Cannabinoid dendrimer compositions for targeted delivery

Abstract

The present invention provides dendrimer formulations comprising one or more cannabinoids and / or cannabinoid derivatives and methods of use thereof for targeted delivery, particularly to neurons and activated microglial cells, or other cells on which the CB1 or CB2 receptor is expressed. The cannabinoid-dendrimer conjugate can be administered to patients in need of treatment of various diseases, the various conditions include central and peripheral nerve diseases, chronic pain, neuropathic pain, neurodegeneration (senescence, dementia, Alzheimer's disease), traumatic brain injury, epileptic seizure, Huntington's disease and multiple sclerosis, epilepsy, inflammatory diseases, and pain.

Description

Field of the Invention

[0001] The present invention relates generally to the field of cannabinoid pharmaceutical formulations, and in particular to dendrimer-cannabinoid conjugates for selective delivery to the cannabinoid receptor family in the CNS and other disease sites.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Application No. 63 / 401,464, filed on August 26, 2022, entitled “CANNABINOID DENDRIMER COMPOSITIONS FOR TARGETED DELIVERY,” the applicant being Johns Hopkins University, the inventors being Kannan Rangaramanujam, Sujatha Kannan, Kunal Parikh, and Anjali Sharma, the entirety of which is hereby incorporated by reference herein.

[0004] Statement Regarding Federally Funded Research

[0005] none. Background of the Invention

[0007] Cannabinoids have beneficial effects on a range of clinical conditions, including pain, inflammation, epilepsy, sleep disorders, symptoms of multiple sclerosis, anorexia, schizophrenia, and other conditions. As a result, the conversion of cannabinoids from herbal preparations to strictly regulated prescription drugs is rapidly developing. Drugs such as cannabinoids are metabolized by liver and intestinal enzymes (first-pass hepatic metabolism), have specific pharmacokinetic requirements, have poor gastrointestinal permeability, and can cause irritation, so alternative approaches to systemic oral administration are needed. Cannabinoids have low oral bioavailability and high lipophilicity, making them difficult to deliver to specific targets. Currently, oils and capsules are more convenient and accurate than juices or teas from the original plant, but are less suitable for pharmacological applications.

[0008] Cannabinoids are highly lipophilic molecules (log P 6-7) with very low water solubility (2-10 μg / mL) and are susceptible to degradation by light and temperature as well as self-oxidation, especially in solution. Formulation plays a crucial role in improving the solubility and physicochemical stability of the drug. Common strategies used in marketed products include salt formation (i.e. pH adjustment), cosolvents (e.g. ethanol, propylene glycol, PEG400, etc.), micellization (e.g. polysorbate 80, cremophor ELP, etc.), (nano)-(micro)emulsification, complexation (e.g. cyclodextrins), and encapsulation in lipid-based formulations (e.g. liposomes) and nanoparticles. Formulations have been proposed for transdermal routes, intranasal administration, and transmucosal adsorption. However, absorption is slow, erratic, and variable. Maximum plasma concentrations are usually reached after 60-120 minutes, but may take longer (up to 6 hours) and may be delayed. In addition, metabolism produces psychoactive metabolites. Extensive first-pass hepatic metabolism further reduces the oral bioavailability of THC, while the duration of action varies from 8 to 20 hours.

[0009] The psychoactive effects of cannabis are primarily mediated by CB1, which is widely distributed throughout the brain but is primarily found in the frontal cortex, basal ganglia, and cerebellum. CB1 is also found in a variety of tissues and organs, including adipose tissue, gastrointestinal tract, spinal cord, adrenal and thyroid glands, liver, reproductive organs, and immune cells. The presence of CB1 receptors on chondrocytes and osteocytes, as well as evidence for CB1 receptors on fibroblast-like synoviocytes, makes CB1-targeted therapy particularly valuable in rheumatic diseases. CB1 activation inhibits adenylate cyclase and reduces cAMP levels and protein kinase A (PKA) activity, resulting in activation of type A potassium channels and reduced cellular potassium levels.

[0010] CB2 is primarily expressed in immune cells but can also be found in various other cell types. It is also present in some neural tissues, such as dorsal root ganglia and microglia. CB2 shares 44% amino acid similarity with CB1 and similarly inhibits adenylate cyclase as well as activates mitogen-activated protein kinase. CB2 activation can increase intracellular calcium levels via phospholipase C. Although both CB1 and CB2 are coupled to G proteins, the transduction pathways they activate may differ, for example in terms of interactions with ion channels. A variant of CB2, called Q63R, has been associated with celiac disease, immune thrombocytopenic purpura, and juvenile idiopathic arthritis. During inflammatory joint disease, the production and release of endocannabinoids is mediated by the production of proinflammatory cytokines such as interferon [IFN]-c, interleukins (IL-12, IL-15, IL-17, IL-18), chemokines, chemical mediators such as nitric oxide synthase (NOS)-2, cyclooxygenase-2 (COX-2), matrix metalloproteinases (MMPs), and various other byproducts of arachidonic acid metabolism.

[0011] There are currently no effective treatments to prevent or reverse neuropathic pain, so current treatments are directed solely at relieving symptoms. Treatment of chronic pain remains an unmet clinical need, and adequate pain relief can be obtained using medications that have adverse effects on the central nervous system. The quality of life of patients with neuropathic pain is often exacerbated by comorbidities such as sleep disturbances, depression, and anxiety. Preclinical studies have shown that cannabinoid receptor agonists block pain in a variety of acute and chronic pain models and reduce inflammation. Both CB1 and CB2 receptor agonists, either alone or in combination, exhibit antinociceptive activity, and CB2 activity is thought to affect microglia, thereby reducing neuroinflammatory mechanisms. CB2 receptors are thought to be particularly important in central neural pain circuits, as agonist activity induces dopamine release in the midbrain region, contributing to reduced pain control.

[0012] THC, the main psychoactive component of cannabis, acts primarily as a partial agonist at CB1 (Ki=53nM) and CB2 (Ki=40nM) receptors, with well-known effects on pain, appetite enhancement, digestion, mood, and processes mediated through the endocannabinoid system. In inflammatory arthritis, the production and release of endocannabinoids is mediated by the production of proinflammatory cytokines (interferon [IFN]-c, interleukins (IL-12, IL-15, IL-17, IL-18), chemokines, chemical mediators such as nitric oxide synthase (NOS)-2, cyclooxygenase-2 (COX-2), matrix metalloproteinases (MMPs), and various other arachidonic acid metabolic byproducts). THC may cause adverse psychoactive events, depending on dose and prior patient tolerance. In contrast, the major nonpsychoactive phytocannabinoid component of CBD, C. sativa, has little affinity for these receptors (Ki of 1.5 and 0.37 μM for human CB1 and CB2, respectively), and acts as a partial antagonist of CB1 and a weak inverse CB2 agonist.

[0013] It is an object of the present invention to provide formulations with improved bioavailability, pharmacokinetics and increased delivery selectivity and reduced side effects. SUMMARY OF THE INVENTION

[0015] Cannabinoid-dendrimer conjugates have been developed. The composition may include one or more endocannabinoids or endocannabinoids. Endocannabinoids are lipid signaling molecules produced from the body that activate cannabinoid receptors and mimic the activity of Δ9-tetrahydrocannabinol, the main psychoactive ingredient of cannabis. Exemplary endocannabinoids and endocannabinoid analogs that may be included in the composition include, but are not limited to, N-arachidonoylethanolamide (arachidonoylethanolamine), 2-arachidonoylglycerol (2-AG), 2-arachidonoylglycerol ether (noradin ether), N-arachidonoyl dopamine (NADA), virodamine (OAE), lysophosphatidylinositol (LPI), 7,10,13,16-docotetraenoylethanolamide and homo-γ-linolenoylethanolamine. Phytocannabinoids or botanicals are a class of structurally diverse, naturally occurring plant chemicals. In some forms, the phytocannabinoids or phytocannabinoid derivatives used in the composition can be derived from a variety of sources, including but not limited to hemp, cannabis, echinacea, oleracea, immortelle, Radulamarginata, kava, black truffles, cloves, rosemary, basil, oregano, black pepper, lavender, cinnamon, malabathrum, ylang-ylang, copaifera spp., and hops. Examples include THC, Δ9-tetrahydrocannabinol; Δ9-THCV, Δ9-tetrahydrocannabinol; CBN, cannabinol; CBDV, cannabidiol; CBG, cannabigerol; CBC and cannachromene.

[0016] The composition may contain one or more phytocannabinoids or phytocannabinoid derivatives belonging to one or more of the following subclasses: cannabidiol (CBD), tetrahydrocannabinol (THC), cannabigerol (CBG), cannabinol (CBN), cannabichromene (CBC), cannabilide (CBE), cannabicyclol (CBL), cannafuran (CBF), cannabitran (CBT), cannabinodivarins (CBV), cannabiripsol (CBR), isocannabinoids and / or their derivatives.

[0017] The dendrimer is preferably a glucose dendrimer, a hydroxyl-terminated PAMAM dendrimer or a sugar-modified dendrimer, most preferably a glucose dendrimer. Preferred glucose dendrimers include G1, G2, G3 glucose dendrimers, and preferred PAMAM dendrimers include G3, G4, G5, G6 hydroxyl-terminated PAMAM dendrimers. The density and loading of cannabinoids on the dendrimers, the manner of attachment of the cannabinoids to the dendrimers, the size and chemical composition of the dendrimers, and the size and composition of the linker to the cannabinoids (if any) determine the rate, selectivity and activity of the delivery site where the cannabinoids bind to the receptor. In a preferred embodiment, a cleavable linker binds the cannabinoid to the dendrimer. In a most preferred embodiment, the cleavable linker is an ester. In some forms, the linker contains a triazole moiety.

[0018] The route of administration, typically oral, intranasal, or applied to other mucosal surfaces, affects the rate and dose of the dendrimer conjugate into the brain. Dendrimers increase brain absorption, solubility, target engagement, PK, and help confine the drug to the correct compartment. In some embodiments, dendrimers are used to retain cannabinoids in peripheral circulation rather than central circulation.

[0019] Cannabinoid-dendrimer conjugates may be used to treat patients with a variety of conditions including neurodegenerative diseases (aging, dementia, Alzheimer's disease), psychosis, acute, chronic and neuropathic pain, cancer-related pain, traumatic brain injury, epilepsy, Huntington's disease and multiple sclerosis, seizures, inflammatory diseases and nausea control during chemotherapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1A and 1B Schematic representation of how different cannabinoid (CB) receptor signaling modes influence neuromodulation in health and disease in specific ways. Figure 1A Key enzymes such as diacylglycerol lipase (DGLα) and phospholipase D (PLD) that generate endogenous ligands anandamide (AEA) and 2-arachidonoylglycerol (2-AG) are shown. These enzymes activate cannabinoid 1 receptor (CB1) receptors in the central nervous system (CNS). The results can include modulation of adenylate cyclase activity to inhibit cAMP accumulation, voltage-gated calcium channels (VGCCs), K+ channels, and neurotransmitter release in presynaptic excitatory and inhibitory synapses. Figure 1B It has been shown that after activation of the CB1 receptor by ligand binding, signaling through G proteins and / or β-arrestins can occur at the plasma membrane, endocytic pits, or endosomes after receptor internalization. G proteins usually bind to unphosphorylated receptors, while β-arrestins bind to receptors phosphorylated by G protein receptor kinases. Figure 1C Schematic representation of key receptors and cells in the brain where dendrimer-cannabinoid conjugates can exert selective effects.

[0022] Figure 2A is a schematic diagram of a stepwise synthetic route to produce exemplary dendrimer-cannabidiol conjugates via click chemistry utilizing enzyme-sensitive ester bonds. Figure 2B is a schematic diagram of a stepwise synthetic route for conjugating an exemplary cannabinoid, cannabidiol, to a dendrimer.

[0023] Figure 3A and 3B Schematic diagram showing the molecular structure of a stepwise synthetic route to produce an exemplary dendrimer-cannabidiol conjugate via click chemistry using a non-cleavable ether bond ( Figure 3A ), and the molecular structure of a stepwise synthetic route for conjugating an exemplary cannabinoid, cannabidiol, to a dendrimer ( Figure 3B ).

[0024] Figure 4A and 4B is a schematic diagram showing a stepwise synthetic route to produce an exemplary dendrimer-HU-308 conjugate by amidation reaction using a phosphatase-cleavable phosphodiester bond ( Figure 4A ), and a stepwise synthetic route for conjugating an exemplary cannabinoid HU-308 to a dendrimer ( Figure 4B ).

[0025] Figure 5A is a schematic diagram of the step-by-step synthetic route to produce an exemplary dendrimer-HU-308 conjugate via a click reaction using an ester bond. Figure 5B Schematic diagram of a step-by-step synthetic route for conjugating an exemplary cannabinoid, HU-308, to a dendrimer. Figure 5C is a schematic diagram of the conjugation of HU-308 to PAMAM-G4-OH dendrimer.

[0026] Fig. 6A and 6B Schematic diagram showing the molecular structure of a stepwise synthetic route for producing an exemplary dendrimer-HU-308 conjugate using a glutathione-sensitive disulfide bond via a click reaction ( Fig. 6A ), and a stepwise synthetic route for conjugating an exemplary cannabinoid HU-308 to a dendrimer to produce a D-HU-308 conjugate with a disulfide linker ( Figure 6B ).

[0027] Fig. 7A Schematic diagram of a stepwise synthetic route to generate exemplary dendrimer-tetrahydrocannabinol (D-THC) conjugates using esterase-sensitive ester bonds via a click reaction. Figure 7B Schematic diagram of a step-by-step synthetic route for conjugating an exemplary cannabinoid, THC, to a dendrimer to generate a D-THC conjugate.

[0028] Fig. 8A and 8B Schematic diagram showing the molecular structure in a stepwise synthetic route to produce an exemplary dendrimer-THC conjugate via a click reaction with a triglycyl peptide linker suitable for lysosomal release ( Fig. 8A ), and a stepwise synthetic route for conjugating an exemplary cannabinoid, THC, to a dendrimer to produce a D-THC conjugate with a triglycyl peptide linker ( Figure 8B ).

[0029] Fig.9A Schematic diagram of the stepwise synthetic route to generate an exemplary dendrimer-arachidonic acid ethanolamine (D-AEA) conjugate via a click reaction. Fig. 9B Schematic representation of a step-by-step synthetic route for conjugating an exemplary cannabinoid anandamide to a dendrimer to generate a D-AEA conjugate.

[0030] Fig. 10A and 10B Schematic diagram of a stepwise synthetic route for producing an exemplary dendrimer-AEA having a non-cleavable ether bond ( Fig. 10A ), and a schematic diagram of a stepwise synthetic route for conjugating an exemplary cannabinoid AEA to a dendrimer to produce a D-AEA conjugate with a non-cleavable ether bond ( Fig. 10B ).

[0031] Fig.11A Schematic diagram of the step-by-step synthetic route to produce an exemplary dendrimer-2-arachidonoylglycerol (D-2-AG) conjugate via a click reaction. Fig. 11B Schematic representation of a step-by-step synthetic route for conjugating an exemplary cannabinoid, D-2-AG, to a dendrimer to produce a D-2-AG conjugate.

[0032] Fig.12 It is a structural table of Hu308 and cannabidiol and their conjugated analogs.

[0033] Figures 13A-13B Shows Fig.12 Synthesis and structure shown; Fig. 13C Binding and release of the compounds are shown.

[0034] Fig.14 Graph showing % binding efficiency versus log (μM) of tryptamine in a 5-HT1A human serotonin GPCR cell-based agonist cAMP assay. DETAILED DESCRIPTION OF THE INVENTION

[0036] I. Definitions

[0037] As used herein, "cannabinoids" refers to lipid-based compounds that have direct or indirect activity at cannabinoid receptors. "Endocannabinoids" are naturally occurring endogenous cannabinoids found in humans and other animals. "Exogenous cannabinoids" include phytocannabinoids, i.e., cannabinoids extracted from the cannabis plant, and their related synthetic derivatives.

[0038] 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 with 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. Analogs are chemically modified active compounds derived from the parent. The term "therapeutic agent" refers to an agent that can be administered to treat one or more symptoms of a disease or condition. 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. Diagnostic agents can label target cells so that these labeled target cells can be subsequently detected or imaged.

[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] "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.

[0054] II. Composition

[0055] Dendrimer-active agent conjugates suitable for delivering one or more cannabinoids and / or cannabinoid derivatives to one or more target cells expressing their receptors, such as neural cells and / or glial cells, have been developed. Typically, the cannabinoids and / or cannabinoid derivatives bind to receptors on the surface of the target cells and / or receptors within the target cells. Exemplary target cells include, but are not limited to, brain cells, such as microglia, astrocytes and / or neurons, for example, cells within a pathological site of the brain or CNS; and / or cells in the peripheral nervous system, such as peripheral neurons and glial cells. The microglia and / or astrocytes to which the cannabinoids and / or cannabinoid derivatives are delivered can be active or inactive microglia and / or astrocytes.

[0056] The cannabinoid and / or cannabinoid derivative 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 cannabinoid and / or cannabinoid derivative binds to the target receptor. In these embodiments, after binding, the agent can be released from the dendrimer or remain conjugated to the dendrimer as an intact dendrimer-active agent conjugate. In some embodiments, the cannabinoid and / or cannabinoid derivative is released from the dendrimer at a location proximal to the target receptor and then binds to the target receptor on the target neural and / or glial cells.

[0057] A. Dendrimers

[0058] 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)).

[0059] 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, dendrimers have a regular dendrimer molecular structure, in other cases, they can be hyperbranched structures with irregular branch lengths.

[0060] 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 an agent bound thereto. In some embodiments, the dendrimer has a diameter effective to be retained in the peripheral circulation to deliver an agent bound thereto to a target cell of the peripheral nervous system.

[0061] 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.

[0062] 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.

[0063] 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. 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. The dendrimers 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 is modified with functional groups to increase the number of surface hydroxyl groups.

[0064] 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 neural cells and macrophages. Alternatively, by appropriately functionalizing the dendrimers (e.g., pegylation), the dendrimer-active agent conjugates can be confined to the peripheral circulation.

[0065] 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 eye, such as neurons and glial cells of the CNS and / or PNS, by using dendrimers of a certain generation, such as PAMAM dendrimers of generation 2 (G2), G3, G4, and G5, and / or glucose dendrimers. In the most preferred embodiments, the dendrimers are made entirely of glucose building blocks. PAMAM dendrimers modified with sugars may also work, but dendrimers made of sugars, especially glucose, are most preferred.

[0066] Monosaccharide-based dendrimers

[0067] In some embodiments, the branching units include monosaccharides. In some embodiments, the monosaccharide branching units are attached to the core or previous monomer layers through a linker such as a polyethylene glycol chain. In some 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 through 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 generating a 1st generation dendrimer, for generating a 2nd generation dendrimer, and for generating a 3rd generation dendrimer, the branching units are glucose-based branching units.

[0068] In some embodiments, the dendrimer has a supercore (e.g., 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 unit.

[0069] In some embodiments, dendrimers synthesized using glucose building blocks, whose surfaces are composed primarily of glucose moieties, allow for specific targeting of 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 within target neural and / or glial cells. In some embodiments, glucose-based dendrimers selectively target or are enriched within 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 within and / or on the surface of damaged, diseased, and / or overactive neurons and / or glial cells.

[0070] 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.

[0071] In some embodiments, the dendrimer is composed of glucose and oligoethylene glycol building blocks. Exemplary glucose dendrimers are shown in structures V and VII.

[0072] 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.

[0073] Dendrimer compositions have been developed that can selectively accumulate within neurons, particularly in the nuclei of injured and / or overactive neurons, termed "glucose dendrimers." These 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. Glucose dendrimers are described by Johns Hopkins University 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.

[0074] 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

[0075]

[0076] or its peracetylated derivatives.

[0077] 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:

[0078]

[0079] In a preferred embodiment, the dendrimer is a second generation dendrimer having the following structure:

[0080]

[0081] 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. In some embodiments, the dendrimer is conjugated to a small molecule, an antibody or antigen-binding fragment thereof, a nucleic acid or a polypeptide. In some embodiments, the therapeutic agent conjugated to the dendrimer is an anti-inflammatory agent, an antioxidant, or an immunomodulatory agent. In other embodiments, 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.

[0082] In some embodiments, the dendrimer and one or more therapeutic, prophylactic or diagnostic 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.

[0083] PAMAM dendrimers

[0084] The term "PAMAM dendrimer" refers to a poly(amidoamine) dendrimer that may contain different cores with amidoamine building blocks and may have carboxylic acid, amine, acetamide and / or hydroxyl termini of any generation, including but not limited to 1st generation PAMAM dendrimers, 2nd generation PAMAM dendrimers, 3rd generation PAMAM dendrimers, 4th generation PAMAM dendrimers, 5th generation PAMAM dendrimers, 6th generation PAMAM dendrimers, 7th generation PAMAM dendrimers, 8th generation PAMAM dendrimers, 9th generation PAMAM dendrimers, or 10th generation PAMAM dendrimers. In some embodiments, the dendrimer is a 4th, 5th, or 6th generation ("G") dendrimer. In some embodiments, the PAMAM dendrimer has hydroxyl termini.

[0085] In general, the overall structure of a dendrimer can be distinguished into a core part followed by radially linked branching units (ie generations), which are further modified with chemical functionalities carrying desired terminal groups on the outer surface of the dendrimer.

[0086] In some embodiments, the dendrimers are in the form of nanoparticles and are described in detail in U.S. Published Application Nos. US 2011 / 0034422, US 2012 / 0003155, and US 2013 / 0136697. For example, the molecular weight of the dendrimers can be varied to prepare polymer nanoparticles that form particles with properties (e.g., drug release rate) optimized for a particular application. 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 preclude their psychoactive effects.

[0087] In some embodiments, different types of dendritic macromolecules can be used as delivery vehicles to bind and deliver one or more active agents, including but not limited to dendrons and tectodendrimers. Dendrons are branch-like wedges with a core containing one function (functional group, f=1) and another function (f=8, 16, 32, etc.) at the periphery. Dendritic macromolecules are usually composed of a central dendritic macromolecule and multiple dendritic macromolecules attached to the periphery.

[0088] 1. Core

[0089] 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.

[0090] Exemplary chemical structures suitable as core moieties include dipentaerythritol, pentaerythritol, 2-(aminomethyl)-2-(hydroxymethyl)propane-1,3-diol, 2-ethyl-2-(hydroxymethyl)propane-1,3-diol, 3,3′,3″,3″′-silanetetrayltetrakis(propane-1-thiol), 3,3-divinylpenta-1,4-diene, 3,3′,3″-nitrilotripropionic acid, 3,3′,3″-nitrilotripropionic acid, 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-1,3,5,215,415,615-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(ethane-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. In some embodiments, the core part is chitosan. Therefore, azide-modified chitosan or alkyne-modified chitosan is suitable for binding to the branch unit using click chemistry.

[0091] 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.

[0092] Table 1. Structural representation of various building blocks (core, branching units, surface functional groups, monomers) used for dendrimer synthesis.

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099] 2. Branch unit

[0100] 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.

[0101] 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.

[0102] 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.

[0103] 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.

[0104] 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 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-1,3,5,215,415,615-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.

[0105] Other examples of chemical structures suitable for forming branching units of dendrimers disclosed herein include, but are not limited to, sugar moieties such as glucose, galactose, mannose, and fructose, and alkylene glycols such as ethylene glycol, and combinations thereof. In some embodiments, the branching unit is chitosan. Thus, azide-modified chitosan or alkyne-modified chitosan is suitable for conjugation to a core moiety or 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).

[0106] 3. Surface functional groups

[0107] 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.

[0108] In some embodiments, the surface functional groups are hydroxyl groups, such as hydroxyl groups of PAMAM dendrimers, hydroxyl groups of 2nd generation PEG 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.

[0109] 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, glucose dendrimers are of generation 1 (G1), G2, G3, G4, and G5.

[0110] 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.

[0111] Glucose dendrimers are preferred. In some embodiments, the dendrimers are 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.

[0112] 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.

[0113] In some embodiments, the amount of surface functional groups (eg, any of the above, eg, hydroxyl groups) of the dendrimer is at least 40%, at least 50%, greater than 40%, greater than 50%, or in the range of greater than 40% to 100%.

[0114] 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 moieties / molecules and / or polyalkylene glycols can be within any of the above hydroxyl ranges. 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 dendrimers are 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 dendrimers are formed from sugar moieties / molecules (e.g., glucose) or include sugar moieties / molecules at the termini, the terminal hydroxyl groups of these dendrimers can 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.

[0115] a. Hydroxyl-terminated dendrimers

[0116] 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.

[0117] 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).

[0118]

[0119] Structure I. Second generation (G2) oligoethylene glycol dendrimers

[0120] 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 , such as at least 10, 15, 20, 25, 30, 35, 40, 45, 50, or greater than 50. 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 3In 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 about 10 kDa, preferably 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 portion of the hydroxyl groups exposed on the outer surface, while the remaining hydroxyl groups are located in the inner core of the dendrimer.

[0121] 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.

[0122] In some embodiments, the dendrimers are capable of specifically targeting specific tissue regions and / or cell types, preferably cells and tissues of the central nervous system (CNS) and the eye. In some embodiments, the dendrimers specifically target neurons of the CNS and the eye. 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 these glucose dendrimers. Glucose dendrimers with terminal glucose monosaccharides and a high density of hydroxyl functional groups effectively target neurons in a generation-dependent manner. The examples show that generation 2 (G2) is effective, and G3 and G4 should be effective. G5 and above are more difficult to use.

[0123] 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.

[0124] In some embodiments, the dendrimers are made from glucose and oligoethylene glycol building blocks. Exemplary glucose dendrimers are shown in the Examples, for example, the first generation dendrimers shown in Structures IV-VI and the Figure 1A and 1B Some exemplary glucose dendrimers include a first generation glucose dendrimer with 24 hydroxyl (-OH) end groups, a second generation glucose dendrimer with 96 hydroxyl (-OH) end groups, a third generation glucose dendrimer with 396 hydroxyl (-OH) end groups, and a fourth generation glucose dendrimer with 1584 hydroxyl (-OH) end groups. In a preferred embodiment, the glucose dendrimer is a second generation glucose based dendrimer with 24 glucose molecules at the periphery and 6 embedded glucose molecules in the backbone, which are held together by PEG segments.

[0125] b. Carbohydrate-modified dendrimers

[0126] 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.

[0127] In some embodiments, hydroxy-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 target nerve cells, glial cells and / or macrophages, allowing one or more active agents conjugated thereto to bind to one or more receptors on / in the target nerve cells and / or glial cells.

[0128] 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.

[0129] Glucose dendrimers or glucose modified dendrimers can be used to obtain selective uptake by target cells. Drugs bound to the dendrimers can bind to receptors or other sites of action. In a preferred embodiment, the dendrimers or functionally modified dendrimers are conjugated to one or more drugs that have affinity and are suitable for binding to one or more cannabinoid receptors (e.g., CB1 receptors, CB2 receptors, and CB3 receptors). In some embodiments, the dendrimers are conjugated to one or more carbohydrate moieties that have affinity and are suitable for binding to one or more non-cannabinoid receptors, such as: G protein-coupled receptors, such as GPR55, GPR18, GPR3, GPR6, GPR12, GPR40, GPR43, GPR41, GPR120, GPR23, GPR92, GPR84, GPR119, or GPR35; adenosine receptors, such as adenosine A3; muscarinic acetylcholine receptors, such as M1 and M4; serotonin receptors, such as 5-HT1A, 5-HT2A; opioid receptors, such as μ- and δ-opioid receptors; and tachykinin NK2 receptors. In some embodiments, the dendrimer is conjugated to one or more carbohydrate moieties, or is made of sugar moieties that have affinity 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 dendrimer is conjugated to one or more glucose and / or glucosamine moieties. In other embodiments, the dendrimer is conjugated to one or more oligosaccharides terminated with glucose and / or glucosamine moieties, i.e., the glucose and / or glucosamine moieties are exposed on the surface of the dendrimer conjugate, suitable for binding to one or more GLUTs, cannabinoid receptors, and / or non-cannabinoid receptors.

[0130] In some embodiments, the dendrimer has multiple carbohydrate moieties / molecules, e.g., monosaccharides, e.g., glucose, or sugar building blocks for the dendrimer 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 , such as at least 10, 15, 20, 25, 30, 35, 40, 45, 50 or more than 50. For example, per nm2 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 the sugar moiety / molecule and is therefore not part of the sugar moiety / molecule, or a combination thereof.

[0131] 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.

[0132] c. Dendrimers modified with polyalkylene glycols

[0133] 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.

[0134] 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, such as at least 10, 15, 20, 25, 30, 35, 40, 45, 50 or greater than 50. 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 / 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 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 , and has a molecular weight of about 100 Da to about 10 kDa.

[0135] 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.

[0136] 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.

[0137] B. Cannabinoids

[0138] Dendrimer compositions typically include one or more cannabinoids and / or cannabinoid derivatives conjugated to dendrimers. Exemplary cannabinoids that may be included in the composition belong to one or more cannabinoid classes, including but not limited to plant-derived cannabinoids, endocannabinoids, and synthetic cannabinoids. The cannabinoids or cannabinoid derivatives contained in the composition typically bind to one or more receptors, thereby modulating signaling in the endocannabinoid system. Many of the effects of cannabinoids and cannabinoid derivatives are mediated by two G protein-coupled receptors (GPCRs), CB1 and CB2, although other receptors may be involved. CB1 receptors are present in higher amounts in several brain regions and in lower amounts in the periphery. CB1 receptors mediate many of the psychoactive effects of cannabinoids. The distribution of CB2 receptors is more restricted, being present in many immune cells and a few neurons. CB1 and CB2 receptors are primarily coupled to inhibitory G proteins and are subject to the same pharmacological effects as other GPCRs. Therefore, partial agonism, functional selectivity, and inverse agonism all play an important role in determining the response of cells to specific cannabinoid receptor ligands. Human cannabinoid type 1 and type 2 receptor agonists and antagonists of interest include, for example, ARAP433912, Parecoxib, Valdecoxib, PSNCBAM-1, GAT 211, GAT 228, GAT-229, (±)19(20)-EDP ethanolamide, Voacamine, LY320135, Falcarinol, Lasofoxifene, ABD459, AM6545, PSB-SB1202, MCHB-1, KM 233, CB-25, CB-52, Leeamine, and Docosahexaenoylethanolamide.

[0139] 1. Endocannabinoids

[0140] The composition may include one or more endogenous cannabinoids or endocannabinoids. Endocannabinoids are lipid signaling molecules produced in the body that activate cannabinoid receptors and mimic the activity of Δ9-tetrahydrocannabinol, the main psychoactive ingredient of cannabis. Exemplary endogenous cannabinoids and endogenous cannabinoid analogs that may be included in the composition include, but are not limited to, N-arachidonoylethanolamide (arachidonoylethanolamine), 2-arachidonoylglycerol (2-AG), 2-arachidonoylglycerol ether (noradin ether), N-arachidonoyl dopamine (NADA), Virodhamine (OAE), lysophosphatidylinositol (LPI), 7,10,13,16-docotetraenoylethanolamide and homo-γ-linolenoylethanolamine. Not all are easy to conjugate, but most can be modified to achieve conjugation.

[0141] In some forms, the endocannabinoids included in the composition bind to the CB1 and CB2 receptors with similar affinity, acting as agonists at both CB1 and CB2 receptors. For example, 2-arachidonylglycerol binds to the CB1 and CB2 receptors with similar affinity, acting as a full agonist at both receptors.

[0142] In some forms, the endocannabinoids included in the composition have high affinity to the CB1 receptor and weak affinity to the CB2 receptor. For example, anandamide, 7,10,13,16-docotetraenoylethanolamine and homo-γ-linolenoylethanolamine bind to the central (CB1) cannabinoid receptor and to a lesser extent to the peripheral (CB2) cannabinoid receptor, where they act as partial agonists. In another embodiment, 2-arachidonoylglycerol ether (noradin ether) mainly binds to the CB1 receptor and is weakly bound to the CB2 receptor, thereby inducing sedation, hypothermia, intestinal stasis, and anti-injury. In a third example, N-arachidonoyl dopamine preferentially binds to the CB1 receptor.

[0143] In some forms, the endocannabinoids included in the composition have a high binding affinity to the CB2 receptor and a weak binding affinity to the CB1 receptor. For example, vildamine or O-arachidonoylethanolamine (OAE) have a strong binding affinity to the CB2 receptor and act as a partial agonist of the CB1 receptor.

[0144] In some forms, the endocannabinoids included in the composition bind to one or more non-CB1 / CB2 receptors. For example, anandamide and NADA can act as potent agonists of the vanilloid receptor subtype 1 (TRPV1), a member of the vanilloid receptor family. In another example, an endogenous lipid metabolite of anandamide (also known as anandamide or AEA) binds to the N-arachidonoylglycine (NAGly) receptor GPR18, an orphan G protein-coupled receptor.

[0145] 2. Phytocannabinoids

[0146] The composition may include one or more phytocannabinoids or phytocannabinoid derivatives. Phytocannabinoids or botanicals are a class of natural chemical components with diverse structures, derived from plants. In some forms, the phytocannabinoids or phytocannabinoid derivatives used in the composition may come from various sources, including but not limited to cannabis (e.g., cannabis stems, cannabis stems, industrial hemp seeds), cannabis (e.g., cannabis flowers, cannabis leaves, cannabis stems, cannabis stems, cannabis seeds), Echinacea, Angustifolia, White Echinacea, European kale, Helichrysum, Edge moss, Kava pepper, black truffles, cloves, rosemary, basil, oregano, black pepper, lavender, true cinnamon, Malabar bay leaf, ylang-ylang, Cuban incense tree (copaiferaspp.) and hops. Examples include THC, Δ9-tetrahydrocannabinol; Δ9-THCV, Δ9-tetrahydrocannabinol; CBN, cannabinol; CBDV, cannabidiol; CBG, cannabigerol; CBC, and cannachromene.

[0147] The composition may contain one or more phytocannabinoids or phytocannabinoid derivatives belonging to one or more of the following subclasses: cannabidiol (CBD), tetrahydrocannabinol (THC), cannabigerol (CBG), cannabinol (CBN), cannabichromene (CBC), cannabichromene (CBE), cannabicycline (CBL), cannafuran (CBF), cannabitran (CBT), cannabivarin (CBV), cannabivarinol (CBR), isocannabinoids and / or their derivatives.

[0148] a. Cannabidiol

[0149] In some forms, the composition may contain one or more cannabidiols (CBD) and / or its derivatives. Suitable examples of cannabidiol (CBD) include but are not limited to cannabidiol (CBD), cannabidiolic acid (CBDA), cannabidiorol (Cannabidiorcol) (CBDC1), cannabidiol-C4 (CBDC4), cannabidiol dimethyl ether (CBDD), cannabidiol monomethyl ether (CBDM), cannabidiol (CBDV), cannabidiolic acid (CBDVA) and derivatives thereof. In some forms, the composition may contain one or more cannabidiols (CBND) and / or its derivatives. Cannabinodiols or cannabidinodiols are usually present in the cannabis plant Cannabis sativa with low concentrations. Cannabidiols (Cannabinodiols) are fully aromatized derivatives of cannabidiol (CBD), can be used as the product of the photochemical conversion of cannabinol (CBN) to appear. In a preferred embodiment, the dendrimer is conjugated to one or more cannabidiol (CBD) or cannabidiol derivatives as shown in Formula I below.

[0150]

[0151] b. Tetrahydrocannabinol

[0152] In some forms, the composition may contain one or more tetrahydrocannabinols (THC). Non-limiting examples of tetrahydrocannabinols include tetrahydrocannabinol (THC), 10-oxo-delta-6a-tetrahydrocannabinol (OTHC), delta-8-tetrahydrocannabinolic acid (Δ8-THCA), delta-8-tetrahydrocannabinol (d8THC, Δ8-THC), delta-9-tetrahydrocannabinol (d9THC, Δ9-THC), delta-9-tetrahydrocannabinol-C4 (THC-C4), delta-9-tetrahydrocannabinolic acid A (Δ9-THCA, THCA-C5), delta-9-tetrahydrocannabinolic acid B (Δ9-THCB, THCA-B), delta-9-tetrahydrocannabinolic acid (Δ9-THCA-C1, THCA-C1), δ-9-tetrahydrocannabinolic acid C4, (Δ9-THCA-C4, THCA-C4), tetrahydrocannabinol (-)-cis-Δ (THC-C5), δ-9-tetrahydrocannabinol (Δ9-THCO-C1), δ-9-tetrahydrocannabinolic acid A (THCOA), δ-9-tetrahydrocannabinol (THCV), δ-9-tetrahydrocannabinolic acid A (THCVA), trihydroxy-Δ-9-tetrahydrocannabinol (TRIOH-THC), δ-10-tetrahydrocannabinol (D10THC, Δ10THC), tetrahydrocannabinol (THCP), THC-O acetate (THCO), hexahydrocannabinol (HHC) and derivatives thereof. In a second preferred embodiment, the dendrimer is conjugated with one or more tetrahydrocannabinol (THC) or THC derivatives as shown in Formula II below.

[0153]

[0154] c. Cannabigerol (CBG)

[0155] In some forms, the composition may contain one or more cannabigerol (CBG) or its derivatives. Suitable examples of cannabigerol include, but are not limited to, cannabigerolic acid (CBGA), cannabigerolic acid A monomethyl ether (CBGAM), cannabigerovarin (CBGV), cannabigerovarin acid (CBGVA), cannabigerovarin monomethyl ether (CBGM) and derivatives thereof. In a third preferred embodiment, the dendrimer is combined with one or more cannabigerol (CBG) or CBG derivatives as shown in Formula III below.

[0156]

[0157] d. Cannabinol (CBN)

[0158] In some forms, the composition may contain one or more cannabinol (CBN) and / or its derivatives. Suitable examples of cannabinol include, but are not limited to, cannabinol (CBN), cannabinolic acid (CBNA), cannabivarin (CBN-Cl), cannabinol-C2 (CBN-C2), cannabivarin (CBN-C3), cannabivarin-C4 (CBN-C4), cannabinodiol (CBND), cannabinodivarin (CBNDC3), cannabinol methyl ether (CBNM-C5), delta-9-cis-tetrahydrocannabinol (CIS-THC) and derivatives thereof. In a fourth preferred embodiment, the dendrimer is conjugated to one or more cannabinol (CBN) or cannabinol derivatives, as shown in Formula IV below.

[0159]

[0160] e. Cannabichromene (CBC)

[0161] In some forms, the composition may include one or more cannabichromenes (CBC) and / or derivatives thereof. Non-limiting examples of cannabichromenes (CBC) that may be included in the composition include cannabichromene (CBC), cannabichromenoic acid A (CBCA), cannabichromenosone (CBCN-C5), cannabichromenosone-C3 (CBCN-C3), cannabichromenosone (CBCON-C5), cannabichromevarin (CBCV), cannabichromenosone acid A (CBCVA) and derivatives thereof. In a fifth embodiment, the dendrimer is combined with one or more cannabichromenes (CBC) or CBC derivatives as shown in Formula V below.

[0162]

[0163]

[0164] f. Cannabidiol (CBL)

[0165] In some forms, the composition may include one or more cannabichromene (CBL) or its derivatives. Cannabichromene (CBE) is a plant cannabinoid metabolite that can be produced by photooxidation of CBD and CBDA, or by biotransformation using tissue culture under normal growth conditions. Non-limiting examples of cannabichromene (CBL) include cannabichromene (CBE-C5), -C3-cannabichromene (CBE-C3), cannabielsoic acid A (CBEA-C5 A), cannabichromene B (CBEA-C5B), C3-cannabichromene B (CBEA-C3 B), cannabichromene C3 (OH-iso-HHCVC3), dehydrocannabichromene (DCBF-C5), cannabichromene (CBF-C5) and its derivatives. In some forms, one or more cannabichromene may be included in the composition to modulate the activity of CB1 receptors, CB2 receptors, non-CB1 receptors, or a combination thereof. In an exemplary embodiment, the dendrimer is conjugated with one or more cannabichromene (CBL) or CBL derivatives, as shown in Formula VI below.

[0166]

[0167] g. Cannabitriol (CBT)

[0168] The composition may include one or more cannabitriols (CBT) and / or derivatives thereof. Cannabitriol is present in lower concentrations in cannabis plants and is an oxidized product of tetrahydrocannabinol (THC). Non-limiting examples of cannabitriols (CBT) include cannabitriol (CBT), cannabitriol variants (CBTV), 10-ethoxy-9-hydroxy-δ-6a-tetrahydrocannabinol, 8,9-dihydroxy-δ-6a-tetrahydrocannabinol (8,9-Di-OH), 9,10-dihydroxyhexahydrocannabinol, cannabiripsol-C5, 6a,7,10a trihydroxy Δ9-tetrahydrocannabinol 10-oxo Δ6a (10a) tetrahydrocannabinol, OTHC, trans-CBT-C5, cannabitriol-C3 (TransCBT-C3), trans-10-O-ethyl cannabitriol (TransCBT-OEt-C5) and derivatives thereof. In some forms, one or more cannabitriols may be included in the composition to modulate the activity of CB1 receptors, CB2 receptors, non-CB1 receptors, or a combination thereof. In an exemplary embodiment, the dendrimer is conjugated with one or more cannabitriols (CBT) or derivatives thereof, as shown in Formula VII below.

[0169]

[0170] h. Other phytocannabinoids

[0171] The composition may include one or more other plant cannabinoids and / or derivatives thereof. In some forms, the composition may include one or more cannabicyclols (CBL) or derivatives thereof. Cannabicyclol (CBL) is a photochemical product derived from the plant cannabinoid cannachromene. Suitable examples of cannabicyclol include, but are not limited to, cannabicyclol (CBL-C5), cannabicyclic acid (CBLA-C5 A,), cannabicycline (CBLV-C3) and derivatives thereof. In some forms, the composition may include one or more cannafurans (CBF) or derivatives thereof. Suitable examples of cannabicyclan include, but are not limited to, cannabicyclan (CBF), dehydrocannabicyclan (DCBF or CBFD) and derivatives thereof. In some forms, the composition may include one or more cannabicyclines (CBV) or derivatives thereof. Suitable examples of cannabicyclines (CBV) include, but are not limited to, cannabicycline (Cannabinodivarin) (CBV), cannabicycline (Cannabinodivarin) (CBVD) and derivatives thereof. In some forms, the composition may include one or more miscellaneous cannabinoids, including, but not limited to, cannabicycline (CBR), isocannabinoids and / or derivatives thereof. Non-limiting examples of isocandinoids include, but are not limited to, iso-tetrahydrocannabinol, iso-tetrahydrocannabinol, iso-tetrahydrocannabinol, and their derivatives.

[0172] i. Synthetic cannabinoids

[0173] The composition may include one or more synthetic cannabinoids and / or derivatives thereof.

[0174] Synthetic cannabinoids (also called cannabinoid receptor agonists) are a class of designer drug molecules that function similarly to Δ9-tetrahydrocannabinol (THC) and bind to one or more cannabinoid and non-cannabinoid receptors in the central and peripheral nervous systems, similar to endocannabinoids and / or phytocannabinoids. However, synthetic cannabinoids are generally regarded as psychoactive substances and their structure differs from synthetic phytocannabinoids (such as THC or CBD obtained by chemical synthesis) and synthetic endocannabinoids. Cannabinoid receptor agonists form a diverse group, but most are lipid-soluble and non-polar, containing from about 22 to about 26 carbon atoms. A common structural feature of synthetic cannabinoids is side chains where more than four and up to nine saturated carbon atoms are required for optimal activity.

[0175] The composition may contain one or more synthetic cannabinoids and / or derivatives thereof belonging to one or more of the following subclasses: classical cannabinoids, non-classical cannabinoids, mixed cannabinoids, aminoalkyl indoles and eicosanoic acid. The composition may contain one or more synthetic cannabinoids and / or derivatives thereof belonging to one or more of the following: naphthoyl indole (e.g., JWH-018, JWH-073 and JWH-398); naphthoyl indole, naphthoyl pyrrole, naphthoyl indole, phenylacetylindole (i.e., benzoyl indole, e.g., JWH-250), cyclohexylphenol (e.g., CP 47,497 and homologues of CP 47,497) and classical cannabinoids (e.g., HU-210). The composition may contain one or more synthetic cannabinoids and / or their derivatives, which are one or more of adamantanoyl indole or indazole amide, benzimidazole, dibenzopyran, eicosanoic acid, naphthylindene, indazole-3-amide, indole-3-amide, indole-3-carboxylic acid or aryloxycarbonyl indole, naphthoyl indazole, pyrazole amide, quinoline ester or aryloxycarbonyl indole, tetramethylcyclopropylcarbonyl indole and / or tetramethylcyclopropylcarbonyl indole.

[0176] In an exemplary embodiment, the dendrimer is conjugated to one or more naphthoyl indole or derivatives thereof, as shown in Formula VIII below, wherein R1, R2, R3 and R4 are positions where substitution variation can occur.

[0177]

[0178] In another exemplary embodiment, the dendrimer is conjugated to one or more naphthylmethylindole or derivatives thereof, as shown in Formula IX below, wherein R1, R2 and R3 are positions where substitution variations can occur.

[0179]

[0180] In another exemplary embodiment, the dendrimer is conjugated with one or more naphthoylpyrroles or derivatives thereof, as shown in Formula X below, wherein R1 and R2 are positions where substitution variations can occur.

[0181]

[0182]

[0183] In another exemplary embodiment, the dendrimer is conjugated to one or more naphthylmethylindene or a derivative thereof, as shown in Formula XI below, wherein R1 and R2 are positions where substitution variations can occur.

[0184]

[0185] In another exemplary embodiment, the dendrimer is conjugated with one or more phenylacetylindole or derivatives thereof, as shown below in Formula XII, wherein R1 and R2 are positions where substitution variations can occur.

[0186]

[0187] In another exemplary embodiment, the dendrimer is conjugated to one or more cyclohexylphenols or derivatives thereof, as shown in Formula XIII below, wherein R1 and R2 are positions where substitution variations can occur.

[0188]

[0189] In another exemplary embodiment, the dendrimer is conjugated to one or more classical cannabinoids (dibenzopyrans) and / or derivatives thereof, as shown below in Formula XIV, wherein R1 and R2 are positions where substituent variations may occur.

[0190]

[0191] C. Coupling Agents and Spacers

[0192] 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.

[0193] 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.

[0194] 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.

[0195] 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.

[0196] 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 end 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 end groups.

[0197] D. Dendrimer-Drug Conjugates

[0198] Dendrimer-active agent conjugates may be formed by covalent conjugation or non-covalent attachment of cannabinoids and / or cannabinoid derivatives to dendrimers, dendrimers or hyperbranched polymers. Methods for conjugating one or more active agents to dendrimers are known, such as those described in U.S. Publication Nos. 2011 / 0034422, 2012 / 0003155 and 2013 / 0136697. Covalent conjugates are preferred, but ionic complexes (cation-anion complexes) may also be used.

[0199] 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 a chloroalkyl bond.

[0200] 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.

[0201] 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.

[0202] The density of active agents covalently bound or non-covalently attached to the dendrimer can be adjusted depending on the specific cannabinoid or cannabinoid derivative 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, a plurality of 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, each nm 2 The surface density of the active agent is greater than 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, such as at least 10, 15, 20, 25, 30, 35, 40, 45, 50 or greater than 50. 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 .

[0203] 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 cannabinoids or cannabinoid derivatives 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 cannabinoids or cannabinoid derivatives conjugated to the dendrimer has a diameter that can effectively 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 cannabinoids or cannabinoid derivatives 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 for the active agent to bind to a targeted receptor on the surface and / or in target neural and / or glial cells.

[0204] 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 cannabinoids or cannabinoid derivatives can affect the surface charge of the dendrimer-active agent conjugate. In some embodiments, the surface charge of the dendrimer conjugated to the cannabinoid or cannabinoid derivative is between -100mV and 100mV, between -50mV and 50mV, between -25mV and 25mV, between -20mV and 20mV, between -10mV and 10mV, between -10mV and 5mV, between -5mV and 5mV, or between -2mV and 2mV. The above ranges include all values ​​from -100mV to 100mV. In preferred embodiments, the surface charge of the dendrimer-active agent conjugate is neutral or nearly neutral, i.e., from about -10mV to about 10mV (including the end values).

[0205] 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).

[0206]

[0207] 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 suitable for providing tailored drug release and receptor binding, preferably containing a triazole moiety; Y can be a bond or 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 carbinol (-CHOH-, -CROH-), a disulfide bond, a phosphodiester group

[0208]

[0209] 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 a cannabinoid or cannabinoid derivative, 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 or 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.

[0210] 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).

[0211] When administered to a subject in need thereof, the cannabinoid and / or cannabinoid derivative 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 cannabinoid and / or cannabinoid derivative 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 cannabinoid and / or cannabinoid derivative X is released from the dendrimer at a location close to the target receptor and then binds to the target receptor.

[0212] 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.

[0213] 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.

[0214] 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

[0215]

[0216] 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.

[0217] 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

[0218]

[0219] 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

[0220]

[0221] 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.

[0222] 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.

[0223] 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.

[0224] 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.

[0225] More specific exemplary dendrimer-active agent conjugates are shown in the Examples below. E. Exemplary cannabinoid-dendrimer conjugates

[0226] In a preferred embodiment, the dendrimer is conjugated to cannabidiol (CBD) as shown below in structures A and B, wherein n is an integer from 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0227]

[0228] In other preferred embodiments, the dendrimer is conjugated to HU-308 as shown in the following structures CE, wherein n is an integer from 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0229]

[0230] In other preferred embodiments, the dendrimer is conjugated to THC as shown below in Structures FG.

[0231]

[0232]

[0233] In other preferred embodiments, the dendrimer is conjugated to AEA as shown in Structure IJ below, wherein n is an integer from 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0234]

[0235]

[0236] In another preferred embodiment, the dendrimer is conjugated to 2-AG as shown in the following structure K, wherein n is an integer from 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0237]

[0238] III. Methods for Preparing Dendrimer Conjugates

[0239] A. Methods of Preparing Dendrimers

[0240] Dendrimers, and in particular glucose 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.

[0241] In some embodiments, dendrimers are prepared using a different approach in which the dendrimer is assembled from a multifunctional core that is extended outward by 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, the dendrimer is first synthesized by coupling AB4 peracetylated β-D glucose-PEG4-azide monomers to a hexapropynylated core. In another example, a PAMAM-NH2 dendrimer is first synthesized by coupling N-(2-aminoethyl)acrylamide monomers to an amine core.

[0242] 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.

[0243] 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.

[0244] 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 reaction to form a triazole moiety.

[0245] 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.

[0246] 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.

[0247] 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.

[0248] 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 a dendrimer. Two dendrimers can be linked by click chemistry, i.e., 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.

[0249] 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.

[0250] 1. Method for preparing glucose dendrimers

[0251] In some embodiments, the glucose-based dendrimers are prepared using a different approach, where the dendrimers are assembled from a multifunctional core that is extended outward through a series of reactions. This strategy involves coupling monomer molecules with reactive and protective groups to the multifunctional core moiety, resulting in a stepwise increase in the number of generations around the core, followed by removal of the protecting groups.

[0252] 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.

[0253]

[0254] 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.

[0255] 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.

[0256] a. Synthesis of AB4 building blocks

[0257] 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.

[0258] 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 allowed to return to room temperature. The reaction is monitored by TLC and quenched after 5 hours by the addition of 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 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 the eluent. The desired compound is obtained in a yield of 60%. The structure of glucose-OAc-TEG-OT is shown below:

[0259]

[0260] In some embodiments, the synthesis of glucose-OAc-TEG-N3 involves the following steps: a solution of glucose-OAc-TEG-OT (6 g, 8.8 mmol) is dissolved in 40 mL of anhydrous DMF, and then sodium azide (2 eq) is added and the reaction mixture is heated to 50 ° C overnight. After completion, the reaction mixture is filtered and the DMF is evaporated. After drying, the crude reaction mixture is passed through combiflash using ethyl acetate: hexane (70: 30) as the eluent. The structure of glucose-OAc-TEG-N3 is as follows:

[0261]

[0262] 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.

[0263]

[0264] b. Synthesis of Glucose Dendrimers

[0265] 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.

[0266] 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.

[0267]

[0268] 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.

[0269]

[0270] 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) was dissolved in anhydrous dimethylformamide (DMF, 50 mL) by ultrasound. Sodium hydride [60% dispersion in mineral oil] (951 mg, 39.65 mmol) was slowly added to the solution in portions under stirring at 0°C. The solution was stirred for another 15 minutes at 0°C. Propargyl bromide (3.85 mL, 34.608 mmol, 80% w / w toluene solution) was then added at 0°C and stirring was continued for another 6 hours at room temperature. The reaction mixture was 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.

[0271]

[0272] 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.

[0273] 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.

[0274] 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.

[0275] The structure of the second generation glucose dendrimer D2-Glu24-OH96 is shown below.

[0276]

[0277] 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.

[0278] 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.

[0279] 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.

[0280] B. Methods of Preparing Dendrimer-Agent Conjugates

[0281] Methods of conjugating pharmaceutical agents to dendrimers are generally known in the art, for example as described in US Published Application Nos. US2011 / 0034422, US2012 / 0003155, and US2013 / 0136697.

[0282] 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.

[0283] 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.

[0284] 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.

[0285] 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.

[0286] 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.

[0287] 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 cannabinoids or cannabinoid derivatives 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 the methods described.

[0288] 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.

[0289] The following examples describe more specific methods for preparing exemplary dendrimer-active agent conjugates.

[0290] IV. Pharmaceutical Preparations

[0291] 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).

[0292] The composition, method and relative amount of the preparation depend on the selected route of administration. The composition can be stored in a disposable vial after lyophilization so that it can be rehydrated immediately before use. Other methods of rehydration and administration are known to those skilled in the art.

[0293] Representative excipients include solvents, diluents, pH regulators, preservatives, antioxidants, suspending agents, 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 individuals without causing adverse biological side effects or unwanted interactions.

[0294] Typically, pharmaceutically acceptable salts 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 the drug with a suitable organic ligand. A list of suitable salts can be found in, for example, 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.

[0295] 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). 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.

[0296] In certain embodiments, the composition is administered topically, for example, by direct injection into the site to be treated. 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. Typically, local administration results in an increase in the local concentration of the composition, which is greater than the concentration that can be achieved by systemic administration.

[0297] Pharmaceutical compositions for administration by parenteral (intramuscular, intraperitoneal, intravenous or subcutaneous injection) and enteral routes of administration are described.

[0298] In some embodiments, the dendrimer conjugates can improve the effectiveness and durability of treatment, thereby reducing the need for repeated dosing to once a week, once a month, once every six months, once a year, or other long-term dosing schedules. Some embodiments can be incorporated into drug delivery systems (e.g., implants, pumps, patches, creams, etc.) to provide controlled, sustained delivery in a manner that reduces compliance and potential for abuse.

[0299] A. Parenteral Administration

[0300] 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 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.

[0301] 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.

[0302] 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)).

[0303] B. Enteral administration

[0304] The dendrimer-active agent conjugate composition can be administered enterally (orally, 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.

[0305] 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.

[0306] 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.

[0307] 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.

[0308] 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.

[0309] 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.

[0310] 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.

[0311] How to use

[0312] In a preferred embodiment, the dendrimer cannabinoid composition crosses the barrier interface of the nervous system and selectively targets specific cells and specific receptors on cells to alleviate symptoms associated with various diseases, disorders and conditions. The method comprises administering to a subject in need thereof an effective amount of the composition to increase the permeability of the cannabinoid agent across the barrier interface of the nervous system and / or to increase the binding of the cannabinoid agent to specific receptors in specific cells, particularly CB1 receptors and CB2 receptors on cells of the central and peripheral nervous systems.

[0313] A. Treatment methods

[0314] The composition is suitable for treating diseases associated with neuropathic pain, inflammation, nausea and vomiting, spasticity and behavioral abnormalities, especially those extending to the central and peripheral nervous systems. For example, see Montero-Oleas et al., "Therapeutic use of cannabis and cannabinoids: an evidence mapping and appraisal of systematic reviews" BMC Complement Med Ther. 20(1): 12 (2020) doi: 10.1186 / s12906-019-2803-2.

[0315] 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. Dendrimers may also include targeting agents for delivery to target tissues and cells in the spinal cord, brain and related areas.

[0316] 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).

[0317] 1. Site-specific targeting

[0318] The compositions and methods are intended to circumvent existing challenges of cannabinoid receptor family-selective, cell-selective drug delivery to the central and peripheral nervous systems and different organs of the body. Cannabinoid receptors are expressed in various cells and organs of the body and as a system regulate many aspects of our function. We take advantage of the small size and unique bio[physical properties of dendrimers to bring these drugs to specific cells and receptors in the body. The compositions and methods can increase drug bioavailability in the central and peripheral nervous systems by one or more of the following: (i) increasing drug levels at specific cells and specific cannabinoid receptors on or within cells, across brain barriers, 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) reducing off-target effects. For example, in some forms, by minimizing brain absorption of the drug, the compositions and methods allow for 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, local anesthesia, traumatic nerve injury, and hereditary and inflammatory neuropathies.

[0319] a. Improve the permeability of drugs across barrier interfaces

[0320] Dendrimer compositions and methods can improve the delivery of cannabinoids across one or more barrier interfaces in the brain and nervous system, particularly the blood-brain barrier (BBB), CSF-blood barrier, and blood-nerve barrier, and to specific receptors on neurons, microglia, or astrocytes. 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. Due to the clinical significance of cannabinoids, dendrimer compositions can be used to deliver cannabinoids with improved permeability across these barrier interfaces to achieve site-specific targeting.

[0321] i. Blood-brain barrier

[0322] 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.

[0323] 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 adjusted in response to various physiological changes in the brain. The dendrimer compositions and methods of the present application overcome the above challenges and are suitable for delivering cannabinoids across the blood-brain barrier via one or more of the above transport mechanisms.

[0324] 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.

[0325] Physicochemical factors that affect BBB permeability include molecular weight, charge, lipid solubility, surface activity, and the relative size of the molecule. 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 cannabinoids across the blood-brain barrier through one or more of the above-mentioned mechanistic pathways.

[0326] ii. Blood-Nerve Barrier (BNB)

[0327] 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.

[0328] 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 cannabinoids across BNBs, thereby improving the delivery of cannabinoids to peripheral nerve targets.

[0329] iii. Blood-CSF Barrier

[0330] The composition can be used to improve the delivery of cannabinoids to target sites across the blood-cerebrospinal fluid barrier (blood-CSF barrier) and the ventricles. The choroid plexus is a vascular tissue present in all ventricles. The functional units of the choroid plexus consist 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 that connect 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.

[0331] The CSF space and brain structures adjacent to 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 multiple sclerosis and related experimental autoimmune encephalitis) or virus-induced neurological diseases (including neuro-AIDS and CMV infection). B cell foci producing potentially harmful antibodies detected in different CNS autoimmune diseases are believed to be primarily located in the pia mater. Therefore, in some forms, dendrimer compositions can be used to deliver cannabinoids to target areas via the blood-CSF space connected to the deep cervical lymph nodes to improve or treat symptoms associated with neuroinflammatory diseases.

[0332] In some forms, dendrimer compositions can be used to deliver cannabinoids to target sites to improve or treat symptoms and conditions associated with vascular degeneration. For example, cerebral amyloid angiopathy induces degenerative vascular changes caused by amyloid beta peptide (Aβ), cystatin c, transthyretin, or gelsolin deposits around infiltrating blood vessels. These deposits are accessible through the interconnected CSF / perivascular spaces.

[0333] In some forms, dendrimer compositions can be used to deliver cannabinoids to target sites to improve or treat symptoms and conditions associated with tumor development. For example, periventricular tumors (including meningiomas, drug-resistant ependymomas, and leptomeningeal metastases from peripheral primary tumors) are in direct contact with the CSF. The blood-tumor barrier is generally considered to be porous due to the reduced efficacy of tight junctions (which allow contrast enhancement in magnetic resonance imaging). However, many cannabinoids are lipophilic and are prevented from crossing the BBB by multidrug resistance (MDR) efflux proteins that control transcellular pathways. In many periventricular tumors (such as ependymomas), MDR proteins are still well expressed on the blood-tumor barrier. Therefore, in some forms, dendrimer compositions can be used to achieve therapeutic concentrations of cannabinoids within tumor tissue using pharmacological pressure from the CSF.

[0334] b. Improve target specific binding

[0335] Dendrimer compositions can be used to deliver cannabinoids with increased binding affinity and specificity to one or more receptors to modulate the endocannabinoid system, including CB1 receptors, CB2 receptors, TRPV receptors, or combinations thereof. As shown in FIG. 1 , dendrimer compositions can be used to deliver cannabinoids that selectively affect target cells and specific receptors on target cells while being cleared from non-target organs, including but not limited to nerve and glial cells in the brain, spinal cord, heart, lungs, kidneys, intestines, and joints. Improved binding to specific receptors on target cells can allow the conjugate to act quickly at lower doses. The degree of binding to the target receptor can be adjusted by the chemical reaction that attaches the drug to the dendrimer. In exemplary embodiments, hydroxyl-terminated PAMAM dendrimers, glucose dendrimers, and other dendrimers can enhance the delivery of cannabinoids to specific receptors (e.g., CB1-specific receptors, CB2-specific receptors, and / or both receptors), thereby improving the performance of these drugs. In other forms, these dendrimer cannabinoid conjugates can be used in combination with other classes of drugs to achieve synergistic effects.

[0336] In some forms, specific ligands (e.g., AEA and 2-AG) can be combined with the dendrimer compositions to provide cell type and receptor type specific modulation of the endocannabinoid system. The dendrimer compositions can also be used to target specific metabolites (e.g., intracellular enzymes in neurons and glial cells) to modulate the synthesis and degradation of endocannabinoid ligands. For example, the dendrimer compositions can be used to target PLA and DAG to modulate the synthesis of endocannabinoid ligands, and to target FAAH and MLGL to modulate the degradation of endocannabinoid ligands.

[0337] In some forms, dendrimer compositions can be designed to deliver cannabinoids via other pathways to modulate cellular activity in the brain and nervous system. For example, dendrimer compositions can be designed to deliver cannabinoids via random diffusion, aggregation to membranes; and lateral diffusion followed by binding to membrane buried ligand binding sites.

[0338] i. Endocannabinoid system in the CNS

[0339] ECS has become one of the key regulatory mechanisms in the brain that controls multiple events such as emotions, pain, learning and memory. It is also believed to provide neuroprotection during traumatic brain injury (TBI) and may be part of the brain's natural compensatory repair mechanism during neurodegeneration. In view of the clinical significance of ECS in drug abuse and dependence, the composition and method are suitable for treating marijuana abuse and narcotic drug abuse for ECS components. In the CNS, eCB acts as a retrograde messenger, mediating feedback inhibition to regulate synaptic plasticity. Activation of CB1 receptors leads to activation of inward rectifier K+ channel conductance, N-type and P / Q-type voltage-operated Ca2+ channel conductance and eCB generation. This can lead to reduced neurotransmitter release at excitatory and inhibitory synapses, resulting in transient effects such as depolarization-induced inhibition (DSI) and depolarization-induced excitation inhibition (DSE), or, for example, long-term inhibition and enhancement (LTP / LTD) during synaptic plasticity. These events make ECS a key regulator of synaptic plasticity.

[0340] Chronic exposure to CB1 receptor agonists leads to a rapid attenuation of behavioral responses, termed tolerance, which has been attributed to a decrease in the receptor's ability to activate effector pathways (i.e., desensitization) and a reduction in the number of receptors expressed on the cell surface (i.e., internalization). At the molecular level, agonist-bound GPCRs become substrates for G protein-coupled receptor kinases (GRKs); these kinases phosphorylate serine and / or threonine residues on the GPCR cytoplasmic domain, which then become high-affinity targets for β-arrestin, whose binding unbinds the G protein and stimulates receptor internalization and β-arrestin-mediated signaling.

[0341] GRK-induced ligand-receptor phosphorylation results in very specific and unique phosphorylation profiles or “barcodes.” These barcodes are finely tuned to determine which signaling cascades are activated, opening up a range of possibilities, often defined as functional selectivity or ligand bias.

[0342] Receptor and β-arrestin interactions and signaling cascades rely on specific phosphorylation sites controlled by unique GRKs. Mutation of the putative GRK site from S426 / S430 to alanine results in reduced α-arrestin2 recruitment and receptor internalization, but enhances interaction with β-arrestin1 and increases β-arrestin1-mediated signaling. β-arrestin-mediated signaling from this biased receptor controls the activation of multiple cascades, including ERK1 / 2, JNK1 / 2 / 3, CREB, and P38α. Notably, these cascades have been previously implicated in the activation of the CB1 receptor. Activation of these cascades by the CB1 receptor and β-arrestin in turn regulates gene expression and protein synthesis.

[0343] These compositions and methods can be used to elucidate the physiological role of beta-arrestin in developing pathway-selective or "biased ligands" with greater therapeutic benefit. These compositions and methods can also be used to study signaling from biased CB1 receptors (such as S426A / S430A), aid in the identification of biased ligands, and provide important tools to elucidate the mechanisms and roles of CB1 receptor signaling.

[0344] The subcellular localization and trafficking of CB1 receptors are highly dynamic and have a significant impact on receptor signaling. CB1-G protein-mediated signaling occurs at both the cell surface and intracellular compartments. At the cell surface, CB1 receptor ligands modulate the interaction between the receptor and β-arrestin as a mechanism to influence β-arrestin-mediated signaling. This interaction begins at the plasma membrane and can continue to the intracellular compartment. Interestingly, these location-specific signaling events appear to be widespread among several GPCRs. For example, LH receptors, β2-adrenergic receptors, and B2 receptors can signal from intracellular compartments through β-arrestin or G proteins through “supercomplexes”, ultimately generating three different spatiotemporal signaling waves. Constitutive activation also plays a role in their trafficking. The localization and trafficking of CB1 receptors are highly dynamic events that are closely related to their signaling.

[0345] ii. Modulation of the endocannabinoid system

[0346] Compositions and methods can be used to pharmacologically manipulate eCB levels or their actions through allosteric modulators, providing better opportunities to modulate the ECS. eCBs are produced on demand, and their synthesis is typically triggered by increases in intracellular Ca2+ at postsynaptic sites in response to sustained synaptic activity. The main eCBs are rapidly inactivated by reuptake mechanisms and degradation enzymes, including fatty acid amide hydrolases (FAAH) and monoacylglycerol lipases. Among the eCBs, derivatives of arachidonic acid, such as AEA and 2-AG, are predominant and orthosteric. These ligands are agonists for both CB1 and CB2 receptors, but have a higher binding affinity to the CB1 receptor (AEA Ki = 89 nM and 321 nM for CB1 and CB2 receptors, respectively; 2-AG Ki = 472 nM and 1400 nM for CB1 and CB2 receptors, respectively; Pertwee et al., 2010). Allosteric eCBs have been identified, including pregnenolone and lipoxin A4, which can modulate CB1 receptor signaling and may have therapeutic value (Pamplona et al., 2012; Vallée et al., 2014; Pertwee, 2015). These compositions and methods can be used to aid in the pharmacological characterization of orthosteric and allosteric modulators to clearly elucidate their physiological roles and modes of action. In certain forms, the compositions and methods can be used to pharmacologically manipulate eCB levels or their effects through allosteric modulators, thereby providing better opportunities to modulate the ECS.

[0347] iii. Improved cannabinoid receptor binding

[0348] Dendrimer compositions can improve binding to one or both cannabinoid receptors, thereby modulating signaling in a cell-specific and tissue-specific manner.

[0349] The identification and cloning of the two major cannabinoid (CB1 and CB2) receptors, as well as the discovery of their endogenous ligands, in the late 1980s and early 1990s sparked a massive effort to understand the mechanisms and physiological actions of the endocannabinoid system (ECS). Because the CB1 receptor is expressed and localized in the central nervous system (CNS), the CB1 receptor and its endogenous ligands (endocannabinoids (eCBs)), as well as the enzymes involved in their synthesis and degradation, have been implicated in a variety of pathophysiological events, including memory deficits, neurodegenerative diseases, and the like. For example, the CB1 receptor is particularly abundant in the frontal cortex, hippocampus, basal ganglia, hypothalamus, cerebellum, spinal cord, and peripheral nervous system. Thus, in certain forms, dendrimer compositions can be used to deliver cannabinoids to ameliorate or treat symptoms of diseases and disorders affecting one or more of these nervous system regions.

[0350] CB1 receptors are also present in inhibitory GABA neurons and excitatory glutamatergic neurons. Therefore, in some forms, the composition can be used to improve or treat the symptoms of diseases and conditions associated with GABAergic and glutamatergic signaling disorders. CB2 receptors are most abundantly present in immune system cells, hematopoietic cells, and glial cells. Under normal healthy conditions, CB2 receptors are mainly expressed in the periphery; under disease or injury conditions, this upregulation occurs in the brain, so CB2 receptors are expressed in the brain under unhealthy conditions. Therefore, in some forms, the composition can be used to improve or treat the symptoms of diseases and conditions associated with damaged immune cells, hematopoietic cells, and glial cells (e.g., macrophages and activated microglia). The neurophysiology of CB1 receptors and CB2 receptors will be described in more detail below.

[0351] a.CB1 receptor

[0352] The CB1 receptor is one of the most abundant G protein-coupled receptors (GPCRs) in the CNS, with particularly high levels in the cerebral cortex, hippocampus, basal ganglia, cerebellum, and brainstem. CB1 receptors are also present in peripheral nerve terminals and in some extraneural sites, such as the testis, eye, vascular endothelium, and spleen. CB1 receptors are highly enriched in presynaptic and axonal compartments, restricting their function to sites of synaptic activity. In addition to being localized on the cell surface, intracellular localization of CB1 receptors has also been reported in heterologous systems and primary cultures. CB1 receptors bind to Δ9-tetrahydrocannabinol (Δ9-THC), the major active ingredient of Cannabis sativa (marijuana), and mediate most of the effects of Δ9-THC on the CNS. In addition, CB1 receptors can bind synthetic cannabinoid mimetics, such as CP55940, JWH-015, WIN55212-2, and the endogenous arachidonic acid derivatives anandamide (AEA) and 2-arachidonylglycerol (2-AG). Following ligand binding and receptor activation, CB1 receptors couple primarily to pertussis toxin (PTX)-sensitive Gi / o-type G proteins, which result in a rapid decrease in cAMP levels by inhibiting adenylate cyclase activity. Coupling to other G proteins, including Gs, can also stimulate adenylate cyclase, albeit with lower efficacy, although the degree of cAMP accumulation is not necessarily a good indicator of G protein coupling. Promiscuous coupling to different G proteins, signaling mediated by β-arrestins, and evidence for signaling from intracellular compartments add another level of complexity, making these receptors as multidimensional as other GPCRs. CB1 receptors exhibit constitutive activity in the absence of agonists that suggests G protein activation, which could mediate their highly polarized localization to axons and presynaptic compartments.

[0353] Figure 1A and 1BSchematic diagram of the key enzymes in the cannabinoid pathway. Diacylglycerol lipase (DGLα) and phospholipase D (PLD) generate the endogenous ligands anandamide (AEA) and 2-arachidonoylglycerol (2-AG; Figure 1A These activate cannabinoid 1 receptors (CB1) in the central nervous system (CNS). Results can include modulation of adenylate cyclase activity to inhibit cAMP accumulation, voltage-gated calcium channels (VGCCs), K+ channels, and neurotransmitter release in presynaptic excitatory and inhibitory synapses ( Figure 1A Following activation of the CB1 receptor by ligand binding, signaling via G proteins and / or β-arrestins may occur at the plasma membrane, in endocytic pits, or in endosomes following receptor internalization ( Figure 1B ). G proteins normally bind to unphosphorylated receptors, whereas β-arrestins bind to receptors phosphorylated by G protein receptor kinases ( Figure 1B ). Figure 1C Schematic representation of how different cannabinoid (cb) receptor signaling patterns influence neuromodulation in health and disease in specific ways.

[0354] b.CB2 receptor

[0355] Compared to CB1 receptors, CB2 receptors have a more defined expression pattern in the brain, being primarily found in cells and tissues of the immune system (Klein, 2005; Mackie, 2006). In the CNS, CB2 receptor expression is associated with inflammation and is primarily localized to microglia, the resident macrophages of the CNS (Mackie, 2008; Palazuelos et al., 2009). This selective localization and the regulatory effects of CB2 receptors on microglial function are particularly important, as microglia play an important role in Alzheimer's disease (AD) and other diseases associated with the basal ganglia (Ramírez et al., 2005; Sagredo et al., 2007; Fernández-Ruiz et al., 2011; Yeh et al., 2016). In addition, CB2 receptors expressed in neurons can control synaptic function and are involved in drug abuse and synaptic plasticity (Xi et al., 2011; Stempel et al., 2016). For example, the selective CB2 receptor agonist JWH133 inhibits dopaminergic firing in the ventral tegmental area and reduces cocaine self-administration (Zhang et al., 2016). In addition, neuronal CB2 receptors act independently of CB1 receptors to regulate inhibitory plasticity and in vivo gamma oscillations in the CA2 / 3 region of the hippocampus (Stempel et al., 2016).

[0356] iv. Improved non-cannabinoid receptor binding

[0357] The dendrimer compositions can improve the binding of cannabinoids to one or more non-cannabinoid receptors, thereby modulating signaling in a cell-specific and tissue-specific manner. For example, cannabinoids also bind to other G protein-coupled receptors, such as GPR55, GPR18, GPR3, GPR6, GPR12 receptors; transient receptor potential channels, such as TRP vanilloids TRPV1 to TRPV4, TRP anchor protein TRPA1, and TRPM member TRPM8 receptors; peroxisome proliferator-activated receptors, such as PPAR2 and PPARγ; monoamine transporters, such as norepinephrine, dopamine, and serotonin 1A receptors, fatty acid amide hydrolases, monoacylglycerol lipases, transporting fatty acid binding proteins, adenosine equilibrative nucleoside transporters, and glycine receptors alpha 1 and alpha 3 receptors. Thus, in certain forms, the dendrimer compositions can be used to improve the binding of cannabinoids to one or more non-cannabinoid receptors to modulate signaling of one or more of the above receptors.

[0358] B. Conditions that require treatment

[0359] The composition is suitable for treating one or more diseases, disorders and injuries in the central and peripheral nervous systems. The composition can also be used to treat a variety of diseases, disorders and injuries, including gastrointestinal diseases, eye diseases and treatment of other tissues where nerves play a role in diseases or disorders. Cannabinoids have been used to treat indications including nausea and vomiting caused by chemotherapy, appetite stimulation in HIV / AIDS, chronic pain, spasticity caused by multiple sclerosis or paraplegia, depression, anxiety, sleep disorders, psychosis, glaucoma or Tourette syndrome.

[0360] The compositions and methods are also suitable for preventive use. For example, the compositions can be administered to patients in need to improve, treat or prevent symptoms associated with inflammatory diseases, neurodegenerative diseases, pain disorders, spasticity and convulsions, bone diseases, mental illnesses, metabolic disorders, and cancer-related neuropathic pain. The compositions are particularly suitable for treating or improving complications associated with chemotherapy, such as nausea and vomiting; symptoms of HIV / AIDs, such as loss of appetite, symptoms associated with multiple sclerosis, such as spasticity; and one or more of sleep disruption and / or sleep disorders, anxiety disorders, mental illnesses, chronic pain, glaucoma, and / or Tourette syndrome.

[0361] The dendrimer conjugate 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 2 The 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.

[0362] a. Relieve chronic pain

[0363] The compositions and methods are useful for treating chronic pain and diseases and conditions associated with chronic pain.

[0364] Chronic pain is a complex sensory, cognitive and emotional experience that imposes a significant personal, psychological and socioeconomic burden on patients. An estimated 1.5 billion people worldwide suffer from chronic pain, which is often difficult to treat and can become resistant to the potent analgesic effects of opioid analgesics. Chronic pain conditions are often resistant to standard treatments, including the use of opioids such as morphine.

[0365] Cannabinoids act primarily through Gi / o protein-coupled cannabinoid CB1 and CB2 receptors, which are found throughout the nervous system. CB1 receptors are located at key nodes in the pain pathway, and their activity controls the sensory and emotional components of pain. CB2 receptors are normally expressed at low levels on microglia, astrocytes, and peripheral immune cells. In chronic pain states, CB2 expression is significantly increased, thereby regulating the activity of these central and peripheral immune cells, with important effects on peripheral pain circuits.

[0366] The compositions and methods can be used to target CB1 or CB2 receptors to control pain and improve pain outcomes for various pain types, including but not limited to musculoskeletal pain, osteoarthritis pain and / or joint pain, and other types of pain caused by inflammation. In some forms, the compositions and methods can be used to control neuropathic pain and / or nociceptive pain, including pain caused by cancer, injury, accident, surgery, inflammation, tissue damage, arthritis (including osteoarthritis and rheumatoid arthritis), joint pain, infection, gastrointestinal pain, diabetes, diabetic neuropathy, postherpetic neuralgia, neuropathic pain, peripheral neuropathy, or multiple sclerosis. In an exemplary embodiment, the compositions and methods regulate microglia function through microglia-neuron interactions in the spinal cord nociceptive circuit to promote cannabinoid-mediated analgesia. In another exemplary embodiment, the compositions and methods can be administered in a supplemental regimen with opioids to provide additional analgesic effects in addition to the analgesic effects provided by opioids. In each of these forms, combination therapy for chronic pain can reduce patients' pain severity, reduce their reliance on (and addiction to) opioids, or both.

[0367] b. Mental health disorders and conditions

[0368] 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, schizophrenia and other psychotic disorders, and substance-related disorders.

[0369] i. Affective or emotional disorders

[0370] 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.

[0371] ii. Anxiety disorders

[0372] 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.

[0373] 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).

[0374] 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.

[0375] 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.

[0376] 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.

[0377] iii. Eating disorders

[0378] 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.

[0379] 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 when 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.

[0380] 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.

[0381] 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.

[0382] The composition is particularly suitable for administration to patients in need to stimulate appetite, reduce nausea and vomiting. For example, loss of appetite is a common symptom in patients with opportunistic infections (such as HIV / AIDS) and patients receiving chemotherapy and / or other cancer treatments. Opportunistic infections are infections that people with weak immune systems are susceptible to. In addition, some drugs may reduce appetite due to side effects, such as tingling around the mouth, nausea and taste changes. In addition, depression caused by terminal illness and / or terminal illness treatment may cause loss of appetite. Therefore, in some forms, the composition can be applied to such patients to stimulate appetite and / or reduce nausea or emaciation syndrome.

[0383] c. Neurological and neurodegenerative diseases

[0384] The compositions and methods are useful for treating symptoms associated with neurological disorders and neurodegenerative diseases.

[0385] Neurodegenerative diseases are chronic progressive neurological diseases that affect neurological and behavioral functions, involving 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, one 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.

[0386] 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).

[0387] The compositions and methods can also be used to deliver active agents to treat neurological 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 neurological or neurodegenerative diseases or disorders or central nervous system disorders. The methods generally include 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.

[0388] Neurodegeneration refers to the progressive loss of neuronal structure or function, including neuronal death. For example, the compositions and methods can be used to treat a subject having a disease or condition, such as Parkinson's disease (PD) and PD-related disorders, Huntington's disease (HD), Amyotrophic Lateral Sclerosis (ALS), Alzheimer's disease (AD) and other dementias, prion diseases, such as Creutzfeldt-Jakob Disease, Corticobasal Degeneration, Frontotemporal Dementia, HIV-related cognitive impairment, Mild cognitive impairment, Motor Neuron Disease (MND), Spinocerebellar Ataxia (SCA), Spinal Muscular Atrophy (SMA), Friedreich's Ataxia, Lewy Body Disease, Alpers' Disease, Batten Disease, Cerebro-oculofacial-skeletal syndrome, Corticobasal Degeneration, Gerstmann-Straussler-Scheinker Disease, Disease), Kuru, Leigh's disease, monosomic muscular dystrophy, multiple system atrophy, multiple system atrophy with orthostatic hypotension (Shy-Drager syndrome), multiple sclerosis (MS), neurodegenerative diseases with brain iron accumulation, opsoclonus, myoclonus, posterior cortical atrophy, primary progressive aphasia, progressive supranuclear palsy, vascular dementia, progressive multifocal leukoencephalopathy, dementia with Lewy bodies (DLB), lacunar syndrome, hydrocephalus, Wernicke-Korsakoff's syndrome, postencephalitic dementia, cancer and chemotherapy-related cognitive impairment and dementia, and dementia and pseudodementia caused by depression.

[0389] In some embodiments, the subject suffers from an excitotoxic disorder. Excitotoxicity is a process in which nerve cells are damaged due to overstimulation. Many diseases are associated with excitotoxicity, including stroke, traumatic brain injury, multiple sclerosis, amyotrophic lateral sclerosis, Alzheimer's disease, and spinal cord injury. Damage to nerve cells can lead to corresponding neurological symptoms, which may vary depending on the damaged cells and the extent of the damage. Once damaged, nerve cells cannot be repaired and the patient may suffer permanent damage. Many drugs have been developed and used in an attempt to interrupt, affect, or temporarily stop the glutamate excitotoxic cascade that causes neuronal damage. One strategy is to "upstream" try to reduce glutamate release. Therefore, in some embodiments, the dendrimer complex includes one or more active agents for treating excitotoxic disorders.

[0390] In some embodiments, the subject suffers from a neurological disease or is in need of neuroprotection.Exemplary conditions and / or subjects include, but are not limited to, subjects who have suffered, are suffering from, or are likely to suffer from a stroke, a traumatic brain injury, a spinal cord injury, post-traumatic stress syndrome, or a combination thereof.

[0391] In some embodiments, the compositions and methods are administered to a subject in need thereof in an effective amount to reduce or prevent one or more molecular or clinical symptoms of a neurodegenerative disease, or one or more mechanisms leading to neurodegeneration.Exemplary neurological diseases and neurodegenerative diseases are further described below.

[0392] i. Multiple sclerosis (MS)

[0393] The compositions and methods can be used to treat and / or alleviate symptoms associated with MS. MS is a major immune-related neurodegenerative disease characterized by demyelination with associated axonal and neuronal loss. Cannabis, Δ9-THC or other CB agonists can improve symptoms of MS such as spasticity, spasticity and pain (Croxford, 2003; Pertwee, 2007; Rog, 2010; Notcutt et al., 2012). Use Nabiximol, a cannabis extract oral mucosal spray containing fixed concentrations of Δ9-THC and cannabidiol (CBD), improves symptoms in MS patients. A meta-analysis of multiple clinical studies showed a reduction in motor dysfunction and pain. Thus, in some forms, the composition can be administered to a patient in need to reduce muscle spasms and pain associated with multiple sclerosis. In some forms, the composition can be administered to a patient in need to reduce fatigue associated with inflammation, which in turn can improve the mobility of MS patients.

[0394] At the molecular level, these improvements are generally associated with agonist activation of both CB1 and CB2 receptors, resulting in dual anti-inflammatory and neuroprotective effects throughout the CNS. These effects include upregulation of pro-survival molecules such as interleukins in astrocytes and reduction of cytotoxic factors such as nitric oxide, reactive oxygen species, and pro-inflammatory cytokines in microglia. The exact mechanism by which the receptors exert their neuroprotective activity may include activation of the phosphatidylinositol 3-kinase / mammalian target of rapamycin complex 1 (mTOR1) pathway and brain-derived neurotrophic factor (BDNF).

[0395] Synthetic CBs attenuated inflammation and neuropathic pain in a mouse model of experimental autoimmune encephalomyelitis (EAE). Similar results were observed with systemic treatment of mice with Theiler's murine encephalomyelitis virus-induced demyelinating disease, a mouse model of chronic progressive MS, using the agonists WIN55212-2, ACEA, and JWH-015. Motor function in mice was improved by modulating the infiltration of microglia and lymphocytes into the spinal cord. Conversely, when an inverse agonist of the CB1 receptor (SR141716A) was applied, EAE could be exacerbated by the release of proinflammatory cytokines in the brain and spinal cord of mice. Together, these results suggest that the neuroprotective effects of CB1 and CB2 receptors may be impaired in MS, and enhancing them could provide new therapeutic approaches.

[0396] Thus, the compositions and methods are effective in treating and / or alleviating one or more symptoms of MS, including neuroinflammation, pain, muscle spasms, fatigue, stiffness and weakness, depression and anxiety, and speech and swallowing difficulties.

[0397] ii. Huntington's disease (HD)

[0398] ECS dysregulation has also been reported in experimental models and HD patients. CB1 receptor expression is at least somewhat reduced before the onset of neurodegenerative HD in mice (e.g., 27% reduction in CB1 receptor mRNA in the striatum) (McCaw et al., 2004). Loss of CB1 receptor expression reduces motor ability and increases the number of aggregates in the striatum of HD mice (Mievis et al., 2011). Significant loss of CB1 receptors has also been reported in HD patients (Glass et al., 2000). Activation of CB1 receptors may help slow the progression of HD. For example, preclinical evidence suggests that the use of CB1 receptors in patients with progressive neurodegenerative diseases such as HD may be beneficial. CB1 receptors can be neuroprotective (Valdeolivas et al., 2012). In addition, selected receptor agonists have neuroprotective potential in cell culture models of HD (Scotter et al., 2010; Laprairie et al., 2016). Ligands that favor β-arrestin-mediated signaling, such as Δ9-THC, reduce cell function and viability in these models, suggesting potential pharmacological properties for therapeutic agonists (Laprairie et al., 2014, 2016). These events are mediated in part by activation of Gαi / o-mediated pathways and may limit glutamate release from cortical neurons and GABA release from striatal medium spiny neurons (Dowie et al., 2010; Laprairie et al., 2016). Results from studies in the R6 / 2 mouse model of HD suggest that CB1 receptor activation parallels BDNF expression, leading to neuroprotection (Báázquez et al., 2015). Overall, the in vivo and in vitro data suggest that CB agonists with specific pharmacological properties (biased towards BDNF upregulation and release) could be developed to treat or ameliorate HD.

[0399] Therefore, by increasing neuroprotection in a subject through one or more of the above mechanisms, these compositions and methods can effectively slow the progression of HD in a subject. These compositions and methods can also effectively treat and / or alleviate one or more symptoms of HD, including but not limited to neuroinflammation, pain, involuntary jerking or twisting movements (chorea), muscle problems such as stiffness or muscle contractures (dystonia), slow or abnormal eye movements, impaired gait, posture and balance, speech or swallowing difficulties, cognitive impairment and abnormal behavior.

[0400] iii. Alzheimer's disease

[0401] The CB1 receptor has also been the focus of intensive research as a potential target for AD. This work has been performed in vitro, in animal models, and in postmortem samples. Altered expression levels of several components of the ECS in postmortem samples from AD patients have been identified, although their role in the pathophysiology of the disease remains unknown. For example, CB1 receptors in the hippocampus of AD patients were not different from age-matched controls. However, levels of the site of action, MAGLs (degrading enzymes for 2-AG), were reduced in these patients, suggesting alterations in eCB signaling and structure (Mulder et al., 2011). Limited positive behavioral outcomes have been observed in small-scale clinical trials and pilot studies using Δ9-THC analogs (Aso and Ferrer, 2014). An analysis of available studies and trials suggests that synthetic CB has significant benefits for certain behavioral and psychological symptoms of dementia (Liu et al., 2015). However, these conclusions are based on short-term and limited studies; further research is needed to evaluate the safety and efficacy of CB in AD. In experimental models of AD, some findings suggest that activation of CB1 and CB2 receptors may have beneficial effects against Aβ toxicity primarily through neuroprotection, as previously suggested in other neurodegenerative diseases (Stumm et al., 2013). CBD also reduces Tau phosphorylation in PC12 cells, thus providing a different mechanism of neuroprotection during AD (Esposito et al., 2006). Since CB1 receptors are unlikely to be directly activated by CBD, the effect on Tau phosphorylation may be through the antioxidant effects of CBD or may be a CB receptor-independent effect. Due to the activation of the immune system and CNS CB system in AD, the reduction of harmful β-amyloid peptides and tau phosphorylation while promoting intrinsic CNS repair mechanisms may occur in succession (Aso and Ferrer, 2014). For example, recent studies of TREM2 receptors in microglia (where CB2 receptors are expressed and control cellular responses) also provide an immune-related mechanism for controlling AD (Yeh et al., 2016).

[0402] Aging is a major risk factor for neurodegenerative diseases, during which neuronal progenitor proliferation is greatly reduced. Notably, CB can stimulate both embryonic and adult neurogenesis (Jiang et al., 2005; Trazzi et al., 2010). Axon guidance, cell migration, synapse formation, and cell survival are also regulated during development. Dysregulation of these processes during development and aging may significantly contribute to multiple CNS diseases.

[0403] iv. Traumatic brain injury

[0404] CB1 / 2 receptors are involved in TBI, and 2-AG is increased after TBI in animal models. There is an "on-demand" signal for the production of eCBs after TBI, which can reduce brain edema and inflammation. These events may have neuroprotective effects, preventing excitotoxicity, inhibiting inflammatory cytokine production, and enhancing stem cell migration and differentiation. In addition, CB1 receptor and CB2 receptor antagonists prevent drug-induced neuroprotection in the mouse TB1 model. However, as previously noted for other diseases, there is limited clinical data available to support the effectiveness and safety of CB during TBI. Therefore, the compositions and methods may also be effective in treating and / or alleviating one or more symptoms of TBI, including but not limited to neuroinflammation, pain, cognitive impairment, behavioral abnormalities, fatigue, speech problems, nausea or vomiting, headaches, sensory impairment, and sleep abnormalities.

[0405] d. Gastrointestinal diseases

[0406] The endocannabinoid system (i.e., endogenous circulating cannabinoids) plays a protective role in the GI tract. Therefore, the compositions and methods can effectively treat and / or alleviate various GI diseases, such as necrotizing enterocolitis (NEC), abdominal sepsis, pneumonia, arthritis, pancreatitis and atherosclerosis, inflammatory bowel disease (especially Crohn's disease), irritable bowel syndrome, and secretion and motility-related diseases. The compositions and methods can also effectively treat and / or alleviate one or more gastrointestinal dysfunction symptoms in the intestines, liver, and pancreas, including but not limited to nausea and vomiting, cannabinoid vomiting syndrome, anorexia, weight loss, and chronic abdominal pain.

[0407] In some embodiments, a single dendrimer complex composition can simultaneously treat and / or diagnose multiple symptoms in two different locations in the human body, including the gastrointestinal tract and the central nervous system. For example, a dendrimer complex composition (including a dendrimer linked to a therapeutic, prophylactic, or diagnostic agent) can be administered enterally to treat the gastrointestinal region while selectively targeting microglia and astrocytes after absorption into the bloodstream. Microglia and astrocytes play a key role in the pathogenesis of NEC.

[0408] e. Glaucoma and visual impairment

[0409] Glaucoma is one of the leading causes of blindness worldwide. While there are a variety of treatments for glaucoma, there is a critical need for new and better glaucoma treatments. Increased intraocular pressure, caused by a blockage in the flow of fluid that helps the eye maintain its rigid shape, is a major risk factor for the development and progression of glaucoma. Normally, this clear fluid, called aqueous humor, circulates between the front of the lens and the back of the cornea. In people with increased intraocular pressure, the outflow of fluid from the front chamber of the eye can be restricted, causing pressure to build up like water behind a dam. Increased intraocular pressure reduces the flow of nutrients to the optic nerve, which can lead to glaucoma.

[0410] Thus, the composition can be administered to a patient in need to reduce intraocular pressure (IOP) and provide neuroprotection. For example, a composition containing THC and / or a THC analog can be administered to reduce IOP, while a composition containing HU-211 can be administered to provide neuroprotection. In certain forms, the composition can be administered to a patient in need to protect or save the optic nerve from damage and / or restore its blood supply.

[0411] f. Sleep disruption and sleep disorders

[0412] CB1 receptors located in the pons and basal forebrain may be involved in sleep induction. This process may be related to the activation of cholinergic neurons located in the basal forebrain and pons through CB1 receptors, thereby assisting in inducing sleep. CB1 receptors enhance the activation of the serotonin system and play a potential regulatory role in the sleep-wake cycle.

[0413] Therefore, the composition can be administered to patients in need to improve sleep structure. In some forms, the composition can increase total sleep time, reduce wakefulness after falling asleep, reduce slow wave sleep and / or reduce or increase REM sleep latency as needed and / or increase sleepiness in patients with insomnia. In some forms, the composition can be administered to improve the alertness of patients (e.g., patients with narcolepsy).

[0414] C. Dosage and Effective Amount

[0415] In some in vivo methods, the dendrimer complex is administered to a subject in a therapeutically effective amount. The term "effective amount" or "therapeutically effective amount" refers to a dose sufficient to treat, inhibit or alleviate one or more symptoms of the disease being treated or otherwise provide the desired pharmacological and / or physiological effect. The exact dose will vary depending on a variety of factors, such as subject-related variables (e.g., age, immune system health, etc.), the disease or condition, and the treatment affected.

[0416] Typically, the dosage of the composition can be from about 0.001 to about 100 mg / kg of the body weight of the subject being treated, from about 0.01 to about 100 mg / kg of body weight, from about 0.1 mg / kg to about 10 mg / kg, and from about 0.5 mg to about 5 mg / kg of body weight. The subject is typically a mammal, most preferably a human.

[0417] For example, the amount of the dendrimer complex composition is effective to deliver one or more active agents to cells at or near the site of inflammation, particularly central nervous system inflammation or ocular inflammation. Thus, in some embodiments, the amount of the dendrimer complex composition comprising one or more active agents is effective to improve inflammation in a subject. In a preferred embodiment, the effective amount of the dendrimer complex composition does not induce significant cytotoxicity in cells of the subject compared to an untreated control subject. Preferably, the amount of the dendrimer complex composition is effective to prevent or reduce inflammation and / or further associated symptoms of a disease or condition in a subject compared to an untreated control.

[0418] In general, the time and frequency of administration will be adjusted to balance the efficacy of a given therapeutic or diagnostic regimen with the side effects of a given delivery system. Exemplary dosing frequencies include continuous infusion, single and multiple administrations, such as hourly, daily or weekly administrations.

[0419] In some embodiments, the dosage is administered once, twice, or three times a day, or once every other day, two days, three days, four days, five days, or six days. In some embodiments, the dosage is administered once or twice a week, once or twice every two weeks, or once every three weeks. In some embodiments, the dosage is administered once or twice a month, once or twice every two months, once or twice every three months, once or twice every four months, once or twice every five months, or once or twice every six months.

[0420] Those 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. The term "chronic" means that the length of the dosing regimen can be hours, days, weeks, months, or possibly years.

[0421] In some embodiments, the regimen includes one or more cycles of treatment followed by a drug holiday (e.g., no drug). The treatment cycle can be, for example, the dosing cycle described above. Likewise, 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.

[0422] The dendrimer complexes may be administered in combination with one or more additional therapeutically active agents known to treat the above-mentioned conditions or diseases.

[0423] D. Control

[0424] The effect of the dendrimer complex composition can be compared to a control. Suitable controls are known in the art and include, for example, untreated cells or untreated subjects. In some embodiments, the control is untreated tissue from a treated subject or from an untreated subject. Preferably, the control cell or tissue is derived from the same tissue as the treated cell or tissue. In some embodiments, the untreated control subject suffers from or is at risk for the same disease or condition as the treated subject.

[0425] E. Combination therapy

[0426] The dendrimer complex composition can be administered alone or in combination with one or more other active agents as part of a therapeutic or prophylactic treatment regimen. The dendrimer complex composition can be administered on the same day as the second active agent or on different days. For example, a composition including the dendrimer complex composition can be administered on the first, second, third, or fourth day, or a combination of these days.

[0427] The term "combination" or "combination" is used to refer to the concomitant, simultaneous or sequential administration of two or more drugs. Thus, the combination can be administered simultaneously (e.g., as a mixture), separately but simultaneously (e.g., through different intravenous lines into the same subject), or sequentially (e.g., one compound or drug is administered first and then the second is administered).

[0428] VI. Kit

[0429] The composition may be packaged in a kit. The kit may include a single or multiple doses of a composition comprising one or more cannabinoids bound or conjugated to 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.

[0430] The invention will be further understood with reference to the following non-limiting examples.

[0431] Example 1: Synthesis of Hydroxyl-Polyamidoamine (PAMAM-OH) Dendrimer Cannabinoid (CBD) Conjugates

[0432] The synthesis of PAMAM-OH-CBD conjugates was achieved using a variety of linking chemistries and linkers (cleavable and non-cleavable). Briefly, the surface hydroxyl groups on PAMAM-OH were modified with linkers to form complementary groups on the surface that could further react with complementary groups on the drug linker. In addition, the target drug was modified with a linker to allow for reaction between the complementary functional groups and the dendrimer linker.

[0433] The linker on the drug is connected by a cleavable or non-cleavable linker. Non-limiting examples of cleavable links include ester bonds sensitive to esterases, disulfide bonds sensitive to glutathione, phosphodiester bonds sensitive to phosphatases, triglycyl peptide linkers (CX) capable of lysosomal release, acid-cleavable hydrazine linkers, etc. Non-limiting examples of non-cleavable links include ether bonds or amide bonds. Amino acids, peptides, polyethylene glycols (n=2-15) and hydrocarbon chains are non-limiting examples of suitable linkers.

[0434] Dendrimer-CBD conjugates are designed to: (1) provide site-specific targeting by directing the drug to activated microglia / macrophages expressing CB2 receptors, thereby reducing dosage, enhancing efficacy, and reducing side effects associated with the free drug; (2) increase the aqueous solubility of the cannabinoid by 10-100-fold compared to the free drug; (3) avoid the systemic and dose-related side effects of the free CBD drug by sustained intracellular release at the target site; and (4) reduce or eliminate the adverse psychoactive effects associated with CB1 agonists by confining the dendrimer-CB1 agonist conjugates to the peripheral circulation using a newer generation of dendrimers.

[0435] The following examples describe non-limiting examples of dendrimer-cannabinoid conjugates in which CBD and CBD derivatives are conjugated to the surface of the dendrimer using various linkers and attachment chemistries.

[0436] Example 2: Synthesis of dendrimer-cannabidiol conjugates

[0437] Synthesis of dendrimer-cannabidiol conjugates by click chemistry using enzyme-sensitive ester bonds.

[0438] The dendrimer cannabidiol was synthesized using an efficient copper(I)-catalyzed alkyne-azide click (CuAAC) chemistry. The synthesis began with the modification of cannabidiol to an orthogonal linker with an azide terminal via a cleavable ester bond ( Figure 2A and 2BThe purpose of the azide group is to participate in the CuAAC reaction together with the alkyne functional groups on the surface of the dendrimer. In the presence of N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC) and 4-(dimethylamino)pyridine (DMAP) as coupling agents, the hydroxyl groups in cannabidiol (l) can react with N3-(OCH2CH2) n -COOH(2) reacts. The crude product is purified by column chromatography to obtain cannabidiol-azide ( Figure 2A ).

[0439] Alternatively, the dendrimer may be modified to attach a linker with a complementary alkyne group ( Figure 2B ). The hydroxyl-terminated PAMAM dendrimer (D-OH; 4) reacts with hexynoic acid in the presence of coupling agents EDC and DMAP to obtain a partially alkynyl-terminated dendrimer (5) to which ten (10) linking groups ( Figure 2B Finally, the dendrimer (5) was subjected to CuAAC click reaction with cannabidiol-azide (3) to obtain D-cannabidiol conjugates with 10 drug molecules attached. The structures and purities of all intermediates and final conjugates were confirmed by 1 H NMR and HPLC were implemented.

[0440] Dendrimer-cannabidiol conjugates were synthesized by click chemistry using a non-cleavable ester linker.

[0441] Dendrimers-cannabidiol can also be obtained by click chemistry using non-cleavable ester linkers ( Figure 3A and 3B The synthesis begins with the modification of cannabidiol by attaching it to an azide-terminal orthogonal linker via a non-cleavable ester bond ( Figure 3A In the presence of potassium carbonate (K2CO3) and dimethylformamide (DMF) as coupling agents, the hydroxyl group in cannabidiol (6) can react with N3-(OCH2CH2) n -Br(7) was reacted. The product was purified by column chromatography to obtain Figure 3A Cannabidiol-azide (8) is shown.

[0442] The dendrimer was also modified to attach a linker with a complementary alkyne group ( Figure 3B ). The hydroxyl-terminated PAMAM dendrimer (D-OH; 9) was reacted with hexynoic acid in the presence of coupling agents EDC and DMAP to obtain a partially alkyne-terminated dendrimer (10) connected with ten (10) linkers ( Figure 3BFinally, a CuAAC click reaction was performed between the dendrimer (9) and cannabidiol-azide (3) to obtain a D-cannabidiol conjugate with 10 drug molecules attached. The final conjugate was purified. The structure confirmation and purity of all intermediates and final conjugates were determined by 1 H NMR and HPLC were implemented.

[0443] Example 3: Synthesis of dendrimer-HU-308 conjugate

[0444] Synthesis of dendrimer-HU-308 conjugates via amidation of phosphatase-cleavable phosphodiester linkages

[0445] Dendrimer-HU-308 conjugates can be obtained by amidation reaction with phosphatase-cleavable phosphodiester linkers ( Figure 4A and 4B The synthesis begins with modification of HU-308 to attach a phosphodiester linker. Figure 4A As shown, the hydroxyl group in HU-308 (1) reacts with diphosphoryl chloride (P2O3Cl4) and tetrahydrofuran (THF) at -40°C to generate compound 1, which is then reacted with FmocHN-CH2CH2-H2PO4 in the presence of triethylamine (Et3N), 1,1′-carbonyldiimidazole (CDI) and DMF at room temperature to generate compound 2. Compound 2 is then reacted with DCM and piperidine at room temperature to obtain compound 3 ( Figure 4A ).

[0446] like Figure 4B As shown, D-OH was reacted with glutaric anhydride in DMF and DMAP to produce compound 4, which was then reacted with compound 3 to produce a D-HU-308 conjugate containing a phosphatase-cleavable phosphodiester linker.

[0447] Synthesis of dendrimer-HU-308 conjugates via click reaction using ester linkers

[0448] The synthesis of dendrimer-HU-308 can also be achieved by modifying HU-308 to attach an orthogonal linker with an azide end via a cleavable ester bond ( Figure 5A and 5B ). The hydroxyl groups in HU-308 can react with N3-(OCH2CH2) in the presence of EDC and DMAP as coupling agents. n -COOH reaction to obtain HU-308-azide ( Figure 5A ).like Figure 5B As shown, D-OH was reacted with hexynoic acid in the presence of coupling agents EDC and DMAP to obtain a partially alkyne-terminated dendrimer with ten (10) linkers ( Figure 5B Finally, a click reaction was performed between the dendrimer and HU-308-azide to obtain a D-HU-308 conjugate with 10 drug molecules attached. The structure confirmation and purity of all intermediates and final conjugates were performed by 1 H NMR and HPLC were implemented. Figure 5C is a schematic diagram of the conjugation of HU-308 to PAMAM-G4-OH dendrimer.

[0449] Synthesis of dendrimer-HU-308 conjugates by click reaction using glutathione-sensitive disulfide linkers

[0450] Dendrimer-HU-308 can also be synthesized by click reaction using glutathione-sensitive disulfide linker ( Fig. 6A and 6B In the presence of EDC and DMAP as coupling agents, the hydroxyl group in HU-308 can react with N3-SS-PEG2-acid to obtain HU-308-azide ( Fig. 6A ).

[0451] On the other hand, D-OH reacts with hexynoic acid in the presence of coupling agents EDC and DMAP to obtain partially alkyne-terminated dendrimers with ten (10) linkers ( Figure 6B Finally, the dendrimer undergoes a click reaction with HU-308-azide to obtain a D-HU-308 conjugate with 10 drug molecules attached ( Figure 6B The structures and purities of all intermediates and final conjugates were confirmed by 1 H NMR and HPLC were implemented.

[0452] Example 4: Synthesis of dendrimer-tetrahydrocannabinol conjugates

[0453] Dendrimer-THC conjugates were synthesized via click reaction using esterase-sensitive ester linkers.

[0454] The synthesis of dendrimer tetrahydrocannabinol (D-THC) conjugates can be achieved by using esterase-sensitive ester linkers via a click reaction ( Fig. 7A and 7B ).like Fig. 7A As shown, in the presence of EDC and DMAP as coupling agents, the hydroxyl group in THC can react with N3-(OCH2CH2) n -COOH reacts to give THC-azide. Then, D-OH reacts with hexynoic acid in the presence of coupling agents EDC and DMAP to give a partially alkyne-terminated dendrimer with ten (10) linkers ( Figure 7BFinally, the dendrimer was subjected to a click reaction with THC-azide to obtain a D-THC conjugate with 10 drug molecules attached. The structure confirmation and purity of all intermediates and final conjugates were performed by 1 H NMR and HPLC were performed.

[0455] Dendrimer-tetrahydrocannabinol conjugates were synthesized via click reaction using triglycyl peptide bonds for lysosomal release.

[0456] The synthesis of dendritic tetrahydrocannabinol (D-THC) conjugates can also be achieved by using a triglycyl peptide linker via a click reaction ( Fig. 8A and 8B The triglycyl peptide linker allows lysosome-mediated cleavage of the dendrimer-THC conjugate. In the presence of DMF and K2CO3 as coupling agents, the hydroxyl groups in THC can react with triglycyl peptide azide to obtain THC-triglycyl peptide azide ( Fig. 8A ).like Figure 8B As shown, D-OH reacted with hexynoic acid in the presence of coupling agents EDC and DMAP to obtain partially alkyne-terminated dendrimers with ten (10) linkers attached. Finally, a click reaction was performed between the dendrimer and THC-triglycyl peptide azide to obtain D-THC conjugates with triglycyl peptide linkers and 10 drug molecules attached. The structures and purities of all intermediates and final conjugates were confirmed by 1 H NMR and HPLC were implemented.

[0457] Example 5: Synthesis of dendrimer-arachidonic acid ethanolamine conjugate

[0458] Dendrimer-arachidonylethanolamine conjugates were synthesized via click reaction using an esterase-sensitive ester linker.

[0459] Dendrimer-arachidonic acid ethanolamine (D-AEA) conjugates can be synthesized by a click reaction using an esterase-sensitive ester linker ( Fig.9A and 9B ). In the presence of EDC and DMAP as coupling agents, the hydroxyl group in AEA can react with N3-(OCH2CH2) n -COOH reaction to give AEA-azide ( Fig.9A ). AEA-azide is then conjugated to the dendrimer (D-OH) via a click reaction to obtain the D-AEA conjugate, such as Fig. 9B shown.

[0460] Synthesis of dendrimer-arachidonylethanolamine conjugates using a noncleavable ether linker.

[0461] The synthesis of dendrimer-arachidonic acid ethanolamine (D-AEA) conjugates can also be obtained by click reaction using non-cleavable ether linkers ( Fig. 10A and 10B ). In the presence of K2CO3 and DMF as coupling agents, the hydroxyl group in AEA can react with N3-(OCH2CH2) n -Br reaction to give AEA-azide ( Fig. 10A ). AEA-azide was conjugated to the dendrimer (D-OH) via a click reaction to obtain a D-AEA conjugate with a non-cleavable linker, such as Fig. 10B shown.

[0462] Example 6: Synthesis of dendrimer-2-arachidonoylglycerol (D-2-AG) conjugates via click reaction using esterase-sensitive ester linkers.

[0463] Dendrimer-2-arachidonoylglycerol (D-2-AG) conjugates can be synthesized by click reaction using non-cleavable esterase-sensitive ester linkers ( Fig.11A and 11B The hydroxyl group in 2-AG can react with N3-(OCH2CH2) in the presence of EDC and DMAP as coupling agents. n -COOH reaction to give 2-AG-azide ( Fig.11A ). 2-AG-azide is conjugated to a dendrimer (D-OH) via a click reaction to obtain a D-2-AG conjugate with an esterase-sensitive ester linker, such as Fig. 11B shown.

[0464] Example 7: Synthesis and conjugation of cannabinoid drugs to hydroxyl terminated PAMAM, glucose dendrimers and hydroxylated bis-MPA dendrimers.

[0465] This approach targets specific receptors on cells (e.g., neurons, microglia, macrophages, and other cells). These receptors can be found anywhere in the body. For example, CB1 / CB2 receptors are found in various organs of the body, including the brain. The conjugates described herein bind to these receptors. Thus, the "intrinsic cell targeting" of these dendrimers, e.g., hydroxy dendrimers are targeted to microglia and / or macrophages, and glucose dendrimers are targeted to neurons, is a secondary factor in binding to and acting on specific receptors.

[0466] Targeting of receptors can be used to achieve the desired results. In one embodiment, the conjugate targets CB1, which can cause agonism, antagonism, inverse or inverse agonism. In another embodiment, the conjugate targets CB2, which can cause agonism, antagonism or inverse agonism, or a combination of activities against CB1 / CB2, TRPV1 and other receptors.

[0467] When combined with the cell targeting capabilities of glucose dendrimers (damaged neurons (primary), microglia / macrophages (secondary) or hydroxy dendrimers (small, microglia / macrophages), the conjugates can be more specifically targeted to specific cannabinoid receptors on specific cells in specific organs.

[0468] The dendrimer conjugates not only aid in targeting, but also improve water solubility by more than 200 times compared to drugs not bound to the dendrimers. This greatly facilitates formulation and delivery.

[0469] The following examples using tryptamines (as non-limiting examples) demonstrate the efficacy of the conjugates for selective targeting, alteration of binding affinity, and Hu-308 for water solubility.

[0470] Materials and Methods

[0471] 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-dimethylchloropropyl)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, serotonin, psilocybin analogs, psilocybin analogs, and cannabidiol drugs were purchased from Cayman Chemicals.

[0472] Synthesis of hydroxy-PAMAM dendrimer drug conjugates.

[0473] 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. 1 H 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.

[0474] Synthesis of D-hexyne

[0475] 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. Then EDC.HCl (1.54 g, 12.6 mmol) was added to the reaction mixture in batches 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, dialyzed against DMF for 12 hours, and then dialyzed against water for 24 hours. The aqueous layer was frozen and lyophilized to give D-hexyne as a hygroscopic white solid (yield of 75%). 1H 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%.

[0476] Synthesis and purification of Hu308-azide

[0477] This method can be used for other cannabinoids. A solution of Hu308 in anhydrous dimethylformamide (DMF) was treated with hexanoic acid-PEG4-azide in anhydrous DMF, DMAP and EDC. The reaction was stirred at room temperature for 24 hours under N2. The reaction was monitored using thin layer chromatography, and upon completion the crude reaction mixture was concentrated, dissolved in saturated sodium bicarbonate and extracted with DCM, followed by washing the organic layer with saturated sodium bicarbonate solution, saturated ammonium chloride solution and brine. The organic layer was then dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was dried under a rotary evaporator and purified on silica gel using CombiFlash chromatography with a 40% ethyl acetate / hexane gradient as the eluent. The desired Hu308-PEG4-azide was obtained as a colorless oil.

[0478] Synthesis of dendrimer-drug conjugates.

[0479] 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.

[0480] Table 2. Physical properties of D-drug conjugates.

[0481]

[0482] G2-glucose dendrimer (GD2)-drug conjugate

[0483] 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.

[0484] Synthesis and characterization of glucose dendrimers (GD2).

[0485] 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 3).

[0486] Table 3: Physicochemical properties of GD2

[0487] size 4.4nm Zeta Potential -6.0 Glucose quantity 24 HPLC purity ＞99％ Molecular weight 12800Da

[0488] Synthesis and characterization of GD2-drug conjugates

[0489] The Hu308 drug was conjugated to the GD2-hexynoic acid dendrimer using a click chemistry strategy. The linker-linked drug moiety was conjugated to the glucose dendrimer using a Cu(I)-catalyzed click (CuAAC) reaction in the presence of a catalytic amount of CuSO4.5H2O and sodium ascorbate to afford the GD2-drug conjugate. Trace copper was removed by dialysis against ethylenediaminetetraacetic acid (EDTA). The final GD2-drug conjugate was characterized by NMR and HPLC.

[0490] Table 4: Physical properties of GD2-drug conjugates

[0491]

[0492] result

[0493] Compounds such as Fig.12 As shown, Figures 13A-13B Show its synthesis and structure, Fig. 13C Binding and release of the compounds are shown.

[0494] Fig.14 Graph showing % binding efficiency versus log (μM) of tryptamine in a 5-HT1A human serotonin GPCR cell-based agonist cAMP assay.

[0495] Both hydroxy and GD dendrimers have OH surface groups. When tryptamines are bound to these dendrimers with the same attachment chemistry, the affinities for serotonin and other receptors vary greatly. Fig. 13C . Binding assays showed that the conjugates had lower affinity than the free drug in a cell-based binding assay (indicative of in vivo efficacy). The lower affinity may allow for less tight binding and possibly 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 active. 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 dendrimer, but may open outward in the hydrophilic interior of the glucose dendrimer.

[0496] The complex can be active with or without drug release and can be designed to release or not release the drug using appropriate linkage chemistry as described herein. Binding affinity measures the activity of the complete conjugate. For example, a Hu-308 conjugate releases the drug over a period of time ( Fig. 13C ), and is active in both conjugated and released forms. Cannabidiol (CBD) conjugates can be designed to not release the drug and are active in conjugated form.

[0497] Example 8. Localization of dendrimer-Hu308 in peripheral nerve injury / sciatic nerve injury:

[0498] Materials and methods

[0499] Animals that had undergone sciatic nerve ligation were given the dendrimer Hu308 labeled with the fluorescent dye Cy5 by intravenous injection 10 days later. The animals were perfused and imaged 24 hours later. Microglia on the injured side are known to overexpress CB2 receptors that regulate neuropathic pain.

[0500] result

[0501] In a rodent model of chronic neuropathic pain caused by sciatic nerve injury, dendrimers conjugated to the CB2 receptor agonist Hu308 bound to CB2 receptors overexpressed in microglia surrounding the injury site. Hu308 binds to the intracellular portion of CB2R, and dendrimer-Hu308 is localized at high concentrations within glial cells, thereby achieving intracellular binding of CB2 receptors.

[0502] The hydroxy-terminated dendrimer, Hu308-Cy5, localized only in microglia surrounding the injured area of ​​the sciatic nerve. These microglia are known to overexpress CB2Rs after injury. On the other hand, the glucose dendrimer, GD, labeled with Cy5, localized in injured neurons along the nerve as well as in neurons and microglia in the dorsal root ganglia ipsilateral to the injury in the acute phase after injury. GD2-Cy5 localized in neurons near the ligation site as well as in neurons and glial cells in the dorsal root ganglia. This suggests that GD-Hu308 can bind to CB2 receptors on injured neurons and glial cells immediately after peripheral nerve injury.

[0503] Modifications and variations of the present invention will be apparent to those skilled in the art and are intended to fall within the scope of the appended claims.

Claims

1. A composition comprising a dendrimer, preferably a hydroxyl-terminated dendrimer, a sugar-terminated dendrimer and / or a sugar-based dendrimer, conjugated to at least one natural cannabis derivative or synthetic cannabinoid compound, optionally wherein the conjugation is covalent.

2. The composition according to claim 1, wherein the compound is covalently conjugated via covalent bonds between modified or unmodified surface groups or internal groups of the dendrimer, the covalent bonds comprising ester bonds, amide bonds, disulfide bonds or ether bonds.

3. A composition according to any one of claims 1 or 2, wherein the covalent bond between the dendrimer and the cannabinoid compound is broken after administration.

4. A composition according to any one of claims 1 to 3, wherein the dendrimer is a polyamidoamine (PAMAM) dendrimer of generation 1 to 7 (e.g., generation 1, 2, 3, 4, 5, 6 or 7), preferably with more than 40% to 100% of its surface groups being hydroxylated.

5. The composition of any one of claims 1 to 4, wherein the dendrimer is a PAMAM dendrimer of generation 1 to 7 (e.g., generation 1, 2, 3, 4, 5, 6, or 7), whose end groups are optionally selected from OOH, NH2, and acetamide.

6. The composition of claim 1, wherein the dendrimer is a generation 1-7 PAMAM dendrimer (e.g., generation 1, 2, 3, 4, 5, 6, or 7) modified with a sugar moiety, wherein the sugar group is selected from the group consisting of glucose, galactose, mannose, and fructose.

7. The composition of claim 1, wherein the dendritic macromolecule is a dendrimer, optionally a hyperbranched polymer.

8. The composition of claim 1, wherein the dendrimer is a PAMAM dendrimer of generation 1 to 7 (eg, a hydroxyl-terminated PAMAM dendrimer), preferably having greater than 50% of the functional groups in the form of hydroxyl (OH) groups.

9. The composition of claim 1, wherein the dendrimer is a glucose dendrimer, preferably a 1 , 2 or 3 generation glucose dendrimer, preferably made of glucose and optionally ethylene glycol building blocks, with >10 surface glucose moieties.

10. The composition of claim 1, wherein the dendrimer is a galactose dendrimer consisting of galactose and optionally ethylene glycol building blocks, having >10 surface sugar moieties.

11. The composition of claim 1, wherein the dendrimer is a sugar dendrimer made of sugar and optionally ethylene glycol building blocks, having >10 surface sugar moieties, optionally wherein the sugar building block is mannose or galactose.

12. A composition according to any one of claims 1-11, wherein the cannabinoid is cannabidiol (CBD) or a CBD analog.

13. The composition of any one of claims 1-11, wherein the cannabinoid is tetrahydrocannabinol (Δ9-THC) or an analog thereof.

14. A composition according to any one of claims 1 to 11, wherein the cannabinoid is a CB1 agonist, preferably selected from the group consisting of anandamide (ACPA) and formylarachidonic acid acetamide.

15. A composition according to any one of claims 1 to 11, wherein the cannabinoid is a CB1 antagonist, preferably selected from the group consisting of rimonabant, AM251 and AM281.

16. The composition according to any one of claims 1 to 11, wherein the cannabinoid is a CB2 agonist, preferably wherein the CB2 agonist is selected from the group consisting of HU-308, JWH-133, AM1241, GW405833 and GW842166X.

17. A composition according to any one of claims 1-11, wherein the cannabinoid is a CB1 and / or CB2 agonist, wherein the cannabinoid is selected from the group consisting of anandamide, Δ9-THC and 2-arachidonoylglycerol.

18. A composition according to any one of claims 1 to 11, wherein the cannabinoid is a TRPV1 agonist, preferably CBD.

19. The composition according to any one of claims 1 to 11, wherein the drug is a CB1 antagonist or a CB2 agonist, preferably tetrahydrocannabinol (THCV) and THCV analogs.

20. The composition of any one of claims 1-11, wherein the cannabinoid is a CB1 antagonist or a CB2 antagonist.

21. The composition of any one of claims 1-11, wherein the drug solubility of the dendrimer-cannabinoid composition is increased at least 10-fold compared to the free cannabinoid.

22. A composition according to any one of claims 1 to 21 wherein the dendrimer-cannabinoid conjugate further comprises a pharmaceutically acceptable excipient and optionally a non-cannabinoid active agent.

23. A composition according to any one of claims 1 to 22, wherein the conjugate is confined to the peripheral circulation by using higher order dendrimers, preferably 4th, 5th or 6th generation PAMAM dendrimers, 2nd, 3rd or higher order glucose dendrimers, or functionalization with PEG.

24. A composition according to any one of claims 1 to 23, wherein at least one natural cannabis derivative or synthetic cannabinoid compound is conjugated to the dendrimer via a spacer comprising a hydrocarbon (e.g. olefin), a diethylene glycol moiety, an oligoethylene glycol chain, a triazole moiety or a combination thereof, preferably wherein the spacer comprises a triazole moiety.

25. A pharmaceutical composition comprising the composition of any one of claims 1 to 24, and a pharmaceutically acceptable carrier or excipient.

26. The pharmaceutical composition according to claim 25, which is in a form selected from hydrogels, nanoparticles, microparticles, suspensions, powders, tablets, capsules, creams, solutions or combinations thereof.

27. A method of administering a dendrimer-cannabinoid conjugate according to any one of claims 1 to 24 to an individual in need thereof, preferably by a route selected from the group consisting of intranasal, intravenous, oral, subcutaneous, inhalation, intraperitoneal, topical and intrathecal.

28. The method of claim 27, wherein the individual suffers from an ocular disease, preferably wherein the dendrimer-cannabinoid conjugate is administered via intravitreal, suprachoroidal, subconjunctival, topical or systemic administration.

29. The method of claim 27, comprising treating a subject suffering from peripheral nerve pain, such as peripheral neuropathy or chemotherapy-induced pain.

30. The method of claim 27, comprising treating an individual in need of treatment for a neurological disorder including epilepsy and seizures, wherein the cannabinoid is cannabidiol (CBD) or a CBD analog.

31. The method of claim 27, comprising treating obesity-related metabolic disorders, psychiatric disorders, liver fibrosis or nicotine addiction in an individual in need thereof with a CB1 antagonist, preferably selected from the group consisting of rimonabant, AM251 and AM281.

32. The method of claim 27, comprising treating a symptom of arthritis, diabetic nephropathy, interstitial renal disease, fibrosis, multiple sclerosis, irritable bowel syndrome, autoimmune disease, traumatic brain injury, acute pain, chronic inflammatory pain, neuropathic pain, eye pain, or an inflammatory disease in a subject in need thereof, wherein the cannabinoid is a CB2 agonist, preferably wherein the CB2 agonist is selected from the group consisting of HU-308, JWH-133, AM1241, GW405833, and GW842166X.

33. The method of claim 27, comprising treating a symptom of neuropathic pain, cancer-related pain, Alzheimer's disease, Parkinson's disease, HIV-related neurological disease in a subject in need thereof, wherein the conjugate is a dendrimer-CB1 / CB2 agonist conjugate, preferably wherein the cannabinoid is selected from the group consisting of anandamide, Δ9-THC and 2-arachidonoylglycerol.

34. The method of claim 27, comprising treating an individual in need of treatment for glaucoma by reducing intraocular pressure or neuroprotection, wherein the cannabinoid is selected from the group consisting of phytocannabinoids, synthetic cannabinoids, endocannabinoids, and combinations thereof.

35. A method of treating symptoms of depression, addiction, renal disease (e.g., AKI), diabetic neuropathy, liver fibrosis, hepatic encephalopathy, or epilepsy in an individual in need thereof comprising administering a dendrimer-CB1 antagonist / CB2 agonist conjugate, preferably tetrahydrocannabinol (THCV) and THCV analogs.

36. The method of any one of claims 27-35, comprising targeting the composition to neurons or neurons in a diseased area by conjugating the CB1 agonist or antagonist to a glucose dendrimer that targets damaged neurons.

37. A method according to any one of claims 27 to 35 comprising administering the cannabinoid via higher generation and / or surface functionalized dendrimers to target peripheral neurons without significant uptake into the brain.

38. A method of targeting a therapeutic, prophylactic or diagnostic agent to neurons in an area affected by a disease or condition in a subject, the method comprising administering the agent together with a CB1 agonist or antagonist conjugated to a dendrimer, preferably a hydroxyl-terminated dendrimer, a sugar-terminated dendrimer and / or a sugar-based dendrimer.

39. The method of claim 38, wherein the dendrimer is a PAMAM dendrimer.

40. A method of targeting a therapeutic, prophylactic or diagnostic agent to neurons in an area affected by a disease or condition in a subject comprising administering the agent to reactive microglia / macrophages for intracellular delivery of the agent, wherein the agent is administered with a conjugate of a CB2 agonist or antagonist and a hydroxyl-terminated PAMAM or glucose dendrimer.

41. The method of any one of claims 38-40, wherein the agent is selected from the group consisting of a non-opioid, a non-cannabinoid, or a cannabinoid, conjugated or unconjugated to the dendrimer.

42. The method of any one of claims 38-41, administered in conjunction with radiation therapy, exercise or other adjunctive treatments.

43. The method of any one of claims 38-41, wherein the agent is an imaging agent or a therapeutic agent of PET, MRI, or radiation.

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