glucose dendrimer with 24 hydroxyl (-OH) end groups, a 2nd generation glucose dendrimer with 96 hydroxyl (-OH) end groups, a 3rd generation glucose dendrimer with 396 hydroxyl (-OH) end groups, and a 4th generation glucose dendrimer with 1584 hydroxyl (-OH) end groups. In a preferred embodiment, the glucose dendrimer is a 2nd generation glucose based dendrimer having 24 glucose molecules at the periphery and 6 embedded glucose molecules in the backbone, which are held together by PEG segments.

[0121] b. Carbohydrate-modified dendrimers

[0122] In some embodiments, the dendrimer comprises one or more carbohydrate molecules at the termini. These terminal carbohydrate molecules can be prepared by conjugating one or more surface functional groups (e.g., amine, carboxyl, or hydroxyl) of the dendrimer to one or more carbohydrate molecules. In a preferred embodiment, prior to carbohydrate conjugation, the dendrimer is a hydroxyl-terminated dendrimer, such as a hydroxyl-terminated PAMAM dendrimer, and one or more hydroxyl groups are conjugated to one or more carbohydrate molecules.

[0123] In some embodiments, hydroxyl-terminated dendrimers modified with surface glucose molecules selectively target central and / or peripheral nerves and / or glial cells in vitro and in vivo; and / or selectively accumulate on the surface and / or inside these targets, so that one or more active agents conjugated thereto bind to one or more receptors on / in the target nerve cells and / or glial cells. In some embodiments, hydroxyl-terminated dendrimers modified with surface glucose molecules selectively target gastrointestinal cells, cardiovascular cells, and / or immune system cells in vivo and in vitro; and / or selectively accumulate on the surface and / or inside these targets, so that one or more active agents conjugated thereto bind to one or more receptors on / in the target gastrointestinal cells, cardiovascular cells, and / or immune system cells.

[0124] In some embodiments, the carbohydrate moiety used to modify one or more surface functional groups of the dendrimer is a monosaccharide. Exemplary monosaccharides suitable for modifying the dendrimer include glucose, glucosamine, galactose, mannose, fructose, dehydroascorbic acid, uric acid, inositol. In some embodiments, the dendrimer is conjugated to glucose and thus comprises glucose as a terminal moiety / molecule. In some embodiments, the hydroxyl-terminated dendrimer is modified with one or more glucose moieties to the dendrimer ("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 (e.g., galactose, mannose, and / or fructose). For example, the carbohydrate moiety is an oligosaccharide, the terminal of which is one or more monosaccharides, including glucose, glucosamine, mannose, fructose, thereby exposing these sugar moieties on the surface for conjugation.

[0125] In preferred embodiments, glucose or hydroxyl terminated PAMAM dendrimers or carbohydrate functionalized dendrimers are conjugated to one or more active agents having affinity for and suitable for direct or indirect binding to one or more serotonin (5HT) receptors, such as 5HT-1A, 5HT-2B, 5HT-2A, 5HT-2B, 5HT-2C, 5HT-3, 5HT-4, 5HT-6, and 5HT-7 receptors. In some embodiments, the dendrimers are conjugated to one or more carbohydrate moieties having affinity for and suitable for binding to one or more norepinephrine (NE) receptors, such as α2A-adrenergic receptors, α2B-adrenergic receptors, α2C-adrenergic receptors, and / or β-adrenergic receptors. In some embodiments, the dendrimer is conjugated to one or more carbohydrate moieties that have affinity for dopamine D1 and D2 receptors and are suitable for direct or indirect binding. 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, such as vesicular monoamine transporter 2 (VMAT2), serotonin reuptake transporter (SERT), norepinephrine 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 AMPA receptors, NMDA receptors, EGFR1 receptors, EGFR2 receptors, histamine (H1) receptors, GABA receptors, and trace amine associated receptor 1 (TAAR1). In some embodiments, dendrimers with or without carbohydrate moieties are bound to one or more active agents that have affinity for and are suitable for transport through one or more of GLUT1, GLUT2, GLUT3, GLUT4, GLUT5, GLUT6, GLUT7, GLUT8, GLUT9, GLUT10, GLUT11, GLUT12, GLUT13, and GLUT14. In further embodiments, the dendrimers are conjugated to one or more glucose and / or glucosamine moieties. In some embodiments, the dendrimers contain carbohydrate moieties that are capable of transporting the active agent to a target cell / receptor, wherein the activity at the target cell or receptor is driven by the active agent. In these embodiments, the carbohydrate and glucose moieties enable better targeting of the drug to the target cell and / or receptor. For example, in some embodiments, the dendrimers are conjugated to one or more glucose and / or glucosamine moieties.In other embodiments, the dendrimer is conjugated to one or more oligosaccharides terminated with a glucose and / or glucosamine moiety, i.e., the glucose and / or glucosamine moiety is exposed on the surface of the dendrimer conjugate, suitable for binding to one or more GLUT, 5HT receptor, NE receptor, DA receptor and / or transporter.

[0126] Fig.13 Schematic representation of the major pharmacological targets of LSD, dimethylhydroxytryptamine phosphate, DMT, MDMA, and ketamine, the signaling cascades involved, hormonal regulation, and the main behavioral consequences following administration in animals and humans.

[0127] In some embodiments, the dendrimer has multiple carbohydrate moieties / molecule, e.g., monosaccharides, e.g., glucose, at its periphery. In some embodiments, the surface density of carbohydrate molecules, e.g., monosaccharides, e.g., glucose, is at least 1 carbohydrate molecule / nm 2 (Number of surface carbohydrate groups / nm 2 In some embodiments, the surface area per nm 2 The surface density of carbohydrate molecules is greater than 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 OH groups / nm 2 , for example at least 10, 15, 20, 25, 30, 35, 40, 45, 50 or more than 50 OH groups / nm 2 For example, per nm 2 The surface density of carbohydrate molecules is greater than 10. In some embodiments, the volume density of surface carbohydrate molecules is from about 1 to about 50 groups / nm 3 , about 5 to about 30 groups / nm 3 or about 10 to about 20 groups / nm 3 In a further embodiment, the surface density of carbohydrate molecules is 2 From about 1 to about 50, from about 5 to about 20 (surface carbohydrate molecules / nm 2 In some embodiments, the surface area of ​​the dendrimer is about 100 Da (about 1000 Da) and the molecular weight of each carbohydrate moiety is about 100 Da to about 1000 Da. In these embodiments, i.e., one or more surface functional groups of the dendrimer are modified to introduce one or more sugar moieties / molecules at the termini, the terminal hydroxyl group can be part of the terminal sugar moiety / molecule or an additional hydroxyl group that is not modified by a sugar moiety / molecule and is therefore not part of a sugar moiety / molecule, or a combination thereof.

[0128] In some embodiments, the weight content of carbohydrate molecules (e.g., monosaccharides, such as glucose) is between about 1% and 40% of the total weight of the glycosylated dendrimer, for example, 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, after conjugation, the carbohydrate moiety is present in an amount of 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. In some embodiments, the conjugation of carbohydrate molecules through one or more surface functional groups occurs through about 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20% or 25% of the total available surface functional groups (preferably hydroxyl groups) of the dendrimer before conjugation. In other embodiments, conjugation of carbohydrate molecules occurs on less than 5%, less than 10%, less than 15%, less than 20%, less than 25%, less than 30%, less than 35%, less than 40% of the total available surface functional groups of the dendrimer prior to conjugation.

[0129] c. Dendrimers modified with polyalkylene glycols

[0130] In some embodiments, the dendrimer contains one or more polyalkylene glycols at the termini. These terminal polyalkylene glycols can be prepared by conjugating one or more surface functional groups (e.g., hydroxyl groups) of the dendrimer with a polyalkylene glycol (e.g., PEG). In some embodiments, prior to conjugation, the dendrimer is a hydroxyl-terminated dendrimer (e.g., a hydroxyl-terminated PAMAM dendrimer), and at least a portion of the surface hydroxyl groups are conjugated with PEG.

[0131] 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 polyalkylene glycols on the surface / nm 2 In some embodiments, each nm 2 The surface density of the polyalkylene glycol is greater than 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 polyalkylene glycols / nm 2 , for example at least 10, 15, 20, 25, 30, 35, 40, 45, 50 or more polyalkylene glycols / nm 2 For example, per nm 2 The surface density of the polyalkylene glycol is greater than 10. In some embodiments, the volume density of the surface polyalkylene glycol is from about 1 to about 50 groups / nm3 , about 5 to about 30 groups / nm 3 or about 10 to about 20 groups / nm 3 In a further embodiment, the surface density of the polyalkylene glycol (eg, PEG) is 2 From about 1 to about 50, from about 5 to about 20 (number of surface polyalkylene glycols / nm 2 The surface area is in units of 100 Da) and has a molecular weight of about 100 Da to about 1000 Da.

[0132] In some embodiments, the weight content of the polyalkylene glycol molecules (e.g., PEG) is about 1% to 40% of the total weight of the PEGylated dendrimer, such as about 2% to 20%, about 5% to 15%, or 9% to 12% of the total weight of the PEGylated dendrimer. For example, in some embodiments, the weight content of the polyalkylene glycol molecules (e.g., PEG) 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.

[0133] In some embodiments, the conjugation of polyalkylene glycol molecules (e.g., PEG) through one or more surface functional groups of the dendrimer occurs through about 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, or 25% of the total available surface functional groups (preferably hydroxyl groups) of the dendrimer before conjugation. In other embodiments, the conjugation of polyalkylene glycol molecules (e.g., PEG) occurs on less than 5%, less than 10%, less than 15%, less than 20%, less than 25%, less than 30%, less than 35%, less than 40% of the total available surface functional groups of the dendrimer before conjugation.

[0134] B. Active Agent

[0135] The dendrimer is conjugated or complexed with one or more hallucinogens. Hallucinogens are a diverse class of drugs that alter a subject's awareness of their surroundings as well as their own thoughts and feelings. Hallucinogens are collectively referred to as "psychedelics" and can be divided into two categories: classic hallucinogens (such as LSD) and dissociative drugs (such as PCP). Both of these hallucinogens can cause hallucinations, or sensations and images that appear real but are not actually real. In addition, dissociative drugs can cause a subject to feel out of control or disconnected from the body and environment. Exemplary hallucinogens that can be conjugated to the dendrimer composition include, but are not limited to, a range of drug classes, including, but not limited to, classic hallucinogens, dissociative agents, and delirium agents. 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 norepinephrine) through selective receptors, thereby increasing the concentration of these specific neurotransmitters in the synaptic cleft. Thus, partial antagonism, functional selectivity, and inverse agonism all play important roles in determining the cellular response to specific neurotransmitter receptor ligands.

[0136] In a preferred embodiment, the dendrimer is conjugated to one or more psychedelic hallucinogens, such as dimethyl-4-hydroxytryptamine, 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), dimethyl-2-hydroxytryptamine phosphate, ibogaine, mescaline, norcyanobacterial toxins, 2C compounds (synthetic hallucinogens belonging to a group of designer agents with structures similar to Ecstasy and MDMA), NBOMes (N-benzylmethoxy derivatives of the 2C family of hallucinogens - 4-iodo-2,5-dimethoxy-N-(2-methoxybenzyl)phenethylamine (25I-NBOMe) for 5-HT 2A / C and 5-HT 1A Serotonin receptors, 3,4-methylenedioxyethylamphetamine (MDE), d-lysergic acid diethylamide (LSD) or their analogs show high binding affinity. In some embodiments, the hallucinogen is functionalized, for example with ester, disulfide, phosphodiester, triglycyl peptide, hydrazine, amide, ether and aminoalkyl linkages, optionally with one or more spacers / linkers, to facilitate conjugation to dendrimers and / or for desired release kinetics.

[0137] Most psychedelic drugs have poor water solubility in the ug / mL range. Thus, in some embodiments, the dendrimer conjugates of these psychedelic drugs can improve water solubility by about 2-fold to about 200-fold (inclusive); about 5-fold to about 150-fold (inclusive); about 10-fold to about 100-fold (inclusive) compared to the free drug (i.e., not conjugated to the dendrimer).

[0138] 1. Classic psychedelics

[0139] The composition may include a dendrimer complexed with one or more classical psychedelics. Classical or serotonergic psychedelic compounds are so named primarily because they interact with the serotonin system and most of them are derived from plants or are semi-synthetic compounds. In some cases, classical psychedelics share part of their chemical structure with the endogenous neurotransmitter serotonin (5-HT) - specifically the indole backbone. However, some of them, such as mescaline, do not have an indole but are still considered serotonergic psychedelics. Typically, classical psychedelics act through serotonin 2A receptor (5HT-2A) agonists.

[0140] The classical hallucinogen conjugated to the dendrimer composition can be one or more of the semisynthetic ergoline LSD, plant-derived tryptamines and / or phenethylamines. Exemplary serotonergic hallucinogens include indolamines, such as dimethyl-2-hydroxytryptamine phosphate and LSD, and phenethylamines, such as mescaline and 2,5-dimethoxy-4-iodoamphetamine (DOI).

[0141] a. Tryptamine

[0142] The classical psychedelic conjugated to the dendrimer composition may be one or more tryptamines. Tryptamines are indoleamine metabolites of the essential amino acid tryptophan. Their chemical structure is defined by an indole (fused benzene and pyrrole rings) and a 2-aminoethyl group on the second carbon (the third aromatic atom, the first being a heterocyclic nitrogen). Tryptamines activate trace amine-related receptors expressed in the brain and modulate the activity of dopaminergic, serotonergic, and glutamatergic systems. In the gut, commensal bacteria convert dietary tryptophan into tryptamines, which activate 5-HT4 receptors and modulate gastrointestinal motility.

[0143] Exemplary psychedelic tryptamines that can be conjugated to the dendrimer include DMT, ethyltryptamine, N,N-diethyltryptamine (DET), dimethyl-4-hydroxytryptamine, and dimethyl-2-hydroxytryptamine phosphate and derivatives thereof.

[0144] i. Dimethyl-2-hydroxytryptamine phosphate

[0145] Dimethyl-2-hydroxytryptamine phosphate is a high-affinity serotonin 5-HT 2Aagonist of 5-HT receptors, which are particularly prominent in the prefrontal cortex. It increases cortical activity due to downstream postsynaptic glutamate effects. 1A , 5-HT 1D and 5-HT 2C Receptors are active, although these are thought to play a minor role in its effects. Dimethyl-2-HT phosphate is well tolerated and has been safe in human studies at oral doses of 8-25 mg and intravenous doses of 1-2 mg.

[0146] Structure of dimethylhydroxytryptamine phosphate

[0147]

[0148] N,N-dimethyltryptamine (DMT or N,N-DMT) is a substituted tryptamine that is both a derivative and a structural analog of tryptamine. DMT has a rapid onset of action, a strong effect, 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 is an agonist for 5-HT1A, 5-HT2A, and 5-HT2C, and has a strong binding affinity for the 5-HT2B receptor. DMT also has affinity for dopamine D1, α1-adrenergic, α2-adrenergic, imidazoline-1, and σ1 receptors. In vitro experiments have also shown that it is a substrate for the cell surface serotonin transporter (SERT) and vesicular monoamine transporter 2 (VMAT2) expressed in platelets. Most of DMT's psychedelic effects can be attributed to functionally selective activation of the 5-HT2A receptor.

[0149] The structure of DMT

[0150]

[0151] b. Lysergic acid diethylamide (LSD)

[0152] The dendrimer composition may be conjugated to one or more lysergic acid amides. The amides of lysergic acid are collectively referred to as lysergic acid amides, and include a variety of compounds that have potent agonist and / or antagonist activity at various serotonin and dopamine receptors. Exemplary lysergic acid amides include, but are not limited to, d-lysergic acid amide (or d-lysergic acid amide; LSA or LAA), lysergic acid diethylamide (LSD), ergometrine, DAM-57, ergotamine, methysergine, 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 lysergic acid amide conjugated to the dendrimer composition is lysergic acid diethylamide (LSD) or a LSD derivative.

[0153] LSD is a semisynthetic ergosterol derived from the natural ergot alkaloid lysergic acid contained in the rye parasite Claviceps sergifer. The mechanism of action of LSD is mainly mediated by activation of serotonin receptors (i.e., 5HT2A receptors or serotonin 2A receptors, 5-HT2ARs) and modulation of 5HT2C and 5HT1A receptors. The interaction between receptor activation and the resulting cognitive impairment and hallucination induction remains less clear. LSD-induced 5-HT2AR activation leads to a breakdown of inhibitory processes in the hippocampal prefrontal cortex. Specifically, LSD decreases brain activity in the right middle temporal gyrus, superior / middle / inferior frontal gyri, anterior cingulate cortex, and left superior frontal and postcentral gyri and cerebellum. Activation of the right hemisphere alters the function of the thalamus and increases activity in the paralimbic structures and frontal cortex, resulting in induced visual imagery.

[0154] The structure of LSD

[0155]

[0156] c. Phenylethylamine

[0157] The dendrimer composition may be conjugated to one or more phenethylamines. Phenylethylamine (PEA) is an organic compound, a natural monoamine alkaloid, and a trace amine that acts as a central nervous system stimulant in humans. In the brain, phenethylamine modulates monoamine neurotransmission by binding to trace amine-associated receptor 1 (TAAR1) and inhibiting vesicular monoamine transporter 2 (VMAT2) in monoamine neurons. To a lesser extent, it also acts as a neurotransmitter in the human central nervous system. Phenylethylamine is produced from the amino acid L-phenylalanine by enzymatic decarboxylation by the enzyme aromatic L-amino acid decarboxylase.

[0158] Phenylethylamine releases norepinephrine and dopamine, induces acetylcholine release through a glutamate-mediated mechanism, and binds to trace amine-associated receptor 1 (TAAR1) as an agonist. Examples of phenethylamines that can be conjugated to the dendrimer composition include mescaline and MDMA.

[0159] i. Mescaline

[0160] Mescaline or mescalin (3,4,5-trimethoxyphenylethylamine) is a naturally occurring psychedelic alkaloid belonging to the class of substituted phenylethylamines and is the active ingredient of psychedelic cacti such as peyote (Lophophora williamsii) and wachuma (Echinopsis pachanoi, also known as San Pedro). Mescaline is biosynthesized from tyrosine, which in turn is generated from phenylalanine by the action of phenylalanine hydroxylase.

[0161] Similar to other classic psychedelics, mescaline is a 5HT 2A / 2C Agonist and one of the most selective serotonergic psychedelics. Mescaline also binds to and modulates the activity of norepinephrine receptors α1 and α2A and TAAR1 receptors. In some forms, the dendrimer composition can be complexed with mescaline in an effective amount of about 300 to about 500 mg. In some forms, the dendrimer composition can be complexed with mescaline in an effective amount of about 6 hours to about 8 hours of hallucinogenic effect.

[0162] Structure of Mescaline

[0163]

[0164] 2. Inducing agents

[0165] Dendrimer compositions may include one or more elicitors. Elicitors are class I monoamine releasers and reuptake inhibitors known to evoke feelings of emotional openness and connection, such as 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA). Exemplary elicitors that may 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-benzodioxol-N-methylbutylamine (MBDB), 6-APB, methyl mephedrone, mephedrone, GBL, αMT, MDAI, and related compounds.

[0166] a.MDMA

[0167] Dendrimer compositions can be complexed with MDMA. MDMA increases the amount of serotonin in the synaptic cleft of serotonergic neurons by inhibiting its entry into neurons and its release directly from neurons. The released serotonin binds to various serotonin receptors and overactivates them, which is the main mechanism of MDMA intoxication. MDMA also induces a large release of norepinephrine.

[0168] Extracellular MDMA binds to the presynaptic serotonin (SERT), norepinephrine (NET), and dopamine transporters (DAT) as a reuptake inhibitor, causing them to take up less of the same monoamine neurotransmitter. MDMA inhibition is most potent for NET and SERT, with much less efficacy for DAT. As a result, more norepinephrine and serotonin remain in the synaptic cleft. These monoamine transporters (SERT, NET, and DAT) reuptake their respective neurotransmitters via ion gradients. Normally, extracellular sodium (Na+) and chloride (Cl–) ion concentrations are high, while potassium (K+) ion concentrations are low, compared to intracellular concentrations. During each reuptake cycle, one monoamine, Na+ and Cl–, is taken up into the cell simultaneously. Then, one intracellular K+ is transported outside the cell. Under experimental conditions, high extracellular K+ concentrations can lead to retrograde transport. MDMA can substitute for K+ for this transport. Therefore, high extracellular MDMA can reverse transport.

[0169] Intracellular MDMA binds to the VMAT2 protein on synaptic vesicles, acting as an inhibitor. Each VMAT2 transports cytoplasmic monoamines into the vesicle by dissipating the proton gradient across the vesicle membrane. VMAT2 inhibition therefore results in more free cytoplasmic monoamines, such as serotonin (in the case of serotonin neurons). These monoamines can then be released via the monoamine transporters, which MDMA reverses. Intracellular MDMA can also bind to monoamine oxidase A (MAO-A), acting as an inhibitor, thereby preventing it from breaking down cytoplasmic serotonin.

[0170] MDMA binds as an agonist to the following receptors: 5-HT1A-, 5-HT2A-, 5-HT2B-, 5-HT2C-serotonin receptors; α1-, α2A-, β-adrenaline receptors, D1 and D2 dopamine receptors, M1 and M2 muscarinic receptors, H1 histamine receptor and TAT (TAAR1) receptor.

[0171] The structure of MDMA

[0172]

[0173] 3. Separation of Anesthetics

[0174] Another class of drugs that have hallucinogenic properties and can be conjugated to the dendrimer compositions are psychedelic or dissociative anesthetics, including arylcyclohexylamines (also known as arylcyclohexylamines). Arylcyclohexylamines are a class of compounds that contain a cyclohexylamine unit and an aryl moiety (usually a benzene ring) attached to the atom to which the amine group is attached. They all exhibit a dissociative effect in that they have antagonistic effects on N-methyl-d-aspartate (NMDA) receptors. Exemplary arylcyclohexylamines that can be conjugated to the dendrimer compositions include phencyclidine (1-(1-phenylcyclohexyl)piperidine; PCP), dextromethorphan (DXM), nitrous oxide, ketamine, and mephedrone (ketamine analogs).

[0175] a. Ketamine

[0176] The dendrimer composition can be conjugated to ketamine (R-ketamine, S-ketamine, (R / S)-ketamine), ketamine analogs, ketamine metabolites, 2-(2-chlorophenyl)-2-(methylamino)-cyclohexanone or analogs thereof (e.g., KEA-1010, mephedrone, norketamine, and 2-fluoronorchloroketamine).

[0177] (Ketamine) is a non-barbiturate dissociative anesthetic. Its beneficial effect is to "dissociate" brainstem functions from higher brain areas, thereby changing the perception of pain and other stimuli during medical procedures and producing amnesia for the event. It is a cyclohexanone derivative with rapid action and significant anesthetic and analgesic effects. Its chemical name is ±)-2-(o-chlorophenyl)-2-(methylamino)cyclohexanone hydrochloride; its structural formula is CHClNO. Ketamine is a non-competitive N-methyl-D-aspartate (NMDA) and glutamate receptor antagonist. It blocks HCN1 receptors. The unique dissociative effect and partial agonism of opioid mu receptors allow painful surgery to be performed under continuous sedation and patient comfort.

[0178] Ketamine's effects on chronic pain and antidepressant effects, as compared to actual drug levels, may be mediated by secondary increases in structural synaptic connectivity mediated by the response of neurons to the ketamine-induced hyperglutamatergic state.

[0179] Ketamine may interact with sigma receptors. It tends to act by reducing central sensitization, end phenomenon (development of persistent, exacerbated, or chronic pain), and pain memory. Cholinergic, aminergic, and opioid systems appear to play both positive and negative regulatory roles in sedation and analgesia. Ketamine may reverse tolerance to opioids. It is metabolized through the hepatic system by N-dealkylation, hydroxylation, conjugation, and dehydration. Ketamine has a half-life of approximately 45 minutes.

[0180] In some forms, the recommended dose of ketamine for induction of anesthesia is approximately 1 to 4.5 mg / kg IV and 6.5 to 13 mg / kg IM, with an off-label recommendation of 0.5 to 2 mg / kg IV and 4 to 10 mg / kg IM, primarily for adjunctive use. For depression, ketamine is most commonly administered at a dose of 0.5 to 0.5 mg / kg IV over 40 minutes, which is below the anesthetic dose.

[0181] Conjugation to dendrimers may further improve the safety and efficacy of ketamine. For example, dendrimer conjugation may alter specific receptor activity and / or alter biodistribution (e.g., using higher generation dendrimers to confine ketamine to the peripheral nervous system to exclude its psychoactive effects).

[0182] Structure of Ketamine

[0183]

[0184] b.PCP

[0185] Phencyclidine (PCP) (also known as phencyclidine) is a hallucinogen, specifically a dissociative anesthetic, that produces a variety of physical and behavioral effects. PCP can cause hallucinations and distorted perception of sounds.

[0186] The most unusual feature of PCP is that oral doses of 5 to 10 mg may induce acute schizophrenia, including agitation, psychosis, visual and auditory hallucinations, paranoid delusions, and catatonia. Doses above 10 mg usually result in coma. More than 50% of adult patients experience typical symptoms of PCP poisoning: violent behavior, nystagmus, tachycardia, hypertension, anesthesia, and analgesia.

[0187] PCP acts primarily on NMDA receptors, an ionotropic glutamate receptor, and is an NMDA receptor antagonist. PCP also inhibits nicotinic acetylcholine receptors (nAChR). In some forms, dendrimer compositions include PCP analogs that differ in potency for nACh receptors and NMDA receptors. PCP-induced interactions between presynaptic nAChRs and NMDA receptors affect postsynaptic maturation of glutamatergic synapses, thereby affecting synaptic development and plasticity in the brain. These effects may result in inhibition of excitatory glutamate activity in certain brain regions, such as the hippocampus and cerebellum.

[0188] PCP, like ketamine, is a potent dopamine D2 receptor partial agonist with affinity for cloned D2High receptors. This activity may be associated with some of the other more severe psychotic features of PCP intoxication, as evidenced by the successful use of D2 receptor antagonists such as haloperidol in the treatment of PCP psychosis.

[0189] The structure of PCP

[0190]

[0191] 4. Ibogaine and its analogs and derivatives

[0192] Ibogaine is an indole alkaloid isolated from the root of the West African shrub Tabernanthe iboga. The therapeutic and dream-interpreting (dream-like) effects of ibogae root have been described for centuries in ethnobotanical literature, where ibogaine root preparations were ingested for ritual and medicinal purposes.

[0193] U.S. Patent No. 4,499,096 (granted to Lots of, HS) describes a method of interrupting a narcotic addiction syndrome by administering ibogaine. Oral ibogaine is described as an effective treatment for opioid detoxification. Subsequent studies have shown that when 500-1000 mg of ibogaine is administered, it can reduce drug cravings and improve depressive symptoms. This dose range appears to be a safe and effective treatment for interrupting opioid addiction syndrome; however, safety and cardiotoxicity issues remain. Similar benefits have been observed in recently withdrawn cocaine abusers attempting to interrupt their intractable drug abuse cycle.

[0194] The molecular structures of ibogaine and neubogaine show that ibogaine undergoes O-demethylation under the action of cytochrome P4502D6 (CYP2D6) to form 12-hydroxyibogamine (neubogaine). Ibogaine is metabolized to neubogaine in the intestinal wall and liver.

[0195] 18-MC is a synthetic derivative of ibogaine, an alpha-3-beta-4 nicotinic receptor antagonist with a different mechanism of action that modulates excessive dopamine fluctuations in the mesolimbic system of the brain. 18-MC is a synthetic organic molecule centered on the coronarone chemical skeleton, which is common to a variety of botanical medicinal compounds such as ibogaine. In preclinical efficacy models, 18-MC has shown potent activity in reducing withdrawal symptoms and self-administration of opioids, stimulants, and other substances of abuse. Extensive preclinical characterization has shown that 18-MC has a strong safety and tolerability profile. 18-MC has the potential to overcome the safety limitations of ibogaine and has not demonstrated proarrhythmic or neurotoxic activity. Other ibogaine derivatives include ME-18-MC, 18-MAC, voacangine, ibogamine, and coronaridine.

[0196] 5. Atypical hallucinogens

[0197] In some embodiments, the active agent is one or more atypical hallucinogens with different mechanisms, such as gamma-9-tetrahydrocannabinol or delta-9-tetrahydrocannabinol (THC) and ibogaine.

[0198] C. Coupling Agents and Spacers

[0199] Dendrimer-active agent conjugates can be formed by covalent conjugation or non-covalent attachment of one or more active agents to a dendrimer. In a preferred embodiment, one or more active agents are covalently conjugated to a dendrimer.

[0200] 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 connect the active agent to the dendrimer. A spacer can be a single chemical entity or two or more chemical entities linked together. Spacers can include any small chemical entity, peptide, or polymer with a thiol, thiopyridine, succinimidyl, maleimide, vinyl sulfone, carbonate, etc.

[0201] In some embodiments, the spacer to which the active agent is conjugated to the dendrimer comprises different bonds, such as disulfide bonds, ester bonds, carbonate bonds, carbamate bonds, thioester bonds, hydrazine bonds, hydrazide bonds, ether bonds, and amide bonds. 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, the conjugation between the active agent and the dendrimer is carried out through an appropriate spacer, which comprises 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 the desired and effective release kinetics in vivo. These spacers can contain cleavable linkages (e.g., esters, disulfide bonds, phosphodiesters, triglycyl peptides, and hydrazines) or non-cleavable linkages (e.g., amides, ethers, and aminoalkyls). The conjugation between the active agent and the dendrimer can be carried out using reactions known in the art, such as click chemistry, acid amine coupling, Steglich esterification, and the like.

[0202] In some embodiments, the conjugation between the active agent and the dendrimer is performed through a spacer comprising a disulfide bond, an ester bond, an ether bond, a thioester bond, a carbamate bond, a carbonate bond, a hydrazine bond, an ether bond, or an amide bond, or a combination thereof. In some embodiments, the conjugation between the active agent and the dendrimer is performed through an appropriate spacer comprising an ester bond or an amide bond between the agent and the dendrimer, depending on the desired release kinetics of the agent.

[0203] The spacer can be selected from compounds terminated by thiol, thiopyridine, succinimidyl, maleimide, vinyl sulfone and carbonate. The spacer can include compounds terminated by thiopyridine, 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 may also include peptides, wherein the peptide is linear or cyclic, essentially having a thiol 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). The spacer may be a thiol acid derivative, such as 3-mercaptopropionic acid, thioglycolic acid, 4-mercaptobutyric acid, thiolane-2-one, 6-mercaptohexanoic acid, 5-mercaptopentanoic acid and other thiol derivatives, such as 2-mercaptoethanol and 2-mercaptoethylamine. The spacer can be thiosalicylic acid and its derivatives, (4-succinimidyloxycarbonyl-methyl-α-2-pyridylthio) toluene, (3-[2-pyridylthio] propionyl hydrazide, the spacer can have a maleimide terminal group, wherein the spacer includes a polymer or a small chemical entity, such as bismaleimido diethylene glycol and bismaleimido triethylene glycol, bismaleimido ethane, bismaleimido hexane. The spacer can include vinyl sulfones, such as 1,6-hexane-bisvinyl sulfone. The spacer can include a thioglycoside, such as thioglucose. The spacer can be a reduced protein, such as bovine serum albumin and human serum albumin, any thiol-terminated compound capable of forming a disulfide bond. The spacer can include polyethylene glycol with maleimide, succinimidyl and / or thiol terminal groups.

[0204] D. Dendrimer-Agent Conjugates or Complexes

[0205] Dendrimer-active agent conjugates can be formed by covalently conjugating or non-covalently attaching an antidepressant and / or antipsychotic to a dendrimer, dendrimer, or hyperbranched polymer. Methods for conjugating one or more active agents to dendrimers are known, such as those described in U.S. Publications 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 alter specific receptor activity and / or alter biodistribution. For example, the use of higher generation dendrimers and / or dendrimers with a molecular weight greater than 24 kDa can confine these agents to the peripheral nervous system to exclude their psychoactive effects.

[0206] In some embodiments, one or more active agents are covalently conjugated to one or more terminal groups (e.g., terminal hydroxyl groups) of the dendrimer. In some embodiments, the dendrimer conjugates include 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 can be cleavable or contain a cleavable chemical bond, for example, cleaved by exposure to an intracellular compartment of a target nerve and / or glial cell or binding to a receptor on the surface or inside of a target nerve and / or glial cell in vivo. Examples of cleavable bonds of spacers that can be used in dendrimer-active agent conjugates include esterase-sensitive ester bonds, glutathione-sensitive disulfide bonds, phosphatase-sensitive phosphodiester bonds, oligopeptides (e.g., triglycyl peptide linkers capable of lysosomal release), acid-cleavable hydrazine bonds, and the like. In some embodiments, the spacer between the dendrimer and the active agent can provide desired and effective release kinetics in vivo. In some embodiments, the spacer between the dendrimer and the active agent may be non-cleavable or comprise a non-cleavable chemical bond, such as an amide bond, an ether bond, and an aminoalkyl bond.

[0207] Typically, the spacer between the dendrimer and the active agent is of sufficient length to enable the active agent conjugated thereto to reach and bind to a target receptor on the surface and / or inside the target cell. For example, the length of the spacer between the dendrimer and the active agent ranges from 50 Da to 2000 Da, depending on the desired release kinetics 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 when located on the cell surface.

[0208] Dendrimers can be of generation 2, 3, 4, 5, 6, up to generation 10. In some embodiments, the dendrimer is conjugated to one or more active agents via a spacer containing a cleavable (ester, disulfide, phosphodiester, triglycine, and hydrazine) or non-cleavable (amide, ether, and aminoalkyl) bond.

[0209] The density of active agents covalently bound or non-covalently attached to the dendrimer can be adjusted depending on the specific antidepressant and / or antipsychotic drug being delivered, the target receptor, the target nerve and / or glial cell, the location of the target nerve and / or glial cell, etc. For example, multiple active agents conjugated to the dendrimer are located at the periphery of the dendrimer and the surface density of the active agents is at least 1 active agent / nm 2 (Number of conjugated active agents / nm 2For example, in some embodiments, per nm 2 The surface density of the active agent is greater than 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 OH / nm 2 , for example at least 10, 15, 20, 25, 30, 35, 40, 45, 50 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 , about 5 to about 30 groups / nm 3 or about 10 to about 20 groups / nm 3 .

[0210] Typically, the dendrimer-active agent conjugate has a hydrodynamic volume in the nanometer range. For example, in some embodiments, the diameter of the glucose dendrimer-active agent conjugate comprising one or more antidepressants and / or antipsychotics conjugated to the dendrimer is from about 2 nm to about 100 nm, or greater than 100 nm, up to 500 nm, depending on the generation of the dendrimer, the chemical composition and amount of the active agent conjugated thereto. In some embodiments, the dendrimer-active agent conjugate comprising one or more antidepressants and / or antipsychotics conjugated to the dendrimer has a diameter that is effective to penetrate brain tissue and remain on the surface and / or in the target neural and / or glial cells for a period of time sufficient for the active agent to bind to a targeted receptor on the surface and / or in the target neural and / or glial cells. In some embodiments, the dendrimer-active agent conjugates comprising one or more antidepressant and / or antipsychotic drugs bound to the dendrimer have a diameter effective to remain in the peripheral circulation and to remain on the surface and / or in target neural and / or glial cells long enough to allow the active agent to bind to a targeted receptor on the surface and / or in target neural and / or glial cells (e.g., neural and / or glial cells of the gastrointestinal system).

[0211] The dendrimer-active agent conjugate can be neutral, have a positive charge or a negative charge. In some embodiments, the dendrimer-active agent conjugate is neutral. The presence of an antidepressant and / or antipsychotic can affect the surface charge of the dendrimer-active agent conjugate. In some embodiments, the surface charge of the 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 ​​from -100 mV to 100 mV. In preferred embodiments, the surface charge of the dendrimer-active agent conjugate is neutral or nearly neutral, ie, from about -10 mV to about 10 mV, inclusive.

[0212] An exemplary dendrimer-active agent conjugate is represented by formula (I). The dendrimer of the exemplary conjugate contains surface hydroxyl groups, wherein one or more surface hydroxyl groups are conjugated to one or more active agents via one or more spacers as shown below in formula (I).

[0213]

[0214] wherein D can be a 1st to 10th generation or a 2nd to 10th generation dendrimer, such as any of the above, such as PAMAM (e.g., a hydroxyl-terminated PAMAM dendrimer) or a glucosyl dendrimer; each occurrence of L can be any suitable chemical moiety, preferably containing a triazole moiety; Y can be a bond or can be selected from a secondary amide (-CONH-), a tertiary amide (-CONR-), a sulfonamide (-S(O)2-NR-), a secondary carbamate (-OCONH-; -NHCOO-), a tertiary carbamate (-OCONR-; -NRCOO-), a carbonate (-OC(O)-O-), a urea (-NHCONH-; -NRCONH-; -NHCONR-, -NRCONR-), a methanol (-CHOH-, -CROH-), a disulfide bond, a phosphodiester group

[0215]

[0216] A linkage of hydrazine, hydrazone, hydrazide, ester (-C(O)-O-), ether (-O-) and oligopeptide (e.g., triglycyl peptide), wherein R is alkyl, aryl or heterocyclic; each occurrence of X can be an antidepressant and / or antipsychotic, wherein the functional group of X (e.g., amino, including primary, secondary or tertiary amino; carboxyl; or hydroxyl) forms part of the linkage Y; n can be an integer from 1 to 100; m can be an integer from 16 to 4096. The dendrimer can be a PAMAM (e.g., hydroxyl-terminated PAMAM) or a glucose dendrimer, which is 100% hydroxyl. m and n depend on the size of the dendrimer D, and n should be such that the weight percentage of the drug in the total conjugate is 5-20%. This range also applies to binding and internalization.

[0217] The oxygen atoms in formula (I) are derived from surface functional groups of the dendrimer, such as surface hydroxyl groups, wherein the surface hydroxyl groups may or may not be part of a terminal sugar moiety / molecule (e.g., glucose). Although not illustrated in formula (I), one or more hydroxyl groups in the dendrimer that are not conjugated to an active agent may be modified with one or more carbohydrates and / or polyalkylene glycols (e.g., PEG).

[0218] When administered to a subject in need thereof, the antidepressant and / or antipsychotic X of formula (I) can bind to a target receptor on the surface of a target cell or inside a target cell. In some embodiments, when the antidepressant and / or antipsychotic 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 remain conjugated to the dendrimer as a complete dendrimer-active agent conjugate. In some embodiments, the antidepressant and / or antipsychotic X is released from the dendrimer at a location close to the target receptor and then binds to the target receptor.

[0219] In some embodiments, each occurrence of L can be represented by -A'-L1-B'-L2-, wherein A' can be a carbonyl (-C(O)-) or a bond (including single bonds, double bonds and triple bonds, such as single bonds); B' can be a bond (including single bonds, double bonds and triple bonds, such as single bonds), amide, ester, ether, thiol, dithiol, aryl, heteroaryl, polyaryl, heteropolyaryl or heterocycle; and L1 and L2 can independently be a bond, alkylene, heteroalkylene, aryl, aralkyl, ether, polyether, thiol, dithiol, thiol ether, polythioether, oligopeptide, polypeptide, oligo(alkylene glycol) or polyalkylene glycol, or L1 and L2 can independently be composed of a combination 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, 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.

[0220] In some embodiments, B' can be a bond (including single, double, and triple bonds, such as a single bond), an amide group, or a heterocyclic group, such as a triazole group.

[0221] In some embodiments, L1 can be a bond; an alkylene group, such as C1-C 10 alkylene, C1-C8 alkylene, C1-C6 alkylene, C1-C5 alkylene, C1-C4 alkylene or C1-C3 alkylene; or oligo- or poly-(alkylene glycol), for example

[0222]

[0223] 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.

[0224] In some embodiments, L2 can be a bond; an alkylene group, such as C1-C 10 alkylene, C1-C8 alkylene, C1-C6 alkylene, C1-C5 alkylene, C1-C4 alkylene or C1-C3 alkylene; oligo- or poly-(alkylene glycol), for example

[0225]

[0226] 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 oligopeptide or polypeptide, such as a triglycyl peptide; a thiol; or a dithiol; or L2 is composed of a combination of two or more of an alkylene, an oligopeptide or poly(alkylene glycol), an oligopeptide or polypeptide, a thiol, and a dithiol. For example, L2 is represented by

[0227]

[0228] 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, a dithiol, an oligopeptide (e.g., a triglycyl peptide) or a polypeptide.

[0229] In some embodiments, Y is the least cleavable linkage in vivo. In some embodiments, Y is a cleavable linkage in vivo. In some embodiments, Y is an amide (-CONH-), an ester (-C(O)-O-), an ether (-O-), a phosphodiester, or a disulfide.

[0230] In some embodiments, L and Y are both single bonds, and D is directly conjugated to X (active agent or analog thereof) through an ether bond.

[0231] In some embodiments, D is a 2nd generation PAMAM dendrimer, a 3rd generation PAMAM dendrimer, a 4th generation PAMAM dendrimer, a 5th generation PAMAM dendrimer, a 6th generation PAMAM dendrimer, a 1st generation glucose dendrimer, a 2nd generation glucose dendrimer, a 3rd generation glucose dendrimer, a 4th generation glucose dendrimer, a 5th generation glucose dendrimer, or a 6th generation glucose dendrimer.

[0232] More specific exemplary dendrimer-active agent conjugates are shown in the Examples below.

[0233] E. Exemplary Dendrimer-Agent Conjugates

[0234] In a preferred embodiment, the dendrimer is conjugated to serotonin or a serotonin analog as shown below in structures AD.

[0235]

[0236] n is an integer from 1 to 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0237] In a preferred embodiment, the dendrimer is conjugated to ketamine or a ketamine analog, as shown in Structures E and F below.

[0238]

[0239] In other preferred embodiments, the dendrimer is conjugated to DMT or a DMT analog as shown below in Structure GJ.

[0240]

[0241]

[0242] In a preferred embodiment, the dendrimer is conjugated to LSD or an LSD analog as shown in structures K and L below.

[0243]

[0244]

[0245] III. Methods for Preparing Dendrimer Conjugates

[0246] Methods of synthesizing dendrimers and making dendrimer nanoparticles are also described.

[0247] A. Methods of Preparing Dendrimers

[0248] Dendrimers can be prepared by a variety of chemical reaction steps. Dendrimers are usually synthesized according to methods that allow control of their structure at every stage of their construction. Dendritic structures are mainly synthesized by two main different methods: divergent or convergent.

[0249] In some embodiments, dendrimers are prepared using a different approach in which the dendrimer is assembled from a multifunctional core that is extended outward through a series of reactions, typically Michael reactions. This strategy involves coupling monomer molecules with reactive and protective groups to a multifunctional core moiety, which results in a stepwise increase in the number 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 amine core.

[0250] In other embodiments, dendrimers are prepared using a convergent approach, where the dendrimer is built from small molecules that eventually reach the surface of the sphere, and the reactions proceed inward, building inward and ultimately attaching to the core.

[0251] There are many other synthetic routes for the preparation of dendrimers, such as the orthogonal method, the accelerated method, the two-stage convergence method or the supercore method, the supermonomer method or the branched monomer method, the double exponential method; the orthogonal coupling method or the two-step method, the double monomer method, and the AB2-CD2 method.

[0252] In some embodiments, the core, one or more branching units, one or more spacers, and / or one or more surface functional groups of the dendrimer may be modified to allow conjugation with further functional groups (branching units, spacers, surface functional groups, etc.), monomers, and / or agents by click chemistry using one or more copper-assisted azide-alkyne cycloadditions (CuAAC), Diels-Alder reactions, thiol-ene and thiol-alkyne reactions, and azide-alkyne reactions (Arseneault M et al., Molecules, 2015 May 20; 20(5): 9263-94). In some embodiments, preformed dendrimers are clicked onto high density hydroxyl polymers. "Click chemistry" involves, for example, the coupling of two different moieties (e.g., a core group and a branching unit; or a branching unit and a surface functional group) through a 1,3-dipolar cycloaddition reaction between an alkyne moiety (or its equivalent) on the surface of the first moiety and an azide moiety (e.g., present on 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 the 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 alkynyl group. The azide and alkynyl groups are then reacted by a 1,3-dipolar cycloaddition to form a triazole moiety.

[0253] In some embodiments, the synthesis of dendrimers relies on one or more reactions such as thiol-ene click reaction, thiol-alkyne click reaction, CuAAC, Diels-Alder click reaction, azide-alkyne click reaction, Michael addition, epoxy ring opening, esterification, silane chemistry, and combinations thereof.

[0254] In some embodiments, the method involves one or more steps of protecting and deprotecting 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, or the addition of therapeutic, prophylactic, or diagnostic agents to generate the desired dendrimer conjugate. In the case of hydroxyl groups, they can be protected by forming ethers, esters, or acetals. Other exemplary protecting groups include Boc and Fmoc.

[0255] Any existing dendritic platform can be used to prepare dendrimers with desired functionality, i.e., dendrimers with a high density of surface hydroxyls by conjugating high hydroxyl content moieties such as 1-thioglycerol or pentaerythritol. Exemplary dendritic platforms such as polyamidoamine (PAMAM), poly(propyleneimine) (PPI), poly-L-lysine, melamine, poly(ether hydroxylamine) (PEHAM), poly(esteramine) (PEA), and polyglycerol can be synthesized and explored.

[0256] Dendrimers can also be prepared by combining two or more dendrimers. A dendrimer is a wedge-shaped portion of a dendrimer with a reactive focal functional group. Many dendrimer scaffolds are commercially available. They are available in 1, 2, 3, 4, 5, and 6 generations, with 2, 4, 8, 16, 32, and 64 reactive groups, respectively. In certain embodiments, one type of agent is linked to one type of dendrimer, and a different type of agent is linked to another type of dendrimer. The two dendrimers are then linked to form the dendrimer. Two dendrimers can be linked by click chemistry, a 1,3-dipolar cycloaddition reaction between an azide moiety on one dendrimer and an alkyne moiety on another dendrimer to form a triazole linker.

[0257] Exemplary methods for preparing dendrimers are described in detail in International Patent Publication Nos. WO2009 / 046446, WO2015168347, WO2016025745, WO20160.25741, WO2019094952, and U.S. Pat. No. 8,889,101.

[0258] 1. Method for preparing glucose dendrimers

[0259] Glucose-based dendrimers are assembled from a multifunctional core that extends outward through a series of reactions. The strategy involves coupling monomer molecules with reactive and protective groups to the multifunctional core moiety, leading to a stepwise increase in the number of generations around the core, followed by the removal of the protecting groups.

[0260] 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 supercore 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 1.

[0261]

[0262] Scheme 1. Synthesis of hypernuclei

[0263] In some embodiments, the branching unit is a supermonomer, i.e., AB n Building units. Exemplary supermonomers include AB3, AB4, AB5, AB6, AB7, AB8 building units. The supermonomer strategy greatly increases the number of available end groups. An exemplary supermonomer is the AB4 orthogonal supermonomer, which includes an azide functional group and four allyl groups and is prepared by reacting dipentaerythritol having five allyl groups with monotosylated triethylene glycol azide.

[0264] In some embodiments, the branching unit is a linear or branched polyglycerol, for example, as shown in Formula III. Other monomers include disaccharides and oligosaccharides, and sugars such as fructose, lactose, and sucrose.

[0265] a. Synthesis of AB4 building blocks

[0266] Some exemplary synthesis methods of supermonomers AB4 are described below. In some embodiments, supermonomers AB4 are based on glucose molecules. In preferred embodiments, supermonomers AB4 are conjugated with polyethylene glycerol (e.g., tetraethylene glycol (PEG4)). In one embodiment, supermonomers AB4 are peracetylated β-D-glucopyranoside tetraethylene glycol azide.

[0267] In some embodiments, the synthesis of glucose-Oac-TEG-Ots involves the following steps: a solution of peracetylated β-D-glucopyranoside (10 g, 25.6 mmol) is dissolved in 50 mL of anhydrous dichloromethane (DCM), followed by the addition of 2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (6.2 g, 17.9 mmol), and the reaction mixture is cooled to 0°C. Boron trifluoride etherate (2.5 eq.) is added and the reaction is returned to room temperature. The reaction is monitored by TLC, and the reaction is quenched after 5 hours by the addition of a saturated sodium bicarbonate solution at 0°C. After stirring for 10 minutes, DCM (300 mL) is added and the organic layer is washed 3 times with a saturated sodium bicarbonate solution until the effervescence is quenched. The reaction mixture is dried over sodium sulfate, filtered, and evaporated under reduced pressure. The crude product is purified by combiflash chromatography using an ethyl acetate / hexane (70:30) mixture as an eluent. The desired compound was obtained with a yield of 60%. The structure of Glucose-Oac-TEG-Ots is shown below:

[0268]

[0269] 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, sodium azide (2 eq) is then added, and the reaction mixture is heated to 50 ° C overnight. After completion, the reaction mixture is filtered and DMF is evaporated. After drying, the crude reaction mixture is passed through combiflash using ethyl acetate: hexane (70:30) as an eluent. The structure of glucose-Oac-TEG-N3 is as follows:

[0270]

[0271] In some embodiments, the synthesis of glucose-OH-TEG-N3 comprises 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 at room temperature overnight, then diluted with methanol, and the pH is adjusted to about 6-7 with Amberlist IR-120+. The reaction mixture is separated by filtration, and the solvent is removed by rotary evaporation. The structure of glucose-OH-TEG-N3 is shown below.

[0272]

[0273] b. Synthesis of Glucose Dendrimers

[0274] 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 hexapropargylated core is linked to the AB4 β-D-glucose-PEG4-azide building unit (2) via a click reaction to obtain a first generation dendrimer.

[0275] In some embodiments, the first generation dendrimer D1-Glu6-Oac24 is prepared according to the following steps: The hexapropargylated compound (0.5 g, 1 mmol) and the azido derivative ((4.1 g, 7.4 mmol), 1.2 equivalents / 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 reactants are irradiated in a microwave at 50°C for 6 hours. The reaction mixture is dialyzed against DMF and then dialyzed against water containing EDTA. EDTA is further removed by dialysis against a large amount of water. The product is lyophilized to obtain D1-Glu6-Oac24. The structure of D1-Glu6-Oac24 is shown below.

[0276]

[0277] In some embodiments, the first generation dendrimer D1-Glu6-OH 24 The preparation of the first generation peracetylated glucose dendrimer (1 g, 0.26 mmol) was as follows: peracetylated first generation glucose dendrimer (1 g, 0.26 mmol) was dissolved in anhydrous methanol and sodium methoxide was added to adjust the pH to about 8.5-9. The reaction was stirred at room temperature overnight and then diluted with methanol and IR-120+ adjusts the pH to about 6-7. The reaction mixture is separated by filtration, the solvent is removed by rotary evaporation, and then dialyzed against water. The structure of the first generation glucose dendrimer D1-Glu6-OH24 is shown below.

[0278]

[0279] In some embodiments, the first generation glucose dendrimer D1-Glu6-OH24 is propargylated to provide D1-Acetylene24 as follows: D1-Glu6-OH24 (2 g, 0.721 mmol) is dissolved in anhydrous dimethylformamide (DMF, 50 mL) by ultrasound. Sodium hydride [60% dispersed in mineral oil] (951 mg, 39.65 mmol) is slowly added to the solution in portions under stirring at 0°C. The solution is stirred for another 15 minutes at 0°C. Propargyl bromide (3.85 mL, 34.608 mmol, 80% w / w toluene solution) is then added at 0°C, and stirring is continued for another 6 hours at room temperature. The reaction mixture is quenched with ice and water, filtered, dialyzed with DMF, and then dialyzed with water to obtain D1-acetylene 24. The structure of D1-acetylene 24 is shown below.

[0280]

[0281] In some embodiments, the first generation dendrimer D1-acetylene 24 is further reacted with AB4β-D-glucose-PEG4-azide to yield a second generation dendrimer with 24 glucose molecules and 96 surface hydroxyl groups.

[0282] The preparation method of the exemplary second generation dendrimer D2-Glu24-Oac96 is 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. CuSO4·5H2O (5 mol% / acetylene, 5 mg) and sodium ascorbate (5 mol% / acetylene, 10 mg) dissolved in a minimum amount of water were added thereto. The reactants were irradiated in a microwave at 50°C for 8 hours. Upon completion, the reaction mixture was dialyzed against DMF and then against water containing EDTA. EDTA was further removed by dialysis against a large amount of water. The product was lyophilized to obtain D2-Glu24-Oac96.

[0283] In some embodiments, the preparation method of the second-generation dendritic macromolecule D2-Glu24-OH96 is as follows: dissolve the peracetylated second-generation glucose dendritic macromolecule D2-Glu24-OH96 in anhydrous methanol, add sodium methoxide to adjust the pH value to about 8.5-9.0, stir the reaction at room temperature overnight, then dilute with methanol, IR-120+ adjusts the pH value to about 6-7. The reaction mixture is filtered to remove the resin, and the filtrate is rotary evaporated and dialyzed against water to obtain an off-white solid product.

[0284] The structure of the second generation glucose dendrimer D2-Glu24-OH96 is shown below.

[0285]

[0286] In some embodiments, the second generation dendrimer D2-Glu24-OH96 is propargylated at one or more terminal hydroxyl groups and is suitable for further conjugation with one or more therapeutic, prophylactic or diagnostic agents. In some embodiments, one or more terminal hydroxyl groups of the second generation dendrimer D2-Glu24-OH96 are propargylated as follows: D2-Glu24-OH96 (5b) (200 mg, 0.016 mmol) was dissolved in anhydrous dimethylformamide (DMF, 10 mL) by ultrasound. Sodium hydride [60% dispersion in mineral oil] (22 mg, 0.934 mmol) was slowly added in portions to the stirred solution at 0°C. The solution was stirred for an additional 15 minutes at 0°C. Propargyl bromide (18.0 μL, 80% weight / weight solution in toluene) was then added at 0°C and stirring was continued for 6 hours at room temperature. The solvent was evaporated using a V10 evaporator system and the mixture was filtered through a PD10 The crude product was purified by G25 M column. The aqueous solution was lyophilized to obtain an off-white solid product.

[0287] In some embodiments, one or more fluorescent dyes (e.g., infrared fluorescent Cy5 dye) are conjugated to the second generation dendrimer D2-Glu24-OH96. In one embodiment, Cy5-D2-Glu24-OH96 ( Figure 1B Compound 7 in (200 mg, 0.016 mmol) was prepared as follows: Compound 6 (200 mg, 0.016 mmol) and Cy5 azide (20.7 mg, 0.02 mmol) were suspended in a 1:1 mixture of DMF and water in a 25 mL round bottom flask equipped with a magnetic stir bar. CuSO4·5H2O (5 mol% / acetylene, 0.3 mg) and sodium ascorbate (10 mol% / acetylene, 0.5 mg) dissolved in a minimum amount of water were added thereto. The reaction was stirred at room temperature for 24 hours. After the reaction was completed, DMF was evaporated with V10, purified with a PD10 Sephadex G25 M column, and the aqueous solution was freeze-dried to obtain a blue solid product.

[0288] In some embodiments, the total number of hydroxyl groups available for further conjugation to active agents (including therapeutic agents and / or diagnostic agents) is about 1-30, 2-20, or 5-10 of the 96 total available hydroxyl groups of an exemplary second generation dendrimer, wherein 24 glucose molecules contain 96 surface hydroxyl groups.

[0289] B. Methods of Preparing Dendrimer-Agent Conjugates

[0290] Methods of conjugating pharmaceutical agents to dendrimers are generally known in the art, for example as described in US2011 / 0034422, US2012 / 0003155 and US2013 / 0136697.

[0291] In some embodiments, one or more pharmaceutical agents are covalently linked to the dendrimer. In some embodiments, the pharmaceutical agent is linked to the dendrimer by a spacer designed to be non-cleavable in vivo. In some embodiments, the pharmaceutical agent is linked to the dendrimer by a spacer designed to be cleavable in vivo. For example, the spacer can be designed to be cleaved by hydrolysis, enzymatically, or a combination of the two, thereby providing sustained release of the pharmaceutical agent in vivo. In some embodiments, the chemical structure of the spacer and its point of attachment to the pharmaceutical agent can be selected so that the cleavage of the spacer releases the pharmaceutical agent or a suitable prodrug thereof. The chemical structure of the spacer can also be selected based on the desired release rate of the pharmaceutical agent.

[0292] In some embodiments, the conjugation between the agent and the dendrimer is achieved through one or more of a disulfide bond, an ester bond, an ether bond, a phosphodiester bond, a triglycyl peptide bond, a hydrazine bond, an amide bond, or an aminoalkyl bond. In some embodiments, the conjugation between the agent and the dendrimer is performed through an appropriate spacer that provides an ester bond or an amide bond between the agent and the dendrimer depending on the desired release kinetics of the agent. In some cases, an ester bond or a disulfide bond is introduced to form a releasable form of the drug. In other cases, an amide bond or an aminoalkyl bond is introduced to form a non-releasable form of the drug.

[0293] The spacer generally contains one or more organic functional groups. Examples of suitable organic functional groups contained in the spacer include secondary amides (-CONH-), tertiary amides (-CONR-), sulfonamides (-S(O)2-NR-), secondary carbamates (-OCONH-; -NHCOO-), tertiary carbamates (-OCONR-; -NRCOO-), carbonates (-OC(O)-O-), ureas (-NHCONH-; -NRCONH-; -NHCONR-, -NRCONR-), methanols (-CHOH-, -CROH-), disulfides, hydrazones, hydrazides, ethers (-O-) and esters (-COO-, -CH2O2C-, CHRO2C-), wherein R is an alkyl, aryl or heterocyclic group. In general, the identity of the one or more organic functional groups in the spacer is selected based on the desired release rate of the agent. In addition, one or more organic functional groups can be selected to facilitate covalent conjugation of the agent to the dendrimer. In some embodiments, the conjugation between the agent and the dendrimer is carried out through an appropriate spacer that provides a disulfide bond between the agent and the dendrimer. In some embodiments, the dendrimer-active agent conjugate is capable of rapidly releasing the agent through a thiol exchange reaction under reducing conditions in vivo.

[0294] In certain embodiments, the spacer comprises a combination of one or more of the above-mentioned organic functional groups and a linking group. The linking group can be composed of any atomic combination, including oligomeric chains and polymer chains; For example, the total number of atoms in the linking group is 3 to 200 atoms, 3 to 150 atoms, 3 to 100 atoms, or 3 to 50 atoms. The example of a suitable linking group includes an alkyl, a heteroalkyl, an alkylaryl, a few and a polyethylene glycol chain, and a few and a poly (amino acid) chain. The change of the linking group provides additional control of the release of the agent in vivo. In the embodiment where the spacer includes a linking group, one or more organic functional groups are usually used to connect the linking group to an anti-inflammatory agent and a dendritic macromolecule.

[0295] Reactions and strategies for covalently attaching agents 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 covalent conjugation method for a given agent can be selected based on the desired linking moiety and the structure of the agent and dendrimer, as this is related to the compatibility of the functional groups, the protection group strategy, and the presence of labile bonds.

[0296] The amount of active agent (drug load) in the dendrimer-active agent conjugate depends on many factors, including the choice of active agent, the structure and size of the dendrimer, and the tissue to be treated. In some embodiments, one or more antidepressants and / or antipsychotics are conjugated to the dendrimer at a concentration of between about 0.01% and about 45%, including between about 0.1% and about 30%, including between about 0.1% and about 20%, including between about 0.1% and about 10%, including between about 1% and about 10%, including between about 1% and about 5%, including between about 3% and about 20%, including between about 3% and about 10%. However, the specific drug load for any given active agent, dendrimer, and target site can be identified by conventional methods, such as those described.

[0297] In some embodiments, conjugation of the agent / spacer occurs through about 1%, 2%, 3%, 4%, or 5% of the total available surface functional groups (e.g., hydroxyl groups) of the dendrimer prior to conjugation. In other embodiments, conjugation of the agent / spacer occurs on less than 5%, less than 10%, less than 15%, less than 20%, less than 25%, less than 30%, less than 35%, less than 40%, less than 45%, less than 50%, less than 55%, less than 60%, less than 65%, less than 70%, less than 75% of the total available surface functional groups of 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 neural and / or glial cells while being conjugated to an effective amount of the agent for treating, preventing, and / or imaging a disease or condition. In some embodiments, the dendrimer-active agent conjugate retains an effective amount of the active agent for targeting to target neural and / or glial cells and binding to target receptors on the surface or within target neural and / or glial cells.

[0298] The following examples describe more specific methods for preparing exemplary dendrimer-active agent conjugates.

[0299] IV. Pharmaceutical Preparations

[0300] Pharmaceutical compositions comprising the dendrimer-active agent conjugates may be formulated in conventional manner using one or more physiologically acceptable carriers, optionally including excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically for oral, mucosal (intranasal, buccal, sublingual, vaginal, rectal or pulmonary), transdermal or injectable (intravenous, subcutaneous, intraperitoneal, intramuscular or intrathecal administration).

[0301] Representative excipients include aqueous buffers, wetting agents, viscosity modifiers, tonicity agents, stabilizers, and combinations thereof. Suitable pharmaceutically acceptable excipients are preferably selected from materials that are generally recognized as safe (GRAS) and can be administered to an individual without causing adverse biological side effects or unwanted interactions.

[0302] Typically, a pharmaceutically acceptable salt of an active substance can be prepared by reacting the free acid or base form of the drug with a stoichiometric amount of an appropriate base or acid in water or an organic solvent or a mixture of the two; typically, 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 and alkaline earth metal salts, and salts (e.g., quaternary ammonium salts) formed by reacting a drug with a suitable organic ligand. A list of suitable salts can be found, for example, in Remington's Pharmaceutical Sciences, 20th edition, Lippincott Williams & Wilkins, Baltimore, Maryland, 2000, p. 704. Examples of ophthalmic drugs sometimes administered in the form of pharmaceutically acceptable salts include timolol maleate, brinzolamide tartrate, and diclofenac sodium.

[0303] The composition is preferably formulated in dosage unit form for ease of administration and uniformity of dosage. The phrase "dosage unit form" refers to physically discrete units of the conjugate suitable for the patient to be treated. However, it should be understood that the total single dose of the composition will be determined by the attending physician within the scope of reasonable medical judgment. The therapeutically effective dose can be initially estimated in cell culture assays or animal models (usually mice, rabbits, dogs or pigs), or inferred from human data. Animal models are also used to achieve the desired concentration range and route of administration. Then, this information should help determine the effective dose and route of administration for humans. The therapeutic effect and toxicity of the conjugate can be determined by standard pharmaceutical procedures in cell culture or experimental animals, for example, ED50 (the dose has a therapeutic effect on 50% of the population) and LD50 (the dose is lethal to 50% of the population). The dose ratio of toxicity to therapeutic effect is the therapeutic index, expressed as the LD50 / ED50 ratio. Pharmaceutical compositions with large therapeutic indices are preferred. Data obtained from cell culture tests and animal studies can be used to formulate dosage ranges for human use.

[0304] In certain embodiments, the composition is administered topically, for example, by direct injection into the site to be treated or by an implant. In some embodiments, the composition is injected, topically applied, or otherwise administered directly into the vascular system, which is located on vascular tissue at or near the site of injury, surgery, or implantation. For example, in embodiments, the composition is topically applied to exposed vascular tissue during surgery, or as a cream, gel, or emulsion. Typically, topical administration results in an increase in the local concentration of the composition, which is greater than the concentration that can be achieved with systemic administration.

[0305] Pharmaceutical compositions for administration by parenteral (intramuscular, intraperitoneal, intravenous or subcutaneous injection) and enteral routes of administration are described.

[0306] A. Parenteral Administration

[0307] The compositions of the dendrimer-active agent conjugates can be administered parenterally. The phrases "parenteral administration" and "intestinal administration" are art-recognized terms that include modes of administration other than enteral and topical administration. The 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 carriers (for subcutaneous, intravenous, intraarterial or intramuscular injection) include, for example, sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's solution, 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, alcohol / aqueous solutions, cyclodextrins, emulsions or suspensions, including saline and buffered media. The dendrimers can also be administered in the form of an emulsion, such as water-in-oil. The example of oil is petroleum, animal oil, vegetable oil or synthetic oil, for example peanut oil, soybean oil, mineral oil, olive oil, sunflower seed oil, cod liver oil, sesame oil, cottonseed oil, corn oil, olive oil, vaseline and mineral oil. Suitable fatty acids for parenteral preparations include for example oleic acid, stearic acid and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters.

[0308] Preparations suitable for parenteral preparations may include antioxidants, buffers, antibacterial agents, and solutes that make the preparation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions that may include suspending agents, solubilizing agents, thickening agents, stabilizers, and preservatives. Intravenous carriers may include fluid and nutrient supplements, electrolyte supplements, such as supplements based on Ringer's dextrose. In general, water, saline, aqueous glucose and related sugar solutions and glycols, such as propylene glycol or polyethylene glycol are preferred liquid carriers, particularly for injectable solutions.

[0309] Injectable pharmaceutical carriers for injectable compositions are well known to those of ordinary skill in the art (see, for example, Pharmaceutics and Pharmacy Practice, JB Lippincott Company, Philadelphia, Pennsylvania, Banker and Chalmers, eds., pp. 238-250 (1982), and ASHP Handbook on Injectable Drugs, Trissel, 15th edition, pp. 622-630 (2009)).

[0310] B. Enteral administration

[0311] The dendrimer-active agent conjugate composition can be administered enterally (orally, sublingually, vaginally, rectally, buccally, intranasally, pulmonary or transdermally). The carrier or diluent can be a solid carrier such as a capsule or tablet or a diluent for a solid formulation, a liquid carrier or a diluent for a liquid formulation, or a mixture thereof.

[0312] 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, alcohol / aqueous solutions, cyclodextrins, emulsions or suspensions, including saline and buffered media.

[0313] The example of oil comprises petroleum, animal oil, vegetable oil or synthetic oil, for example peanut oil, soybean oil, mineral oil, olive oil, sunflower seed oil, cod liver oil, sesame oil, cottonseed oil, corn oil, olive, vaseline and mineral oil. The fatty acid suitable for parenteral preparation comprises for example oleic acid, stearic acid and isostearic acid. Ethyl oleate and isopropyl myristate are the examples of suitable fatty acid esters.

[0314] The carrier includes for example sodium chloride solution, Ringer's dextrose, glucose and sodium chloride, lactated Ringer's solution and fixed oil.Preparation includes for example aqueous and non-aqueous isotonic sterile injection solution, it can contain antioxidant, buffer, antibacterial agent and make the preparation and the solute of the blood isotonic of the expected recipient, and aqueous and non-aqueous sterile suspension, it can include suspending agent, solubilizing agent, thickening agent, stabilizer and preservative.Carrier can include for example fluid and nutritional supplement, electrolyte supplement, for example supplement based on Ringer's dextrose.In general, water, saline, aqueous glucose and relevant sugar solution are preferred liquid carriers.These also can be prepared together with protein, fat, carbohydrate and other components of infant formula.

[0315] Oral preparations can be in the form of chewing gum, gel strips, tablets, capsules or lozenges. Encapsulating materials used to prepare enteric coated oral preparations include cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate and methacrylate copolymers. Solid oral preparations such as capsules or tablets are preferred. Elixirs and syrups are also well-known oral preparations.

[0316] Formulations for administration to mucosal surfaces (e.g., the nasal cavity, oral surfaces, or the lungs) typically contain pharmaceutically acceptable excipients (e.g., excipients for parenteral administration), alone or in combination with various surfactants, penetration enhancers, and the like.

[0317] The compositions can also be formulated as aerosols (i.e., they can be "nebulized") for administration by inhalation. Aerosols can be placed in pressurized acceptable propellants, such as dichlorodifluoromethane, propane, nitrogen and air. For administration by inhalation, the compound is delivered in the form of an aerosol spray from a pressurized pack or nebulizer, using a suitable propellant.

[0318] V. How to use

[0319] The present invention describes methods of using dendrimer-psychedelic drug combinations. The methods can treat or alleviate one or more symptoms of psychological, cognitive, behavioral and / or emotional disorders. The composition is administered in an amount effective to provide a therapeutic effect but insufficient to produce adverse effects.

[0320] In a preferred embodiment, the dendrimer composition crosses the barrier interface of the central and peripheral nervous systems and selectively targets specific cells and specific receptors on cells to treat various diseases, disorders, injuries and conditions. The method comprises administering to a subject in need thereof an effective amount of the composition to increase the permeability of the psychedelic compound across the barrier interface of the central and peripheral nervous systems and / or to increase the binding of the psychedelic compound to specific receptors in specific cells, particularly serotonin receptors, dopamine receptors, adrenaline and / or monoamine transporters in cells in the central nervous system, peripheral nervous system and / or peripheral circulation (e.g., neurons, glial cells, gastrointestinal cells and / or immune cells).

[0321] A. Treatment methods

[0322] The compositions can be administered to prevent, treat and / or manage symptoms of various disorders, diseases and conditions, including but not limited to anxiety, mood disorders, eating disorders, personality disorders, stress disorders and / or psychosis. In certain forms, when the dendrimer is complexed with one or more hallucinogens, the compositions can be administered to treat one or more neurological diseases, such as mental health disorders (e.g., mood disorders, anxiety disorders, eating disorders, substance-related disorders) and stress disorders (e.g., post-traumatic stress disorder), learning disorders (e.g., autism), and pain disorders (e.g., neuropathic pain). In other forms, the dendrimer-hallucinogen conjugates can be administered to subjects in need of mood stabilization (e.g., bipolar disorder), reduction of anxiety in anxiety disorders, and reduction of cancer-related mental distress. In other forms, the dendrimer is complexed with one or more hallucinogens to treat non-neurological diseases (e.g., autoimmune diseases).

[0323] Typically, an effective amount of a dendrimer complex comprising a dendrimer in combination with one or more therapeutic, prophylactic and / or diagnostic agents is administered to an individual in need thereof. The dendrimer may also contain targeting agents, but as shown in the examples, these are not necessary for delivery to injured tissues of the spinal cord, brain and related areas.

[0324] In some embodiments, the dendrimer complex includes an agent attached or conjugated to the dendrimer that is capable of preferentially releasing the drug at a target receptor. The agent may be covalently attached or dispersed or encapsulated within the molecule. The amount of the dendrimer complex administered to the subject is selected to deliver an effective amount to reduce, prevent or otherwise alleviate one or more clinical or molecular symptoms of the disease or condition being treated as compared to a control (e.g., a subject treated with the active agent without the dendrimer).

[0325] B. Conditions and diseases requiring treatment

[0326] 1. Site-specific targeting

[0327] The compositions and methods are intended to circumvent existing challenges in selective drug delivery to the central and peripheral nervous systems. The compositions and methods can increase the bioavailability of drugs in the central and peripheral nervous systems by one or more of the following: (i) increasing the density of drugs across the brain barrier (particularly the blood-brain barrier and the blood-cerebrospinal fluid barrier), (ii) increasing drug solubility, (iii) promoting target binding, i.e., increasing site-specific binding, (iv) improving drug pharmacokinetics, and (v) increasing brain distribution. For example, in some forms, the compositions and methods allow for the selective delivery of compounds to the peripheral nervous system, thereby increasing the potential of the compositions for selective treatment of peripheral specific diseases and conditions, including but not limited to neuropathic pain, traumatic nerve injury, and inflammatory diseases.

[0328] A. Improve drug permeability across barrier interfaces

[0329] Dendrimer compositions and methods can improve the ability of hallucinogenic compounds to cross 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 substances and help maintain water homeostasis and an environment suitable for neuronal function in the blood. Since hallucinogenics are of clinical interest in the treatment of mental health disorders, dendrimer compositions can be used to deliver hallucinogenic compounds with improved permeability across these barrier interfaces to achieve site-specific targeting.

[0330] i. Blood-brain barrier

[0331] 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 impedes drug delivery to the central nervous system (CNS), in part because (1) systemic delivery of drugs intended for the brain may result in excessive peripheral drug concentrations and (2) the complex interactions between cells and molecules that make up 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.

[0332] Brain microvascular endothelial cells, pericytes, astrocytes, tight junctions, neurons, and basement membranes constitute physically tight brain capillaries within the BBB. Brain capillary endothelial cells have no pores, which restricts the diffusion of small molecules and proteins. Interendothelial junctions connect endothelial cells to a continuous barrier, severely limiting the permeation of water-soluble substances. Pericytes, astrocytes, and basement membranes surround endothelial cells, ultimately forming an impermeable BBB. In addition, efflux transporters are located in brain capillary endothelial cells, which further prevents substances from entering the brain. The permeability of the BBB is mainly controlled by interendothelial junctions, which are protein complexes such as adherens junctions, tight junctions, and gap junctions. Adherens junctions mainly regulate the permeability of the endothelial barrier. Tight junctions play a crucial role in maintaining the permeability barrier of epithelial and endothelial cells, which controls tissue homeostasis. Gap junctions are composed of six connexin molecules and allow direct electrical and chemical communication between endothelial cells. Finally, the composition of the BBB does not have a static structure, but is constantly adapting in response to various physiological changes in the brain. The dendrimer compositions and methods overcome the above challenges and are suitable for delivering hallucinogenic compounds across the blood-brain barrier via one or more of the above transport mechanisms.

[0333] Molecules cross the BBB via the paracellular pathway (between adjacent cells) or the transcellular pathway (across cells). For the paracellular pathway, ions and solutes cross the BBB by passive diffusion using concentration gradients. The transcellular pathway includes different mechanisms such as passive diffusion, receptor-mediated transport, and transcytosis.

[0334] Physicochemical factors that affect BBB permeability include molecular weight, charge, lipid solubility, surface activity, and relative size of molecules. BBB permeability may also be affected by physiological factors, such as efflux transporters, such as P-glycoprotein (P-gp), enzyme activity, plasma protein binding, and cerebral blood flow. Hydrophilic molecules (such as proteins and peptides) enter the brain through specific and saturable receptor-mediated transport mechanisms (such as glucose transporter-1 (GLUT-1), insulin transporter, and transferrin transporter). These endogenous transporters are expressed on the luminal and extraluminal endothelial cell membranes. Among these transport mechanisms, receptor-mediated transcytosis has been widely studied for drug delivery to the brain. The dendrimer compositions and methods of the present application are suitable for delivering hallucinogen compounds across the blood-brain barrier through one or more of the above-mentioned mechanistic pathways.

[0335] ii. Blood-Nerve Barrier (BNB)

[0336] The blood-nerve barrier (BNB) defines the physiological space in which the axons, Schwann cells, and other related cells of the peripheral nerve function, thereby ensuring the normal function of the peripheral nerve and maintaining the homeostasis of the endoneurial environment. The BNB is composed of endoneurial microvessels and perineurium within the nerve bundle. Tight junctions between endothelial cells and pericytes in the endoneurial vessels isolate the endoneurium from the blood, thereby preventing the uncontrolled leakage of molecules and ions from the circulatory system into the peripheral nerve. In addition, there is a diffusion barrier within the perineurium, which is formed by tight junctions between adjacent perineurial cells and the basement membrane surrounding each perineurial cell layer. Endoneurial capillaries and perineurial channels are restrictive barriers that separate the endoneurial extracellular environment of peripheral nerves from the space around the epineurial bundle and the systemic circulation, thereby protecting the endoneurial microenvironment from drastic concentration changes in blood vessels and other extracellular spaces.

[0337] For drug targets located in peripheral nerves, BNBs may be problematic because they may limit or prevent the drug from reaching its site of action, negatively affecting drug efficacy. In addition, the expression profiles of transporters in peripheral nerves may be very different from those in the central nervous system. The dendrimer compositions of the present application may be used to improve the permeability of hallucinogenic compounds across BNBs, thereby improving the delivery of hallucinogenic compounds to peripheral nerve targets.

[0338] iii. Blood-CSF Barrier

[0339] The composition can be used to improve the delivery of hallucinogens to target sites through the blood-cerebrospinal fluid barrier (blood-CSF barrier) and the ventricles of the brain. The choroid plexus is a vascular tissue present in all ventricles of the brain. The functional units of the choroid plexus are composed of capillaries wrapped by a layer of differentiated ependymal epithelium. Unlike the capillaries that form the blood-brain barrier, the choroid plexus capillaries have holes and no tight junctions. Therefore, the endothelium does not form a barrier to the movement of small molecules. In contrast, the blood-CSF barrier of the choroid plexus is formed by epithelial cells and the tight junctions connecting them. Another part of the blood-CSF barrier is the arachnoid membrane that wraps the brain. The cells of this membrane are also connected by tight junctions.

[0340] The CSF space and brain structures near the CSF compartment are pharmacological targets for CNS diseases. For example, the subarachnoid space, perivascular or periventricular spaces are areas where pathogenic lymphocytes, monocytes and neutrophils accumulate in neuroinflammatory diseases (such as autoimmune diseases, eosinophilic inflammation and / or asthma). B cell foci producing potentially harmful antibodies detected in different CNS autoimmune diseases are believed to be mainly located in the pia mater. Therefore, in some forms, dendrimer compositions can be used to deliver antidepressants and / or antipsychotics to target areas through the blood-CSF space connected to deep cervical lymph nodes to improve or treat symptoms associated with neuroinflammatory diseases, such as changes in cytokine responses after stroke and allergic attacks.

[0341] In some forms, the dendrimer compositions can be used to deliver hallucinogens to target sites to improve or treat psychedelic and inflammatory symptoms associated with tumor development. Thus, in some forms, the dendrimer compositions can be used to achieve therapeutic concentrations of hallucinogen compounds in the tumor microenvironment using pharmacological pressure from CSF to reduce inflammatory responses and tumor development.

[0342] b. Improve target specific binding

[0343] The dendrimer compositions are useful for delivering psychedelic compounds with increased binding affinity and specificity to one or more receptors to modulate serotonin (5HT) receptors, such as 5HT-1A, 5HT-2B, 5HT-2A, 5HT-3B, 5HT-2C, 5HT-3, 5HT-4, 5HT-6, and 5HT-7 receptors and / or one or more dopamine receptors, such as dopamine D1 and D2 receptors. The dendrimer compositions can also be used to deliver psychedelic compounds for direct or indirect modulation of one or more norepinephrine (NE) receptors, such as α 2A -Adrenergic receptors, alpha 2B -Adrenergic receptors, alpha 2C-adrenergic receptors, beta adrenergic receptors, monoamine transporters, such as serotonin reuptake transporter (SERT), dopamine transporter (DAT) and / or vesicular monoamine transporter (VMAT2). In addition, hallucinogenic compounds having affinity and 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 agents with increased 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.

[0344] i. Improve serotonin receptor binding

[0345] Dendrimer compositions can improve binding to one or both serotonin receptors, thereby modulating signaling in a cell-specific and tissue-specific manner.

[0346] In the central nervous system (CNS), serotonin is produced almost exclusively by neurons in the raphe nuclei located in the midline of the brainstem. These serotonin-producing neurons constitute the largest and most complex efferent system in the human brain. The most caudal raphe nuclei innervate the spinal cord, while the more anterior raphe nuclei, dorsal raphe nuclei, and medial raphe nuclei innervate much of the rest of the CNS via diffuse projections. Nearly every cell in the brain is in close proximity to serotonin fibers, and nearly all behaviors, as well as many other brain functions, are regulated by serotonin.

[0347] Serotonin exerts its effects by interacting with 13 serotonin G protein-coupled receptors (GPCRs). Serotonin receptors are found throughout the body and regulate a range of different processes, including but not limited to learning and memory, control of sleep / wake cycles, temperature regulation, appetite, sexual behavior in men and women, pain, motor activity, and aspects of autonomic function such as arterial blood pressure and heart rate.

[0348] 5-HT2Ars are widely expressed in the cerebral cortex - especially layers I and IV-V, piriform and entorhinal cortices, claustrum, internal piriform nucleus and olfactory bulb / anterior olfactory nucleus, brainstem, as well as the limbic system and basal ganglia; especially the nucleus accumbens and caudate nucleus. Therefore, dysfunction of the serotonin system is associated with a variety of diseases and conditions, such as anxiety and depression, migraine, personality disorders, obsessive-compulsive disorder, drug addiction and neurodegenerative diseases. Activation of the 5-HT2A receptor is a prerequisite for the action of "classical" psychedelics (such as LSD, dimethyl-4-hydroxytryptamine and mescaline), which can act as full or partial agonists of this receptor and represent the three major classes of 5-HT2A agonists, namely ergoline, tryptamine and phenylethylamine, respectively. Therefore, in some embodiments, the dendrimer-psychedelic composition can be used to activate the 5-HT2A receptor to improve symptoms associated with one of the above diseases. For example, the dendrimer-R-DOI conjugate can be administered to a subject in need of reducing anti-inflammatory responses associated with neurodegenerative diseases or anxiety disorders, optionally, through functional selectivity. Functional selectivity refers to the process by which different ligands induce slightly different conformations of the receptor to recruit different sets of effector pathways. In a second example, the composition can activate 5-HT2A receptors located on the apical dendrites of pyramidal cells in the prefrontal cortical region, thereby inducing hallucinogenic activity. In a third example, the composition can be used to deliver classic psychedelics to a subject in need of modulation of the activity of the receptor heterodimer 5-HT2A–mGlu2 and dopamine receptors to enhance PFC activity and improve learning, memory, and attention deficits.

[0349] 5-HT2A receptors are also widely distributed in peripheral tissues, and serotonin disorders are also associated with peripheral tissue diseases, such as pulmonary hypertension, cholangiocarcinoma, chronic renal failure, and inflammatory bowel disease. In some forms, the composition can be used to regulate 5-HT2A receptor activity in immune-related tissues (e.g., spleen, thymus, and circulating lymphocytes). In some forms, the composition is used to regulate 5-HT2A receptor protein levels in peripheral blood mononuclear cells (PBMCs), eosinophils, and T cells to regulate innate and / or adaptive immune responses. For example, the composition can be administered to a subject who needs to reduce inflammation and eosinophil infiltration caused by asthma caused by allergies.

[0350] ii. Improve dopamine receptor binding

[0351] There are five types of dopamine receptors, including D1, D2, D3, D4 and D5, each with different functions. Dendrimer compositions can improve binding to one or more dopamine receptors, preferably D1 and / or D2 receptors, modulating signal transduction in a cell-specific and tissue-specific manner. D1 receptors are associated with memory, attention, impulse control, renal function regulation, movement, and couple to G stimulatory sites and activate adenylate cyclase. Activation of adenylate cyclase leads to the production of the second messenger cAMP, which leads to the production of protein kinase A (PKA), which leads to further transcription in the nucleus. On the other hand, D2 receptors are associated with movement, attention, sleep, memory and learning, and couple to G inhibitory sites, inhibit adenylate cyclase and activate K+ channels.

[0352] Dopamine receptors are expressed in the central nervous system, particularly in the dentate gyrus and subventricular zone of the hippocampus. D1 receptors are found in higher densities in the striatum, nucleus accumbens, olfactory bulb, and substantia nigra. These receptors regulate reward systems, motor activity, memory, and learning. In addition to stimulating adenylate cyclase, D1 receptors activate phospholipase C, which induces intracellular calcium release and activates protein kinase C. Protein kinase C is a calcium-dependent protein kinase. Calcium also regulates neurotransmitter release through exocytosis. D2 receptors are primarily expressed in the striatum as well as in the external globus pallidus, core nucleus accumbens, hippocampus, amygdala, and cerebral cortex and regulate postsynaptic receptor-mediated extrapyramidal activity. D2 receptors play an important role in signaling for dopamine neuron survival and neuronal development in humans. Dopamine receptors are also expressed in the periphery, more prominently in the kidney and blood vessels. For example, in the kidney, D1 receptors inhibit Na / K ATPase via the PKA and PKC pathways, thereby increasing electrolyte excretion and renal vasodilation.

[0353] Thus, in certain forms, the compositions can be used to deliver LSD alone or in combination with L-DOPA to modulate D1 and D2 receptor activity and improve Parkinson's disease-induced psychotic symptoms such as depression, anxiety, limb pain, fatigue, sleep disruption, and cognitive impairment.

[0354] iii. Improve norepinephrine receptor binding

[0355] Dendrimer compositions can improve binding to one or both norepinephrine receptors, thereby modulating signaling in a cell-specific and tissue-specific manner.

[0356] Norepinephrine, also known as noradrenaline, is a neurotransmitter of the brain that plays an important role in regulating arousal, attention, cognitive function, and stress responses. It also functions as a peripheral hormone in the "fight or flight" response as part of the sympathetic nervous system. In states of stress or anxiety, norepinephrine and epinephrine are released and bind to adrenergic receptors throughout the body, exerting effects such as dilation of the pupils and bronchioles, increased heart rate and vasoconstriction, increased renal renin secretion, and inhibition of peristalsis. The norepinephrine system plays a role in the pathogenesis of several major neuropsychiatric disorders and has emerged as an important pharmacological target for a variety of psychiatric, neurological, and cardiopulmonary diseases.

[0357] The central noradrenergic system is composed of two main ascending projections originating from the brainstem: the dorsal noradrenergic bundle (DNB) and the ventral noradrenergic bundle (VNB). The DNB originates from the A6 locus coeruleus located on the dorsal side of the pons and is mainly composed of noradrenergic neurons. It is the main site of noradrenaline production in the central nervous system. It sends out projections to specifically control the cerebral cortex, hippocampus and cerebellum, and its projections overlap with the projections from the VNB to control the areas of the amygdala, hypothalamus and spinal cord. The VNB originates from the nuclei in the pons and medulla oblongata, and sends out projections to control the areas of the amygdala, hypothalamus and midbrain and medulla oblongata.

[0358] The sympathetic nervous system and neuroendocrine chromaffin cells (located in the adrenal medulla) are primarily responsible for the synthesis and release of norepinephrine and other catecholamines into the circulation. These hormones act on alpha and beta adrenergic receptors on smooth muscle cells and adipose tissue throughout the body.

[0359] After the action potential enters the presynaptic terminal, voltage-gated calcium channels are stimulated, allowing calcium ions to flow from outside the cell into the cell. This influx causes norepinephrine (stored in vesicles) to bind to the cell membrane and be released into the synaptic cleft by exocytosis. Norepinephrine can then bind to three major receptors: alpha 1 (α-1), alpha 2, and beta receptors. These receptors are G protein-coupled receptors that have either inhibitory or excitatory properties and differ in their binding affinity to norepinephrine.

[0360] The alpha-1 receptor is further subdivided into alpha-1a, alpha-1b, and alpha-1d receptors. These receptors are located in postsynaptic areas of the brain, including the locus coeruleus, olfactory bulb, cerebral cortex, dentate gyrus, amygdala, and thalamus. The alpha-1 receptor has a moderate binding affinity for norepinephrine and is coupled to the Gq protein signaling pathway. In this pathway, phospholipase C (PLC) is activated, converting phosphatidylinositol 4,5-bisphosphate (PIP2) on the cell membrane to inositol 1,4,5-triphosphate (IP3) and diacylglycerol (DAG). IP3 is released into the cytosol and binds to transmembrane IP3 receptors located on the endoplasmic reticulum (ER), which functions as a calcium channel. Upon binding, the receptor undergoes a conformational change that results in the release of calcium ions from the ER into the cytoplasm. DAG remains on the cell membrane and actively regulates protein kinase C (PKC), which phosphorylates other proteins. These combined actions produce excitatory cellular effects.

[0361] The alpha-2 receptors are subdivided into alpha-2a, alpha-2b, and alpha-2c receptors. These receptors are located in presynaptic and postsynaptic areas of the brain, including the locus coeruleus, amygdala, and hypothalamus. These receptors have the highest binding affinity for norepinephrine and are coupled to the Gi / o protein signaling pathway. In this pathway, cAMP levels are reduced, which results in decreased adenylate cyclase activity, producing an inhibitory cellular effect. Presynaptic noradrenergic terminals contain alpha-2 autoreceptors that prevent further norepinephrine release.

[0362] Beta receptors are subdivided into beta-1, beta-2, and beta-3 receptors. These receptors are located in different areas of the brain, with beta-1 and beta-2 receptors being most common in the cerebral cortex. These receptors have the lowest binding affinity for norepinephrine and are coupled to the Gs protein signaling pathway. In this pathway, elevated cAMP levels lead to activation of protein kinase A (PKA), which in turn phosphorylates other proteins within the cell and leads to excitatory cellular effects. Beta-2 receptors are also coupled to the Gi protein signaling pathway. Beta-3 receptors are found in adipose tissue.

[0363] In the adrenal medulla, acetylcholine stimulates the release of epinephrine and norepinephrine. Acetylcholine binds to nicotinic receptors located on adrenal chromaffin cells, generating an action potential maintained by voltage-gated sodium and potassium channels. This action potential triggers an influx of calcium into the cytosol, causing norepinephrine vesicles to bind to the cell membrane, resulting in the release of norepinephrine into the circulation, which then binds to alpha and beta receptors on smooth muscle and adipocytes.

[0364] Norepinephrine can be degraded intracellularly or in the synaptic cleft by the enzymes monoamine oxidase (MAO) or catechol-O-methyltransferase (COMT). MAO oxidizes norepinephrine, while COMT metabolizes deaminated norepinephrine via 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 degradation of norepinephrine to vanillylmandelic acid (VMA).

[0365] C. Conditions requiring treatment

[0366] The composition is suitable for treating one or more diseases, disorders and injuries in the central and peripheral nervous systems. The composition can be used to treat various diseases, disorders and injuries, including mental health disorders, gastrointestinal diseases and / or other tissues in which the treatment nerve plays a role in diseases or disorders. The composition and method are also suitable for preventive purposes. For example, the composition can be applied to patients in need to improve, treat or prevent symptoms related to various disorders, diseases and conditions, including but not limited to mental health and nervous system disorders, such as depression (including major depressive disorder, refractory depression and postpartum depression), post-traumatic stress disorder, panic disorder, social anxiety disorder, anorexia nervosa, suicidal ideation, obsessive-compulsive disorder, premenstrual dysphoria, anorexia, substance abuse disorders, epilepsy, autism spectrum disorder, attention deficit hyperactivity disorder, schizophrenia, cluster headache, migraine, epileptic seizures, fibromyalgia, narcolepsy, obesity, Alzheimer's disease, Tourette syndrome, pain such as neuropathic pain and chronic pain, phobia and cardiovascular disease); pain disorders such as neuropathic pain and / or gastrointestinal diseases. In other forms, the composition may be administered to a subject in need of stabilization of mood (eg, bipolar disorder), reduction of anxiety in anxiety disorders, and inflammation in Parkinson's disease.

[0367] The dendrimer complex composition preferably has a diameter less than 20 nm and a hydroxyl surface density of at least 0.8 OH groups / nm 2 , preferably with a diameter of less than 10 nm and a hydroxyl surface density of at least 1 OH group / nm 2 , more preferably less than 6 nm in diameter, with a hydroxyl surface density of at least 1 OH group / nm 2 , most preferably with a diameter between 5 nm and a hydroxyl surface density of at least 1.5 OH groups / nm 2The invention relates to a method for delivering a therapeutic, prophylactic or diagnostic agent to selectively target microglia and astrocytes, which play a key role in the pathogenesis of many diseases and conditions, including neurodevelopment, neurodegenerative diseases, neuropsychiatric diseases and chronic pain. Thus, the dendrimer complex is administered in a dosage unit amount effective to treat or alleviate a condition associated with a pathological condition of the central and peripheral nervous systems. For example, the dendrimer complex is administered in an effective dosage unit to treat or alleviate a condition associated with a pathological condition affecting neurons, microglia and astrocytes. In general, by targeting these cells, the dendrimer can specifically deliver an agent to treat neuroinflammation.

[0368] In preferred embodiments, the compositions may include glucose or hydroxy dendrimers having a diameter of 5 nm or less and are combined with antidepressants, antipsychotics, or other drugs that work by modulating monoaminergic neurotransmission for the treatment of mental health and CNS diseases. In other preferred embodiments, the compositions may include glucose or hydroxy dendrimers having a diameter greater than 5 nm and are combined with antidepressants, antipsychotics, or other drugs that work by modulating monoaminergic neurotransmission for the treatment of peripheral nervous system diseases.

[0369] a. Mental Health Disorders and Conditions

[0370] These compositions and methods are useful 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.

[0371] i. Affective or emotional disorders

[0372] Affective or mood disorders are marked disturbances of mood (low moods are called depression, and high moods are called hypomania or mania). These disorders include bipolar disorder, cyclothymia, hypomania, major depressive disorder, disruptive mood dysregulation disorder, persistent depressive disorder, premenstrual dysphoric disorder, seasonal affective disorder, medical condition-related depression, and depression caused by substance or medication use.

[0373] In some embodiments, the compositions and methods are useful for treating symptoms associated with depression, refractory depression, and suicidal ideation. Up to 30% of patients with depression do not respond effectively to antidepressant treatment, in part because of biological differences between patients and the time it takes to respond to medication, resulting in low compliance with treatment regimens. Therefore, there is an urgent need to expand the types of medications available to patients with depression.

[0374] In an exemplary embodiment, the composition is used to deliver dimethyl-4-hydroxytryptamine, dimethyl-2-hydroxytryptamine phosphate and / or DMT to a subject with treatment-resistant depression to establish long-term behavioral outcomes, such as improved coping strategies and enhanced cognitive function (e.g., improved associative learning, a cognitive function that is often impaired in major depressive disorder (MDD)).

[0375] ii. Anxiety disorders

[0376] Anxiety disorders are different from normal nervousness or anxiety, which involve excessive fear or worry. Anxiety disorders include generalized anxiety disorder, panic disorder, social anxiety disorder, and various phobia-related disorders.

[0377] Generalized anxiety disorder (GAD) generally involves persistent anxiety or fear, which interferes with daily life. It is different from occasionally worrying about things or feeling anxious because of stressful life events. People with GAD often feel anxious, which lasts for months or even years. These compositions and methods 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 unexplained pain, excessive worry, sleep problems (such as difficulty falling asleep or difficulty staying asleep).

[0378] Panic disorder is an anxiety disorder characterized by sudden, recurring attacks of intense fear accompanied by physical symptoms, including chest pain, palpitations, shortness of breath, dizziness or abdominal discomfort, or a feeling of being out of control even when there is no apparent danger or trigger. People with panic disorder often worry about when the next attack will occur and actively try to prevent future attacks by avoiding places, situations, or behaviors associated with panic attacks. Panic attacks may occur as frequently as several times a day or as infrequently as a few times a year. These compositions and methods are useful for treating one or more panic attack symptoms, including, but not limited to, palpitations, excessive sweating, shaking or tingling, chest pain, and feelings of being out of control, such as feelings of impending doom and feelings of being out of control.

[0379] Social anxiety disorder is an intense and persistent fear of being watched and judged by others. For people with social anxiety disorder, the fear of social situations can be so intense that it seems beyond their control. For some people, this fear can prevent them from going to work, school, or doing everyday things. The compositions and methods are useful for treating one or more symptoms of social anxiety disorder, including, but not limited to, excessive blushing, sweating or shaking, heart palpitations, stomach aches, stiff body posture or soft voice, and self-consciousness or fear of negative judgment.

[0380] A phobia is an intense fear or aversion to a particular object or situation. Although the anxiety felt is real in certain situations, the fear felt by a person with a phobia is disproportionate to the actual danger posed by the situation or object. These compositions and methods are useful for treating one or more phobia symptoms, including, but not limited to, unreasonable or excessive worry about encountering the phobic object or situation, immediate intense anxiety upon encountering the phobic object or situation, and tolerating the unavoidable objects and situations with intense anxiety.

[0381] In an exemplary embodiment, the composition can be used to deliver MDMA to a subject in need of improving symptoms associated with a trauma-related disorder (e.g., PTSD), such as reducing pathological fear responses, social disconnection, and emotional numbness, 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.

[0382] iii. Eating disorders

[0383] Eating disorders are serious and often fatal conditions involving severe disturbances in a person's eating behaviours and related thoughts and emotions. Eating disorders involve an obsessive focus on food, weight and body shape. Common eating disorders include anorexia nervosa, bulimia nervosa and binge eating disorder.

[0384] Anorexia nervosa is a disorder in which people refuse to eat, severely restrict food, or eat only very small amounts of certain foods. They may also weigh themselves repeatedly. They may think they are overweight even though they are severely underweight. There are two subtypes of anorexia nervosa: restricting and binge-purge. People with restricting anorexia nervosa severely restrict the amount and types of food they eat. People with binge-purge anorexia nervosa also greatly restrict the amount and types of food they eat. In addition, they may have binge eating and purging episodes—eating large amounts of food in a short period of time and then vomiting or using laxatives or diuretics to get rid of the food they ate.

[0385] Symptoms of anorexia nervosa include, but are not limited to, thinning bones (osteopenia or osteoporosis), mild anemia and muscle weakness, brittle hair and nails, dry, yellow skin, growth of fine hair all over the body (lanugo), severe constipation, low blood pressure, slowed breathing and pulse, impaired heart structure and function, brain damage, multiple organ failure, a drop in internal body temperature causing a person to feel cold all the time, lethargy, lack of energy or tired all the time, and infertility.

[0386] Bulimia nervosa is a disorder in which the patient repeatedly and frequently eats large amounts of food and feels unable to control these eating. After a binge, the patient engages in behaviors to compensate for the overeating, such as forced 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, wear of tooth enamel and tooth sensitivity and decay due to exposure to stomach acid, acid reflux and other gastrointestinal problems, intestinal discomfort and irritation from laxative abuse, severe dehydration due to purging of fluids, and electrolyte imbalances (too low or too high levels of sodium, calcium, potassium and other minerals), which can lead to a stroke or heart attack.

[0387] In an exemplary embodiment, the composition can be used to deliver dimethyl-2-hydroxytryptamine phosphate, LSD, and / or ayahuasca to a subject in need thereof to (1) increase connectivity between neuronal networks and create the potential to transcend the subject's self-imposed limitations that are debilitating to the subject, and / or (2) promote ideal brain states that may accelerate the healing process, e.g., increase neuroplasticity and neurogenesis, improve mood, reduce fear responses, and promote acceptance and compassion for oneself and others.

[0388] b. Neurological and neurodegenerative diseases

[0389] The compositions and methods are useful for treating symptoms associated with neurological disorders and neurodegenerative diseases.

[0390] Neurodegenerative diseases are chronic progressive neurological disorders that affect neurological and behavioral functions and involve biochemical changes that lead to different histopathological and clinical syndromes (Hardy H et al., Science. 1998; 282: 1075–9). Abnormal proteins that are resistant to cellular degradation mechanisms accumulate within cells. The pattern of neuronal loss is selective, that is, a group of neurons is affected while other neurons remain intact. Usually, there is no clear inciting event for the disease. Classic neurodegenerative diseases include Alzheimer's disease, Huntington's disease, and Parkinson's disease.

[0391] Neuroinflammation mediated by activated microglia and astrocytes is a major hallmark of various neurological diseases, making them potential therapeutic targets (Hagberg, H et al., Annals of Neurology 2012, 71, 444; Vargas, DL et al., Annals of Neurology 2005, 57, 67; Pardo, CA et al., International Review of Psychiatry 2005, 17, 485). Several scientific reports have shown that reducing early neuroinflammation by targeting these cells can delay the onset of the disease, thereby 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). It is a challenging task to deliver therapeutic agents across the blood-brain barrier. Neuroinflammation can lead to damage of the blood-brain barrier (BBB). The damaged BBB in neuroinflammatory diseases can be used to deliver drug-loaded nanoparticles to various parts of 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).

[0392] The compositions and methods can also be used to deliver active agents to treat nerves or neurodegenerative diseases or disorders or central nervous system disorders. In preferred embodiments, the compositions and methods can effectively treat and / or alleviate neuroinflammation associated with nerves or neurodegenerative diseases or disorders or central nervous system disorders, or peripheral symptoms caused by nervous system or neurodegenerative diseases or disorders. The method generally includes administering to the subject an effective amount of the composition to increase cognition or reduce cognitive decline, increase cognitive function or reduce cognitive function decline, increase memory or reduce memory decline, increase learning ability or talent or reduce learning ability or talent decline, or a combination thereof.

[0393] Neurodegeneration refers to the gradual loss of neuronal structure or function, including neuronal death. For example, the compositions and methods can be used to treat subjects with diseases or conditions, such as Parkinson's disease (PD) and PD-related conditions, Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD) and other dementias, multiple sclerosis (MS), post-encephalitic dementia, cancer and chemotherapy-related cognitive impairment and dementia, and dementia and pseudodementia caused by depression.

[0394] In an exemplary embodiment, the composition can be used to deliver one or more psychedelic compounds to a subject in need thereof to improve symptoms associated with a neurodegenerative disease, such as stimulating neurogenesis, neuroprotection, and neuroplasticity, reducing neuronal and oligodendrocyte loss, reducing oxidative stress and BBB disruption, and reducing inflammation and endoplasmic reticulum damage.

[0395] c. Pain

[0396] The compositions and methods are suitable for treating neuropathic and / or non-neuropathic pain associated with various diseases, such as complex regional pain syndrome, peripheral neuropathy, multiple sclerosis, and pain caused by cancer. For example, dimethyl-2-hydroxytryptamine phosphate and LSD are effective in treating neuropathic (chronic) neuralgia. Chronic neuralgia, also known as neuropathic pain, is caused by nerve damage or other nerve problems and is generally unresponsive to conventional analgesics (e.g., paracetamol). In some forms, the compositions can be used to deliver dimethyl-2-hydroxytryptamine phosphate and LSD to subjects who need to reduce the severity of headaches and migraines and extend the remission period between headaches and migraines.

[0397] The mechanisms of chronic pain are not fully understood, but may involve complex interactions between somatic and visceral afferent inputs, peripheral and central sensitization, emotional states, and behavior and cognition. Distraction and mood changes can have a huge impact on pain perception. Therefore, in some embodiments, the composition can be used to deliver dimethyl-2-hydroxytryptamine phosphate to subjects who need to relieve pain caused by cancer-related anxiety and depression.

[0398] In some forms, the composition can be used to treat chronic pain that does not involve nerves (non-neuropathic pain). For example, the dendrimer composition can be compounded with LSD or MDMA for the treatment of 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 back pain, and chronic neck pain. For example, the composition can be used to deliver dimethyl-2-hydroxytryptamine phosphate or LSD to a subject who needs relief of symptoms associated with fibromyalgia and / or chronic pelvic pain, including, but not limited to, relief of pain and stiffness in the muscles, abdomen, neck and / or back, fatigue, sleep disorders, headaches, and migraines. The most important indications are major depressive disorder, refractory 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, headaches, epilepsy, fibromyalgia, narcolepsy, obesity, Alzheimer's disease, and Tourette syndrome.

[0399] D. Dosage and effective amount

[0400] The dosage and administration regimen will depend on the severity and site of the disease or injury and / or the method of administration, as well as the specific agent being delivered. This can be determined by one skilled in the art.

[0401] In some embodiments, dosages are expressed in mg / kg, particularly when expressed as in vivo dosages of dendrimer gene editing compositions.

[0402] Typically, the dosage range is from micrograms / kg to about 100 mg / kg body weight. The dosage can 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, or about 25, 30, 35, 40, 45, 50, 55, 60, 65, 70 or 75 mg / kg per dose.

[0403] Preferably, the composition of dendrimer gene editing agents will not target or otherwise genetically modify non-targeted or healthy cells that are not in or associated with diseased tissue, or that are present at lower levels than cells associated with a disease or condition such as cancer and / or a proliferative disease. Thus, byproducts and other side effects associated with the composition are reduced. Thus, in preferred embodiments, the administration of the dendrimer composition is in an amount that results in improved or enhanced function in an individual suffering from a disease or condition such as cancer and / or a proliferative disease.

[0404] The actual effective amount of the composition will vary depending on the specific agent being administered, the specific composition being formulated, the mode of administration, and the age, weight, condition, and route of administration and disease or condition of the subject being treated. Generally, for intravenous injection or infusion, the dosage will be lower than for oral administration.

[0405] Dosages may vary and may be administered once or multiple times per day for one or more days. In some embodiments, the dendrimer conjugates may increase the effectiveness and durability of the treatment, which may reduce the need for repeated dosing to weekly, monthly, six-monthly, annual, or other long-term dosing regimens. Some embodiments may be incorporated into drug delivery systems (e.g., implants, pumps, patches, creams, etc.) to provide controlled, sustained delivery in a manner that reduces compliance and the potential for abuse. Guidance on appropriate dosing for a given class of drug products can be found in the literature. The optimal dosing regimen can be calculated by measuring drug accumulation in the subject or patient. One of ordinary skill in the art can readily determine the optimal dose, method of administration, and repetition rate. The optimal dose may vary depending on the relative potency of the individual drug compositions and can generally be estimated based on effective doses in in vitro and in vivo animal models. It is noteworthy that in some cases, minimizing or preventing hallucinations caused by psychedelic compounds may be a desirable option. This can be achieved by peripheral confinement of the dendrimer composition or by controlled and / or sustained activity of the dendrimer composition.

[0406] Also provided are dosage forms of pharmaceutical compositions comprising the dendrimer compositions. "Dosage form" refers to the physical form, such as a capsule or vial, of a dose of a therapeutic compound for administration to a patient. The term "dosage unit" refers to the amount of a therapeutic compound to be administered to a patient in a single dose.

[0407] In general, the timing and frequency of dosing will be adjusted to balance the efficacy of a given therapeutic or diagnostic regimen with the side effects of a given delivery system.

[0408] In some embodiments, the dose is administered daily, biweekly, weekly, biweekly or less frequently, in an amount that effectively increases the blood level of the therapeutic agent. When administration is not by oral route, the composition can be delivered over a period of more than one hour, e.g., 3-10 hours, to produce a therapeutically effective dose within 24 hours. Alternatively, the composition can be formulated for controlled release, wherein the composition is administered in a single dose and is administered repeatedly according to a regimen of once a week or less.

[0409] One of ordinary skill in the art will appreciate that the dosing regimen can be any length of time sufficient to treat the subject's disease. In some embodiments, the regimen includes one or more treatment cycles followed by a drug holiday (e.g., no drug). The drug holiday 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.

[0410] E. Control

[0411] The results of treatment with a composition comprising one or more gene editing compositions associated or conjugated to a dendrimer can be compared to 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 symptoms of a subject before and after administration of a glucose dendrimer composition. The condition or symptom can be a biochemical, molecular, physiological or pathological readout. For example, the effect of the composition on a particular symptom, pharmacological or physiological index can be compared to an untreated subject or the condition of the subject before treatment. In some embodiments, the symptom, pharmacological or physiological index is measured in the subject before treatment and measured again one or more times after the start of treatment. In some embodiments, the control is a reference level or average value determined based on measuring one or more symptoms, pharmacological or physiological indexes of subjects (e.g., healthy subjects) who do not suffer from the disease or condition to be treated. In some embodiments, the treatment effect is compared to conventional treatments known in the art. In some embodiments, the untreated control subject suffers from the same disease or condition as the treated subject.

[0412] In some embodiments, a control includes an equivalent amount of a gene editing composition delivered alone or bound to a dendrimer without a glucose-based branching unit, such as a dendrimer of similar generation, molecular weight, and / or surface group density (eg, hydroxyl groups).

[0413] VI. Kit

[0414] The composition may be packaged in a kit. The kit may include a single or multiple doses of a composition comprising one or more psychedelic drugs bound or combined with a dendrimer (e.g., one or more hydroxyl-terminated PAMAM dendrimers or glucose dendrimers as described in the Examples), and instructions for administering the composition. Specifically, the instructions direct administration of an effective amount of the dendrimer composition to an individual suffering from a particular disease / disorder. The composition may be formulated with reference to a particular treatment method as described above, and may be packaged in any convenient manner.

[0415] The invention will be further understood with reference to the following non-limiting examples. Example

[0416] Example 1: Synthesis of hydroxy-polyamidoamine (PAMAM-OH) dendrimer psychedelic conjugates

[0417] The synthesis of PAMAM-OH-psychedelic conjugates is achieved through a variety of linking chemistries and linkers (cleavable and non-cleavable). In short, the surface hydroxyl groups on the PAMAM-OH are modified with a linker to produce a complementary group on the surface that can further react with a complementary group on the drug linker. The drug compound is modified by a linker to produce a complementary functional group to react with the dendrimer linker. The linker on the drug is connected by a cleavable or non-cleavable linker. Examples of cleavable links include esterase-sensitive ester bonds, glutathione-sensitive disulfide bonds, phosphatase-sensitive phosphodiester bonds, triglycyl peptide linkers (CX) capable of lysosomal release, and acid-cleavable hydrazine links. Examples of non-cleavable links include ether bonds, aminoalkyl bonds, or amide bonds. The linker can be an amino acid, a peptide, polyethylene glycol (n=2-15), or a hydrocarbon chain.

[0418] 1. Synthesis of dendrimer-dimethyl-4-hydroxytryptamine conjugates with cleavable ester linkages

[0419] according to Figure 1A and 1B The reaction shown prepares a dendrimer-dimethyl-4-HT conjugate with a cleavable ester. Figure 1A and 1B is a schematic diagram showing the stepwise synthetic route for conjugating dimethyl-4-hydroxytryptamine to dendrimers using click chemistry.

[0420] 2. Synthesis of dendrimers with non-cleavable amide linkages - dimethyl-4-hydroxytryptamine analogs

[0421] according to Figure 2A and 2B The reaction shown synthesizes a dendrimer-dimethyl-4-HT conjugate having a non-cleavable amide linkage. Figure 2A and 2B Schematic representation of the stepwise synthetic route for conjugating dimethyl-4-HT analogs to dendrimers using click chemistry.

[0422] 3. Synthesis of Ketamine, a Dendrimer with Non-Cleavable Amino-Alkyl Linkers

[0423] according to Figure 3A and 3B The reaction shown synthesizes a dendrimer-ketamine conjugate with a non-cleavable amino-alkyl linkage. Figure 3A and 3Bis a schematic diagram of an exemplary synthetic route for conjugating ketamine to dendrimers using copper-catalyzed alkyne azide click chemistry ( Figure 3B ).

[0424] 4. Synthesis of Dendrimer-DMT Analogs with Non-cleavable Amide Linkers

[0425] according to Figure 4A and 4B The reaction shown synthesizes a dendrimer-N,N-dimethyltryptamine (dendrimer-DMT) conjugate with a non-cleavable amide linkage. Figure 4A and 4B is a schematic diagram of an exemplary step-by-step synthetic route for conjugating DMT to dendrimers using copper-catalyzed alkyne azide click chemistry.

[0426] 5. Synthesis of dendrimer-DMT analogs with non-cleavable amino-alkyl linkages

[0427] according to Figure 5A and 5B The reaction shown also forms a dendrimer-DMT conjugate with a non-cleavable amino-alkyl linkage. Figure 5A and 5B is a schematic diagram of an exemplary stepwise synthetic route for conjugating DMT to dendrimers using copper-catalyzed alkyne azide click chemistry.

[0428] 6. Synthesis of dendrimer-LSD with non-cleavable amino-alkyl linkages

[0429] according to Fig. 6A and 6B The reaction shown synthesizes a dendrimer-lysergic acid diethylamide (dendrimer-LSD) conjugate having a non-cleavable amino-alkyl linkage. Fig. 6A and 6B is a schematic diagram of an exemplary step-by-step synthetic route for conjugating LSD to dendrimers using copper-catalyzed alkyne azide click chemistry.

[0430] in conclusion

[0431] Dendrimer conjugates direct drugs to target sites, thereby providing site-specific targeting, lower doses, enhanced efficacy, and reduced side effects associated with free drugs. The PAMAM-OH platform has extremely high water solubility (>300mg / mL). Most psychedelics have poor water solubility in the ug / mL range. Dendrimer conjugates will increase water solubility 10-100 times compared to free drugs. Sustained intracellular release at the target will avoid systemic and dose-related side effects of free psychedelic drugs. Dendrimer conjugates can significantly reduce the time required for drugs to take effect.

[0432] Example 2: Synthesis of Glucose Dendrimer (GD) Psychedelic Conjugate

[0433] The synthesis of glucose dendrimer-psychedelic conjugates is achieved using a combination of linking chemistries and linkers (cleavable and non-cleavable). Briefly, surface hydroxyl groups on the glucose dendrimer are modified with linkers to promote the reaction of complementary groups on the surface with complementary groups on the drug linker. The drug compound is also modified by the linker to promote the reaction between the complementary functional group and the dendrimer linker. The linker on the drug compound is connected by a cleavable or non-cleavable linker. Examples of cleavable links include esterase-sensitive ester bonds, glutathione-sensitive disulfide bonds, phosphatase-sensitive phosphodiester bonds, triglycyl peptide linkers (CX) capable of lysosomal release, and acid-cleavable hydrazine bonds. Examples of non-cleavable links include ether bonds or amide bonds. The linker can be an amino acid, a peptide, polyethylene glycol (n=2-15), or a hydrocarbon chain.

[0434] 1. Synthesis of dimethyl-4-hydroxytryptamine, a dendrimer with cleavable ester linkages

[0435] according to Figure 7 The reaction shown in the figure is used to prepare a glucose dendrimer-dimethyl-4-hydroxytryptamine conjugate having a cleavable ester linkage. The synthesis process of dimethyl-4-hydroxytryptamine-azide is as follows Figure 1A The synthesis of the glucose dendrimer and dimethyl-4-hydroxytryptamine conjugate is to partially modify the OH group of the glucose dendrimer using the complementary group on the surface of the glucose dendrimer, and then react with the complementary linker containing azide connected to dimethyl-4-hydroxytryptamine to generate a glucose dendrimer-dimethyl-4-hydroxytryptamine conjugate ( Figure 7 ).

[0436] 2. Synthesis of Dendrimer-Dimethyl-4-HT Analog Conjugates with Non-cleavable Amide Linkers

[0437] according to Figure 8 The reaction shown in the figure is used to prepare a glucose dendrimer-dimethyl-4-hydroxytryptamine conjugate with a non-cleavable amide linkage. The synthetic scheme of dimethyl-4-hydroxytryptamine analog-azide is as follows Figure 2A The synthesis of the glucose dendrimer and dimethyl-4-hydroxytryptamine analog conjugate is to partially modify the OH group of the glucose dendrimer using the complementary group on the surface of the glucose dendrimer, and then react with the complementary linker containing azide connected to the dimethyl-4-hydroxytryptamine analog to generate the glucose dendrimer-dimethyl-4-hydroxytryptamine analog conjugate ( Figure 8 ).

[0438] 3. Synthesis of Ketamine Dendrimers with Non-Cleavable Amino-Alkyl Linkers

[0439] according to Fig. 9 The glucose dendrimer-ketamine conjugate with a cleavable amino-alkyl linkage was prepared by the reaction shown in FIG. 1 . Ketamine hydrochloride (1) was first modified with an alkyne, such as Figure 3A An exemplary synthetic route for conjugating glucose dendrimers to ketamine is shown in Fig. 9 shown.

[0440] 4. Synthesis of Dendrimer-DMT Analogs with Non-cleavable Amide Linkers

[0441] according to Fig.10 The glucose dendrimer-DMT conjugate with a non-cleavable amide linkage is prepared by the reaction shown in the figure. First, N,N-dimethyltryptamine analog (DMT analog) is linked to a linker with an azide moiety via an amide linkage ( Figure 4A ). An exemplary synthetic route of glucose dendrimer and DMT conjugate is as follows Fig.10 shown.

[0442] 5. Synthesis of Dendrimer-DMT with Non-cleavable Amino-Alkyl Linkers

[0443] according to Fig.11 The glucose dendrimer-DMT conjugate with a non-cleavable amino-alkyl linkage is prepared by the reaction shown in FIG. First, an N,N-dimethyltryptamine analog (DMT analog) is modified with an alkynyl group, such as Figure 5A The synthetic route of glucose dendrimer and DMT conjugate is shown in Fig.11 shown.

[0444] 6. Synthesis of glucose dendrimers with non-cleavable amino-alkyl linkages - LSD

[0445] according to Fig.12 The glucose dendrimer-lysergic acid diethylamide (LSD) conjugate with a non-cleavable amino-alkyl linkage was prepared by the reaction shown in FIG. First, LSD was modified with an alkyne group, such as Fig. 6A An exemplary synthesis route of glucose dendrimer and LSD conjugate is shown in Fig.12 shown.

[0446] in conclusion

[0447] Dendrimer conjugation provides site-specific targeting by directing drugs to target sites, lowering dosage, enhancing efficacy, and reducing side effects associated with free drugs. The glucose dendrimer platform has very high water solubility (>500mg / mL). Most psychedelic drugs have poor water solubility in the ug / mL range. Dendrimer conjugation will increase water solubility 10-100 fold compared to unconjugated free drug. Sustained intracellular release at the target will avoid systemic and dose-related side effects of free psychedelic drugs. Dendrimer conjugation can significantly reduce the time required for drug activity to begin.

[0448] These compositions can deliver drugs to receptors on specific cells (neurons, glial cells, macrophages), including targets on the cell surface and inside the cell. These formulations can enhance the effectiveness of these drugs, thereby reducing dosages, bringing new mechanistic insights, reducing side effects, and improving solubility, formulation, and pharmacokinetics.

[0449] Overall, dendrimer-based psychedelics will significantly improve the safety, efficacy, reproducibility, and ease of implementation of these molecules.

[0450] Example 3: Synthesis and Binding Properties of Dendrimer Conjugates

[0451] New chemical entities of psychedelic drugs and norketamine with hydroxyl-terminated PAMAM and glucose dendrimers have been synthesized and studies have been conducted to determine in vivo efficacy and targeting data for norketamine / ketamine and binding affinity data for D-tryptamine.

[0452] Materials and Methods

[0453] Fig.14A Schematic diagram of the synthesis of PAMAM dendrimer-norketamine conjugate. Fig. 14B Schematic diagram of the synthesis of glucose dendrimer-norketamine conjugate.

[0454] Unless otherwise stated, reactions were performed in flame-dried glassware under a positive pressure of nitrogen 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)-monoNHS ester was purchased from Amersham Bioscience-GE Healthcare. Deuterated solvents dimethyl sulfoxide (DMSO-d6), water (D2O), and chloroform (CDCl3) were purchased from Cambridge Isotope Laboratories Inc. (Andover, MA). Ethylenediamine core polyamidoamine (PAMAM) dendrimers, generation 4.0, hydroxyl surface (G4-OH; diagnostic grade; consisting of 64 hydroxyl end groups), methanol solution (13.75% w / w) were 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, norketamine, 5-hydroxytryptamine, dimethyl-2-hydroxytryptamine phosphate analog, dimethyl-2-hydroxytryptamine phosphate analog, and cannabidiol drugs were purchased from Cayman Chemicals.

[0455] Synthesis of hydroxy-PAMAM dendrimer drug conjugates.

[0456] The PAMAM-G4-OH(D4-OH) dendrimer consisting of approximately 64 terminal hydroxyl groups was used in the synthesis. After each synthetic step, the product was purified by dialysis in DMF for 24 h to eliminate small molecule impurities, followed by dialysis against water to remove DMF. 1H NMR (in DMSO-d6 and D2O) and analytical HPLC were used to confirm the formation and purity of the intermediates and final products. The monofunctional D4-OH was functionalized with alkyne groups to afford the D-hexyne difunctional dendrimer by treating 5-hexynoic acid with EDC.HCl and 4-DMAP in DMF at room temperature under standard esterification conditions for 36 hours. The number of alkyne groups on the surface of the dendrimer was selected to be maintained at ~10-15 to maintain the overall water solubility of the conjugate. The crude product was dialyzed against ultrapure water using a 1 kDa membrane for 24 hours to remove low molecular weight impurities by selective diffusion across the semipermeable dialysis membrane. 1 HNMR and analytical HPLC confirmed product formation and the purity of the intermediates and final products. These were shown in Fig.12 A-12C.

[0457] Synthesis of D-hexyne

[0458] 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), DMAP (2.41 g, 12.6 mmol) and stirred at room temperature for 5 minutes. EDC.HCl (1.54 g, 12.6 mmol) was then added to the reaction mixture in portions over 5 minutes. The reaction mixture was stirred at room temperature for 36 hours. The crude product was transferred to a 1 kD MW cut-off cellulose dialysis tubing and dialyzed against DMF for 12 hours and then against water for 24 hours. The aqueous layer was frozen and lyophilized to give D-hexyne as a hygroscopic white solid (yield 75%). 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 -99%.

[0459] Synthesis of dendrimer-drug conjugates.

[0460] A solution of D-hexyne and drug azide in DMF (5 mL) was treated with copper sulfate pentahydrate (CuSO4.5H2O) and sodium ascorbate aqueous solution. The reaction mixture was stirred in a microwave synthesizer and heated at 50°C for 10 hours. Upon completion, the reaction mixture was dialyzed against DMF in a 1 KDa cutoff cellulose dialysis tubing. To this solution was added EDTA (50 μL, 0.5 M) solution to remove copper by chelation. Dialysis against DMF was followed by dialysis against water overnight. Drug loading was calculated by proton integration, where the peaks corresponding to the dendrimer and drug were compared.

[0461] G2-glucose dendrimer (GD2)-drug conjugate

[0462] The second generation of glucose dendrimers (GD2) consist of 24 glucose molecules (96 surface hydroxyl groups) for conjugation. Glucose dendrimers are primarily composed of glucose moieties consisting of a central core of dipentaerythritol and one or more branching units of monosaccharide glucose molecules. Unlike hydroxyl-terminated PAMAM dendrimers, glucose dendrimers are primarily taken up by damaged neurons and were found to specifically target overexcitable neurons in culture and in vivo mouse models.

[0463] Synthesis and characterization of glucose dendrimers (GD2).

[0464] The GD synthesis begins with the reaction of the hexapropargylated core with AB4 and β-D-glucose-PEG4-azide to generate the first generation of glucose dendrimers (GD1) via a click reaction. The OH groups on GD1 are propargylated to give GD1-acetylene 24, which reacts with β-D-glucose-PEG4-azide to give the second generation (GD2) with 24 glucose moieties, providing 96 surface hydroxyl groups. In addition, a Cy5 fluorescent tag is attached to GD2 by propargylation of ~2-3 hydroxyl groups to give the alkyne-containing GD2 dendrimer. The GD intermediates and final products are purified using dialysis and used 1 The physicochemical properties of GD2 dendrimers were also evaluated (Table 2).

[0465] Table 2: Physicochemical properties of GD2

[0466] size 4.4nm Zeta Potential -6.0 Glucose quantity 24 HPLC purity >99％ Molecular weight 12800Da

[0467] Synthesis and characterization of GD2-drug conjugates

[0468] A click chemistry strategy was used to conjugate the drugs of norketamine, tryptamine, venlafaxine, and Hu308 to GD2-hexynoic acid dendrimers. The linker-linked drug moiety was conjugated to the glucose dendrimer using a Cu(I)-catalyzed click (CuAAC) reaction in the presence of catalytic amounts of CuSO4.5H2O and sodium ascorbate to afford the GD2-drug conjugates. Trace amounts of copper were removed by dialysis against ethylenediaminetetraacetic acid (EDTA). The final GD2-drug conjugates were characterized by NMR and HPLC.

[0469] Table 3: Drugs and their combinations

[0470]

[0471] Table 4. Physical properties of D-drug conjugates.

[0472]

[0473] *https: / / go.drugbank.com / metabolites / DBMET00189 Example 4: Binding assay of dendrimer-ketamine conjugates

[0474] Materials and methods

[0475] Human Serotonin 5-HT2A Receptor (Agonist Radioligand):

[0476] Objective: To evaluate the affinity of compounds for the human 5-HT2A receptor in transfected HEK-293 cells by radioligand binding assay.

[0477] Experimental protocol: Cell membrane homogenates (30 μg of protein) were incubated with 0.1 nM [125I]DOI 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 in the presence or absence of test compounds.

[0478] Nonspecific binding was determined in the presence of 1 μM DOI.

[0479] After incubation, the samples were quickly filtered under vacuum through glass fiber filters (GF / B, Packard) pre-soaked with 0.3% PEI and rinsed several times with ice-cold 50 mM Tris-HCl using a 96-sample cell harvester (Unifilter, Packard). The filters were dried and the radioactivity was counted in a scintillation counter (Topcount, Packard) using scintillation cocktail (Microscint 0, Packard).

[0480] Results are expressed as percent inhibition of specific binding of control radioligand.

[0481] The standard reference compound is DOI, which is tested in each experiment at several concentrations to obtain competition curves from which its IC50 is calculated.

[0482] See Bryant et al. (1996), A novel class of 5-HT2A receptor antagonists: aryl aminoguanidines, Life Sci., 15:1259.

[0483] Delta (DOP) Human Opioid GPCR Cell Antagonist cAMP Assay:

[0484] Purpose: To evaluate the potency (IC50) and efficacy (maximum response) of compounds against the human delta (DOP) receptor in stably transfected CHO-K1 cells. The assay principle is a cAMP cell-based assay.

[0485] Experimental Protocol A total volume of 20 μL of cells was seeded into a white-walled 384-well microplate and incubated overnight at 37° C. Prior to testing, the cell seeding medium was replaced with 10 uL of assay buffer (HBSS+10 mM HEPES).

[0486] Briefly, intermediate dilutions were made to the sample stock to generate 4X samples in assay buffer. 5 μL of 4X sample was added to the cells and incubated at 37°C for 30 minutes. 5 uL of 4X EC80 was added to 4X Forskolin reagent and cells were incubated at 37°C for 30 minutes. The final assay vehicle concentration was 1%. Results are expressed as percent inhibition of the control ligand.

[0487] General information

[0488]

[0489] TA1 Human Trace Amine GPCR Cellular Agonist cAMP Assay

[0490] Objective: To evaluate the potency (EC50) and efficacy (maximal response) of compounds at the human TA1 receptor in stably transfected CHO-K1 cells using a GPCR cell-based cAMP assay.

[0491] Experimental Protocol: Cells in a total volume of 20 μL were seeded into white-walled 384-well microplates and incubated overnight at 37 °C prior to testing.

[0492] Prior to testing, the cell seeding medium was replaced with 15 μL of assay buffer (HBSS + 10 mM HEPES). Briefly, intermediate dilutions were made to the sample stock solution to generate 4X samples in assay buffer. 5 μL of 4X samples were added to the cells and incubated at 37°C for 30 minutes. The final assay vehicle concentration was 1%.

[0493] Results are expressed as percent efficacy relative to the maximal response of the control ligand.

[0494] General information

[0495]

[0496] result

[0497] Table 5: Results of the binding test

[0498]

[0499] Fig.15A Figure 1 is a graph of the NMDAR 1A / 2B antagonist assays of glucose dendrimer-ketamine (IC50 = 4.54 μM), hydroxy dendrimer-ketamine (IC50>100), and norketamine (IC50 = 6.96 μM). Fig. 15B is the percent binding efficiency of the micromolar log concentration of the compound in the D2L human dopamine GPCR cell agonist cAMP assay. Norketamine (closed circles), glucose dendrimer-ketamine EC50 = 13.08 micromolar (open circles), and hydroxy dendrimer-ketamine EC50 = 4.263 micromolar (triangles). Fig. 15C is the percent binding efficiency of micromolar log concentrations of ketamine in the TA1 human microamine GPCR cell agonist cAMP assay. Norketamine (closed circles), glucose dendrimer-ketamine EC50 = 13.08 micromolar (open circles), and hydroxy dendrimer-ketamine EC50 = 4.263 micromolar (triangles).

[0500] Significant efficacy data have been obtained for a clinically important ketamine derivative (norketamine), which has been tested in the treatment of depression and addiction. Hydroxy dendrimer drugs target these receptors in microglia / macrophages, while glucose dendrimer drugs target these receptors in neurons and microglia.

[0501] The conjugate can be active with or without drug release. Binding affinity is a measure of the activity of the intact conjugate. Often, conjugates of psychedelic drugs are prepared in a non-releasing form (e.g., tryptamine, dimethyl-4-hydroxytryptamine, dimethyl-2-hydroxytryptamine phosphate, ketamine) and are intended to be active in the intact form. This allows the intact conjugate to be released through the kidney without toxicity from drug release. Alternatively, they can be designed to be released through an analog.

[0502] Both GD-ketamine and HD-ketamine exhibit unique effects that may be beneficial for neuropsychiatric disorders. Both dendrimer-ketamine conjugates bind to opioid μ, opioid k, and sigma 2 receptors with greater affinity than norketamine, suggesting that the dendrimer conjugates have a positive effect on binding to these receptors. Norketamine does not bind to opioid μ receptors (Ki 53 μM for GD-ket and 40 μM for HD-ket). Norketamine has a Ki of 381 uM for opioid k receptors, while this constant is approximately 4-fold higher for GD-ket (Ki = 82 μM) and 7-fold higher for HD-Ket (Ki = 556 μm). The antidepressant effects of ketamine are thought to be mediated through opioid k receptors, and dendrimer binding may increase affinity for these receptors.

[0503] Binding to sigma 2 receptors was also improved after dendrimer conjugation. A stronger ability of HD ketamine to bind to sigma 2 receptors indicates a stronger neuroprotective effect. The neuroprotective mechanism may be related to increased production of NGF and BDNF. Activation of sigma 2 receptors is also beneficial in the treatment of neuropsychiatric disorders such as schizophrenia and psychosis. Sigma 2 receptor targeted therapy can reduce anxiety and enhance antidepressant effects. Ketamine binds to sigma 2 receptors at mM concentrations (Pergolizi, 2023; Bonaventura, 2021).

[0504] Only dendrimer-conjugated ketamine has the ability to bind to dopamine 2 receptors. Free norketamine does not appear to bind to dopamine 2 receptors. Likewise, ketamine does not show any affinity for dopamine 2 receptors. The dopaminergic effects of ketamine are thought to be indirect. However, studies have shown that glucose dendrimer-ketamine has an RC50 of 13.07 μM at D2 receptors, while hydroxy dendrimer-ketamine is more effective with an RC50 of 4.3 μM. The antidepressant and neuroprotective effects of ketamine are primarily mediated through these and dopamine receptors. Ketamine, norketamine, and ketamine metabolites do not appear to have a direct effect on dopamine receptors and do not bind to dopamine receptors. The increased D2 receptor activity observed in functional assays suggests that dendrimer-ketamine will be more effective as an antidepressant.

[0505] GD-ketamine and HD-ketamine exhibited different profiles. Binding to hydroxy dendrimers (HD) appeared to alter ketamine function compared to glucose dendrimers (GD), as expected. HD-ketamine did not exhibit NMDAR 1A / 2B ion channel blockade, while GD-ketamine showed that the type of dendrimer bound to ketamine was critical for the functional differences between them. GD-ketamine also exhibited agonist activity against 5HT1A receptors, while HD-ketamine did not.

[0506] These results suggest that dendrimer conjugation provides unexpected benefits on receptor binding of free drug and is dependent on the structure of the dendrimer. The microglial targeting of hydroxy dendrimers and the additional neuronal targeting of glucose dendrimers provide unexpected results with clinical implications.

[0507] These conjugates target specific receptors on any cell they are on. This includes neurons, microglia, macrophages, and other cells. These receptors may be located anywhere in the brain. Examples include serotonin receptors such as 5HT1A, 5HT2A, NMDA, etc. Serotonin receptors can also be found in microglia. These compounds bind to and act on these receptors. The "intrinsic cell targeting" of these dendrimers (hydroxy dendrimers-microglia / macrophages and glucose dendrimers-neurons) is somewhat secondary to the action on specific receptors because these compositions target specific receptors. Important receptors include serotonin receptors [5HT1A, 5HT2A (agonist, antagonist, inverse or inverse agonist)] and NMDA receptors. In many cases, using dendrimers to bind to specific receptors without binding to other receptors can enhance the binding efficacy and reduce the side effects of these drugs.

[0508] When combined with the cell targeting capabilities of glucose dendrimers, e.g. to damaged neurons (primary), microglia / macrophages (secondary), or hydroxy dendrimers (microglia / macrophages), dendrimer conjugation can increase water solubility by more than 5-200 fold, these conjugates offer distinct benefits: modifiable binding, improved targeting selectivity, easier formulation and delivery, and reduced side effects.

[0509] In the case of tryptamine, the results showed that the binding affinity of drugs conjugated to these dendrimers had unexpected properties.

[0510] Both hydroxy and glucose dendrimers have OH surface groups. When tryptamines were conjugated to these dendrimers with the same attachment chemistry, the affinities for serotonin and other receptors varied greatly. In cell-based binding assays (indicative of in vivo efficacy), the conjugates exhibited lower affinity than the free drug. The lower affinity may make the binding less tight and may allow us to modulate the undesirably strong effects of the drug on this receptor. Second, the hydroxy dendrimer conjugates were not active, but the glucose dendrimer conjugates were. This was unexpected and may be due to the different internal structures of the glucose and hydroxy dendrimers. The drug may fold into the hydrophobic core of the hydroxy dendrimers, but may open outward in the hydrophilic interior of the glucose dendrimers.

[0511] Example 5: Treatment of Rett syndrome animal model with ketamine and ketamine-dendrimer conjugates

[0512] Rett syndrome (RTT) is an inherited female neurodevelopmental disorder that affects 1 in 10,000-15,000 births. Affected females develop normally for 6-18 months, but then lose voluntary motor skills, including speech and hand skills. Most people with RTT are heterozygous for mutations in the X-linked gene MECP2, which encodes a protein that binds to methylated sites in genomic DNA and promotes gene silencing. The symptoms, progression, and severity of Rett syndrome vary from person to person. A variety of disabilities may be associated with Rett syndrome. Symptoms typically develop in stages. RTT is typically characterized by a period of normal development after birth, followed by regression of speech and hand movements, abnormal gait, irregular hand movements, and slowed head growth. Other diagnostic criteria for RTT include irregular breathing, gastrointestinal and musculoskeletal problems, seizures, poor sleep, decreased response to physical pain, and behavioral problems.

[0513] Ketamine is a well-established anesthetic that produces “dissociative anesthesia” and exerts central and peripheral effects, including hypnotic, analgesic, and sympathomimetic effects, resulting in hypertension and tachycardia. The primary mechanism is thought to be due to its action as an N-methyl-d-aspartate (NMDA) receptor antagonist. This can result in rapid action and response in the treatment of refractory depression, MDD, and suicidal ideation, whereas SSRIs and SNRIs only show delayed effects in the control of depression. Ketamine is also effective in the treatment of chronic pain, again due to its effects on NMDAR inhibition. Increased dopamine release and its effects on However, ketamine also exerts neuroprotective effects through non-NMDAR-mediated mechanisms, such as increasing BDNF and mTOR. Ketamine also has effects on opioid receptors, which can increase dopamine. These effects are beneficial in chronic pain, refractory depression, MDD, and refractory epilepsy / seizures.

[0514] However, ketamine has several short-term and long-term side effects. High doses of ketamine can cause respiratory depression and may result in systemic side effects such as increased heart rate, hypertension, hyperthermia, loss of coordination, dizziness, nausea, vomiting, sensory perceptual disturbances, and high rates of auditory and visual hallucinations. More than half of patients treated with ketamine experience a phenomenon when ketamine wears off, characterized by euphoria, vivid dreams, hallucinations, illusions, distortions of body image and objects, and delirium, which can be very disturbing and may lead to self-harm. Some of these symptoms are associated with symptoms of schizophrenia. Long-term use of ketamine can lead to impaired memory and decreased executive function. Ketamine also causes tolerance and has addictive properties, leading to withdrawal and dependence. Due to these significant side effects, ketamine should only be used in controlled environments.

[0515] Ketamine has been shown to be effective in a mouse model of Rett syndrome, with high-dose ketamine given IP at 8 mg / kg daily for 40 days (total dose 320 mg / kg) increasing survival by 50% at 80 days postnatally (Patrizi et al., 2016)

[0516] Materials and methods

[0517] Dendrimer conjugates were prepared as described above.

[0518] Guy et al. reported that deletion mutations in the Mecp2 gene in mice can cause neurological symptoms similar to Rett syndrome (Nat Genet. 2001 Mar; 27(3):322-6. doi:10.1038 / 85899). PMID: 11242117 Mice with Mecp2 gene deletion and mice with Mecp2 gene deletion in the brain both develop severe neurological symptoms at about six weeks of age.

[0519] 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. MID:26410354; PMCID:PMC7410367) report a systematic, randomized preclinical trial in which Mecp2-deficient mice were chronically administered low-dose (8 mg / kg, intraperitoneally) ketamine, an NMDAR antagonist, starting early in development or at the onset of the RTT phenotype. Mice were treated with 8 mg ketamine / kg / day intraperitoneally from day 15 to day 55 or from day 30 to day 55 (total dose of 320 mg / kg for 40 days, or 200 mg / kg for 25 days). Treatment starting from day 30 was ineffective, while treatment starting from day 15 showed some effect at a total dose of 320 mg / kg.

[0520] Experimental Paradigm for MECP2 Knockout ("KO") Mice:

[0521] Treatment was started at 28 days of age (4 weeks of age, symptomatic). Untreated animals died after approximately 55-60 days.

[0522] Mice were treated with ketamine or ketamine conjugated to dendrimers. Controls were wild-type mice, which were treated with saline; knockout mice were treated with saline.

[0523] Animals were given 2.5 mg / kg intraperitoneally twice weekly. The total dose at the time of evaluation (60 days of age) was approximately 22.5 mg / kg.

[0524] Dendrimer ketamine was tested in the Rett syndrome mouse model at a dose of 2.5 mg / kg (ketamine) IP every two weeks for 8 weeks (total ketamine dose 40 mg / kg).

[0525] Mecp2 KO (gene knockout) at 4 weeks old were randomized and treated with saline or 2.5mg / Kg intraperitoneal injection of ketamine or D-ketamine every two weeks for 8 weeks (WT-saline group, KO-saline group, KO-ketamine group, KO D-ketamine group). Neurobehavioral performance was assessed weekly by recording their composite neurobehavioral score (NBS), which is based on a scale that includes assessment of mobility, gait, paw clenching, tremor, and respiration, with scores of 0-3 for each scale; the higher the score, the worse the phenotype. The D-ketamine treated group showed a slowed progression of the disease phenotype and better neurobehavioral scores after treatment, while the untreated KO did not.

[0526]

[0527] Open Field Test: Long-term behavioral changes in saline (WT), ketamine, and D-ketamine groups compared to the KO-saline group at week 8.5 of treatment. Motor function was assessed by recording mice in an open field arena (10.5" x 19" x 8"). The room where the mice were recorded was the same as where they were housed to avoid stress and changes during testing. 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 open field 5 cm away from the longer walls in the same manner.

[0528] result

[0529] Animals were evaluated for survival, neurobehavioral scores, and activity rates (total distance traveled, speed, and time spent in a corner).

[0530] Fig.16A and 16B is the composite neurobehavioral score of wild-type, knockout saline (control) and knockout mice treated with dendrimer-ketamine conjugate ( Fig.16A ) and postnatal survival ( Fig. 16B ) in the table below. Fig. 16C is a graph of distance travelled (m); Fig.16D is a graph of the speed at which mice travel; Fig.16E is a graph of the time spent in the corner.

[0531] Dendrimers conjugated to ketamine improve efficacy and increase binding to NMDARs without the associated side effects. The doses used were significantly lower than free ketamine, which reduced side effects. This was tested in a Rett syndrome mouse model as proof of concept, as Rett syndrome is a disease with increased glutamate production and increased NMDAR expression / activation.

[0532] Animals treated with D-ketamine showed significantly improved survival, with 100% survival up to 90 days, compared to untreated animals and animals treated with free ketamine. Previous data published by others in a similar model showed only 50% survival after treatment with 320mg / kg ketamine (an 8-fold higher dose).

[0533] After administration of D-ketamine, motor function was significantly improved and behavior was similar to that of normal healthy controls.

[0534] There was a significant difference in the total distance traveled between WT and KO mice in the 10-min test in the open field. In the open field test, the total distance traveled increased in KO mice treated with D-ketamine, and motor function was also significantly improved. The maximum speed and time spent in the corner were significantly improved in the D-ketamine group compared to the KO saline group.

[0535] The video showed that their phenotype had improved significantly and was close to that of healthy mice.

Claims

1. A composition comprising a hydroxyl-terminated dendrimer, a sugar-terminated dendrimer and / or a sugar-based dendrimer covalently conjugated to at least one or more hallucinogens or psychedelics, optionally via a spacer, wherein the hallucinogens or psychedelics are not cannabinoids.

2. The composition of claim 1, wherein the dendrimer is a 1st, 2nd, 3rd, 4th, 5th, 6th, 7th or 8th generation dendrimer.

3. The composition according to claim 1 or 2, wherein the dendrimer is a glucose dendrimer, preferably a 1st, 2nd or 3rd generation glucose dendrimer.

4. A composition according to claim 2, wherein the dendrimer is a hydroxyl terminated poly(amidoamine) (PAMAM) dendrimer, preferably having greater than 40% or 50% hydroxyl surface groups.

5. The composition of any one of claims 1-4, wherein the dendrimer is a 3rd, 4th, 5th, 6th or 7th generation dendrimer.

6. A composition according to any one of claims 1, 2 or 5 comprising sugar-capped dendrimers comprising on their surface one or more monosaccharides selected from the group consisting of glucose, galactose, glucosamine, galactose, mannose and fructose, preferably greater than ten surface monosaccharide moieties.

7. A composition according to any one of claims 1 to 3, 5 or 6, wherein the dendrimer is made from monosaccharide and optionally ethylene glycol building blocks, preferably from galactose or glucose and optionally ethylene glycol and has more than 10 surface sugar moieties.

8. The composition according to claims 1-3 or 5-7, wherein the monosaccharide building blocks are one or more selected from glucose, galactose, glucosamine, galactose, mannose and fructose, wherein the dendrimer is preferably a glucose dendrimer.

9. The composition of any one of claims 1-8, wherein the dendrimer is conjugated to one or more hallucinogens via a spacer having a cleavable bond or a non-cleavable bond.

10. The composition of claim 9, wherein the cleavable bond is selected from the group consisting of an ester bond, a disulfide bond, a phosphodiester bond, a triglycine bond, and a hydrazine bond.

11. The composition of claim 9, wherein the non-cleavable bond is selected from the group consisting of an amide bond, an ether bond, and an aminoalkyl bond.

12. A composition according to any one of claims 1 to 11, wherein the spacer linking the dendrimer and the hallucinogen comprises a hydrocarbon such as an olefin, a diethylene glycol moiety and / or an oligoethylene glycol chain.

13. The composition of any one of claims 1 to 12, wherein the spacer comprises a triazole moiety.

14. A composition according to any one of claims 1-13, wherein the hallucinogen is selected from the group consisting of classical serotonergic hallucinogens, for example wherein the drug is an inducing agent, a dissociative anesthetic, or an atypical hallucinogen.

15. The composition of any one of claims 1-14, wherein at least one or more hallucinogens or psychedelics comprise dimethyl-4-hydroxytryptamine, ketamine (R-ketamine, S-ketamine, (R / S)-ketamine), norketamine, ketamine metabolites, N,N-dimethyltryptamine (DMT), 4-acetoxy-N,N-dimethyltryptamine, 5-methoxy-DMT, 5-chloro-DMT, lysergic acid monourea (LSD), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), dimethyl-2-hydroxytryptamine phosphate, ibogaine, mescaline, mianserin, 2,5-dimethoxy-4-iodoamphetamine (DOI), ayahuasca, 1-(1-phenylcyclohexyl)piperidine (PCP), norcyanobacterial toxins, or analogs of derivatives of these agents.

16. The composition of claim 14, wherein the classical serotonin hallucinogen is selected from the group consisting of dimethyl-2-hydroxytryptamine phosphate, dimethyl-4-hydroxytryptamine, lysergic acid diethylamide (LSD), mescaline, mianserin, and 2,5-dimethoxy-4-iodoamphetamine (DOI).

17. The composition of claim 14, wherein the inducing agent is selected from 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA).

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

19. The composition of claim 1, wherein the composition provides sustained release to produce an effective amount over a period of 24 hours.

20. The composition of any one of claims 1-19, wherein the concentration of one or more hallucinogens conjugated to the dendrimer is from about 0.01% to about 30%, preferably from about 1% to about 20%, more preferably from about 5% to about 20% by weight.

21. The composition according to any one of claims 1-20, wherein the composition further comprises one or more diagnostic agents, preferably wherein the diagnostic agent is selected from the group consisting of fluorescent dyes, near infrared dyes, SPECT imaging agents, PET imaging agents and radioisotopes.

22. A composition according to any one of claims 1-21 which provides sustained delivery of an agent or a level of sustained receptor activity to achieve clinical benefit without hallucinations, or to maintain a psychotic state at a relatively constant dose, preferably at a therapeutically effective level for 24 hours.

23. A pharmaceutical preparation comprising the dendrimer according to any one of claims 1 to 22, and a pharmaceutically acceptable carrier or excipient.

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

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

26. The pharmaceutical formulation of claim 23, wherein the formulation is formulated for oral, mucosal (intranasal, buccal, rectal, vaginal, sublingual, pulmonary), intramuscular, intravenous, subcutaneous, transdermal or intrathecal administration.

27. The pharmaceutical composition according to any one of claims 23 to 26, in a form selected from the group consisting of hydrogels, nanoparticles or microparticles, suspensions, powders, tablets, capsules, creams and solutions.

28. A method for treating one or more psychological, cognitive, behavioral and / or emotional disorders in a subject in need thereof, the method comprising administering to the subject an effective amount of a composition of any one of claims 1-22 or a pharmaceutical composition of any one of claims 23-26 to treat, alleviate and / or prevent one or more symptoms associated with one or more psychological, cognitive, behavioral and / or emotional disorders.

29. The method of claim 28, wherein one or more of the psychological, cognitive, behavioral and / or emotional disorders are selected from apathy, low motivation, attention disorders, executive function and / or cognitive engagement disorders, obsessive compulsive disorder, neurocognitive disorders.

30. The method of claim 28 or 29, wherein the pharmaceutical composition is administered in an effective amount to provide improved motivation, attention, accuracy, reaction speed, persistence and / or cognitive engagement without adverse side effects.

31. The method of any one of claims 28-30, wherein the pharmaceutical composition is administered in an amount effective to provide binding of one or more hallucinogens covalently conjugated to the dendrimer to one or more receptors on the surface or within a target cell.

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

33. The method of claim 31, wherein the receptor is selected from the group consisting of 5-HT receptor subtypes, adrenergic receptors, dopaminergic receptors, and histamine receptors.

34. The method of claim 33, wherein the receptor is selected from the group consisting of 5-HT2, 5-HT6, 5-HT7, adrenergic alpha 2, D1 and D2 receptors.

35. The method of claim 31, wherein the receptor is 5-HT 2A .

36. A method according to any one of claims 28-35 comprising treating a psychiatric disorder, such as obsessive compulsive disorder, eating disorders, attention deficit and hyperactivity disorder and schizophrenia.

37. The method of any one of claims 28-35, comprising treating a patient suffering from depression, such as treatment-resistant depression, anxiety disorders, and post-traumatic stress disorder (PTSD).

38. A method according to any one of claims 28-35 comprising treating a patient suffering from drug and / or alcohol dependence.

39. The method of any one of claims 28-35 comprising treating a patient suffering from Alzheimer's disease, dementia, multiple sclerosis or other neurodegenerative disease.

40. The method of any one of claims 28-35 comprising treating patients suffering from chronic inflammatory and autoimmune diseases such as rheumatoid arthritis, atherosclerosis, Parkinson's disease, Alzheimer's disease and multiple sclerosis.

41. The method of any one of claims 28-35, comprising treating pain, including cluster headaches, migraines, chronic pain, postoperative pain, and cancer-related pain.

42. The method of any one of claims 28-41, wherein the composition provides efficacy for at least 24 hours after administration.

43. The method of any one of claims 28-42, wherein the composition comprises a glucose dendrimer-ketamine conjugate.

Images