Cannabinoid dendrimer compositions for targeted delivery

Abstract

Dendrimer formulations containing one or more cannabinoids and / or cannabinoid derivatives and methods of use thereof are used, inter alia, for targeted delivery to neurons and activated microglia, or other cells that express CB1 or CB2 receptors on their surface. The cannabinoid-dendrimer conjugates can be administered to patients in need of treatment for a variety of conditions, including central and peripheral neuropathies, chronic pain, neuropathic pain, neurodegeneration (aging, dementia, Alzheimer's), traumatic brain injury, stroke, Huntington's disease and multiple sclerosis, epilepsy, inflammatory disorders, and pain.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention is generally in the field of cannabinoid drug formulations, specifically dendrimer-cannabinoid conjugates, for the selective delivery of the cannabinoid family to receptors in the CNS and other sites of disease.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 401,464, filed August 26, 2022, entitled "CANNABINOID DENDRIMER COMPOSITIONS FOR TARGETED DELIVERY," by Johns Hopkins University, listed inventors Kannan Rangaramanujam, Sujatha Kannan, Kunal Parikh, and Anjali Sharma, which is incorporated herein by reference in its entirety.

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

[0004] Background of the Invention Cannabinoids offer benefits for a range of clinical conditions, including pain, inflammation, epilepsy, sleep disorders, symptoms of multiple sclerosis, anorexia, schizophrenia, and other conditions. Therefore, rapid progress is being made in converting cannabinoids derived from herbal preparations into highly regulated prescription drugs. Drugs such as cannabinoids, which are metabolized by liver and gastrointestinal enzymes (first-pass hepatic metabolism), have unique pharmacokinetic requirements, have demonstrated poor gastrointestinal permeability, and cause irritation, thus necessitating alternatives to systemic oral delivery. The low oral bioavailability and highly lipophilic nature of cannabinoids make them difficult to deliver to selected targets. While oils and capsules currently allow for more convenient and precise administration than juices or teas derived from plant sources, they are not fully suited for pharmacological use.

[0005] Cannabinoids are highly lipophilic molecules (log P 6-7) with very low aqueous solubility (2-10 μg / mL), making them susceptible to degradation by light and temperature, as well as autooxidation, especially in solution. Formulation plays an important role in increasing drug solubility and physicochemical stability. Strategies commonly used in marketed products include salt formation (i.e., pH adjustment), cosolvency (e.g., ethanol, propylene glycol, PEG 400, 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 for transdermal, intranasal, and transmucosal administration have been proposed. However, absorption is slow, erratic, and variable. Maximum plasma concentrations are usually reached after 60-120 minutes, but this can be longer (up to 6 hours) and can be delayed. Further metabolism produces psychoactive metabolites. Extensive first-pass hepatic metabolism further reduces the oral bioavailability of THC, but the duration of effectiveness varies from 8 to 20 hours.

[0006] The psychotropic effects of cannabis are primarily mediated by CB1, which is widely distributed throughout the brain, primarily in the frontal cortex, basal ganglia, and cerebellum. CB1 is also present in several tissues and organs, including adipose tissue, the gastrointestinal tract, spinal cord, adrenal and thyroid glands, liver, reproductive organs, and immune cells. Evidence for the presence of CB1 receptors on the surface of chondrocytes and bone cells, as well as on fibroblast-like synoviocytes, makes targeting CB1 particularly effective in rheumatic diseases. CB1 activation inhibits adenylate cyclase and reduces cAMP levels and protein kinase A (PKA) activity, leading to activation of A-type potassium channels and a decrease in cellular potassium levels.

[0007] CB2 is primarily expressed in immune cells but can also be found in various other cell types. CB2 is also present in some neural tissues, such as dorsal root ganglia and microglial cells. CB2 shares 44% amino acid similarity with CB1 and similarly inhibits adenylate cyclase and activated mitogen-activated protein kinase. CB2 activation can increase intracellular calcium levels via phospholipase C. Both CB1 and CB2 couple to G proteins, but the transduction pathways that activate them may differ, for example, in terms of their interaction with ion channels. A CB2 variant known as Q63R has been associated with celiac disease, immune thrombocytopenic purpura, and juvenile idiopathic arthritis. During inflammatory joint disease, endocannabinoid production and release are 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 by-products of arachidonic acid metabolism.

[0008] There are no effective treatments for preventing or reversing neuropathic pain, and current treatments are limited to symptomatic relief. Treatment of chronic pain remains an unmet clinical need, and adequate pain relief can be achieved through the use of drugs with adverse effects on the central nervous system. The quality of life of patients with neuropathic pain is often compromised by comorbid conditions such as sleep disorders, depression, and anxiety. Preclinical studies have shown that cannabinoid receptor agonists block pain and reduce inflammation in various acute and chronic pain models. Both CB1 and CB2 receptor agonists, whether used alone or in combination with CB2 activators thought to act on microglial cells, have demonstrated antinociceptive activity, thereby reducing neuroinflammatory mechanisms. The CB2 receptor is believed to be particularly important in central neuronal pain circuits, as agonist activity induces dopamine release in the midbrain, contributing to descending pain control.

[0009] THC, the primary psychoactive component of cannabis, acts primarily as a partial agonist at CB1 (Ki = 53 nM) and CB2 (Ki = 40 nM) receptors, with well-known effects on pain, appetite, digestion, mood, and processes mediated by the endocannabinoid system. Endocannabinoid release is mediated during inflammatory joint disease by the production of proinflammatory cytokines (interferon [IFN]-c, interleukins (IL-12, IL-15, IL-17, IL-18), chemokines), nitric oxide synthase (NOS)-2, cyclooxygenase-2 (COX-2), matrix metalloproteinases (MMPs), and various other by-products of arachidonic acid metabolism. Adverse psychoactive events can be induced by THC, depending on the dose and prior patient tolerance. In contrast, CBD, the major non-psychoactive phytocannabinoid component of C. sativa, has little affinity for these receptors (Ki of 1.5 μM and 0.37 μM for human CB1 and CB2, respectively), acting as a partial CB1 antagonist and a weak inverse CB2 agonist.

[0010] It is an object of the present invention to provide formulations with improved bioavailability, pharmacokinetics, and increased selectivity of delivery, and reduced side effects. Summary of the Invention [Means for solving the problem]

[0011] Summary of the Invention A cannabinoid-dendrimer conjugate has been developed. The composition can contain one or more endogenous cannabinoids or endocannabinoids. Endocannabinoids are lipid-signaling molecules produced within the body that activate cannabinoid receptors and mimic the activity of Δ9-tetrahydrocannabinol, the main psychoactive component of cannabis. Exemplary endocannabinoids and endocannabinoid analogs that can be included in the composition include, but are not limited to, N-arachidonoylethanolamide (anandamide), 2-arachidonoylglycerol (2-AG), 2-arachidonylglyceryl ether (noladin ether), N-arachidonoyldopamine (NADA), virodamine (OAE), lysophosphatidylinositol (LPI), 7,10,13,16-docosatetraenoylethanolamide, and homo-γ-linolenoylethanolamine. Phytocannabinoids or herbicides are structurally diverse classes of natural chemical compounds derived from plants.In some forms, the phytocannabinoids or phytocannabinoid derivatives used in the present compositions can be derived from various sources, including but not limited to, hemp, cannabis, Echinacea, Acmella oleracea, Helichrysum umbraculigerum, Radula marginata, kava, black truffle, Syzygium aromaticum (clove), Rosmarinus officinalis, basil, oregano, black pepper, lavender, true cinnamon, Malabathurum, Cananga odorata, Copaifera spp. and hops.Examples include THC, Δ9-tetrahydrocannabinol; Δ9-THCV, Δ9-tetrahydrocannabivarin; CBN, cannabinol; CBDV, cannabidivarin; CBG, cannabigerol; CBC and cannabichromene.

[0012] 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), cannabielsoin (CBE), cannabicyclol (CBL), cannabifuran (CBF), cannabitran (CBT), cannabinodivalin (CBV), cannabilipsol (CBR), isocannabinoids and / or derivatives thereof.

[0013] 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, and G3 glucose dendrimers, while preferred PAMAM dendrimers include G3, G4, G5, and G6 hydroxyl-terminated PAMAM dendrimers. The density and loading of the cannabinoid on the dendrimer, the means of attachment of the cannabinoid to the dendrimer, the size and chemical composition of the dendrimer, and the size and composition of the linker to the cannabinoid, if present, govern the rate, selectivity, and activity of the cannabinoid to the site of delivery where the cannabinoid binds to a receptor. In a preferred embodiment, the cannabinoid is attached to the dendrimer by a cleavable linker. In a most preferred embodiment, the cleavable linker is an ester. In some embodiments, the linker comprises a triazole moiety.

[0014] The route of administration, usually oral, intranasal, or applied to other mucosal surfaces, affects the rate and dosage of dendrimer conjugates to the brain. Dendrimers increase brain uptake, solubility, target engagement, and PK, helping to retain the drug in the correct compartment. In some embodiments, dendrimers are used to retain cannabinoids in the peripheral circulation rather than the central circulation.

[0015] Cannabinoid-dendrimer conjugates can be administered to patients in need of treatment for a variety of conditions, including neurodegeneration (aging, dementia, Alzheimer's), psychiatric disorders, acute, chronic and neuropathic pain, cancer-related pain, traumatic brain injury, stroke, Huntington's disease and multiple sclerosis, epilepsy, inflammatory disorders, and the control of nausea in chemotherapy. [Brief explanation of the drawings]

[0016] [Figure 1-1] Figures 1A and 1B provide a schematic representation of how various cannabinoid (CB) receptor signaling modalities can influence neuroregulation in health and disease through unique pathways. Figure 1A shows key enzymes, such as diacylglycerol lipase (dglα) and phospholipase d (PLD), which produce the endogenous ligands arachidonylethanolamine (AEA) and 2-arachidonylglycerol (2-AG), which activate the cannabinoid 1 receptor (CB1) in the central nervous system (CNS). The results can include modulation of cAMP accumulation, voltage-gated calcium channels (VGCCs), K+ channels, and adenylate cyclase activity, which inhibits neurotransmitter release, at presynaptic excitatory and inhibitory synapses. Figure 1B shows that after CB1 receptor activation by ligand binding, signaling through G proteins and / or β-arrestins can occur at the plasma membrane in endocytic pits or endosomes after receptor internalization. While G proteins normally bind to unphosphorylated receptors, β-arrestins bind to receptors phosphorylated by G protein receptor kinases. Figure 1C shows a schematic diagram of key receptors and cells in the brain where dendrimer-cannabinoid conjugates can exert selective effects. [Figure 1-2] Same as above.

[0017] [Figure 2-1]Figure 2A is a schematic diagram of a stepwise synthetic route for producing an exemplary dendrimer-cannabidiol conjugate with an enzyme-sensitive ester linking group via click chemistry. Figure 2B is a schematic diagram of a stepwise synthetic route for conjugating an exemplary cannabinoid, cannabidiol, to a dendrimer. [Figure 2-2] Same as above.

[0018] [Figure 3] 3A and 3B are schematic diagrams showing molecular structures in a stepwise synthetic route for producing an exemplary dendrimer-cannabidiol conjugate with a non-cleavable ether linking group via click chemistry (FIG. 3A), and for conjugating an exemplary cannabinoid, cannabidiol, to a dendrimer (FIG. 3B).

[0019] [Figure 4] 4A and 4B are schematic diagrams showing a stepwise synthetic route for generating an exemplary dendrimer-HU-308 conjugate having a phosphatase-cleavable phosphodiester linker via an amidation reaction (FIG. 4A), and a stepwise synthetic route for conjugating an exemplary cannabinoid, HU-308, to a dendrimer (FIG. 4B).

[0020] [Figure 5-1] Figure 5A is a schematic diagram of a stepwise synthetic route for producing an exemplary dendrimer-HU-308 conjugate with an ester linking group via a click reaction. Figure 5B is a schematic diagram of a stepwise 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 a PAMAM-G4-OH dendrimer. [Figure 5-2] Same as above.

[0021] [Figure 6]6A and 6B are schematic diagrams showing molecular structures for a stepwise synthetic route for generating an exemplary dendrimer-HU-308 conjugate with a glutathione-sensitive disulfide linking group via a Click reaction (FIG. 6A), and for conjugating an exemplary cannabinoid, HU-308, to a dendrimer to generate a D-HU-308 conjugate with a disulfide linker (FIG. 6B).

[0022] [Figure 7] Figure 7A is a schematic diagram of a stepwise synthetic route for producing an exemplary dendrimer-tetrahydrocannabinol (D-THC) conjugate with an esterase-sensitive ester linking group via a click reaction. Figure 7B is a schematic diagram of a stepwise synthetic route for conjugating the exemplary cannabinoid THC to a dendrimer to produce a D-THC conjugate.

[0023] [Figure 8] 8A and 8B are schematic diagrams showing molecular structures for a stepwise synthetic route for producing an exemplary dendrimer-THC conjugate with a triglycyl peptide linker suitable for lysosomal release via a click reaction (FIG. 8A), and for conjugating the exemplary cannabinoid THC to a dendrimer to produce a D-THC conjugate with a triglycyl peptide linker (FIG. 8B).

[0024] [Figure 9] Figure 9A is a schematic diagram of a stepwise synthetic route for producing an exemplary dendrimer-anandamide (D-AEA) conjugate via a click reaction. Figure 9B is a schematic diagram of a stepwise synthetic route for conjugating an exemplary cannabinoid, anandamide, to a dendrimer to produce a D-AEA conjugate.

[0025] [Figure 10]10A and 10B are schematic diagrams of a stepwise synthetic route for producing an exemplary dendrimer-AEA with a non-cleavable ether linking group (FIG. 10A), and a stepwise synthetic route for conjugating an exemplary cannabinoid, AEA, to a dendrimer to produce a D-AEA conjugate with a non-cleavable ether linker (FIG. 10B).

[0026] [Figure 11] Figure 11A is a schematic diagram of a stepwise synthetic route for producing an exemplary dendrimer-2-arachidonoylglycerol (D-2-AG) conjugate via a click reaction. Figure 11B is a schematic diagram of a stepwise synthetic route for conjugating an exemplary cannabinoid, D-2-AG, to a dendrimer to produce the D-2-AG conjugate.

[0027] [Figure 12] FIG. 12 is a table of the structures of Hu308 and cannabidiol, as well as their conjugable analogs.

[0028] [Figure 13-1] Figures 13A-13B show the synthesis and structures shown in Figure 12. Figure 13C shows the binding and release of the compounds. [Figure 13-2] Same as above. [Figure 13-3] Same as above. [Figure 13-4] Same as above. [Figure 13-5] Same as above.

[0029] [Figure 14] FIG. 14 is a graph of % binding efficiency versus log tryptamine (μM) in the 5-HT1A human serotonin GPCR cell-based agonist cAMP assay. DETAILED DESCRIPTION OF THE INVENTION

[0030] Detailed Description of the Invention I. Definition As used herein, "cannabinoid" refers to lipid-based compounds that have direct or indirect activity at cannabinoid receptors. "Endocannabinoids" are endocannabinoids that are naturally produced in the body and are found in humans and other animals. "Exogenous cannabinoids" include phytocannabinoids, which are cannabinoids derived from the cannabis plant, and related synthetic derivatives thereof.

[0031] The terms "active agent" or "biologically active agent" are used interchangeably to refer to a chemical or biological compound that induces a desired pharmacological and / or physiological effect, which may be prophylactic, therapeutic, or diagnostic. These may be nucleic acids, nucleic acid analogs, small molecules with a molecular weight of less than 2 kD, more typically less than 1 kD, peptidomimetics, proteins or peptides, carbohydrates or sugars, lipids, or combinations thereof. The term also encompasses pharmaceutically acceptable, pharmacologically active derivatives of drugs, including, but not limited to, salts, esters, amides, prodrugs, active metabolites, and analogs. 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 disorder. The term "diagnostic agent" generally refers to an agent that can be administered to locate, precisely target, and confirm the location of a pathological process. Diagnostic agents can label target cells, allowing for subsequent detection or imaging of these labeled target cells.

[0032] 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 specific targeting construct being administered, the size of the subject, or the severity of the disease or condition. One of ordinary skill in the art can empirically determine the effective amount of a particular compound without necessitating undue experimentation. In some embodiments, the term "effective amount" refers to the amount of a therapeutic or prophylactic agent that relieves or reduces one or more disease symptoms.

[0033] The terms "inhibit" or "reducing," in the context of inhibition, refer to a decrease or reduction in activity and amount. This can be complete inhibition or reduction, or partial inhibition or reduction, of activity or amount. Inhibition or reduction can be compared to a control or standard level. Inhibition can be 5, 10, 25, 50, 75, 80, 85, 90, 95, 99, or 100%. For example, a dendrimer composition comprising one or more inhibitors can inhibit or reduce the activity and / or amount of affected neurons by about 10%, 20%, 30%, 40%, 50%, 75%, 85%, 90%, 95%, or 99% from the activity and / or amount of the same cells in a comparable tissue of a subject not administered or treated with the dendrimer composition. In some embodiments, inhibition and reduction are compared at the level of mRNA, protein, cells, tissues, and organs. For example, inhibition and reduction of the rate of neuronal loss, the rate of brain weight loss, or the rate of hippocampal volume loss compared to untreated control subjects.

[0034] The terms "treating" or "preventing" refer to improving, reducing, or otherwise halting the development or progression of a disease, disorder, or condition in an animal that may be susceptible to, but has not yet been diagnosed with, a disease, disorder, and / or condition; inhibiting, e.g., slowing the progression of, a disease, disorder, or condition; and alleviating, e.g., causing regression of, a disease, disorder, and / or condition. Treating a disease or condition includes improving at least one symptom of a particular disease or condition even when the underlying pathophysiology is unaffected, such as treating a subject's pain by administering an analgesic drug even when such a drug does not treat the cause of the pain. Desired effects of treatment include slowing the rate of disease progression, improving or alleviating the disease state, and remission or improved prognosis. For example, an individual is successfully "treated" when one or more symptoms associated with depression are reduced or eliminated, including, but not limited to, reduced levels of anxiety, agitation, or restlessness; amelioration of feelings of sadness, fear, emptiness, or hopelessness; an increase in the quality of life of an individual affected by the disease; a reduction in the dosage of other medications required to treat the disease; and a slowing of the progression of the disease.

[0035] The phrases "pharmaceutically acceptable" or "biocompatible" refer to compositions, polymers, and other materials and / or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. The phrase "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable substance, composition, or vehicle, such as a liquid or solid filler, diluent, solvent, or encapsulating substance, that is involved in carrying or transporting a subject composition from one organ or body part to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the subject composition and not injurious to the patient.

[0036] The term "biodegradable" generally refers to a material that, under physiological conditions, breaks down or erodes into smaller units or species that can be metabolized in vivo, excreted, or excreted. The degradation time is a function of composition and geometry.

[0037] The term "dendrimer" includes, but is not limited to, a molecular architecture having an inner core, an inner layer or "generation" of repeating units regularly attached to this initial core, and an outer surface of terminal groups attached to the outermost generation.

[0038] The term "functionalize" means to modify a compound or molecule in a manner that results in the attachment of a functional group or moiety. For example, a molecule can be functionalized by introducing a molecule that makes it a strong nucleophile or a strong electrophile.

[0039] The term "targeting moiety" refers to a moiety that is localized to or distant from a specific location. The moiety can be, for example, a protein, a nucleic acid, a nucleic acid analog, a carbohydrate, or a small molecule. The location can be a tissue, a specific cell type, an intracellular compartment, or a molecule such as a receptor.

[0040] The term "prolonged residence time" refers to an increase in the time required for a drug to clear from a patient's body or from an organ or tissue of the patient. In certain embodiments, "prolonged residence time" refers to a drug that is cleared with a half-life that is 10%, 20%, 50%, or 75% longer than a comparative standard, such as a comparable drug that is not conjugated to a delivery vehicle, such as a dendrimer. In certain embodiments, "prolonged residence time" refers to a drug that is cleared with a half-life that is 2, 5, 10, 20, 50, 100, 200, 500, 1000, 2000, 5000, or 10,000 times longer than a comparative standard, such as a comparable drug that does not carry a dendrimer that specifically targets a particular cell type.

[0041] The terms "incorporated" and "encapsulated" refer to incorporating, formulating, or otherwise including an agent within and / or on the surface of a composition that allows for release, such as sustained release, of the agent in a desired application. Drugs or other substances can be incorporated into dendrimers by binding (by covalent, ionic, or other bonding interactions) to one or more surface functional groups of such dendrimers, by physical admixture, by enveloping the agent within the dendrimer structure, and / or by encapsulating the agent within the dendrimer structure.

[0042] As used herein, the central nervous system ("CNS") includes the brain and spinal cord. As used herein, the peripheral nervous system ("PNS") refers to nerves other than the brain and spinal cord.

[0043] "Hydroxyl-terminated" in reference to a dendrimer refers to a dendrimer having hydroxyl groups on its surface that are not attached to the termini of the dendrimer via a sugar moiety (such as a saccharide moiety).

[0044] "Sugar-terminated" in reference to a dendrimer refers to a dendrimer that contains sugar moieties (such as saccharide moieties) on its surface, but not within its core.

[0045] "Sugar-based" in reference to dendrimers refers to dendrimers that contain sugar moieties (such as saccharide moieties) in their core, or in their core and on their surface.

[0046] An "analog" in relation to a given compound refers to another compound that is structurally similar, functionally similar, or both to the specified compound. Structural similarity can be determined using any criterion known in the art, such as the Tanimoto coefficient, which provides a quantitative measure of similarity between two compounds based on their molecular descriptors. Preferably, molecular descriptors are 2D properties such as fingerprints, topological indices, and maximum common substructures, or 3D properties such as overall shape and molecular field. The Tanimoto coefficient ranges between 0 for a pair of dissimilar molecules and 1 for identical molecules (inclusive). A compound can be considered an analog of the specified compound if it has a Tanimoto coefficient between 0.5 and 1.0 (inclusive), preferably between 0.7 and 1.0 (inclusive), and most preferably between 0.85 and 1.0 (inclusive) relative to the specified compound. A compound is functionally similar to the specified compound if it induces the same pharmacological or physiological effect, or both, as the specified compound. "Analog" can also refer to modifications, including, but not limited to, hydrolysis, reduction, or oxidation products of a compound. Hydrolysis, reduction, and oxidation reactions are known in the art. II. Composition

[0047] 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 nervous system cells and / or glial cells, have been developed. Generally, the cannabinoids and / or cannabinoid derivatives bind to receptors on the surface of the target cells and / or to receptors inside the target cells. Exemplary target cells include, but are not limited to, brain cells such as microglia, astrocytes, and / or neurons, e.g., cells within the site of pathology in the brain or CNS, and / or cells in the peripheral nervous system, such as peripheral neurons and glia. The microglia and / or astrocytes to which the cannabinoids and / or cannabinoid derivatives are delivered may be activated or inactivated microglia and / or astrocytes.

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

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

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

[0051] Typically, dendrimers have a diameter between about 1 nm and about 60 nm, more preferably between about 1 nm and about 50 nm, about 1 nm and about 40 nm, about 1 nm and about 30 nm, about 1 nm and about 20 nm, about 1 nm and about 10 nm, or about 1 nm and about 5 nm. In some embodiments, the diameter is between about 1 nm and about 2 nm. The preferred size of dendrimers for crossing the blood-brain barrier ("BBB") is less than 5 nm, while those that do not cross the BBB and remain in the peripheral circulation are greater than 5 nm. In some embodiments, the dendrimers have a diameter effective to penetrate the BBB and remain in proximity to or within target nervous system cells and / or glial cells for delivery of agents conjugated thereto. In some embodiments, the dendrimer has a diameter effective to penetrate the BBB and internalize into target nervous system cells and / or glial cells, such as, for example, neurons, oligodendrocytes, astrocytes, microglial cells, and neuroglial supporting cells, for delivery of an agent conjugated thereto. In some embodiments, the dendrimer has a diameter effective to penetrate a barrier interface, such as the blood-nerve barrier ("BNB"), and internalize into nervous system cells and glial cells of the peripheral nervous system, such as, for example, neurons, Schwann cells, satellite cells, and neuroglial supporting cells, for delivery of an agent conjugated thereto. In some embodiments, the dendrimer has a diameter effective to remain in the peripheral circulation for delivery of an agent conjugated thereto to a target cell in the peripheral nervous system.

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

[0053] In some embodiments, the dendrimer has a hypercore (e.g., dipentaerythritol) and one or more monosaccharide branching units. In some embodiments, the monosaccharide branching units are conjugated to the core or previous layer of monomer via a linker, such as a polyethylene glycol chain. In a preferred embodiment, the hypercore is dipentaerythritol, and the monosaccharide branching units are glucose-based branching units, such as those shown in Structures II-IV. In the most preferred embodiment, the dendrimer is made entirely from glucose building blocks. While sugar-modified PAMAM dendrimers are also functional, dendrimers made from sugars, particularly glucose, are most preferred. Particularly preferred glucose dendrimers are G1-G3 glucose dendrimers, such as G1, G2, and / or G3 glucose dendrimers.

[0054] Dendrimer scaffolds suitable for use in the conjugates include, but are not limited to, poly(amidoamine), also known as PAMAM, or STARBURST™ dendrimers; polypropylamine (POPAM), polyethyleneimine, polylysine, polyesters, iptycene, aliphatic poly(ether), aromatic polyether dendrimers, sugar (e.g., glucose, galactose, mannose, fructose, etc.) dendrimers and copolymers thereof, such as copolymers of sugars and alkylene glycols (e.g., dendrimers formed with glucose and ethylene glycol building blocks). Dendrimers can have multiple surface functional groups, such as carboxylic acid, amine, hydroxyl, and / or acetamide. The terms "surface functional group" and "end group" are used interchangeably herein. In some embodiments, dendrimers have surface hydroxyl groups. In some embodiments, one or more of these surface functional groups are further modified with other molecules, such as sugars (e.g., glucose, galactose, mannose, fructose, etc.) and / or polyalkylene glycols, e.g., polyethylene glycol, thus having sugar molecules and / or polyalkylene glycols as terminal moieties / molecules. Preferred PAMAM dendrimers include hydroxyl-terminated PAMAM dendrimers, particularly G3-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, first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, or tenth generation. In some embodiments, dendrimers are used as platforms, and are PAMAM dendrimers modified with functional groups to increase the number of surface hydroxyl groups.

[0055] In some embodiments, dendrimer-active agent conjugates can be restricted to the peripheral circulation by using higher generation dendrimers (such as fourth, fifth, or sixth generation PAMAM dendrimers, second, third, or higher generation glucose-based dendrimers), specifically targeting particular tissue regions and / or cell types, such as peripheral nervous system cells and macrophage cells. Additionally or alternatively, dendrimer-active agent conjugates can be restricted to the peripheral circulation by appropriate functionalization of the dendrimer (such as PEGylation).

[0056] In some embodiments, dendrimers may be used to specifically target specific tissue regions and / or cell types in the central nervous system (CNS), peripheral nervous system (PNS), and / or eye, such as neurons and glia in the CNS and / or PNS, by using certain generations of dendrimers, such as second generation (G2), G3, G4, and G5 PAMAM dendrimers and / or glucose dendrimers. In the most preferred embodiment, the dendrimers are made entirely from glucose building blocks. Although sugar-modified PAMAM dendrimers can also function, dendrimers made from sugars, especially glucose, are most preferred. Monosaccharide-based dendrimers

[0057] In some embodiments, the branching unit comprises a monosaccharide. In some embodiments, the monosaccharide branching unit is conjugated to the core or to a previous layer of monomers via a linker such as a polyethylene glycol chain. In some embodiments, the monosaccharide branching unit is a glucose-based branching unit. In some embodiments, the branching unit can include a PEG and / or alkyl chain linker between different dendrimer generations. For example, glucose layers are connected by a PEG linker and a triazole ring. In some embodiments, the branching unit is the same for each generation of dendrimers generated from the core. Thus, for example, the branching unit is a glucose-based branching unit for generating a first-generation dendrimer, a second-generation dendrimer, and a third-generation dendrimer.

[0058] In some embodiments, the dendrimer has a hypercore (e.g., dipentaerythritol) and one or more monosaccharide branching units. In some embodiments, the hypercore is dipentaerythritol and the monosaccharide branching units are glucose-based branching units. In further embodiments, the spacer molecule is alkyl(CH2). n - It can also be a hydrocarbon-like unit.

[0059] In some embodiments, dendrimers synthesized using glucose building blocks and having surfaces made primarily of glucose moieties enable specific targeting of cells, including injured neurons, ganglion cells, and other nervous system cells, in the brain, eye, and / or peripheral nervous system. In some embodiments, glucose-based dendrimers selectively target or are concentrated within target nervous system cells and / or glial cells. In some embodiments, glucose-based dendrimers selectively target or are concentrated on the surface of target nervous system cells and / or glial cells. In some embodiments, glucose-based dendrimers selectively target or are concentrated within target nervous system cells and on the surface of target nervous system cells and / or glial cells. In some embodiments, glucose-based dendrimers selectively target or are concentrated within damaged, diseased, and / or overactive neurons and / or glial cells.

[0060] In some cases, the dendrimer comprises a number of sugar molecules and terminal groups, such as glucose and / or hydroxyl groups, effective for targeting one or more neurons and / or glia in the CNS, PNS, and / or eye. The terminal hydroxyl groups of these dendrimers may be terminal glucose molecules, or extra hydroxyl groups that are not part of glucose molecules, or a combination thereof. In some embodiments, all of the terminal hydroxyl groups are part of terminal glucose molecules. In some embodiments, the number of sugar molecules on the end of the dendrimer is determined by the generation number.

[0061] In some embodiments, the dendrimers are made from glucose and oligoethylene glycol building blocks. Exemplary glucose dendrimers are shown in structures V and VII.

[0062] Some exemplary glucose dendrimers include a first-generation glucose dendrimer with 24 hydroxyl (-OH) end groups, a second-generation glucose dendrimer with 96 hydroxyl (-OH) end groups, a third-generation glucose dendrimer with 396 hydroxyl (-OH) end groups, and a fourth-generation glucose dendrimer with 1584 hydroxyl (-OH) end groups. For example, the glucose dendrimer is a second-generation glucose-based dendrimer with 24 glucose molecules on the periphery and 6 embedded glucose molecules in the backbone held together by PEG segments.

[0063] Dendrimer compositions, termed "glucose dendrimers," have been developed that can selectively accumulate within neurons, particularly within the nuclei of damaged and / or overactive neurons. These dendrimers can also accumulate at high levels within activated microglia. However, compared to hydroxyl dendrimers, which primarily accumulate within microglia, these dendrimers primarily target neurons. Glucose dendrimers are described in U.S. Patent Application No. 63 / 327,610, "Dendrimer Compositions for Targeted Delivery of Therapeutics to Neurons," filed April 5, 2022, by Johns Hopkins University, inventors Kannan Rangaramanujam, Rishi Sharma, Anjali Sharma, Sujatha Kannan, Nirnath Sah, Mira Sachdeva, and Siva P. Kambhampati.

[0064] Glucose dendrimers comprise (a) a central core, (b) one or more branching units based on the monosaccharide glucose, optionally conjugated with linkers, and, optionally, (c) one or more therapeutic, prophylactic, and / or diagnostic agents. Generally, the one or more branching units are conjugated to the central core, and the surface groups of the dendrimer are monosaccharide glucose molecules. In some embodiments, the central core is dipentaerythritol or a hexa-propargylated derivative thereof. In some embodiments, the branching units are conjugated to the central core via a linker, such as a hydrocarbon or oligoethylene glycol chain. In a preferred embodiment, the branching unit is β-D-glucopyranoside tetraethylene glycol azide, having the following structure: [ka] or a peracetylated derivative thereof.

[0065] In some embodiments, the glucose dendrimer is a first, second, third, fourth, fifth or sixth generation dendrimer. In one embodiment, the dendrimer has the following structure: [ka] It is a first generation dendrimer having the following structure:

[0066] In a preferred embodiment, the dendrimer has the following structure: [ka] It is a second generation dendrimer having the following structure:

[0067] In some embodiments, one or more therapeutic, prophylactic, and / or diagnostic agents are encapsulated within, associated with, and / or conjugated to the dendrimer at a concentration of between about 0.01% and about 30% by weight, preferably between about 1% and about 20% by weight, and more preferably between about 5% and about 20% by weight. 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 a fluorescent dye, a near-infrared dye, a SPECT imaging agent, a PET imaging agent, and a radioisotope.

[0068] In some embodiments, the dendrimer and therapeutic, prophylactic, or diagnostic agent are conjugated by one or more linkers or coupling agents, such as one or more hydrocarbon or oligoethylene glycol chains. Exemplary linking groups are disulfide, ester, ether, thioester, and amide linking groups. PAMAM Dendrimer

[0069] The term "PAMAM dendrimer" refers to a poly(amidoamine) dendrimer of any generation, including, but not limited to, a first-generation PAMAM dendrimer, a second-generation PAMAM dendrimer, a third-generation PAMAM dendrimer, a fourth-generation PAMAM dendrimer, a fifth-generation PAMAM dendrimer, a sixth-generation PAMAM dendrimer, a seventh-generation PAMAM dendrimer, an eighth-generation PAMAM dendrimer, a ninth-generation PAMAM dendrimer, or a tenth-generation PAMAM dendrimer, which may contain different cores having amidoamine building blocks, and which may have carboxylic acid, amine, acetamide, and / or hydroxyl termini. In some embodiments, the dendrimer is a fourth-, fifth-, or sixth-generation ("G") dendrimer. In some embodiments, the PAMAM dendrimer has hydroxyl termini.

[0070] In general, the complete architecture of a dendrimer can be distinguished into an interior core moiety followed by radially linked branching units (i.e., generations) that are further modified with chemical functional groups bearing desired terminal groups on the exterior surface of the dendrimer.

[0071] In some embodiments, the dendrimers are in the form of nanoparticles, as described in detail in U.S. Published Application Nos. US2011 / 0034422, US2012 / 0003155, and US2013 / 0136697. For example, the molecular weight of the dendrimer can be varied to prepare polymer nanoparticles that form particles with properties, such as drug release rate, optimized for specific applications. In general, conjugation to dendrimers can further improve the safety and efficacy of these drugs. For example, dendrimer conjugates can alter specific receptor activity and / or modify biodistribution. For example, the use of higher generation dendrimers and / or dendrimers with molecular weights greater than 24 kDa can restrict these drugs to the peripheral nervous system to prevent their psychoactive effects.

[0072] In some embodiments, different variants of dendrimers, including but not limited to dendrons and tectodendrimers, can be used as delivery vehicles to conjugate and deliver one or more active agents. Dendrons are dendritic wedges that contain one type of functional group at the core (functional group, f=1) and another functional group at the periphery (f=8, 16, 32, etc...). Tectodendrimers generally consist of a central dendrimer to which multiple dendrimers are attached at its periphery. 1. Core

[0073] In some embodiments, dendrimers are prepared using methods in which the dendrimer is assembled from a multifunctional core that is extended outward by a series of reactions, allowing for the stepwise addition of branching units (i.e., generations) around the core.

[0074] Exemplary chemical structures suitable as core moieties include dipentaerythritol, pentaerythritol, 2-(aminomethyl)-2-(hydroxymethyl)propane-1,3-diol, 2-ethyl-2-(hydroxymethyl)propane-1,3-diol, 3,3',3",3"'-silanetetrayltetrakis(propane-1-thiol), 3,3-divinylpenta-1,4-diene, 3,3',3"-nitrilotripropionic acid, 3,3',3"-nitrilotris(N-(2-aminoethyl)propanamide), 3,3',3"-nitrilotripropionic acid, ... ",3'''-(ethane-1,2-diylbis(azanetriyl))tetrapropanamide, 3-(carboxymethyl)-3-hydroxypentanedioic acid, 2,2'-((2,2-bis((2-hydroxyethoxy)methyl)propane-1,3-diyl)bis(oxy))bis(ethan-1-ol), tetrakis(3-(trichlorosilyl)propyl)silane, 1-thioglycerol, 2,2,4,4,6,6-hexachloro-1,3,5,2l5,4l5,6l5-triazatriphosphinine, 3-(hydroxymethyl)-5,5 -Dimethylhexane-2,4-diol, 4,4',4"-(ethane-1,1,1-triyl)triphenol, 2,4,6-trichloro-1,3,5-triazine, 5-(hydroxymethyl)benzene-1,2,3-triol, 5-(hydroxymethyl)benzene-1,3-diol, 1,3,5-tris(dimethyl(vinyl)silyl)benzene, carbosiloxane core, nitrilotrimethanol, ethylenediamine, propane-1,3-diamine, butane-1,4-diamine, 2,2',2"-nitrilotris(ethane-1- Examples of suitable core moieties include cyclodextrins, alpha cyclodextrins, beta cyclodextrins, gamma cyclodextrins, cucurbituril, benzene-1,2,3,4,5,6-hexathiol, monosaccharides, disaccharides, trisaccharides, oligosaccharides, or azide- or alkyne-modified portions thereof. In some embodiments, the core moiety is chitosan. Thus, azide- or alkyne-modified chitosans are suitable for conjugation to branching units using click chemistry.

[0075] In some embodiments, the core moiety is ethylenediamine or tetra(ethylene oxide). In some embodiments, the core moiety is dipentaerythritol. Exemplary chemical structures suitable for use as the core moiety are shown in Table 1 below. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] 2. Branching Unit

[0076] Exemplary chemical structures suitable as branching units include monosaccharides. In some embodiments, the monosaccharide branching units are conjugated to the core or previous layer of monomers via a linker, such as a polyethylene glycol chain. In a preferred embodiment, the monosaccharide branching units are glucose-based branching units. Exemplary glucose-based branching units are shown in Structures II-IV. These are spacer molecules and, therefore, can also be alkyl(CH)n-hydrocarbon-like units.

[0077] The branching units are PEG or alkyl chain linkers between different dendrimer generations, e.g., glucose layers are attached via PEG linkers and triazole rings.

[0078] In a preferred embodiment, the branching units are the same for each generation of dendrimer produced from the core. Thus, in one embodiment, the branching units are glucose-based branching units to produce the first generation dendrimers shown in structures V-VII.

[0079] In some embodiments, the branching unit is a hypermonomer, i.e., AB n These are building blocks. Exemplary hypermonomers include the AB4, AB5, AB6, AB7, and AB8 building blocks. The hypermonomer strategy dramatically increases the number of available end groups. An exemplary AB4 hypermonomer is peracetylated β-D-glucopyranoside tetraethylene glycol azide, shown in Structure III.

[0080] The chemical structures listed in Table 1 are also suitable as building blocks for forming branching units of dendrimers. For example, branching units of dendrimers include dipentaerythritol, pentaerythritol, 2-(aminomethyl)-2-(hydroxymethyl)propane-1,3-diol, 2-ethyl-2-(hydroxymethyl)propane-1,3-diol, 3,3',3",3'"-silanetetrayltetrakis(propane-1-thiol), 3,3-divinylpenta-1,4-diene, 3,3',3"-nitrilotripropionic acid, 3,3',3"-nitrilotris(N-(2-aminoethyl)propane)), 3,3',3"-nitrilotriisopropyl ether, ... )propanamide), 3,3',3",3'''-(ethane-1,2-diylbis(azanetriyl))tetrapropanamide, 3-(carboxymethyl)-3-hydroxypentanedioic acid, 2,2'-((2,2-bis((2-hydroxyethoxy)methyl)propane-1,3-diyl)bis(oxy))bis(ethan-1-ol), tetrakis(3-(trichlorosilyl)propyl)silane, 1-thioglycerol, 2,2,4,4,6,6-hexachloro-1,3,5,2l5, 4l5,6l5-triazatriphosphinine, 3-(hydroxymethyl)-5,5-dimethylhexane-2,4-diol, 4,4',4"-(ethane-1,1,1-triyl)triphenol, 2,4,6-trichloro-1,3,5-triazine, 5-(hydroxymethyl)benzene-1,2,3-triol, 5-(hydroxymethyl)benzene-1,3-diol, 1,3,5-tris(dimethyl(vinyl)silyl)benzene, carbosiloxane core, nitrilotrimethanol, Formed by ethylenediamine, propane-1,3-diamine, butane-1,4-diamine, 2,2',2"-nitrilotris(ethan-1-ol), alpha cyclodextrin, beta cyclodextrin, gamma cyclodextrin, cucurbituril, benzene-1,2,3,4,5,6-hexathiol, monosaccharides, disaccharides, trisaccharides, oligosaccharides, or their azide-modified moieties, alkyne-modified moieties, or combinations thereof.

[0081] Other examples of chemical structures suitable for forming the branching units of the 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 additional, same or different, branching units using click chemistry. In some embodiments, the branching unit is methyl acrylate or ethylenediamine, or a combination thereof. In some embodiments, the branching unit is linear or branched polyethyleneglycerol. In some embodiments, the branching unit is a copolymer of an alkylene glycol (such as ethylene glycol) and a sugar moiety such as glucose, galactose, mannose, and / or fructose. 3.Surface functional groups

[0082] The surface functional groups / molecules of the dendrimer include, but are not limited to, primary amine end groups, hydroxyl end groups, carboxylic acid end groups, acetamide end groups, sugar molecules, oligo- or poly-alkylene glycols, and / or thiol end groups. In some embodiments, the desired terminal functional groups can be added by one of the conjugation methods to the core and branching units.

[0083] In some embodiments, the surface functional groups are hydroxyl groups, such as the hydroxyl groups of the PAMAM dendrimer of the second generation PEG dendrimer shown in Structure I, or the terminal glucose hydroxyl groups of dendrimers prepared with glucose-based branching units shown in Structures V and VII. In some embodiments, the desired surface functional groups can be modified or added by one of the conjugation methods to the core and branching units. Exemplary surface functional groups include hydroxyl, amine, carboxylic acid, acetamide, and thiol end groups, and combinations thereof.

[0084] In some embodiments, dendrimers may specifically target particular tissue regions and / or cell types, such as cells and tissues of the central nervous system (CNS), peripheral nervous system (PNS), and / or eye. In some embodiments, dendrimers specifically target neurons and / or glia of the CNS. In some embodiments, dendrimers specifically target neurons and / or glia of the PNS. In some embodiments, glucose dendrimers are of first generation (G1), G2, G3, G4, and G5.

[0085] In some embodiments, the dendrimer comprises a number of terminal glucose and / or hydroxyl groups effective to target one or more neurons and / or glia of the CNS, PNS and / or eye.

[0086] Glucose dendrimers are preferred. In some embodiments, dendrimers are made from glucose and oligoethylene glycol building blocks. Exemplary first-generation glucose dendrimers are shown in structure VI, and second-generation glucose dendrimers are shown in structure VIII.

[0087] In some embodiments, dendrimers have a plurality of surface functional groups (also referred to herein as surface or peripheral functional groups), such as hydroxyl (-OH), amine, acetamide, and / or carboxyl groups, at the periphery of the dendrimer. In some embodiments, the surface density of such peripheral functional groups is at least 1 group / nm 2 (number of surface functional groups / surface area (nm 2 For example, in some embodiments, the surface density of surface functional groups, such as hydroxyl groups, is at least 10, 15, 20, 25, 30, 35, 40, 45, 50, or more than 50 OH groups / nm 2 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more OH groups / nm 2In some embodiments, the volume density of surface functional groups, such as hydroxyl groups, is from about 1 to about 50 groups / nm 3 Between about 5 and about 30 groups / nm 3 Between about 10 and about 20 groups / nm 3 In a further embodiment, the surface density of surface functional groups, such as hydroxyl groups, is between about 1 and about 50 groups / nm 2 Preferably, between 5 and 20 groups / nm 2 (number of surface functional groups / surface area (nm 2 )), while each surface functional group moiety has a molecular weight between about 100 Da and about 10 kDa, preferably between about 100 Da and 1000 Da.

[0088] In some embodiments, the amount of surface functional groups on the dendrimer, such as any one of the above, e.g., hydroxyl groups, is at least 40%, at least 50%, greater than 40%, greater than 50%, or in the range of greater than 40% to 100%.

[0089] In some embodiments, one or more of the surface functional groups, such as any one of those listed above, on the outer periphery of the dendrimer are further modified by conjugation with one or more carbohydrate molecules and / or one or more polyalkylene glycols, such as polyethylene glycol. In these embodiments, the surface density of the terminal carbohydrate moieties / molecules and / or polyalkylene glycols can be in any of the ranges described above for hydroxyl groups. Hydroxyl-terminated PAMAM dendrimers, PAMAM dendrimers surface-modified with sugar moieties (having >10% of surface groups modified with sugars, particularly glucose), and glucose dendrimers (where the dendrimer is made from glucose building blocks) are preferred. For delivery to the brain, constructs with a total molecular weight of <30,000 Da are preferred. For primarily peripheral circulation restriction, constructs with a total molecular weight of >50,000 Da are preferred. When dendrimers are formed from or include terminal sugar moieties / sugar molecules, such as glucose, the terminal hydroxyl groups of these dendrimers may be part of the terminal sugar moiety / sugar molecule, or extra hydroxyl groups that are not part of the sugar moiety / sugar molecule, or a combination thereof. In some embodiments, all of the terminal hydroxyl groups are part of the terminal sugar moiety / sugar molecule. a. Hydroxyl-terminated dendrimers

[0090] In some embodiments, the dendrimer contains multiple hydroxyl groups. Some exemplary densely hydroxyl-containing dendrimers include commercially available polyester dendritic polymers, such as hyperbranched 2,2-bis(hydroxyl-methyl)propionic acid polyester polymers (e.g., hyperbranched bis-MPA polyester-64-hydroxyl, 4th generation), and dendritic polyglycerols. In some embodiments, the hydroxyl-terminated dendrimers include hydroxyl-terminated PAMAM dendrimers, particularly G3-G6 hydroxyl-terminated PAMAM dendrimers, such as G3, G4, G5, and G6 hydroxyl-terminated PAMAM dendrimers.

[0091] In some embodiments, the dense hydroxyl-containing dendrimer is an oligoethylene glycol (OEG)-like dendrimer. For example, the second-generation OEG dendrimer (D2-OH-60) shown in Structure I can be synthesized using highly efficient, robust, and atom-economical chemical reactions, such as Cu(I)-catalyzed alkyne-azide click and photocatalytic thiol-ene click chemistry. Highly dense polyol dendrimers of very low generation with minimal reaction steps can be achieved by using an orthogonal hypermonomer and hypercore strategy, as described, for example, in WO2019094952. In some embodiments, the dendrimer backbone has non-cleavable polyether bonds throughout the structure (non-biodegradable), avoiding dendrimer collapse in vivo and enabling the excretion of such dendrimers from the body as a single entity. [ka]

[0092] In some embodiments, the dendrimer has a plurality of hydroxyl (-OH) groups at the periphery of the dendrimer. In some embodiments, the surface density of the hydroxyl (-OH) groups is at least 1 OH group / nm 2 (number of surface hydroxyl groups / surface area (nm 2 For example, in some embodiments, nm 2 The surface density of hydroxyl groups per nm 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 more than 50 OH groups / nm 2 In some embodiments, the volume density of hydroxyl groups is from about 1 to about 50 groups / nm 3 Between about 5 and about 30 groups / nm 3 Between about 10 and about 20 groups / nm 3 In a further embodiment, the surface density of hydroxyl (—OH) groups is between about 1 and about 50 OH groups / nm 2 or 5-20 OH groups / nm2 (number of surface hydroxyl groups / surface area (nm 2 )), while having a molecular weight between about 100 Da and about 10 kDa, preferably between about 100 Da and 1000 Da. In some embodiments, the amount of surface hydroxyl groups on the dendrimer is preferably greater than 35%, at least 40%, at least 50%, greater than 40%, greater than 50%, or in the range of greater than 40% to 100%. In some embodiments, the dendrimer may have a percentage of hydroxyl groups exposed on the outer surface, with other hydroxyl groups present in the interior core of the dendrimer.

[0093] In some embodiments, the dendrimer specifically targets a particular tissue region and / or cell type after administration to the body. In some embodiments, the dendrimer specifically targets a particular tissue region and / or cell type without a targeting moiety. In some embodiments, the dendrimer contains a number of hydroxyl groups effective to target CNS and / or PNS cells, such as microglia, astrocytes, and / or neurons, associated with a disease, disorder, or injury of the central or peripheral nervous system. In some embodiments, the dendrimer specifically targets a particular tissue region and / or cell type without a targeting moiety, and the active agent conjugated to the dendrimer binds directly to receptors on and / or within the target nervous system cells and / or glial cells.

[0094] In some embodiments, dendrimers may specifically target specific tissue regions and / or cell types, preferably cells and tissues of the central nervous system (CNS) and eye. In some embodiments, dendrimers specifically target neurons of the CNS and eye. Unmodified PAMAM dendrimers with hydroxyl terminal groups are as abundant as these glucose dendrimers, but are not enriched in brain neurons and / or retinal ganglion cells (RGCs) of the eye. Glucose dendrimers with terminal glucose monosaccharides and a high density of hydroxyl functional groups effectively target neurons in a generation-dependent manner. Examples demonstrate that the efficacy of using second generation (G2), G3, and G4 is effective. G5 and above are more difficult to use.

[0095] In a preferred embodiment, the dendrimer comprises a number of terminal glucose and / or hydroxyl groups effective for targeting one or more neurons in the CNS or eye. The hydroxyl groups on the surface of the dendrimer are part of glucose molecules. There are no extra hydroxyls other than the glucose molecules on the surface. The number of sugar molecules on the surface is determined by the number of generations. It is expected that all generations will target neurons.

[0096] In some embodiments, dendrimers are made from glucose and oligoethylene glycol building blocks. Exemplary glucose dendrimers, such as the first-generation dendrimers shown in Structures IV-VI and the second-generation dendrimers shown in Figures 1A and 1B, are shown in the Examples. Some exemplary glucose dendrimers include a first-generation glucose dendrimer with 24 hydroxyl (-OH) end groups, a second-generation glucose dendrimer with 96 hydroxyl (-OH) end groups, a third-generation glucose dendrimer with 396 hydroxyl (-OH) end groups, and a fourth-generation glucose dendrimer with 1584 hydroxyl (-OH) end groups. In a preferred embodiment, the glucose dendrimer is a second-generation glucose-based dendrimer with 24 glucose molecules on the periphery and 6 embedded glucose molecules in the backbone, held together by PEG segments. B. Carbohydrate-modified dendrimers

[0097] In some embodiments, the dendrimer contains one or more carbohydrate molecules at its terminal end. These terminal carbohydrate molecules can be prepared by conjugating one or more surface functional groups of the dendrimer, such as amine, carboxyl, or hydroxyl groups, with one or more carbohydrate molecules. In a preferred embodiment, the dendrimer prior to carbohydrate conjugation is a hydroxyl-terminated dendrimer, such as a hydroxyl-terminated PAMAM dendrimer, and one or more of the hydroxyl groups are conjugated to one or more carbohydrate molecules.

[0098] In some embodiments, hydroxyl-terminated dendrimers modified with surface glucose molecules selectively target central and / or peripheral nervous system cells and / or glial cells in vitro and in vivo; and / or selectively accumulate on and / or within these target nervous system cells, glial cells, and / or macrophage cells, such that active agent(s) conjugated to said dendrimers bind to one or more receptors on / in the target nervous system cells and / or glial cells.

[0099] In some embodiments, the carbohydrate moiety used to modify one or more surface functional groups of a dendrimer is a monosaccharide. Exemplary monosaccharides suitable for modifying a dendrimer include glucose, glucosamine, galactose, mannose, fructose, dehydroascorbic acid, urate, and myo-inositol. In some embodiments, the dendrimer is conjugated to glucose and thus contains glucose as a terminal moiety / molecule. In some embodiments, hydroxyl-terminated dendrimers are modified with one or more glucose moieties ("D-Glu"). In some embodiments, the dendrimer is conjugated to galactose. In some embodiments, the dendrimer is conjugated to mannose. In some embodiments, the dendrimer is conjugated to fructose. In some embodiments, the dendrimer is conjugated to one or more monosaccharides other than glucose, such as galactose, mannose, and / or fructose. For example, the carbohydrate moiety may be an oligosaccharide terminated with one or more monosaccharides including glucose, glucosamine, mannose, fructose, thus exposing these sugar moieties on the surface for attachment.

[0100] The use of glucose dendrimers or glucose-modified dendrimers allows selective uptake by target cells.The drugs conjugated to dendrimers bind to receptors or other sites of action.In a preferred embodiment, dendrimers or dendrimers modified by functional groups are conjugated to one or more drugs that have affinity to and are suitable for binding to one or more cannabinoid receptors, such as CB1 receptor, CB2 receptor and CB3 receptor. In some embodiments, the dendrimer is conjugated to one or more carbohydrate moieties that have affinity for, and are suitable for binding to, one or more non-cannabinoid receptors such as G-protein coupled receptors, e.g., GPR55, GPR18, GPR3, GPR6, GPR12, GPR40, GPR43, GPR41, GPR120, GPR23, GPR92, GPR84, GPR119, or GPR35; adenosine receptors, such as adenosine A3; muscarinic acetylcholine receptors, e.g., M1 and M4; serotonin receptors, e.g., 5-HT1A, 5-HT2A; opioid receptors, e.g., μ- and δ-opioid receptors; and tachykinin NK2 receptors. In some embodiments, the dendrimer is conjugated to one or more carbohydrate moieties or made from sugar moieties suitable for affinity to and transport by 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 and are suitable for binding to one or more of GLUTs, cannabinoid receptors, and / or non-cannabinoid receptors.

[0101] In some embodiments, the dendrimer has a plurality of carbohydrate moieties / molecules, such as monosaccharides, e.g., glucose, or sugar building blocks for dendrimers, at the outer periphery of the dendrimer. In some embodiments, the surface density of carbohydrate molecules, such as monosaccharides, e.g., glucose, is at least one carbohydrate molecule / nm 2 (number of surface carbohydrate groups / surface area (nm 2 In some embodiments, nm 2 The surface density of carbohydrate molecules per nm 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 more than 50 OH groups / nm 2 For example, nm 2 In some embodiments, the surface density of carbohydrate molecules per nanometer is greater than 10. In some embodiments, the volume density of surface carbohydrate molecules is from about 1 to about 50 groups / nm. 3 Between about 5 and about 30 groups / nm 3 Between about 10 and about 20 groups / nm 3 In a further embodiment, the surface density of carbohydrate molecules is between nm 2 Between about 1 and about 50, and between about 5 and about 20 per surface carbohydrate molecule (number of surface carbohydrate molecules / surface area (nm 2 )) while each carbohydrate moiety has a molecular weight between about 100 Da and about 1000 Da. That is, in these embodiments, one or more surface functional groups of the dendrimer are modified at the terminus to introduce one or more sugar moieties / molecule(s), and the terminal hydroxyl groups may be part of a terminal sugar moiety / molecule, or may be part of an extra hydroxyl group that is not modified by a sugar moiety / molecule and therefore is not part of a sugar moiety / molecule, or a combination thereof.

[0102] In some embodiments, the carbohydrate molecule, such as a monosaccharide, e.g., glucose, is present in an amount by weight that is between about 1% and 40% of the total weight of the glycosylated dendrimer, e.g., between about 2% and 20%, between about 5% and 15%, or between 9% and 12% of the total weight of the glycosylated dendrimer. For example, in some embodiments, the carbohydrate moiety is present in an amount that is about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% of the total weight of the glycosylated dendrimer after conjugation. In some embodiments, conjugation of carbohydrate molecules to one or more surface functional groups occurs with about 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, or 25% of the total number of surface functional groups, preferably hydroxyl groups, available prior to conjugation of the dendrimer. In other embodiments, conjugation of carbohydrate molecules occurs with less than 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40% of the total number of available surface functional groups on the dendrimer prior to conjugation. c. Dendrimers modified with polyalkylene glycol

[0103] In some embodiments, the dendrimer contains one or more terminal polyalkylene glycols. These terminal polyalkylene glycols can be prepared by conjugating one or more of the dendrimer's surface functional groups, such as hydroxyl groups, with a polyalkylene glycol, such as PEG. In some embodiments, the dendrimer prior to conjugation is a hydroxyl-terminated dendrimer, such as a hydroxyl-terminated PAMAM dendrimer, and at least a portion of the surface hydroxyl groups are conjugated with PEG.

[0104] In some embodiments, the dendrimer has a plurality of polyalkylene glycols, such as PEG, at the periphery of the dendrimer. In some embodiments, the surface density of the polyalkylene glycol, such as PEG, is at least 1 polyalkylene glycol / nm 2(number of surface polyalkylene glycols / surface area (nm 2 In some embodiments, nm 2 The surface density of polyalkylene glycol per nm 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 more than 50. For example, 2 In some embodiments, the surface polyalkylene glycol volume density is from about 1 to about 50 groups / nm 3 Between about 5 and about 30 groups / nm 3 Between about 10 and about 20 groups / nm 3 In a further embodiment, the surface density of the polyalkylene glycol, such as PEG, is between nm 2 The number of surface polyalkylene glycols per surface area (nm) is between about 1 and about 50, and between about 5 and about 20 per surface area (nm 2 )) On the other hand, it has a molecular weight between about 100 Da and about 10 kDa.

[0105] In some embodiments, the polyalkylene glycol molecule, such as PEG, may be present in an amount by weight that is between about 1% and 40% of the total weight of the PEGylated dendrimer, e.g., between about 2% and 20%, between about 5% and 15%, or between 9% and 12% of the total weight of the PEGylated dendrimer. For example, in some embodiments, the polyalkylene glycol molecule, such as PEG, is present in an amount that is about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% of the total weight of the PEGylated dendrimer after conjugation.

[0106] In some embodiments, conjugation of polyalkylene glycol molecules such as PEG via one or more surface functional groups of the dendrimer is achieved with about 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, or 25% of the total number of surface functional groups, preferably hydroxyl groups, available prior to conjugation of the dendrimer. In other embodiments, conjugation of polyalkylene glycol molecules such as PEG is achieved with less than 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40% of the total number of available surface functional groups of the dendrimer prior to conjugation. B. Cannabinoids

[0107] Dendrimer compositions typically contain 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 classes of cannabinoids, including, but not limited to, plant-derived cannabinoids, endogenous cannabinoids, and synthetic cannabinoids. The cannabinoids or cannabinoid derivatives included in the composition typically bind to one or more receptors, thereby modulating signal transduction 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 additional receptors may be involved. CB1 receptors are present at high levels in some brain regions and in lower amounts in the periphery. CB1 receptors mediate many of the psychoactive effects of cannabinoids. CB2 receptors have a more strict distribution and are found in some immune cells and several types of neurons. Both CB1 and CB2 receptors are primarily coupled to inhibitory G proteins and are subject to the same pharmacological influences as other GPCRs. Therefore, partial agonism, functional selectivity, and inverse agonism all play important roles in determining cellular responses 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, GAT211, GAT228, 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. 1. Endocannabinoids

[0108] The composition can 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 component of cannabis. Exemplary endocannabinoids and endocannabinoid analogs that can be included in the composition include, but are not limited to, N-arachidonoylethanolamide (anandamide), 2-arachidonoylglycerol (2-AG), 2-arachidonylglyceryl ether (noladin ether), N-arachidonoyldopamine (NADA), virodamine (OAE), lysophosphatidylinositol (LPI), 7,10,13,16-docosatetraenoylethanolamide, and homo-γ-linolenoylethanolamine. Although not all can be easily conjugated, most can be modified to allow conjugation.

[0109] In some forms, the endocannabinoids contained in the compositions bind to both the CB1 and CB2 receptors with similar affinity and act as agonists at both the CB1 and CB2 receptors, for example, 2-arachidonoylglycerol binds to both the CB1 and CB2 receptors with similar affinity and acts as a full agonist at both receptors.

[0110] In some embodiments, the endocannabinoids contained in the compositions bind with high affinity to CB1 receptors and weaker affinity to CB2 receptors. For example, anandamide, 7,10,13,16-docosatetraenoylethanolamide, and homo-γ-linolenoylethanolamine bind to central (CB1) cannabinoid receptors and less to peripheral (CB2) cannabinoid receptors, acting as partial agonists at the latter. In another example, 2-arachidonyl glyceryl ether (noladin ether) binds primarily to CB1 receptors and only weakly to CB2 receptors, thereby inducing sedation, hypothermia, slowed intestinal motility, and antinociception. In yet a third example, N-arachidonoyldopamine preferentially binds to CB1 receptors.

[0111] In some forms, the endocannabinoids contained in the compositions bind with high affinity to the CB2 receptor and low affinity to the CB1 receptor. For example, virodamine, or O-arachidonoyl-ethanolamine (OAE), binds with high affinity to the CB2 receptor and acts as a partial agonist at the CB1 receptor.

[0112] In some embodiments, the endocannabinoid contained in the composition binds to one or more non-CB1 / CB2 receptors.For example, anandamide and NADA act as strong agonists for vanilloid receptor subtype 1 (TRPV1), a member of the vanilloid receptor family.In another example, the endogenous lipid metabolite of anandamide (also known as arachidonoylethanolamide or AEA) binds to the N-arachidonoylglycine (NAGly) receptor GPR18, an orphan G protein-coupled receptor. 2. Phytocannabinoids

[0113] The composition may include one or more phytocannabinoids or phytocannabinoid derivatives. Phytocannabinoids or herbicides are a structurally diverse class of naturally occurring chemical constituents derived from plants. In some forms, the phytocannabinoids or phytocannabinoid derivatives used in the compositions may be derived from a variety of sources, including, but not limited to, hemp (e.g., hemp stalk, hemp stalk, hemp seed), cannabis (e.g., cannabis flower, cannabis leaf, cannabis stalk, cannabis stalk, cannabis seed), Echinacea purpurea, Echinacea angustifolia, Echinacea pallida, Acmella oleracea, Helichrysum umbraculigerum, Radula marginata, kava, black truffle, Syzygium aromaticum (clove), Rosmarinus oficinalis, basil, oregano, black pepper, lavender, true cinnamon, Malabathum, cananga odorata, copaifera spp., and hops. Examples include THC, Δ9-tetrahydrocannabinol; Δ9-THCV, Δ9-tetrahydrocannabivarin; CBN, cannabinol; CBDV, cannabidivarin; CBG, cannabigerol; CBC and cannabichromene.

[0114] The compositions 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), cannabielsoin (CBE), cannabicyclol (CBL), cannabifuran (CBF), cannabidtolan (CBT), cannabinodivalin (CBV), cannabilipsol (CBR), isocannabinoids and / or derivatives thereof. Cannabidiol

[0115] In some embodiments, the composition may contain one or more cannabidiol (CBD) and / or derivatives thereof. Suitable examples of cannabidiol (CBD) include, but are not limited to, cannabidiol (CBD), cannabidiolic acid (CBDA), cannabidiolcol (CBDCl), cannabidiol-C4 (CBDC4), cannabidiol dimethyl ether (CBDD), cannabidiol monomethyl ether (CBDM), cannabidivarin (CBDV), cannabidivaric acid (CBDVA), and derivatives thereof. In some embodiments, the composition may contain one or more cannabinodiols (CBND) and / or derivatives thereof. Cannabinodiols or cannabidinodiols are usually present in low concentrations in the Cannabis sativa plant. Cannabinodiol is a fully aromatized derivative of cannabidiol (CBD) and can arise as a product of the photochemical conversion of cannabinol (CBN). In a preferred embodiment, the dendrimer is conjugated to one or more cannabidiol (CBD) or cannabidiol derivatives, as shown in Formula I below: [ka] b. Tetrahydrocannabinol

[0116] In some forms, the compositions 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 (Delta-9-tetrahydrocannabinolic acid), (Δ9-THCA-C1, THCA-C1), Delta-9-tetrahydrocannabinolic acid C4, (Δ9-THCA-C4, THCA-C4), tetrahydrocannabinol (-)-cis-Δ(THC-C5), Delta-9-tetrahydrocannabiorcol (Δ9-THCO-C1), Delta-9-tetrahydrocannabigerolic acid A A) (THCOA), delta-9-tetrahydrocannabivarin (THCV), delta-9-tetrahydrocannabivarin A (THCVA), trihydroxy-delta-9-tetrahydrocannabinol (TRIOH-THC), delta-10-tetrahydrocannabinol (D10THC, Δ10THC), tetrahydrocannabiphorol (THCP), THC-O-acetate (THCO), hexahydrocannabinol (HHC) and derivatives thereof. In a second preferred embodiment, the dendrimer is conjugated to one or more tetrahydrocannabinol (THC) or THC derivatives, as shown in Formula II below. [ka] c. Cannabigerol (CBG)

[0117] In some embodiments, the composition may contain one or more cannabigerols (CBG) or derivatives thereof. Suitable examples of cannabigerols include, but are not limited to, cannabigerolic acid (CBGA), cannabigerolic acid A monomethyl ether (CBGAM), cannabigerovarin (CBGV), cannabigerovaric acid (CBGVA), cannabigerol monomethyl ether (CBGM), and derivatives thereof. In a third preferred embodiment, the dendrimer is conjugated to one or more cannabigerols (CBG) or CBG derivatives, as shown in Formula III below. [ka] d. Cannabinol (CBN)

[0118] In some embodiments, the composition may contain one or more cannabinols (CBN) and / or derivatives thereof. Suitable examples of cannabinols include, but are not limited to, cannabinol (CBN), cannabinolic acid (CBNA), cannabidiolcol (CBN-C1), cannabinol-C2 (CBN-C2), cannabivarin (CBN-C3), cannabinol-C4 (CBN-C4), cannabinodivalin (CBND), cannabinodivalin (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 cannabinols (CBN) or cannabinol derivatives, as shown in Formula IV below. [ka] e. Cannabichromene (CBC)

[0119] In some embodiments, the composition may contain 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), cannabichromenic acid A (CBCA), cannabichromanone (CBCN-C5), cannabichromanone-C3 (CBCN-C3), cannabicoumaronone (CBCON-C5), cannabichromevarin (CBCV), cannabichromevaric acid A (CBCVA), and derivatives thereof. In a fifth embodiment, the dendrimer is conjugated to one or more cannabichromenes (CBC) or CBC derivatives, as shown in Formula V below. [ka] f.Cannabielsoin (CBL)

[0120] In some embodiments, the composition may include one or more cannabielsoins (CBLs) or derivatives thereof. Cannabielsoin (CBE) is a plant cannabinoid metabolite that can be produced by photooxidation from CBD and CBDA or by biotransformation using tissue culture under normal growth conditions. Non-limiting examples of cannabielsoins (CBLs) include cannabielsoin (CBE-C5), -C3-cannabielsoin (CBE-C3), cannabielsonic acid A (CBEA-C5 A), cannabielsonic acid B (CBEA-C5 B), C3-cannabielsonic acid B (CBEA-C3 B), cannabiglendol C3 (OH-iso-HHCVC3), dehydrocannabifuran (DCBF-C5), cannabifuran (CBF-C5), and derivatives thereof. In some forms, one or more cannabielsoins may be included in the compositions to modulate the activity of CB1 receptors, CB2 receptors, non-CB1 receptors, or combinations thereof. In an exemplary embodiment, the dendrimer is conjugated to one or more cannabielsoins (CBLs) or CBL derivatives, as shown in Formula VI below: [ka] g.Cannabidiol (CBT)

[0121] The composition may include one or more cannabidiol (CBT) and / or derivatives thereof. Cannabidiol is present in low concentrations in the cannabis plant and is an oxidation product of tetranhydrocannabinol (THC). Non-limiting examples of cannabidiol (CBT) include cannabidiol (CBT), cannabitriolvarin (CBTV), 10-ethoxy-9-hydroxy-delta-6a-tetrahydrocannabinol, 8,9-dihydroxy-delta-6a-tetrahydrocannabinol (8,9-di-OH), 9,10-dihydroxyhexahydrocannabinol, cannabilipsol, (cannabilipsol-C5), 6a,7,10a trihydroxyΔ9-tetrahydrocannabinol, 10-oxoΔ6a(10a)tetrahydrocannabinol, OTHC, Trans-cannabidiol (Trans-CBT-C5), cannabidiol-C3 (TransCBT-C3), Trans10-O-ethylcannabidiol (TransCBT-OEt-C5), and derivatives thereof. In some forms, one or more cannabidiols may be included in the compositions to modulate the activity of CB1 receptors, CB2 receptors, non-CB1 receptors, or combinations thereof. In an exemplary embodiment, the dendrimer is conjugated to one or more cannabidiol (CBT) or derivatives thereof, as shown in Formula VII below. [ka] h. Other phytocannabinoids

[0122] The composition may contain one or more other plant cannabinoids and / or their derivatives. In some embodiments, the composition may include one or more cannabicyclols (CBLs) or derivatives thereof. Cannabicyclols (CBLs) are photochemical products derived from the plant cannabinoid cannabichromene. Suitable examples of cannabicyclols include, but are not limited to, cannabicyclol (CBL-C5), cannabicyclolic acid (CBLA-C5 A), cannabicyclovalin (CBLV-C3), and derivatives thereof. In some embodiments, the composition may include one or more cannabifurans (CBFs) or derivatives thereof. Suitable examples of cannabifurans include, but are not limited to, cannabifuran (CBF), dehydrocannabifuran (DCBF or CBFD), and derivatives thereof. In some embodiments, the composition may include one or more cannabinodivalins (CBVs) or derivatives thereof. Suitable examples of cannabinodivarins (CBVs) include, but are not limited to, cannabinodivarin (CBV), cannabinodivarin (CBVD), and derivatives thereof. In some forms, the compositions may include one or more of a variety of cannabinoids, including, but not limited to, cannabilipsol (CBR), isocannabinoids, and / or derivatives thereof. Non-limiting examples of isocannabinoids include, but are not limited to, isotetrahydrocannabinol, isotetrahydrocannabivarin, isotetrahydrocannabivarin, and derivatives thereof. i. Synthetic cannabinoids

[0123] The composition may include one or more synthetic cannabinoids and / or derivatives thereof.

[0124] Synthetic cannabinoids (also called cannabinoid receptor agonists) are a class of designer drug molecules functionally similar to Δ9-tetrahydrocannabinol (THC) that 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 considered psychoactive substances and are structurally distinct 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, non-polar, and contain about 22 to about 26 carbon atoms. A common structural feature of synthetic cannabinoids is the side chain, which requires more than four, and up to nine, saturated carbon atoms for optimal activity.

[0125] 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, hybrid cannabinoids, aminoalkyl indoles, and eicosanoids. The composition may contain one or more synthetic cannabinoids and / or derivatives thereof belonging to one or more of the following subclasses: naphthoyl indoles (e.g., JWH-018, JWH-073, and JWH-398); naphthylmethyl indoles, naphthoyl pyrroles, naphthylmethyl indenes, phenylacetyl indoles (i.e., benzoyl indoles, e.g., JWH-250), cyclohexylphenols (e.g., CP 47,497 and homologs of CP 47,497), and classical cannabinoids (e.g., HU-210). The composition may contain one or more synthetic cannabinoids and / or derivatives thereof belonging to one or more of the following classes: adamantyoly indole or indazole carboxamide, benzimidazole, dibenzopyran, eicosanoid, naphtylindene, indazole-3-carboxamide, indole-3-carboxamide, indole-3-carboxylate or aryloxycarbonylindole, naphthoylindazole, pyrazole carboxamide, quinolinyl ester or aryloxycarbonylindole, tetramethylcyclo-propylcarbonylindazole and / or tetramethylcyclo-propylcarbonylindole.

[0126] In an exemplary embodiment, the dendrimer is conjugated to one or more naphthoylindoles or derivatives thereof, as shown in Formula VIII below, where R1, R2, R3, and R4 are positions where substituent variations are possible. [ka]

[0127] In another exemplary embodiment, the dendrimer is conjugated to one or more naphthylmethylindoles or derivatives thereof, as shown in Formula IX below, where R1, R2, and R3 are positions where substituent variations are possible. [ka]

[0128] In another exemplary embodiment, the dendrimer is conjugated to one or more naphthoylpyrroles or derivatives thereof, as shown in formula X below, where R1 and R2 are positions where substituent variations are possible. [ka]

[0129] In another exemplary embodiment, the dendrimer is conjugated to one or more naphthylmethylindenes or derivatives thereof, as shown in Formula XI below, where R1 and R2 are positions where substituent variations are possible. [ka]

[0130] In another exemplary embodiment, the dendrimer is conjugated to one or more phenylacetylindoles or derivatives thereof, as shown in Formula XII below, where R1 and R2 are positions where substituent variations are possible. [ka]

[0131] In another exemplary embodiment, the dendrimer is conjugated to one or more cyclohexylphenols or derivatives thereof, as shown in Formula XIII below, where R1 and R2 are positions where substituent variations are possible. [ka]

[0132] In another exemplary embodiment, the dendrimer is conjugated to one or more classical cannabinoids (dibenzopyrans) and / or derivatives thereof, as shown in Formula XIV below, where R1 and R2 are positions where substituent variations are possible. [ka] C. Coupling Agents and Spacers

[0133] The dendrimer-active agent conjugates can be formed from one or more active agents covalently conjugated to the dendrimer or non-covalently bound to the dendrimer. In a preferred embodiment, the one or more active agents are covalently conjugated to the dendrimer.

[0134] Optionally, one or more active agents are conjugated to the dendrimer via one or more spacers. The term "spacer" includes chemical moieties and functional groups used to link the active agent to the dendrimer. A spacer can be either a single chemical entity or two or more chemical entities linked together. A spacer can include any small chemical entity, peptide, or polymer with sulfhydryl, thiopyridine, succinimidyl, maleimide, vinyl sulfone, carbonate, etc.

[0135] In some embodiments, the spacer through which the active agent is conjugated to the dendrimer contains various linking groups, such as disulfide, ester, carbonate, carbamate, thioester, hydrazine, hydrazide, ether, and amide linking groups. The spacer between the dendrimer and the active agent can be designed to provide a releasable or non-releasable form of the dendrimer conjugate in vivo. In some embodiments, conjugation between the active agent and the dendrimer is via an appropriate spacer, including an ester bond between the active agent and the dendrimer. In some embodiments, one or more spacers between the dendrimer and the active agent can provide a desired effective release rate in vivo. These spacers may contain cleavable linking groups (e.g., ester, disulfide, phosphodiester, triglycyl peptide, hydrazine) or non-cleavable linking groups (e.g., amide, ether, and aminoalkyl). Conjugation between the active agent and the dendrimer can be performed using reactions known in the art, such as click chemistry, acid-amine coupling, and Steglich esterification.

[0136] In some embodiments, conjugation between the active agent and the dendrimer is via a spacer containing a disulfide, ester, ether, thioester, carbamate, carbonate, hydrazine, ether, or amide linking group, or a combination thereof. In some embodiments, conjugation between the active agent and the dendrimer is via a suitable spacer, including an ester or amide linking group between the agent and the dendrimer, depending on the desired release rate of the agent.

[0137] The spacer can be selected from the classes of compounds terminated with sulfhydryl, thiopyridine, succinimidyl, maleimide, vinyl sulfone, and carbonate groups. The spacer can include a thiopyridine-terminated compound such as dithiodipyridine, N-succinimidyl 3-(2-pyridyldithio)-propionate (SPDP), succinimidyl 6-(3-[2-pyridyldithio]-propionamido)hexanoate LC-SPDP, or sulfo-LC-SPDP. The spacer can also include peptides, linear or cyclic, inherently containing a sulfhydryl group, such as glutathione, homocysteine, cysteine ​​and its derivatives, arg-gly-asp-cys (RGDC), cyclo(Arg-Gly-Asp-d-Phe-Cys) (c(RGDfC)), cyclo(Arg-Gly-Asp-d-Tyr-Cys), cyclo(Arg-Ala-Asp-d-Tyr-Cys), etc. The spacer can also be a mercapto acid derivative, such as 3-mercaptopropionic acid, mercaptoacetic acid, 4-mercaptobutyric acid, thiolan-2-one, 6-mercaptohexanoic acid, 5-mercaptovaleric acid, and other mercapto derivatives, such as 2-mercaptoethanol and 2-mercaptoethylamine. The spacer may be thiosalicylic acid and its derivatives, (4-succinimidyloxycarbonyl-methyl-alpha-2-pyridylthio)toluene, (3-[2-pyridylthio]propionyl hydrazide). The spacer may have a maleimide terminus, in which case the spacer comprises a polymer or a small chemical entity such as bis-maleimidodiethylene glycol and bis-maleimidotriethylene glycol, bis-maleimidoethane, bismaleimidohexane, etc. The spacer may comprise a vinyl sulfone such as 1,6-hexane-bis-vinyl sulfone. The spacer may comprise a thioglycoside such as thioglucose. The spacer may be a reduced protein such as bovine serum albumin and human serum albumin, or any thiol-terminated compound capable of forming a disulfide bond.The spacer can include maleimide, succinimidyl and / or thiol-terminated polyethylene glycol. D. Dendrimer-Drug Conjugates

[0138] Dendrimer-active agent conjugates can be formed from cannabinoids and / or cannabinoid derivatives covalently or non-covalently conjugated to dendrimers, dendritic polymers, or hyperbranched polymers. Methods for conjugating one or more active agents to dendrimers are known, such as those described in U.S. Publication Nos. 2011 / 0034422, 2012 / 0003155, and 2013 / 0136697. While covalent conjugation is preferred, ionic complexes (cation-anion complexes) may also be used.

[0139] In some embodiments, one or more active agents are covalently conjugated to one or more terminal groups of the dendrimer, such as a terminal hydroxyl group. In some embodiments, the dendrimer conjugate comprises one or more active agents conjugated to the dendrimer via one or more spacers. The spacer between the dendrimer and the active agent can be designed to provide a releasable or non-releasable form of the dendrimer conjugate in vivo. For example, the spacer may be cleavable or may contain a chemical linking group that is cleavable, for example, upon exposure to the intracellular compartment of a target nervous system cell and / or glial cell, or upon binding to a receptor on or within a target nervous system cell and / or glial cell in vivo. Examples of cleavable linking groups that can be used in the spacer of a dendrimer-active agent conjugate include an esterase-sensitive ester bond, a glutathione-sensitive disulfide bond, a phosphatase-sensitive phosphodiester bond, an oligopeptide such as a triglycyl peptide linker capable of lysosomal release, an acid-cleavable hydrazine linking group, and the like. In some embodiments, the spacer between the dendrimer and the active agent can provide a desired effective release rate in vivo, hi some embodiments, the spacer between the dendrimer and the active agent can be non-cleavable or can include non-cleavable chemical linking groups such as amide, ether, and aminoalkyl linking groups.

[0140] Generally, the spacer between the dendrimer and the active agent is long enough to allow the active agent conjugated to the spacer to reach and bind to the target receptor on the surface and / or inside the target cell. For example, the spacer between the dendrimer and the active agent can have a length ranging from 50 Da to 2000 Da, depending on the desired release rate and the desired receptor binding flexibility. The length of the spacer can vary depending on the location of the target receptor (e.g., on the cell surface, in the cytoplasm of the cell, or in the intercellular compartment of the cell) and / or the density of the receptor if located on the cell surface.

[0141] Dendrimers can be second, third, fourth, fifth, sixth and up to tenth generation. In some embodiments, dendrimers are conjugated to one or more active agents via spacers containing cleavable (ester, disulfide, phosphodiester, triglycyl peptide and hydrazine) or non-cleavable (amide, ether and aminoalkyl) linking groups.

[0142] The density of active agents covalently conjugated to the dendrimer or non-covalently bound to the dendrimer can be adjusted based on the particular cannabinoid or cannabinoid derivative to be delivered, the target receptor, the target nervous system cells and / or glial cells, the location of the target nervous system cells and / or glial cells, etc. For example, multiple active agents conjugated to the dendrimer may be present on the outer periphery of the dendrimer, and the surface density of the active agents may be at least 1 active agent / nm 2 (number of conjugated active agents / surface area (nm 2 )), preferably 3 to 10. For example, in some embodiments, nm 2 In some embodiments, the surface density of active agent per nanometer 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 more than 50. In some embodiments, the volume density of active agent per nanometer is greater than about 1 to about 50 groups / nm. 3 Between about 5 and about 30 groups / nm 3 Between about 10 and about 20 groups / nm 3 It is between.

[0143] Typically, dendrimer-active agent conjugates have hydrodynamic volumes in the nanometer range. For example, in some embodiments, glucose dendrimer-active agent conjugates comprising one or more cannabinoids or cannabinoid derivatives conjugated to the dendrimer have diameters of about 2 nm to about 100 nm or greater than 100 nm, up to 500 nm, depending on the generation of the dendrimer and the chemical composition and amount of active agent conjugated thereto. In some embodiments, dendrimer-active agent conjugates comprising one or more cannabinoids or cannabinoid derivatives conjugated to the dendrimer have a diameter effective to penetrate brain tissue and retain the active agent on and / or within target nervous system cells and / or glial cells for a period of time sufficient for the active agent to bind to target receptors on and / or within the target nervous system cells and / or glial cells. In some embodiments, the dendrimer-active agent conjugates comprising one or more cannabinoids or cannabinoid derivatives conjugated to a dendrimer have a diameter effective to be retained in the peripheral circulation and to be retained on and / or within target nervous system cells and / or glial cells for a period of time sufficient for the active agent to bind to a target receptor on and / or within the target nervous system cells and / or glial cells.

[0144] The dendrimer-active agent conjugate may be neutral, positively charged, or negatively charged. In some embodiments, the dendrimer-active agent conjugate is neutral. The presence of a cannabinoid or cannabinoid derivative can affect the surface charge of the dendrimer-active agent conjugate. In some embodiments, the surface charge of a dendrimer conjugated to a cannabinoid or cannabinoid derivative is between -100 mV and 100 mV, between -50 mV and 50 mV, between -25 mV and 25 mV, between -20 mV and 20 mV, between -10 mV and 10 mV, between -10 mV and 5 mV, between -5 mV and 5 mV, or between -2 mV and 2 mV. The above ranges include all values ​​between -100 mV and 100 mV. In a preferred embodiment, the surface charge of the dendrimer-active agent conjugate is neutral or near neutral, ie, from about −10 mV to about 10 mV, inclusive.

[0145] An exemplary dendrimer-active agent conjugate is represented by formula (I): The dendrimer of the exemplary conjugate contains surface hydroxyl groups, one or more of which are conjugated to one or more active agents via one or more spacers, as shown in formula (I) below. [ka] wherein D can be a 1st to 10th generation or a 2nd to 10th generation dendrimer such as any one of those described above, for example, PAMAM (such as a hydroxyl-terminated PAMAM dendrimer) or a glucose-based dendrimer; L, at each occurrence, can be any suitable chemical moiety suitable for providing modulation of drug release and receptor binding, preferably containing a triazole moiety; and Y is a bond, or a secondary amide (-CONH-), a tertiary amide (-CONR-), a sulfonamide (-S(O)-NR-), a secondary carbamate (-OCONH-; -NHCOO-), a tertiary carbamate (-OCONR-; -NRCOO-), a carbonate (-OC(O)-O-), a urea (-NHCONH-; -NRCONH-; -NHCONR-, -NRCONR-), a carbinol (-CHOH-, -CROH-), a disulfide group, a phosphodiester group. [ka] R may be a linking group selected from a hydrazino group, a hydrazone, a hydrazide, an ester (-C(O)-O-), an ether (-O-), and an oligopeptide (e.g., a triglycyl peptide); R is an alkyl group, an aryl group, or a heterocyclic group; X, at each occurrence, may be a cannabinoid or cannabinoid derivative; the functional group of X (such as an amino group, including a primary amino, secondary amino, or tertiary amino group, a carboxyl group, or a hydroxyl group) forms part of the linking group Y; n may be an integer between 1 and 100; and m may be an integer between 16 and 4096. The dendrimer may be a PAMAM or glucose dendrimer that is 100% hydroxyl. m and n depend on the size of dendrimer D; n should be such that the weight percent of the drug in the total conjugate is 5-20%. This range is also appropriate for binding and internalization.

[0146] The oxygen atoms shown in formula (I) are derived from surface functional groups of the dendrimer, such as surface hydroxyl groups, which may or may not be part of a terminal sugar moiety / molecule (e.g., glucose). Although not depicted in formula (I), one or more hydroxyl groups of the dendrimer that are not conjugated to an active agent may be modified with one or more carbohydrates and / or polyalkylene glycols, such as PEG.

[0147] 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 or inside a target cell. In some embodiments, when the cannabinoid and / or cannabinoid derivative X binds to the target receptor, the drug X remains conjugated to the dendrimer. In these embodiments, after binding, the drug X may be released from the dendrimer or may remain conjugated to the dendrimer as an intact dendrimer-active agent conjugate. In some embodiments, the cannabinoid and / or cannabinoid derivative X is released from the dendrimer in close proximity to the target receptor and then binds to the target receptor.

[0148] In some embodiments, L, at each occurrence, can be represented by -A'-L1-B'-L2-, where A' can be carbonyl (-C(O)-) or a bond (including single, double, and triple bonds, e.g., a single bond), B' can be a bond (including single, double, and triple bonds, e.g., a single bond), an amide, an ester, an ether, a thiol, a dithiol, an aryl, a heteroaryl, a polyaryl, a heteropolyaryl, or a heterocyclic, and L1 and L2 can be independently a bond, an alkylene, a heteroalkylene, an aryl, an aralkyl, an ether, a polyether, L1 and L2 may be a thiol, dithiol, thiol ether, polythioether, oligopeptide, polypeptide, oligo(alkylene glycol) or polyalkylene glycol, or L1 and L2 may independently be composed of combinations of these groups, such as a combination of alkylene and polyether, a combination of alkylene and thiol or dithiol, a combination of alkylene and oligopeptide, a combination of alkylene, polyether and thiol or dithiol, or a combination of polyether and thiol or dithiol. In some forms, L1-B'-L2- together form a chemical moiety selected from -alkylene-triazole-di(alkylene glycol)-, -di(alkylene glycol)-triazole-alkylene-, -alkylene-triazole-oligo(alkylene glycol)-, -oligo(alkylene glycol)-triazole-alkylene-, -alkylene-triazole-poly(alkylene glycol)-, -poly(alkylene glycol)-triazole-alkylene-, -alkylene-triazole-ether-, -alkylene-triazole-alkylene-, -alkylene-amide-alkylene-, and combinations thereof.

[0149] In some embodiments, B' can be a bond (including single, double and triple bonds, eg, single bonds), an amide group, or a heterocyclic group such as a triazole group.

[0150] In some embodiments, L1 is a bond, C1-C 10alkylene, such as C1-C8 alkylene, C1-C6 alkylene, C1-C5 alkylene, C1-C4 alkylene or C1-C3 alkylene, or [ka] wherein p is an integer of 1 to 20, 1 to 18, 1 to 16, 1 to 14, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 or 2.

[0151] In some embodiments, L2 is a bond, C1-C 10 alkylene such as alkylene, C1-C8 alkylene, C1-C6 alkylene, C1-C5 alkylene, C1-C4 alkylene or C1-C3 alkylene; [ka] wherein p is an integer from 1 to 20, 1 to 18, 1 to 16, 1 to 14, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 or 2), an oligo- or poly-peptide such as triglycyl peptide, a thiol, or a dithiol, or L2 is comprised of a combination of two or more of alkylene, oligo- or poly-(alkylene glycol), oligo- or poly-peptide, thiol, and dithiol. For example, L2 can be [ka] wherein p, q, r, s, t, and u are independently integers from 0 to 10, 0 to 8, 0 to 6, 0 to 5, 0 to 4, 0 to 3, or 0 to 2, such as 0, 1, or 2, and G' is a thiol, dithiol, oligo-peptide (such as triglycyl peptide), or poly-peptide.

[0152] In some embodiments, Y is a linking group that is minimally cleavable in vivo. In some embodiments, Y is a linking group that is cleavable in vivo. In some embodiments, Y is an amide (-CONH-), ester (-C(O)-O-), ether (-O-), phosphodiester, or disulfide group.

[0153] In some embodiments, both L and Y are single bonds and D is conjugated directly to X (the active agent or analog thereof) through an ether linking group.

[0154] In some embodiments, D is a second generation PAMAM dendrimer, a third generation PAMAM dendrimer, a fourth generation PAMAM dendrimer, a fifth generation PAMAM dendrimer, a sixth generation PAMAM dendrimer, a first generation glucose dendrimer, a second generation glucose dendrimer, a third generation glucose dendrimer, a fourth generation glucose dendrimer, a fifth generation glucose dendrimer, or a sixth generation glucose dendrimer.

[0155] More specific exemplary dendrimer-active agent conjugates are shown in the Examples below. E. Exemplary Cannabinoid-Dendrimer Conjugates

[0156] In a preferred embodiment, the dendrimer is conjugated to cannabidiol (CBD), as shown in structures A and B below, where n is an integer from 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. [ka]

[0157] In other preferred embodiments, the dendrimer is conjugated to HU-308 as shown in structures C to E below, where n is an integer from 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. [ka]

[0158] In other preferred embodiments, the dendrimer is conjugated to THC, as shown in structures F-G below. [ka] [ka]

[0159] In other preferred embodiments, the dendrimer is conjugated to an AEA as shown in structures IJ below, where n is an integer from 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. [ka] [ka]

[0160] In another preferred embodiment, the dendrimer is conjugated to 2-AG, as shown in structure K below, where n is an integer from 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. [ka] III. Methods of Making Dendrimer Conjugates A. Methods for Making Dendrimers

[0161] Dendrimers, especially glucose dendrimers, can be prepared by various chemical reaction steps. Dendrimers are usually synthesized according to methods that allow control of their structure at each stage of construction. Dendritic structures are mostly synthesized by two main different methods: divergent or convergent.

[0162] In some embodiments, dendrimers are prepared using a divergent method, in which dendrimers are assembled from a multifunctional core that is extended outward through a series of reactions, typically the Michael reaction. This strategy involves coupling the multifunctional core moiety with monomer molecules bearing reactive and protecting groups, resulting in the stepwise addition of generations around the core, followed by removal of the protecting groups. For example, dendrimers are first synthesized by coupling AB4 peracetylated β-D glucose-PEG4-azide monomers to a hexapropargylated core. In another example, PAMAM-NH2 dendrimers are first synthesized by coupling N-(2-aminoethyl)acrylamide monomers to an ammonia core.

[0163] In other embodiments, dendrimers are prepared using convergent methods, where dendrimers are built from small molecules that terminate on the surface of a sphere, and the reaction proceeds inward, building inward and finally attaching to the core.

[0164] Many other synthetic routes exist for the preparation of dendrimers, such as the orthogonal, accelerated, two-step convergent or hypercore approach, the hypermonomer or branched monomer approach, the double exponential, the orthogonal coupling or two-step approach, the two-monomer approach, and the AB2-CD2 approach.

[0165] In some embodiments, the dendrimer core, one or more branching units, one or more spacers, and / or one or more surface functional groups can be modified to allow conjugation to additional functional groups (such as branching units, spacers, surface functional groups), monomers, and / or drugs via click chemistry using one or more of copper-assisted azide-alkyne cycloaddition (CuAAC), Diels-Alder reaction, thiol-ene reaction, thiol-yne reaction, and azide-alkyne reaction (Arseneault M et al., Molecules 2015 May 20;20(5):9263-94). In some embodiments, prefabricated dendrons are clicked onto high-density hydroxyl polymers. "Click chemistry" involves coupling two different moieties (e.g., a core group and a branching unit, or a branching unit and a surface functional group) via, for example, a 1,3-dipolar cycloaddition reaction between an alkyne moiety (or its equivalent) on the surface of a first moiety and an azide moiety (e.g., present in a triazine composition or its equivalent) or any reactive end group (e.g., a primary amine end group, a hydroxyl end group, a carboxylic acid end group, a thiol end group, etc.) on a second moiety. In some embodiments, one or more hydroxyl groups on the surface of a dendrimer (e.g., 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 (e.g., a hydroxyl-terminated PAMAM dendrimer or a glucose dendrimer) are modified to contain an azide group, and the drug is modified to contain an alkyne group. The azide and alkyne then react via a 1,3-dipolar cycloaddition reaction to form the triazole moiety.

[0166] In some embodiments, dendrimer synthesis relies on one or more reactions such as thiol-ene click reaction, thiol-yne click reaction, CuAAC, Diels-Alder click reaction, azide-alkyne click reaction, Michael addition, epoxy ring-opening, esterification, silane chemistry, and combinations thereof.

[0167] In some embodiments, the method includes one or more protection and deprotection steps of functional groups (e.g., hydroxyl groups) on the central core, branching units, and / or therapeutic, prophylactic, or diagnostic agents to facilitate the addition of branching units to generate the desired dendrimer molecule, or the addition of the therapeutic, prophylactic, or diagnostic agent to generate the desired dendrimer conjugate. In the case of hydroxyl groups, the hydroxyl group may be protected by the formation of an ether, ester, or acetal. Other exemplary protecting groups include Boc and Fmoc.

[0168] Any existing dendritic platform can be used to create dendrimers with the desired functionality, i.e., a high density of surface hydroxyl groups, by conjugating higher hydroxyl-containing moieties such as 1-thio-glycerol or pentaerythritol. Exemplary dendritic platforms such as polyamidoamine (PAMAM), poly(propyleneimine) (PPI), poly-L-lysine, melamine, poly(etherhydroxylamine) (PEHAM), poly(esteramine) (PEA), and polyglycerol can be synthesized and explored.

[0169] Dendrimers can also be prepared by combining two or more dendrons. A dendron is a wedge-shaped section of a dendrimer that carries a reactive focal group. Numerous dendron scaffolds are commercially available. These dendrons have appeared in the first, second, third, fourth, fifth, and sixth generations, with 2, 4, 8, 16, 32, and 64 reactive groups, respectively. In certain embodiments, one type of drug is linked to one type of dendron, and a different type of drug is linked to another type of dendron. The two dendrons are then linked to form a dendrimer. The two dendrons can be linked via click chemistry, i.e., a 1,3-dipolar cycloaddition reaction, between an azide moiety on one dendron and an alkyne moiety on another dendron to form a triazole linker.

[0170] Exemplary methods of making dendrimers are described in detail in International Patent Publication Nos. WO2009 / 046446, WO2015168347, WO2016025745, WO2016025741, WO2019094952, and U.S. Pat. No. 8,889,101. 1. Method for preparing glucose dendrimers

[0171] In some embodiments, glucose-based dendrimers are prepared using a divergent method in which dendrimers are assembled from a multifunctional core that is extended outward through a series of reactions. This strategy involves coupling monomer molecules bearing reactive and protecting groups to the multifunctional core moiety, resulting in the stepwise addition of generations around the core, followed by removal of the protecting groups.

[0172] In some embodiments, glucose dendrimers are synthesized by coupling AB4 peracetylated β-D glucose-PEG4-azide monomers to a hexapropargylated core. In a preferred embodiment, the hypercore is prepared from dipentaerythritol, for example, by propargylating dipentaerythritol to obtain a hexapropargylated core. An exemplary scheme for preparing such glucose dendrimers is shown in Scheme I. [ka]

[0173] In some embodiments, the branching unit is a hypermonomer, i.e., AB n These are building blocks. Exemplary hypermonomers include the AB3, AB4, AB5, AB6, AB7, and AB8 building blocks. The hypermonomer strategy dramatically increases the number of available end groups. An exemplary hypermonomer is the orthogonal hypermonomer AB4, which contains one azide functional group and four allyl groups and is prepared by reacting dipentaerythritol, which has five allyl groups, with monotosylated triethylene glycol azide.

[0174] In some embodiments, the branching unit is a linear or branched polyethyleneglycerol, for example, as shown by Formula III. Other monomers include disaccharides and oligosaccharides, as well as saccharides such as fructose, lactose, and sucrose. a. Synthesis of AB4 building blocks

[0175] Some exemplary synthesis methods for hypermonomer AB4 are described below. In some embodiments, hypermonomer AB4 is based on a glucose molecule. In a preferred embodiment, hypermonomer AB4 is conjugated to polyethyleneglycerol, e.g., tetraethyleneglycol (PEG4). In one embodiment, hypermonomer AB4 is peracetylated β-D-glucopyranoside tetraethyleneglycol azide.

[0176] In some embodiments, the synthesis of glucose-OAc-TEG-OTs comprises the following steps: a solution of peracetylated β-D-glucopyranoside (10 g, 25.6 mmol) was dissolved in 50 mL of anhydrous dichloromethane (DCM), and then 2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (6.2 g, 17.9 mmol) was added, and the reaction mixture was cooled to 0 °C. Boron trifluoride diethyl etherate (2.5 equivalents) was added, and the reaction was allowed to reach room temperature. The reaction was monitored by TLC and, after 5 h, quenched by adding saturated sodium bicarbonate solution at 0 °C. After stirring for 10 min, DCM (300 mL) was added, and the organic layer was washed three times with saturated sodium bicarbonate solution until foaming disappeared. The reaction mixture was dried over sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The crude product was purified by combiflash® chromatography using an ethyl acetate / hexane (70:30) mixture as the eluent. The desired compound was obtained in 60% yield. The structure of Glucose-OAc-TEG-OTs is shown below: [ka]

[0177] In some embodiments, the synthesis of glucose-OAc-TEG-N3 involves the following steps: a solution of glucose-OAc-TEG-OTs (6 g, 8.8 mmol) is dissolved in 40 mL of anhydrous DMF, then sodium azide (2 equivalents) is added, and the reaction mixture is heated to 50° C. overnight. Upon completion, the reaction mixture is filtered and the DMF is evaporated. Once dry, the crude reaction mixture is passed through a combiflash® column using ethyl acetate:hexane (70:30) as the eluent. The structure of glucose-OAc-TEG-N3 is shown below: [ka]

[0178] In some embodiments, the synthesis of glucose-OH-TEG-N3 involves the following steps: dissolving peracetylated β-D-glucopyranoside tetraethylene glycol azide in anhydrous methanol and adding sodium methoxide to adjust the pH to about 8.5-9. The reaction is stirred overnight at room temperature, then diluted with methanol and adjusted to about pH 6-7 with Amberlist IR-120+. The reaction mixture is isolated by filtration, and the solvent is removed by rotary evaporation. The structure of glucose-OH-TEG-N3 is shown below. [ka] b. Synthesis of glucose dendrimers

[0179] In some embodiments, glucose dendrimers are synthesized by coupling AB4 peracetylated β-D-glucose-PEG-azide monomers to a hexapropargylated core. In a preferred embodiment, the hexapropargylated core is linked to the AB4 β-D-glucose-PEG-azide building block (2) via a click reaction to yield a first-generation dendrimer.

[0180] In some embodiments, the first-generation dendrimer D1-Glu6-OAc24 is prepared as follows: The hexapropargylated compound (0.5 g, 1 mmol) and the azide derivative (4.1 g, 7.4 mmol, 1.2 equivalents per acetylene) are suspended in a 1:1 mixture of DMF and water in a 20 mL microwave vial equipped with a magnetic stir bar. CuSO4·5H2O (5 mol% / acetylene, 75 mg) and sodium ascorbate (5 mol% / acetylene, 60 mg) dissolved in a minimum amount of water are added. The reaction is microwaved at 50 °C for 6 h. The reaction mixture is dialyzed against DMF followed by dialysis against water containing EDTA. The EDTA is further removed by extensive water dialysis. The product is lyophilized to afford D1-Glu6-OAc24. The structure of D1-Glu6-OAc24 is shown below. [ka]

[0181] In some embodiments, the first generation dendrimer D1-Glu6-OH 24 is prepared as follows: peracetylated first-generation glucose dendrimer (1 g, 0.26 mmol) is dissolved in anhydrous methanol and sodium methoxide is added to adjust the pH to approximately 8.5-9. The reaction is stirred overnight at room temperature, then diluted with methanol and the pH adjusted to approximately 6-7 with AMBERLIST® IR-120+. The reaction mixture is separated by filtration, the solvent removed by rotary evaporation, and then subjected to water dialysis. The structure of first-generation glucose dendrimer D1-Glu6-OH24 is shown below. [ka]

[0182] In some embodiments, first-generation glucose dendrimer D1-Glu6-OH24 was propargylated to give D1-acetylene 24 as follows: D1-GLu6-OH24 (2 g, 0.721 mmol) was dissolved in anhydrous dimethylformamide (DMF, 50 mL) by sonication. To this solution, sodium hydride [60% dispersion in mineral oil] (951 mg, 39.65 mmol) was slowly added in small portions with stirring at 0°C. The solution was stirred for an additional 15 minutes at 0°C. After this, propargyl bromide (3.85 mL, 34.608 mmol, 80% w / w solution in toluene) was added at 0°C, and stirring was continued for an additional 6 hours at room temperature. The reaction mixture was quenched with ice and water, filtered, and dialyzed against DMF, followed by water dialysis, to give D1-acetylene 24. The structure of D1-acetylene 24 is shown below. [ka]

[0183] In some embodiments, the first generation dendrimer D1-acetylene 24 is further reacted with AB4β-D-glucose-PEG4-azide to give a second generation dendrimer with 24 glucose molecules, containing 96 surface hydroxyl groups.

[0184] An exemplary second-generation dendrimer, D2-Glu24-OAc96, was prepared as follows: D1-acetylene dendrimer 24 (0.5 g, 0.13 mmol) and glucose-OAc-TEG-azide (2.2 g, 4 mmol) were suspended in a 1:1 mixture of DMF and water in a 20 mL microwave vial equipped with a magnetic stir bar. To this was added CuSO4·5H2O (5 mol% / acetylene, 5 mg) and sodium ascorbate (5 mol% / acetylene, 10 mg) dissolved in a minimal amount of water. The reaction was microwaved at 50 °C for 8 h. Upon completion, the reaction mixture was dialyzed against DMF followed by water dialysis containing EDTA. EDTA was further removed by extensive water dialysis. The product was lyophilized to yield D2-Glu24-OAc96.

[0185] In some embodiments, second-generation dendrimer D2-Glu24-OH96 is prepared as follows: peracetylated second-generation glucose dendrimer D2-Glu24-OH96 is dissolved in anhydrous methanol and sodium methoxide is added to adjust the pH to about 8.5-9.0. The reaction is stirred overnight at room temperature, then diluted with methanol and the pH adjusted to about 6-7 with AMBERLIST® IR-120+. The reaction mixture is filtered to remove the resin, and the filtrate is rotary evaporated, followed by water dialysis to give the product as an off-white solid.

[0186] The structure of the second generation glucose dendrimer D2-Glu24-OH96 is shown below. [ka]

[0187] In some embodiments, the second-generation dendrimer D2-Glu24-OH96 is propargylated at one or more terminal hydroxyl groups suitable for further conjugation to one or more therapeutic, prophylactic, or diagnostic agents. In some embodiments, one or more terminal hydroxyl groups of the second-generation dendrimer D2-Glu24-OH96 are propargylated as follows: D2-Glu24-OH96 (5b) (200 mg, 0.016 mmol) is dissolved in anhydrous dimethylformamide (DMF, 10 mL) by sonication. To this stirred solution, sodium hydride (60% dispersion in mineral oil) (22 mg, 0.934 mmol) is added slowly in small portions at 0° C. The solution is further stirred at 0° C. for 15 minutes. After this, propargyl bromide (18.0 μL, 80% w / w solution in toluene) is added at 0° C., and stirring is continued for an additional 6 hours at room temperature. The solvent is evaporated using a V10 evaporator system and the crude product is purified by passing through a PD10 SEPHADEX® G25M column. The aqueous solution is lyophilized to give the product as an off-white solid.

[0188] In some embodiments, one or more fluorescent dyes, such as infrared-fluorescent Cy5 dyes, are conjugated to the second-generation dendrimer D2-Glu24-OH96. In one embodiment, Cy5-D2-Glu24-OH96 (compound 7 in Figure 1B) is prepared as follows: Compound 6 (200 mg, 0.016 mmol) and Cy5 azide (20.7 mg, 0.02 mmol) are suspended in a 1:1 mixture of DMF and water in a 25 mL round-bottom flask equipped with a magnetic stir bar. To this, CuSO4·5H2O (5 mol% / acetylene, 0.3 mg) and sodium ascorbate (10 mol% / acetylene, 0.5 mg) dissolved in a minimum amount of water are added. The reaction is stirred at room temperature for 24 hours. Upon completion, the DMF is evaporated using a V10 column, and purification is performed using a PD10 Sephadex G25 M column. The aqueous solution is lyophilized to give the product as a blue solid.

[0189] In some embodiments, the total hydroxyl groups available for further conjugation to active agents, including therapeutic and / or diagnostic agents, are about 1-30, 2-20, or 5-10 of the total 96 hydroxyl groups available for an exemplary second generation dendrimer having 24 glucose molecules containing 96 surface hydroxyl groups. B. Methods of Making Dendrimer-Drug Conjugates

[0190] Methods for conjugating drugs to dendrimers are generally known in the art, for example, as described in U.S. Published Application Nos. US2011 / 0034422, US2012 / 0003155, and US2013 / 0136697.

[0191] In some embodiments, one or more drugs are covalently attached to the dendrimer. In some embodiments, the drugs are attached to the dendrimer via a spacer designed to be non-cleavable in vivo. In some embodiments, the drugs are attached to the dendrimer via a spacer designed to be cleavable in vivo. For example, the spacer can be designed to be cleaved by hydrolysis, enzymes, or a combination thereof to achieve sustained release of the drug in vivo. In some embodiments, both the chemical structure of the spacer and its point of attachment to the drug can be selected so that cleavage of the spacer releases either the drug or a suitable prodrug thereof. The chemical structure of the spacer can also be selected taking into account the desired release rate of the drug.

[0192] In some embodiments, the conjugation between the drug and the dendrimer is via one or more of a disulfide, ester, ether, phosphodiester, triglycyl peptide, hydrazine, amide, or aminoalkyl linking group. In some embodiments, the conjugation between the drug and the dendrimer is via a suitable spacer that results in an ester or amide bond between the drug and the dendrimer, depending on the desired release rate of the drug. In some cases, an ester or disulfide bond is introduced for a releasable form of the drug. In other cases, an amide or aminoalkyl bond is introduced for a non-releasable form of the drug.

[0193] The spacer generally contains one or more organic functional groups. Examples of suitable organic functional groups contained in the spacer include secondary amide (-CONH-), tertiary amide (-CONR-), sulfonamide (-S(O)2-NR-), secondary carbamate (-OCONH-; -NHCOO-), tertiary carbamate (-OCONR-; -NRCOO-), carbonate (-OC(O)-O-), urea (-NHCONH-; -NRCONH-; -NHCONR-, -NRCONR-), carbinol (-CHOH-, -CROH-), disulfide group, hydrazone, hydrazide, ether (-O-) and ester (-COO-, -CHOC-, CHROC-), where R is an alkyl group, an aryl group or a heterocyclic group. Generally, the one or more organic functional groups in the spacer are selected taking into account the desired release rate of the drug. Additionally, one or more organic functional groups can be selected to facilitate covalent conjugation of a drug to the dendrimer. In some embodiments, conjugation between the drug and the dendrimer is via a suitable spacer that results in a disulfide bridge between the drug and the dendrimer. In some embodiments, the dendrimer-active agent conjugate is capable of rapid release of the drug in vivo by a thiol exchange reaction under reducing conditions found in the body.

[0194] In certain embodiments, the spacer comprises one or more of the organic functional groups described above in combination with a linking group. The linking group may be composed of any collection of atoms, including oligomeric and polymeric chains, for example, where the total number of atoms in the linking group is between 3 and 200 atoms, between 3 and 150 atoms, between 3 and 100 atoms, or between 3 and 50 atoms. Examples of suitable linking groups include alkyl groups, heteroalkyl groups, alkylaryl groups, oligoethylene glycol chains and polyethylene glycol chains, and oligo(amino acid) chains and poly(amino acid) chains. Variations in the linking group provide further control over in vivo drug release. In embodiments in which the spacer comprises a linking group, one or more organic functional groups are typically used to link the linking group to both the anti-inflammatory agent and the dendrimer.

[0195] Reactions and strategies useful for covalently conjugating drugs to dendrimers are known in the art. See, for example, March, "Advanced Organic Chemistry," 5th Edition, 2001, Wiley-Interscience Publication, New York) and Hermanson, "Bioconjugate Techniques," 1996, Elsevier Academic Press, USA. The appropriate method for covalently conjugating a given drug involves functional group compatibility, protecting group strategy, and the presence of labile bonds, and can be selected taking into account the desired linking moiety, as well as the structure of the drug and dendrimer.

[0196] The amount of active agent (drug loading) in the dendrimer-active agent conjugate depends on a number of factors, including the choice of active agent, the structure and size of the dendrimer, and the tissue to be treated. In some embodiments, the one or more cannabinoids or cannabinoid derivatives are conjugated to the dendrimer at a concentration of between about 0.01% and about 45% by weight (inclusive), between about 0.1% and about 30% by weight (inclusive), between about 0.1% and about 20% by weight (inclusive), between about 0.1% and about 10% by weight (inclusive), between about 1% and about 10% by weight (inclusive), between about 1% and about 5% by weight (inclusive), between about 3% and about 20% by weight (inclusive), or between about 3% and about 10% by weight (inclusive). However, the specific drug loading for any given active agent, dendrimer and targeting site can be determined by routine methods such as those described.

[0197] In some embodiments, drug / spacer conjugation occurs on about 1%, 2%, 3%, 4%, or 5% of the total number of available surface functional groups, such as hydroxyl groups, on the dendrimer prior to conjugation. In other embodiments, drug / spacer conjugation occurs on less than 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of the total number of available surface functional groups on the dendrimer prior to conjugation with the active agent. In some embodiments, the dendrimer-active agent conjugate retains an effective amount of surface functional groups for targeting to target nervous system cells and / or glial cells while simultaneously being conjugated to an effective amount of drug for treating, preventing, and / or imaging a disease or disorder. In some embodiments, the dendrimer-active agent conjugate carries an amount of active agent effective to target the target nervous system cells and / or glial cells and to bind to a target receptor on or within the target nervous system cells and / or glial cells.

[0198] More specific methods for preparing exemplary dendrimer-active agent conjugates are described in the Examples below. IV. Pharmaceutical Preparations

[0199] Pharmaceutical compositions comprising the dendrimer-active agent conjugates can be formulated in a conventional manner using one or more physiologically acceptable carriers, optionally including additives and adjuvants that facilitate processing into preparations of the active compound that can be pharmaceutically used for oral, mucosal (intranasal, buccal, sublingual, vaginal, rectal or pulmonary), transdermal or injectable (intravenous, subcutaneous, intraperitoneal, intramuscular or intrathecal administration) administration.

[0200] The composition, method and relative amounts of the formulation depend on the selected route of administration. The composition can be stored lyophilized in a single-use vial for rehydration immediately before use. Other means for rehydration and administration are known to those skilled in the art.

[0201] Representative additives include solvents, diluents, pH modifiers, preservatives, antioxidants, suspending agents, wetting agents, viscosity modifiers, tonicity agents, stabilizers, and combinations thereof. Suitable pharmaceutically acceptable additives are preferably selected from generally recognized as safe (GRAS) substances and can be administered individually without causing undesired biological side effects or undesired interactions.

[0202] Generally, pharmaceutically acceptable salts can be prepared by reacting a drug in the form of a free acid or free base with a stoichiometric amount of an appropriate base or acid in water or an organic solvent, or a mixture of the two; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Pharmaceutically acceptable salts include drug salts derived from inorganic acids, organic acids, alkali metal salts, alkaline earth metal salts, and salts formed by the reaction of a drug with a suitable organic ligand (e.g., quaternary ammonium salts). A list of suitable salts can be found, for example, in Remington's Pharmaceutical Sciences, 20th ed., Lippincott Williams & Wilkins, Baltimore, MD, 2000, p. 704. Examples of ophthalmic drugs that are sometimes administered in the form of pharmaceutically acceptable salts include timolol maleate, brimonidine tartrate, and diclofenac sodium.

[0203] The compositions are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. The phrase "dosage unit form" refers to a physically discrete unit of the conjugate appropriate for the patient being treated. However, it will be understood that the total number of single doses of the composition will be decided by the attending physician within the scope of sound medical judgment. The therapeutically effective dose can be estimated initially either in cell culture assays or in animal models, usually mice, rabbits, dogs, or pigs. Animal models are also used to obtain a desirable concentration range and route of administration. Such information can then be used to determine effective doses and routes for administration in humans. The therapeutic efficacy and toxicity of the conjugates can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, such as determining the ED50 (the dose therapeutically effective in 50% of the population) and the LD50 (the dose lethal to 50% of the population). The dose ratio of toxic to therapeutic effects is the therapeutic index, and the LD 50 / ED 50Pharmaceutical compositions that exhibit large therapeutic indices are preferred. The data obtained from cell culture assays and animal studies can be used in formulating a range of dosage for human use.

[0204] In certain embodiments, the composition is administered locally, for example, by injection directly into the site to be treated. In some embodiments, the composition is injected, topically applied, or otherwise administered directly to the vasculature on the site of injury, surgery, or implantation, or on vascular tissue adjacent to the site. For example, in embodiments, the composition is applied locally to vascular tissue exposed during a surgical procedure. Typically, local administration results in an increased local concentration of the composition, which is higher than can be achieved by systemic administration.

[0205] Pharmaceutical compositions formulated for administration by parenteral (intramuscular, intraperitoneal, intravenous or subcutaneous injection) and enteral routes of administration are described.

[0206] In some embodiments, the dendrimer conjugates may increase the efficacy and duration of treatment, thereby reducing the need for repeat administration for weekly, monthly, 6-monthly, yearly, or other longer-term dosing regimens. Some embodiments may be incorporated into drug delivery systems (e.g., implants, pumps, patches, creams, etc.) to achieve controlled, sustained delivery in a manner that reduces compliance requirements and the potential for abuse. A. Parenteral Administration

[0207] The dendrimer-active agent conjugate composition can be administered parenterally. The terms "parenteral administration" and "parenterally administered" are art-recognized terms and include modes of administration other than enteral and topical administration. Dendrimers can be administered orally, intranasally, subcutaneously, intraperitoneally, or intramuscularly. For liquid formulations, pharmaceutically acceptable carriers can be, for example, aqueous or non-aqueous solutions, suspensions, emulsions, or oils. Parenteral vehicles (for subcutaneous, intravenous, intraarterial, or intramuscular injection) include, for example, sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's, and fixed oils. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, and injectable organic esters such as ethyl oleate. Aqueous carriers include, for example, water, alcoholic / aqueous solutions, emulsions or suspensions, including cyclodextrins, saline, and buffered media. Dendrimers can also be administered in emulsion, for example, in water in oil.Examples of oils are those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, olive oil, sunflower oil, fish liver oil, sesame oil, cottonseed oil, corn oil, olive, petrolatum and inorganic.The fatty acids suitable for use in parenteral formulations include, for example, oleic acid, stearic acid and isostearic acid.Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters.

[0208] Formulations suitable for parenteral administration can include aqueous and non-aqueous sterile suspensions, which can contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, as well as suspending agents, solubilizers, thickeners, stabilizers, and preservatives. Intravenous vehicles can include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer's dextrose). In general, water, saline, aqueous dextrose and related sugar solutions, and glycols such as propylene glycol or polyethylene glycol are preferred liquid carriers, particularly for injectable solutions.

[0209] Injectable pharmaceutical carriers for injectable compositions are well known to those skilled in the art (see, e.g., Pharmaceutics and Pharmacy Practice, J.B. Lippincott Company, Philadelphia, PA, Banker and Chalmers, eds., pages 238-250 (1982), and ASHP Handbook on Injectable Drugs, Trissel, 15th ed., pages 622-630 (2009)). B. Enteral Administration

[0210] The dendrimer-active agent conjugate compositions may be administered enterally (oral, vaginal, rectal, buccal, nasal, pulmonary, or transdermally). The carrier or diluent may be a solid carrier such as a capsule or tablet or diluent for solid formulations, a liquid carrier or diluent for liquid formulations, or a mixture thereof.

[0211] For liquid preparations, pharmaceutically acceptable carriers can be, for example, aqueous or non-aqueous solutions, suspensions, emulsions or oils.Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, and injectable organic esters such as ethyl oleate.Aqueous carriers include, for example, water, alcoholic / aqueous solutions, cyclodextrins, emulsions or suspensions including saline and buffered media.

[0212] Examples of oils include those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, olive oil, sunflower oil, fish-liver oil, sesame oil, cottonseed oil, corn oil, olive, petrolatum, and inorganic. Fatty acids suitable for use in parenteral formulations include, for example, oleic acid, stearic acid, and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters.

[0213] Vehicles include, for example, sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's, and fixed oils. Formulations include, for example, aqueous and non-aqueous isotonic sterile injection solutions (which may contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient), and aqueous and non-aqueous sterile suspensions (which may contain suspending agents, solubilizers, thickeners, stabilizers, and preservatives). Vehicles may include, for example, fluid and nutrient supplements, electrolyte supplements (such as those based on Ringer's dextrose). In general, water, saline, aqueous dextrose, and related sugar solutions are preferred liquid carriers. They may also be formulated with proteins, fats, saccharides, and other ingredients of infant formula.

[0214] Oral preparations may be in the form of chewing gum, gel strips, tablets, capsules or lozenges. Encapsulating materials for preparing enteric-coated oral preparations include cellulose acetate phthalate, polyvinyl phthalate acetate, hydroxypropylmethylcellulose phthalate and methacrylic acid ester copolymers. Solid oral preparations such as capsules or tablets are preferred. Elixirs and syrups are also well-known oral preparations.

[0215] Formulations for administration to mucosal surfaces such as the nose, oral surfaces or lungs will usually contain pharmaceutically acceptable excipients such as those used for parenteral administration, alone or in combination with various surfactants, penetration enhancers, etc.

[0216] The present compositions can also be made into aerosol formulations (i.e., they can be "nebulized") to be administered by inhalation. Aerosol formulations can be placed into pressurized acceptable propellants, such as dichlorodifluoromethane, propane, nitrogen, and air. For administration by inhalation, the compounds are delivered in the form of an aerosol spray supply from a pressurized pack or nebulizer, using a suitable propellant. V. How to use

[0217] In preferred embodiments, the dendrimer cannabinoid compositions cross the barrier interfaces of the nervous system and selectively target specific cells and specific receptors on the cell surface to alleviate symptoms associated with various diseases, disorders and conditions. The method comprises administering to a subject in need thereof an amount of the composition effective to increase the permeability of the cannabinoid agent across the barrier interfaces of the nervous system and / or to increase the binding of the cannabinoid agent to specific receptors on specific cells, particularly CB1 and CB2 receptors on the surface of cells of the central and peripheral nervous system. A. Treatment Method

[0218] The compositions are suitable for treating disorders associated with neuropathic pain, inflammation, nausea and vomiting, spasticity, and behavioral abnormalities, particularly those that extend to the central and peripheral nervous systems. See, for example, 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.

[0219] Typically, an effective amount of a dendrimer complex comprising a combination of a dendrimer and one or more therapeutic, prophylactic and / or diagnostic active agents is administered to an individual in need thereof. The dendrimer may also include a targeting agent for delivery to target tissues and cells in the spinal cord, brain and related regions.

[0220] In some embodiments, the dendrimer complex comprises a drug bound to or conjugated to a dendrimer, which is capable of preferentially releasing the drug at the target receptor. The drug can be covalently bound or dispersed or encapsulated within the molecule. The amount of dendrimer complex administered to a subject is selected to deliver an amount effective to reduce, prevent, or otherwise alleviate one or more clinical or molecular symptoms of the disease or disorder being treated, compared to a control, for example, a subject treated with an active agent without a dendrimer. 1. Site-specific targeting

[0221] The present compositions and methods are designed to overcome the existing challenges in receptor-selective and cell-selective delivery of cannabinoid family drugs to the central and peripheral nervous systems and various organs of the body. Cannabinoid receptors are expressed in various cells and organs throughout the body and act as a system for regulating many aspects of human function. The present inventors utilize the small size of dendrimers and their unique biophysical properties to deliver these drugs to specific cells and receptors in the body. The present compositions and methods can enhance the bioavailability of drugs in the central and peripheral nervous systems by one or more of the following: (i) increasing drug levels at specific cells of interest and specific cannabinoid receptors on or within those cells that cross the brain barrier, particularly the blood-brain and blood-spinal fluid barriers; (ii) increasing drug solubility; (iii) promoting target engagement, i.e., increasing site-specific binding; (iv) improving pharmacokinetics; and (v) reducing off-target effects. For example, in some forms, the compositions and methods enable the selective delivery of compounds to the peripheral nervous system by minimizing drug uptake in the brain, thereby enhancing the potential of the compositions to be used to selectively treat peripherally specific diseases and disorders, including, but not limited to, neuropathic pain, local anesthesia, traumatic nerve injury, and hereditary and inflammatory neuropathies. a.Improved drug permeability through the barrier interface

[0222] The present dendrimer compositions and methods can improve the delivery of cannabinoids to specific receptors on the surface of neurons, microglia, or astrocytes by crossing one or more barrier interfaces in the brain and nervous system, particularly the blood-brain barrier (BBB), CSF-blood barrier, and blood-nerve barrier. These barrier interfaces normally protect neurons from blood-borne infectious agents and help maintain water homeostasis and an appropriate environment for neuronal function in the blood. Due to the clinical significance of cannabinoids, the present dendrimer compositions can be used to deliver cannabinoids with improved permeability across these barrier interfaces for site-specific targeting. i. Blood-brain barrier

[0223] The "blood-brain barrier" (BBB) ​​is a continuous endothelial membrane that, together with pericytes and other components of the neurovascular unit, restricts the entry of toxins, pathogens, and blood cells into the brain. However, the BBB also poses an obstacle to drug delivery to the central nervous system (CNS), in part because (1) delivery of drugs targeted to the brain via systemic routes can result in unacceptably high drug levels in the periphery, and (2) the complex interactions of cells and molecules that contribute to the structure and function of the BBB make it difficult to determine drug permeability across the BBB, drug distribution in the brain, and target engagement in the brain.

[0224] Brain microvascular endothelial cells, pericytes, astrocytes, tight junctions, neurons, and basement membrane structures physically stiffen brain capillaries at the BBB. Brain capillary endothelial cells lack fenestrae, limiting the diffusion of small molecules and proteins. Interendothelial junctions connect endothelial cells to form a continuous barrier, severely restricting the penetration of water-soluble substances. Pericytes, astrocytes, and basement membranes surround endothelial cells, ultimately forming the impermeable BBB. Furthermore, efflux transporters are located in brain capillary endothelial cells, further obstructing the entry of substances into the brain. BBB permeability is primarily controlled by interendothelial junctions, which are protein complexes including adherens junctions, tight junctions, and gap junctions. Adherens junctions primarily regulate the permeability of the endothelial barrier. Tight junctions play a key role in maintaining the permeability barrier between epithelial and endothelial cells, thereby controlling tissue homeostasis. Gap junctions, composed of six connexin molecules, direct electrical and chemical communication between endothelial cells. Finally, instead of having a static structure, the components of the BBB continuously adapt in response to various physiological changes in the brain. The dendrimer compositions and methods of the present application overcome the aforementioned challenges and are suitable for delivering cannabinoids across the blood-brain barrier by one or more of the transport mechanisms described above.

[0225] Molecules cross the BBB via the paracellular route (between adjacent cells) or the transcellular route (through cells). In the paracellular route, ions and solutes cross the BBB by passive diffusion, taking advantage of their concentration gradient. The transcellular route involves various mechanisms, such as passive diffusion, receptor-mediated transport, and transcytosis.

[0226] Physicochemical factors affecting BBB permeability include molecular weight, charge, lipid solubility, surface activity, and relative size of molecules. BBB permeability can also be affected by physiological factors such as efflux transporters, e.g., P-glycoprotein (P-gp), enzyme activity, plasma protein binding, and cerebral blood flow. Hydrophilic molecules, such as proteins and peptides, enter the brain via specific receptor- and saturable receptor-mediated transport mechanisms, such as glucose transporter-1 (GLUT-1), insulin transporter, and transferrin transporter. These endogenous transporters are expressed in the luminal and abluminal endothelial cell membranes. Among these transport mechanisms, receptor-mediated transcytosis has been extensively studied for delivering drugs to the brain. The dendrimer compositions and methods of the present application are suitable for delivering cannabinoids across the blood-brain barrier via one or more of the above-mentioned mechanistic pathways. ii. Blood-nerve barrier (BNB)

[0227] The blood-nerve barrier (BNB) defines a physiological space within which axons, Schwann cells, and other associated cells of peripheral nerves function, ensuring proper peripheral nerve function and maintaining homeostasis of the endoneurial environment. The BNB consists of endoneurial microvessels within nerve bundles and the surrounding perineurium. Tight junctions between endothelial cells and pericytes in the endoneurial vasculature isolate the endoneurial membrane from the blood, preventing uncontrolled leakage of molecules and ions from the circulatory system into peripheral nerves. Additionally, a diffusion barrier exists within the perineurium, formed by tight junctions between adjacent perineurial cells and the basement membrane surrounding each perineurial cell layer. Endoneurial capillaries and perineurial passages are strict barriers that isolate the endoneurial extracellular environment of peripheral nerves from both the epineurial perifascicular space and the systemic circulation, thus protecting the endoneurial microenvironment from dramatic concentration changes in the vascular space and other extracellular spaces.

[0228] For drug targets located in peripheral nerves, BNBs can be difficult to resolve, as they can restrict or prevent drugs from reaching their sites of action, thus negatively affecting drug efficacy.Furthermore, the expression profile of transporters in peripheral nerves can be very different from that in the central nervous system.The dendrimer compositions of the present application can be used to improve the permeability of cannabinoids through BNBs, thereby improving the delivery of cannabinoids to peripheral nerve targets. iii. Blood-CSF barrier

[0229] The composition may be used to improve the delivery of cannabinoids to target sites through the blood-cerebrospinal fluid barrier (blood-CSF barrier) and ventricles. The choroid plexus is a vascular tissue found in all ventricles. The functional unit of the choroid plexus is composed of capillaries surrounded by a layer of differentiated ependymal epithelium. Unlike the capillaries that form the blood-brain barrier, choroid plexus capillaries are fenestrated and do not have tight junctions. Therefore, the endothelium does not form a barrier to the movement of small molecules. Instead, the blood-CSF barrier in the choroid plexus is formed by epithelial cells and the tight junctions that connect them. The other part of the blood-CSF barrier is the arachnoid membrane that surrounds the brain. The cells of this membrane are also connected by tight junctions.

[0230] The CSF space and brain structures adjacent to the CSF compartment are pathological targets of interest in CNS disorders. For example, the subarachnoid perivascular or periventricular space is an area where pathogenic lymphocytes, monocytes, and neutrophils accumulate in neuroinflammatory disorders such as multiple sclerosis and related experimental autoimmune encephalitis or viral-induced neuropathies (including neuroAIDS and CMV infection). Nests of potentially harmful antibody-producing B cells found in various CNS autoimmune diseases are thought to be primarily located in the pia mater. Therefore, in some forms, the dendrimer compositions can be used to deliver cannabinoids to areas of interest via the blood-CSF space connected to deep cervical lymph nodes to ameliorate or treat symptoms associated with neuroinflammatory disorders.

[0231] In some embodiments, dendrimer compositions can be used to deliver cannabinoids to target sites to improve or treat the symptoms and conditions associated with vascular degeneration.For example, cerebral amyloid angiopathy induces degenerative vascular changes driven by the deposition of amyloid beta (Aβ) peptide, cystatin c, transthyretin or gelsolin around penetrating blood vessels.The deposits are accessible from the interconnected CSF / perivascular space.

[0232] In some embodiments, 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 myeloma, drug-resistant ependymoma, and leptomeningeal tumor metastases derived from peripheral primary tumors, all directly contact CSF.As a result of the reduced effectiveness of tight junctions, which allow contrast enhancement in magnetic resonance imaging, the blood-tumor barrier is often considered leaky.However, many cannabinoids are lipophilic and are prevented from crossing the BBB by multidrug resistance (MDR) efflux proteins that control the transcellular pathway.In some periventricular tumors, such as ependymoma, MDR proteins are still fully expressed in the blood-tumor barrier.Therefore, in some embodiments, dendrimer compositions can be used to utilize pharmacological pressure from CSF to achieve therapeutic concentrations of cannabinoids in tumor tissue. b. Improved target-specific binding

[0233] The dendrimer compositions may be used to deliver cannabinoids with high 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 , the dendrimer compositions may be used to deliver cannabinoids that can selectively affect target cells and specific receptors on the target cell surface while being cleared from non-target organs, including, but not limited to, nervous system cells and glial cells in the brain, spinal cord, heart, lungs, kidneys, gastrointestinal tract, and joints. Improved binding to specific receptors on the target cell surface may enable conjugates to achieve rapid onset of efficacy at lower doses. The degree of binding to the target receptor can be controlled by the chemical nature of the drug linkage to the dendrimer. In exemplary embodiments, hydroxyl-terminated PAMAM dendrimers, glucose dendrimers, and other dendrimers may enhance the delivery of cannabinoids specific to specific receptors, such as CB1-specific, CB2-specific, and / or both, thereby enabling enhanced 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.

[0234] In some forms, specific ligands, such as AEA and 2-AG, can be conjugated to dendrimer compositions for the cell type and receptor type specific modulation of endocannabinoid system.The dendrimer compositions can also be used to target specific metabolites, such as intracellular enzymes, in nervous system cells and glial cells to modulate the synthesis and degradation of endocannabinoid ligands.For example, dendrimer compositions can be used to target PLA and DAG to modulate the synthesis of endocannabinoid ligands, and FAAH and MLGL to modulate the degradation of endocannabinoid ligands.

[0235] In some forms, the dendrimer compositions may be engineered to deliver cannabinoids by other routes to modulate cellular activity in the brain and nervous system, for example, the dendrimer compositions may be engineered to deliver cannabinoids by random diffusion, aggregation into membranes, and lateral diffusion, and then bind to membrane-embedded ligand binding sites. i. The endocannabinoid system in the CNS

[0236] The ECS has emerged as one of the key regulatory mechanisms in the brain, controlling multiple events, including mood, pain perception, learning, and memory, among others. It is also thought to play a neuroprotective role during traumatic brain injury (TBI) and may be part of the brain's natural compensatory repair mechanism during neurodegeneration. Given the clinical importance of the ECS in drug abuse and addiction, the present compositions and methods are suitable for targeting components of the ECS to treat cannabis and opioid abuse. In the CNS, eCB acts as a retrograde messenger mediating feedback inhibition, modulating synaptic plasticity. Activation of CB1 receptors leads to activation of inwardly rectifying K+ channel conductance, a decrease in N-type and P / Q-type voltage-gated Ca2+ channel conductance, and eCB production. This leads to a decrease in neurotransmitter release at excitatory and inhibitory synapses, resulting in transient effects such as depolarization-induced disinhibition (DSI) and depolarization-induced deexcitation (DSE) during synaptic plasticity, or sustained effects such as long-term depression and activation (LTP / LTD). These events make the ECS a key modulator of synaptic plasticity.

[0237] Chronic exposure to CB1 receptor agonists results in a rapid decline in behavioral responses, termed tolerance, which is attributed to both a decreased ability of the receptor to activate effector pathways (i.e., desensitization) and a decrease 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 in the GPCR cytoplasmic domain, which then become high-affinity targets for β-arrestin. β-arrestin binding uncouples the G protein, stimulating receptor internalization and β-arrestin-mediated signaling.

[0238] Ligand-induced receptor phosphorylation by GRKs can yield highly specific and distinct phosphorylation profiles or "barcodes." These barcodes can be finely tuned to define which signaling cascades are activated, thus expanding the range of possibilities often defined as functional group selectivity or ligand bias.

[0239] The interaction of receptors with β-arrestins and the signaling cascade depend on specific phosphorylation sites regulated by intrinsic GRKs. Mutation of the putative GRK sites S426 / S430 to alanine results in reduced β-arrestin2 recruitment and receptor internalization, but enhances interaction with β-arrestin1, resulting in increased β-arrestin1-mediated signaling. β-arrestin-mediated signaling from such polarized receptors controls the activation of several cascades, including ERK1 / 2, JNK1 / 2 / 3, CREB, and P38α. It is important to note that these cascades are already linked to CB1 receptor activation. Activation of these cascades by CB1 receptors and β-arrestins then regulates gene expression and protein synthesis.

[0240] The compositions and methods can be used to elucidate the physiological role of β-arrestins in the development of pathway-selective or "biased ligands" with greater therapeutic benefit. The compositions and methods can also be used to examine signaling from biased CB1 receptors, such as S426A / S430A, aid in the identification of biased ligands, and provide an important tool for elucidating the mechanisms and roles of CB1 receptor signaling.

[0241] The intracellular localization and trafficking of CB1 receptors are highly dynamic and have a profound impact on receptor signaling. CB1-G protein-mediated signaling occurs at the cell surface and in intracellular compartments. At the cell surface, CB1 receptor ligands modulate the interaction between the receptor and β-arrestin as a mechanism for influencing β-arrestin-mediated signaling. This interaction can initiate at the plasma membrane and continue into intracellular compartments. Interestingly, these location-specific signaling events appear to extend to several GPCRs. For example, the LH receptor, β2-adrenergic receptor, and CB2 receptor transmit signals from intracellular compartments via "supercomplexes" via either β-arrestin or G proteins, ultimately resulting in three distinct spatiotemporal signaling waves. Constitutive activation also plays a role in their trafficking. The location and trafficking of CB1 receptors are highly dynamic events closely linked to their signaling. ii. Regulation of the endocannabinoid system

[0242] The present compositions and methods can be used to pharmacologically manipulate eCB levels or their functions with allosteric modulators, thereby improving opportunities to regulate 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. Primary eCBs are rapidly inactivated by reuptake mechanisms and degradative enzymes, including fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase. Among eCBs, derivatives of arachidonic acid, such as AEA and 2-AG, are predominant and orthosteric. These ligands are agonists of both CB1 and CB2 receptors, but bind to the CB1 receptor with higher affinity (AEA has a Ki of 89 nM and 321 nM for CB1 and CB2 receptors, respectively, and 2-AG has a Ki of 472 nM and 1400 nM for CB1 and CB2 receptors, respectively; Pertwee et al., 2010). Allosteric eCBs, including pregnenolone and lipoxin A4, have been identified that modulate CB1 receptor signaling and may have therapeutic value (Pamplona et al., 2012; Vallee et al., 2014; Pertwee, 2015). The present compositions and methods can be used to aid in the pharmacological characterization of orthosteric and allosteric modulators and to clearly elucidate their physiological roles and modes of action. In some forms, the present compositions and methods can be used to pharmacologically manipulate eCB levels or their effects with allosteric modulators, thereby improving opportunities to regulate the ECS. iii. Improved cannabinoid receptor binding

[0243] The present dendrimer compositions can improve binding to one or both cannabinoid receptors and modulate signal transduction in a cell- and tissue-specific manner.

[0244] The identification and cloning of the two major cannabinoid receptors (CB1 and CB2), along with the discovery of their endogenous ligands, in the late 1980s and early 1990s led to extensive efforts aimed at understanding the mechanisms and physiological roles of the endocannabinoid system (ECS). Due to the expression and localization of both receptors in the central nervous system (CNS), the CB1 receptor (along with its endogenous ligand, endocannabinoid (eCB))) and the enzymes involved in their synthesis and degradation have been implicated in multiple pathophysiological events ranging from memory deficits to neurodegenerative disorders, among others. For example, CB1 receptors are particularly abundant in the frontal cortex, hippocampus, basal ganglia, hypothalamus, cerebellum, spinal cord, and peripheral nervous system. Thus, in some forms, the present dendrimer compositions may be used to deliver cannabinoids to ameliorate or treat symptoms of diseases and disorders affecting one or more of these nervous system regions.

[0245] CB1 receptors are also present in both inhibitory GABAergic neurons and excitatory glutamatergic neurons. Therefore, in some embodiments, the present compositions may be used to improve or treat symptoms of diseases and disorders associated with dysregulated GABAergic and glutamatergic signaling. CB2 receptors are most abundantly found in immune system cells, hematopoietic cells, and glial cells. Under normal, healthy conditions, CB2 receptors are primarily expressed in the periphery. In disease or injury, such upregulation occurs in the brain, and thus CB2 receptors are expressed in the brain under unhealthy conditions. Therefore, in some embodiments, the present compositions may be used to improve or treat symptoms of diseases and disorders associated with damaged immune cells, hematopoietic cells, and glial cells, such as macrophages and activated microglia. The neurophysiology of CB1 and CB2 receptors is described in more detail below. CB1 receptor

[0246] CB1 receptors are one of the most abundant G protein-coupled receptors (GPCRs) in the CNS, with particularly high levels found in the neocortex, hippocampus, basal ganglia, cerebellum, and brainstem. CB1 receptors are also found in peripheral nerve endings and several extraneuronal sites, such as the testes, eyes, 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 their location on the cell surface, the subcellular localization of CB1 receptors has also been reported in heterogeneous systems and primary cultures. CB1 receptors bind Δ9-tetrahydrocannabinol (Δ9-THC), the primary active component of Cannabis sativa, and mediate most of Δ9-THC's CNS effects. Additionally, CB1 receptors bind synthetic cannabinoid-mimetic compounds, such as CP55940, JWH-015, and WIN55212-2, as well as the endogenous arachidonic acid derivatives arachidonylethanolamine (AEA) and 2-arachidonylglycerol (2-AG). Upon ligand binding and receptor activation, CB1 receptors primarily couple to pertussis toxin (PTX)-sensitive G-type G proteins, which rapidly reduce cAMP levels by inhibiting adenylate cyclase activity. Binding to other G proteins, including G, can also stimulate adenylate cyclase, albeit with lower potency, but the extent of cAMP accumulation is not necessarily a good indicator of G protein coupling. Evidence of contingent binding to different G proteins, the role of β-arrestin-mediated signaling, and signaling from intracellular compartments adds another level of complexity, making these receptors, like other GPCRs, multidimensional. CB1 receptors exhibit constitutive activation in the absence of agonist, indicative of G protein activation, which may mediate highly polarized localization to axonal and presynaptic compartments.

[0247] Figures 1A and 1B are schematic diagrams of key enzymes in the cannabinoid pathway. Diacylglycerol lipase (DGLα) and phospholipase D (PLD) produce the endogenous ligands arachidonylethanolamine (AEA) and 2-arachidonylglycerol (2-AG; Figure 1A). These activate the cannabinoid 1 receptor (CB1) in the central nervous system (CNS). The consequences can include modulation of cAMP accumulation, voltage-gated calcium channels (VGCCs), K+ channels, and adenylate cyclase activity at presynaptic excitatory and inhibitory synapses, inhibiting neurotransmitter release (Figure 1A). After CB1 receptor activation by ligand binding, signaling via G proteins and / or β-arrestins can occur at the plasma membrane in endocytic pits or endosomes after receptor internalization (Figure 1B). While G proteins normally bind to unphosphorylated receptors, β-arrestins bind to receptors phosphorylated by G protein receptor kinases (Figure 1B). Figure 1C is a schematic representation of the impact of various cannabinoid (CB) receptor signaling modalities on neuroregulation in health and disease in specific pathways. b.CB2 receptor

[0248] CB2 receptors exhibit a more distinct expression pattern in the brain than CB1 receptors and are found primarily 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, resident macrophages of the CNS (Mackie, 2008; Palazuelos et al., 2009). This selective localization, along with the regulatory effects of CB2 receptors on microglial function, is particularly relevant because microglial cells play an important role in Alzheimer's disease (AD) and other disorders involving the basal ganglia (Yeh et al., 2016). Furthermore, CB2 receptors expressed on neurons can regulate 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 blocks dopaminergic firing from the ventral tegmental area and reduces cocaine self-administration (Zhang et al., 2016). Furthermore, neuronal CB2 receptors act independently of CB1 receptors to modulate inhibitory plasticity and gamma oscillations in the CA2 / 3 region of the hippocampus in vivo (Stempel et al., 2016). iv. Improved non-cannabinoid receptor binding

[0249] The dendrimer composition can improve the binding of cannabinoids to one or more non-cannabinoid receptors, modulating signal transduction 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, and GPR12 receptors; transient receptor potential channels, such as TRP vanilloids TRPV1 to TRPV4, TRP ankyrins TRPA1 and TRPM members TRPM8 receptors; peroxisome proliferator-activated receptors, such as PPAR2 and PPARγ; monoamine transporters, such as norepinephrine, dopamine, and serotonin 1A receptors, fatty acid amide hydrolase, monoacylglycerol lipase, transport fatty acid binding protein, adenosine equilibrative nucleoside transporter, and glycine receptor α1 and α3 receptors. Thus, in some forms, the present dendrimer compositions can be used to improve binding of cannabinoids to one or more non-cannabinoid receptors to modulate signal transduction at one or more of the receptors listed above. B. The condition to be treated

[0250] This composition is suitable for treating one or more diseases, conditions and injuries in the central nervous system and peripheral nervous system.This composition can also be used to treat various diseases, disorders and injuries, including gastrointestinal disorders, eye diseases, and other tissues in which nerves play a role in disease or injury.Cannabinoids have been tested for the treatment of indications including chemotherapy-induced nausea and vomiting, appetite stimulation in HIV / AIDS, chronic pain, spasticity due to multiple sclerosis or paraplegia, depression, anxiety disorders, sleep disorders, psychiatric disorders, glaucoma or Tourette's syndrome.

[0251] The present composition and method are also suitable for prophylactic use.For example, the present composition can be administered to patients who need it to improve, treat or prevent the symptoms associated with inflammatory disease, neurodegenerative disease, pain disorder, convulsive and spasmodic disorders, bone disorder, psychiatric disorder, metabolic disorder and cancer-related neuropathic pain.The present composition is particularly useful for treating or improving one or more of the following complications associated with chemotherapy, such as nausea and vomiting; HIV / AIDs symptoms, such as appetite loss; multiple sclerosis-related symptoms, such as spasticity; and sleep disturbance and / or sleep disorder, anxiety disorder, psychosis, chronic pain, glaucoma and / or Tourette's syndrome.

[0252] deliver therapeutic, prophylactic or diagnostic agents, preferably less than 20 nm in diameter and at least 0.8 OH groups / nm 2 hydroxyl group surface density, preferably less than 10 nm in diameter and at least 1 OH group / nm 2 hydroxyl group surface density of 0.01, more preferably less than 6 nm in diameter and at least 1 OH group / nm 2 and most preferably a diameter of between 5 nm and at least 1.5 OH groups / nm 2 Dendrimer conjugate compositions having a hydroxyl group surface density of 0.05% selectively target microglia and astrocytes, which play an important role in the pathogenesis of numerous disorders and conditions, including neurodevelopmental disorders, neurodegenerative disorders, neuropsychiatric disorders, and chronic pain. Thus, the dendrimer complexes are administered in dosage units effective to treat or alleviate conditions associated with pathologies in the central and peripheral nervous systems. For example, the dendrimer complexes are administered in dosage units effective to treat or alleviate conditions associated with pathologies affecting neuronal microglia and astrocytes. Generally, by targeting these cells, the dendrimers specifically deliver agents to treat neuroinflammation. Relief from chronic pain

[0253] The compositions and methods are suitable for treating chronic pain and diseases and disorders associated with chronic pain.

[0254] 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 be resistant to the powerful pain-relieving effects of opioid analgesics. Chronic pain conditions are often resistant to standard treatments, including treatment with opioid medications such as morphine.

[0255] Cannabinoids exert their effects primarily through Gi / o-protein-coupled cannabinoid CB1 and CB2 receptors, which are expressed throughout the nervous system. CB1 receptors are found at key nodes along pain pathways, and their activity regulates both 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 increases significantly, modulating the activity of these central and peripheral immune cells with important consequences for peripheral pain circuits.

[0256] The present compositions and methods can be used to target CB1 or CB2 receptors to manage pain and improve pain outcomes in 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 present compositions and methods can be used to manage neuropathic 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 present compositions and methods modulate microglial function to promote cannabinoid-mediated analgesia through microglia-neuron interactions within the nociceptive circuitry of the spinal cord. In another exemplary embodiment, the present compositions and methods can be administered in a complementary regimen with an opioid to provide additional pain relief in addition to the regimen provided by the opioid. In these forms, combination therapy for chronic pain can have the benefit of reducing the patient's pain level, reducing their dependence (and addiction) to opioids, or both. b. Mental health disorders and conditions

[0257] The compositions and methods are suitable for treating a variety of mental health disorders and conditions, including, but not limited to, affective or mood disorders, anxiety disorders, childhood disorders, eating disorders, personality disorders, schizophrenia and other psychotic disorders, and substance-related disorders. i. Affective or mood disorders

[0258] Affective or mood disorders are described by marked disruptions of emotion (severe low spirits called depression or severe elevated mood called hypomania or mania). These include bipolar disorder, cyclothymia, hypomania, major depressive disorder, severe mood dysregulation disorder, persistent depressive disorder, premenstrual dysphoric disorder, seasonal affective disorder, depression associated with a medical illness, and depression induced by substance use or medication. ii. Anxiety disorders

[0259] Anxiety disorders involve excessive fear or apprehension as opposed to normal feelings of tension or apprehension, and include generalized anxiety disorder, panic disorder, social anxiety disorder, and various phobia-related disorders.

[0260] Generalized anxiety disorder (GAD) usually involves persistent feelings of anxiety or fear, which can interfere with daily life. It is not the same as occasionally worrying about things or feeling anxious due to stressful life events. People living with GAD experience frequent anxiety for months, sometimes even years. The composition and method are suitable for treating one or more symptoms of GAD, including but not limited to restlessness, fatigue, difficulty concentrating, irritability, headache, muscle pain, stomach pain, or pain of unknown origin, excessive worry, and sleep problems, such as difficulty falling asleep or staying asleep.

[0261] Panic disorder is an anxiety disorder characterized by unexpected and repeated episodes of intense fear accompanied by physical symptoms, which can include chest pain, palpitations, shortness of breath, dizziness or abdominal distress, or a sense of losing control even when there is no obvious danger or trigger.Persons with panic disorder often worry about when the next attack will occur, and actively try to prevent future attacks by avoiding the places, situations or actions that are associated with panic attacks.Panic attacks can occur frequently, such as several times a day, or infrequently, such as several times a year.The present composition and method are suitable for treating one or more symptoms of panic attack, including but not limited to palpitations, excessive sweating, trembling or tingling, chest pain, and difficulty in controlling emotions, such as feelings of impending doom and uncontrollable emotions.

[0262] Social anxiety disorder is an intense and persistent fear of being observed and judged by others.For people with social anxiety disorder, the fear of social situations can be so intense that it seems uncontrollable.For some people, this fear can prevent them from going to work, going to school or doing everyday things.The composition and method are suitable for treating one or more symptoms of social anxiety disorder, including but not limited to excessive blushing, sweating or trembling, palpitations, stomach pain, rigid body posture or speaking in an excessively low voice, and self-consciousness or fear of negative judgment.

[0263] Phobia is the strong fear or aversion to certain objects or situations.In some situations, feeling anxiety may be realistic, but the fear that phobia people feel is disproportionate to the actual danger caused by that situation or object.The composition and method are suitable for treating one or more symptoms of phobia, including but not limited to the irrational or excessive worry about encountering the object or situation of fear, the strong immediate anxiety when encountering the object or situation of fear, and the endurance of unavoidable objects and situations with strong anxiety. iii. Eating disorders

[0264] Eating disorders are serious, often fatal, illnesses that involve severe disturbances in a person's eating behavior and related thoughts and emotions. Eating disorders involve preoccupations with food, weight, and body shape. Common eating disorders include anorexia nervosa, bulimia nervosa, and binge eating disorder.

[0265] Anorexia nervosa is a condition in which a person avoids food, severely restricts food, or eats only small amounts of certain foods. They may repeatedly weigh themselves. They may view themselves as overweight even when dangerously thin. There are two subtypes of anorexia nervosa: the restrictive subtype and the bulimic subtype. People with the restrictive subtype of anorexia nervosa severely restrict the amount and type of food they consume. People with the bulimic subtype of anorexia nervosa also severely restrict the amount and type of food they consume. In addition, they may have episodes of binge eating and vomiting, eating a large amount of food in a short period of time followed by vomiting or using laxatives or diuretics to expel the food.

[0266] Symptoms of anorexia nervosa include, but are not limited to, thinning of bones (osteopenia or osteoporosis), mild anemia, and muscle wasting and weakness, brittle hair and nails, dry, yellowish skin, thin hair growth (vellus hair) all over the body, severe constipation, low blood pressure, slowed breathing and pulse rate, damage to the structure and function of the heart, brain damage, multiple organ failure, a drop in internal body temperature, feeling constantly cold, lethargy, sluggishness or constant fatigue, and infertility.

[0267] Bulimia nervosa is a condition in which a person has repeated and frequent episodes of eating abnormally large amounts of food and feels unable to control these episodes. This binge eating is followed by behaviors to counteract the binge eating, such as compulsive vomiting, excessive use of laxatives or diuretics, fasting, excessive exercise, or a combination of these behaviors. People with bulimia nervosa may be slightly underweight, normal weight, or overweight. Symptoms of bulimia nervosa include chronic inflammation and pain in the throat, swelling of the salivary glands in the neck and jaw area, worn tooth enamel and increased tooth sensitivity and tooth decay as a result of exposure to stomach acid, acid reflux and other gastrointestinal problems, intestinal discomfort and irritation due to laxative abuse, severe dehydration due to fluid expulsion, and electrolyte imbalances (too low or too high levels of sodium, calcium, potassium, and other minerals) that can lead to stroke or heart attack.

[0268] This composition is particularly useful for stimulating appetite and reducing nausea and vomiting in patients who need it. For example, loss of appetite is a common symptom in patients with opportunistic infections, such as HIV / AIDS, and patients undergoing cancer chemotherapy and / or other treatments. Opportunistic infections are infections that people with weakened immune systems are prone to. In addition, some medications can reduce appetite due to side effects such as tingling around the mouth, nausea, and changes in taste. In addition, depression caused by terminal illness and / or the treatment of terminal illness can cause loss of appetite. Therefore, in some forms, this composition can be administered to such patients to stimulate appetite and / or alleviate nausea or wasting syndrome. c. Neurological and neurodegenerative diseases

[0269] The compositions and methods are suitable for treating symptoms associated with neurological and neurodegenerative diseases.

[0270] Neurodegenerative diseases are chronic, progressive disorders of the nervous system that affect neurological and behavioral functions and involve biochemical changes that result in distinct histopathological and clinical syndromes (Hardy H, et al., Science. 1998;282:1075-9). Abnormal proteins that resist cellular degradation accumulate within cells. The pattern of neuronal loss is selective, in the sense that one group is affected while others remain intact. In many cases, there is no clear precipitating event for the disease. Diseases traditionally described as neurodegenerative include Alzheimer's disease, Huntington's disease, and Parkinson's disease.

[0271] Neuroinflammation, mediated by activated microglia and astrocytes, is a major hallmark of various neurological disorders and represents a potential therapeutic target (Hagberg, H et al., Annals of Neurology 2012, 71, 444; Vargas, DL et al., Annals of Neurology 2005, 57, 67; and Pardo, CA et al., International Review of Psychiatry 2005, 17, 485). Several scientific reports suggest that targeting these cells early in the disease process can reduce neuroinflammation, potentially delaying disease onset and providing a longer therapeutic window for treatment (Dommergues, MA et al., Neuroscience 2003, 121, 619; Perry, VH et al., Nat Rev Neurol 2010, 6, 193; Kannan, S et al., Sci. Transl. Med. 2012, 4, 130ra46; and Block, ML et al., Nat Rev Neurosci 2007, 8, 57). Delivering therapeutic agents across the blood-brain barrier (BBB) ​​is challenging. Neuroinflammation causes disruption of the BBB. BBB damage in neuroinflammatory disorders can be exploited to transport drug-loaded nanoparticles through the brain (Stolp, HB et al., Cardiovascular Psychiatry and Neurology 2011, 2011, 10; and Ahishali, B et al., International Journal of Neuroscience 2005, 115, 151).

[0272] The present compositions and methods can also be used to deliver active agents for treating neurological or neurodegenerative diseases, or neurological or neurodegenerative disorders, or central nervous system disorders.In a preferred embodiment, the present compositions and methods are effective for treating and / or improving the neuroinflammation associated with neurological or neurodegenerative diseases, or neurological or neurodegenerative disorders, or central nervous system disorders.The present methods generally include administering an effective amount of the composition to the subject to increase cognition or reduce cognitive decline, increase cognitive function or reduce cognitive decline, increase memory or reduce memory decline, increase learning ability or learning capacity, or reduce decline in learning ability or learning capacity, or a combination thereof.

[0273] 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 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-associated cognitive impairment, mild cognitive impairment, motor neuron disease (MND), spinocerebellar degeneration (SCA), spinal muscular atrophy (SMA), Friedreich's ataxia, Lewy body disease, Alpers' disease, Batten disease, cerebro-oculofacial syndrome, corticobasal degeneration, Gerstmann-Straussler-Scheinker syndrome, kuru disease, Leigh's disease. It can be used to treat subjects with diseases or disorders such as unilateral upper limb muscular atrophy, multiple system atrophy, multiple system atrophy with orthostatic hypotension (Shy-Drager syndrome), multiple sclerosis (MS), neurodegeneration with cerebral 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 syndrome, post-encephalitic dementia, cancer- and chemotherapy-associated cognitive impairment and cancer- and chemotherapy-associated dementia, and depression-induced dementia, and pseudodementia.

[0274] In some embodiments, the subject has an excitotoxicity disorder. Excitotoxicity is a process in which nerve cells become damaged due to overstimulation. Several conditions, including stroke, traumatic brain injury, multiple sclerosis, amyotrophic lateral sclerosis, Alzheimer's disease, and spinal cord injury, are linked to excitotoxicity. Damage to nerve cells leads to corresponding neurological symptoms, which can vary depending on which cells are damaged and how extensive the damage is. Once damaged, nerve cells cannot be repaired, and patients may suffer permanent damage. Several drugs have been developed and used in an attempt to interfere with, affect, or temporarily stop the glutamate excitotoxicity cascade leading to nerve damage. One strategy is to attempt "upstream" to reduce glutamate release. Therefore, in some embodiments, the dendrimer complex comprises one or more active agents for treating excitotoxic disorders.

[0275] In some embodiments, the subject has a nervous system disorder or is in need of neuroprotection. Exemplary conditions and / or subjects include, but are not limited to, subjects who have suffered from, have had, or are at risk of developing, or are suffering from a stroke, traumatic brain injury, spinal cord injury, post-traumatic stress syndrome, or a combination thereof.

[0276] In some embodiments, the compositions and methods are administered to a subject in need thereof in an amount effective to reduce or prevent one or more molecular or clinical symptoms of neurodegenerative disease, or one or more mechanisms that cause neurodegeneration.Exemplary neurological and neurodegenerative disorders are further described below. i.Multiple sclerosis (MS)

[0277] The present compositions and methods may be useful for treating MS and / or alleviating symptoms associated with MS. MS is a major immune-mediated neurodegenerative disease characterized by demyelination accompanied by axonal and neuronal loss. Cannabis, Δ9-THC, or other CB agonists improve spasticity, convulsions, and pain, among other symptoms of MS (Croxford, 2003; Pertwee, 2007; Rog, 2010; Notcutt et al., 2012). The use of Sativex® (nabiximols), an oromucosal spray of cannabis extract containing fixed concentrations of Δ9-THC and cannabidiol (CBD), has resulted in symptomatic improvement in patients with MS. A meta-analysis of several clinical trials has observed a reduction in motor dysfunction and pain. Therefore, in some forms, the present compositions may be administered to patients in need thereof to reduce muscle spasms and pain associated with multiple sclerosis. In some forms, the compositions may be administered to a patient in need thereof to reduce fatigue associated with inflammation, which may result in improved exercise capacity in MS patients.

[0278] At the molecular level, these improvements are generally linked to the activation of both CB1 and CB2 receptors by agonists, resulting in their dual anti-inflammatory and neuroprotective effects throughout the CNS. These effects include upregulation of proinflammatory molecules such as interleukins in astroglia and reduction of cytotoxic factors such as nitric oxide, reactive oxygen species, and proinflammatory cytokines in microglia. The precise mechanism by which the receptors exert their neuroprotective activity may involve activation of the phosphatidylinositol 3-kinase / mammalian target of rapamycin complex 1 (mTOR1) pathway and brain-derived neurotrophic factor (BDNF).

[0279] Synthetic CB1 receptors attenuate inflammation and neuropathic pain in a mouse model of experimental autoimmune encephalomyelitis (EAE). Similar results were observed in mice with established Theiler's murine encephalomyelitis virus-induced demyelinating disease, a mouse model of chronic progressive MS, after systemic treatment with the agonists WIN55212-2, ACEA, and JWH-015. The mice's motor function improved by modulating the infiltration of microglia and lymphocytes into the spinal cord. In contrast, application of an inverse agonist of the CB1 receptor (SR141716A) exacerbated EAE in the mouse brain and spinal cord, likely through the release of proinflammatory cytokines. Collectively, these results suggest that the neuroprotective role of CB1 and CB2 receptors may be impaired in MS, and that enhancing these receptors may provide a novel therapeutic approach.

[0280] Thus, the present compositions and methods may be 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 difficulty speaking and swallowing. ii. Huntington's disease (HD)

[0281] Dysregulation of the ECS has also been reported in experimental models and patients with HD. CB1 receptor expression is reduced, at least to some extent, in mice before the onset of neurodegenerative HD symptoms (e.g., a 27% decrease in CB1 receptor mRNA in the striatum) (McCaw et al., 2004). Loss of CB1 receptor expression reduces motor function and increases the amount of CB1 receptor aggregates in the striatum of HD mice (Mievis et al., 2011). Significant loss of CB1 receptors has also been reported in patients with HD (Glass et al., 2000). Activation of CB1 receptors may help alleviate the progression of HD. For example, preclinical evidence suggests the use of CBs, such as Sativex®, for neuroprotection in patients with progressive neurodegenerative conditions such as HD (Valdeolivas et al., 2012). Furthermore, selective receptor agonists have the potential to be neuroprotective in cell culture models of HD (Scotter et al., 2010; Laprairie et al., 2016). Ligands that bias β-arrestin-mediated signaling, such as Δ9-THC, reduce cell function and viability in these models, suggesting a potential pharmacological profile for therapeutic agonists (Laprairie et al., 2014, 2016). These events are mediated in part by activation of Gαi / o-mediated pathways, which can limit glutamate release from cortical neurons and GABA release from striatal medium spiny neurons (Dowie et al., 2010; Laprairie et al., 2016). Results from the R6 / 2 mouse model of HD indicate that CB1 receptor activation, paralleling BDNF expression, confers neuroprotection (Blázquez et al., 2015). In general, in vivo and in vitro data suggest that CB agonists with specific pharmacological profiles (biased towards BDNF upregulation and release) may be developed to treat or ameliorate HD.

[0282] Thus, the present compositions and methods may be effective in reducing the progression of HD in a subject by increasing neuroprotection in the subject through one or more of the above mechanisms. The present compositions and methods may also be effective in treating and / or alleviating one or more symptoms of HD, including, but not limited to, neuroinflammation, pain, involuntary spasms or writhing movements (chorea), muscle problems (such as stiffness or muscle contractures (dystonia)), slow or abnormal eye movements, gait, posture and balance disorders, difficulty speaking or swallowing, cognitive impairment and behavioral abnormalities. iii. Alzheimer's disease

[0283] The CB1 receptor has also been the focus of intense research as a potential target in AD. This research has been conducted in vitro, in animal models, and in postmortem samples. Although altered expression levels of several components of the ECS have been identified in postmortem samples from AD patients, the role of these components in the pathophysiology of the disorder remains unknown. For example, CB1 receptors in the hippocampus from patients with AD were not different from those in age-matched controls. However, levels of MAGL, the enzyme that degrades 2-AG, were reduced in these patients at the site of action, suggesting alterations in eCB signaling and architecture (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). Analysis of available studies and trials suggests significant benefits from synthetic CBs for some behavioral and psychological symptoms of dementia (Liu et al., 2015). However, these conclusions are based on short-term and limited studies. Further studies are needed to evaluate the safety and efficacy of CBs in AD. In experimental models of AD, several findings suggest that activation of both CB1 and CB2 receptors may have beneficial effects, primarily through neuroprotection against Aβ toxicity, as previously noted for other neurodegenerative disorders (Stumm et al., 2013). Tau protein phosphorylation is also reduced by CBD in PC12 cells, providing a distinct neuroprotective mechanism during AD (Esposito et al., 2006). Because CB1 receptors are unlikely to be directly activated by CBD, the effect on tau phosphorylation may be due to CBD's antioxidant effects or, perhaps, a CB receptor-independent effect. The reduction of harmful β-amyloid peptides and tau phosphorylation may occur constitutively through activation of the immune and CNS CB systems in AD, while promoting intrinsic CNS repair mechanisms (Aso and Ferrer, 2014).For example, recent investigations into TREM2 receptors in microglia, where CB2 receptors are expressed and regulate cellular responses, also suggest that there are immune-related mechanisms regulating AD ( Yeh et al., 2016 ).

[0284] Aging is a major risk factor for neurodegenerative diseases, and the proliferation of neuronal progenitor cells is significantly reduced during this process. Notably, CBs can stimulate fetal and adult neurogenesis (Jiang et al., 2005; Trazzi et al., 2010). Axon guidance, cell migration, synaptogenesis, and cell survival are also modulated during development. Dysregulation of these processes during development and aging may significantly contribute to multiple disorders of the CNS. iv. Traumatic brain injury

[0285] CB1 / 2 receptors are involved in TBI, and in animal models, 2-AG is increased after TBI. Following TBI, there is an "on-demand" signal that generates eCB, which can reduce brain edema and inflammation. These events can be neuroprotective, preventing excitotoxicity, inhibiting inflammatory cytokine production, and increasing stem cell migration and differentiation. Furthermore, CB1 and CB2 receptor antagonists prevent drug-induced neuroprotection in mouse models of TBI. However, as has been shown in other disorders, limited clinical data are available supporting the efficacy and safety of CB during TBI. Therefore, the present 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, headache, sensory impairment, and sleep abnormalities. d. Gastrointestinal disorders

[0286] The endocannabinoid system (i.e., endogenous circulating cannabinoids) exerts protective activity in the GI tract.Therefore, the present compositions and methods can be effective in treating and / or alleviating various GI conditions, such as necrotizing enterocolitis (NEC), abdominal sepsis, pneumonia, arthritis, pancreatitis and atherosclerosis, inflammatory bowel disease (especially Crohn's disease), irritable bowel syndrome, and secretory and motility-related disorders.The present compositions and methods can also be effective in treating and / or alleviating one or more symptoms of gastrointestinal dysfunction in the gastrointestinal tract, liver, and pancreas, including, but not limited to, nausea and vomiting, cannabinoid hyperemesis syndrome, anorexia, weight loss, and chronic abdominal pain.

[0287] In some embodiments, a single dendrimer complex composition can simultaneously treat and / or diagnose multiple conditions in two separate 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 treat a region of the gastrointestinal tract by enteral administration while selectively targeting microglia and astrocytes after absorption into the bloodstream. Microglia and astrocytes play important roles in the pathogenesis of NEC. e. Glaucoma and vision problems

[0288] Glaucoma is one of the leading causes of blindness worldwide. Despite diverse therapeutic options, new and better treatments for glaucoma are highly desirable. Elevated intraocular pressure, resulting from obstruction of the flow of fluid that helps the eye maintain its rigid shape, is a primary 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 elevated intraocular pressure, fluid has limited outflow from the anterior chamber of the eye, causing increased pressure, like water behind a dam. Increased pressure in the eye contributes to glaucoma by reducing the flow of nutrients to the optic nerve.

[0289] Therefore, the composition can be administered to a patient in need thereof to reduce intraocular pressure (IOP) and provide neuroprotection.For example, a composition containing THC and / or a THC analogue can be administered to reduce IOP, while a composition containing HU-211 can be administered to provide neuroprotection.In some forms, the composition can be administered to a patient in need thereof to protect or rescue the optic nerve from damage and / or restore its blood supply. f. Sleep Disturbances and Sleep Disorders

[0290] CB1 receptors located in the pons and basal forebrain may be involved in sleep induction. This process is likely related to the activity of cholinergic neurons located in the basal forebrain and pons via CB1 receptors, which support sleep induction. CB1 receptors enhance the activation of the serotonergic system, potentially playing a regulatory role in the sleep-wake cycle.

[0291] Therefore, this composition can be administered to the patient who needs it to improve sleep architecture.In some forms, this composition can increase total sleep time, reduce wakefulness after falling asleep, reduce slow wave sleep, and / or shorten or increase the latency to REM sleep as needed, and / or increase the sleepiness of the patient who suffers from insomnia.In some forms, this composition can be administered to improve wakefulness in the patient, for example, the patient who suffers from narcolepsy. C. Dosage and Effective Amount

[0292] In some in vivo procedures, the dendrimer complex is administered to a subject in a therapeutically effective amount. The term "effective amount" or "therapeutically effective amount" refers to a dosage sufficient to treat, inhibit, or alleviate one or more symptoms of the disorder being treated, or to achieve other desired pharmacological and / or physiological effect. The exact dosage will vary according to various factors, such as subject-dependent variables (e.g., age, immune system health, etc.), the disease or disorder, and the treatment being performed.

[0293] Generally, the dosage of the composition may be about 0.001 to about 100 mg / kg, about 0.01 to about 100 mg / kg, about 0.1 mg to about 10 mg / kg, or about 0.5 mg to about 5 mg / kg of body weight of the subject to be treated. The subject is typically a mammal, most preferably a human.

[0294] For example, the dendrimer conjugate composition may be in an amount effective to deliver one or more active agents to cells at or near the site of inflammation, particularly inflammation of the central nervous system or inflammation of the eye. Thus, in some embodiments, the dendrimer conjugate composition comprising one or more active agents is in an amount effective to ameliorate inflammation in a subject. In preferred embodiments, an effective amount of the dendrimer conjugate composition does not induce significant cytotoxicity in cells of a subject compared to an untreated control subject. Preferably, the amount of the dendrimer conjugate composition is effective to prevent or reduce inflammation and / or further associated symptoms of a disease or disorder in a subject compared to an untreated control.

[0295] Generally, the timing and frequency of administration will be adjusted to balance the efficacy of a given treatment or diagnostic schedule with the side effects of a given delivery system. Exemplary administration frequencies include continuous infusion, single administration, and multiple administrations, such as hourly, daily, or weekly administration.

[0296] In some embodiments, the dosage is administered to a human once, twice, or three times daily, or every other day, every two days, every three days, every four days, every five days, or every six days. In some embodiments, the dosage is administered about once or twice every week, every two weeks, or every three weeks. In some embodiments, the dosage is administered about once or twice every month, every two months, every three months, every four months, every five months, or every six months.

[0297] It will be understood by those skilled in the art that the dosage regimen can be of any length of time sufficient to treat the disorder in the subject. The term "long-term" means that the length of time of the dosage regimen can be hours, days, weeks, months, or possibly years.

[0298] In some embodiments, the regimen includes one or more cycles of treatment followed by a rest period (e.g., drug-free). The treatment round may be, for example, a round of administration as discussed above. Similarly, the rest period may 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.

[0299] The dendrimer complexes may be administered in combination with one or more additional therapeutically active agents known to be capable of treating the conditions or diseases discussed above. D. Control

[0300] The effect of the dendrimer conjugate composition can be compared to a control. Suitable controls are known in the art and include, for example, untreated cells or an untreated subject. In some embodiments, the control is an untreated tissue derived from a treated or untreated subject. Preferably, the control cells or tissue are derived from the same tissue as the treated cells or tissue. In some embodiments, the untreated control subject suffers from or is at risk of suffering from the same disease or condition as the treated subject. E. Combination Therapy

[0301] The dendrimer complex composition can be administered alone or in combination with one or more additional 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 a different day. For example, a composition containing the dendrimer complex composition can be administered on day 1, day 2, day 3, day 4, or a combination thereof.

[0302] The terms "combination" or "combined" are used to refer to either simultaneous, parallel, or sequential administration of two or more agents. Thus, combinations can be administered simultaneously (e.g., as an admixture), separately but in parallel (e.g., through separate intravenous lines to the same subject), or sequentially (e.g., one of the compounds or agents is administered first, followed by the second). VI. Kit

[0303] The composition can be packaged in a kit.The kit can include a single dose or multiple doses of a composition comprising one or more cannabinoids associated with or conjugated to a dendrimer (for example, one or more hydroxyl-terminated PAMAM dendrimers or glucose dendrimers described in the examples), and instructions for administering the composition.Specifically, the instructions instruct to administer an effective amount of the dendrimer composition to an individual with a specific disease / disorder that is indicated.The composition can be formulated as described above with reference to a specific treatment method, and can be packaged in any convenient manner.

[0304] The present invention will be further understood by reference to the following non-limiting examples. [Example]

[0305] Example 1 Synthesis of hydroxyl-polyamidoamine (PAMAM-OH) dendrimer-cannabinoid (CBD) conjugates The synthesis of PAMAM-OH-CBD conjugates can be achieved using a variety of linking chemistries and linkers (both cleavable and non-cleavable). Briefly, the surface hydroxyl groups of PAMAM-OH are modified with a linker to provide complementary groups on the surface that can further react with complementary groups on the drug-linker. Similarly, the drug of interest is modified with a linker to allow reaction between the dendrimer-linker and the complementary functional groups.

[0306] The linker on the drug is attached by a cleavable or non-cleavable linking group. Non-limiting examples of cleavable linking groups include esterase-sensitive ester bonds, glutathione-sensitive disulfide bonds, phosphatase-sensitive phosphodiester bonds, triglycyl peptide linkers (CX) that enable lysosomal release, and acid-cleavable hydrazine linkers. Non-limiting examples of non-cleavable linking groups include ether or amide bonds. Non-limiting examples of suitable linkers include amino acids, peptides, polyethylene glycol (n=2-15), and hydrocarbon chains.

[0307] Dendrimer-CBD conjugates are designed to (1) achieve site-specific targeting by directing the drug to activated microglia / macrophages that express CB2 receptors, thereby reducing dosage, enhancing efficacy, and reducing side effects associated with the free drug; (2) improve the aqueous solubility of cannabinoids by 10-100 times compared to the free drug; (3) avoid the systemic and dose-related side effects of free CBD drugs through sustained intracellular release at the target site; and (4) reduce or eliminate the undesirable psychotropic effects associated with CB1 agonists by restricting the dendrimer-CB1 agonist conjugates to the peripheral circulation using high-generation dendrimers.

[0308] Non-limiting examples of dendrimer-cannabinoid conjugates in which CBD and CBD derivatives are conjugated to the dendrimer surface using various linkers and linking chemistries are described in the Examples below. Example 2 Synthesis of dendrimer-cannabidiol conjugates Synthesis of dendrimer-cannabidiol conjugates with enzyme-sensitive ester linkers via click chemistry

[0309] The synthesis of dendrimer-cannabidiol is achieved using highly efficient copper(I)-catalyzed alkyne-azide click (CuAAC) chemistry. The synthesis begins with the modification of cannabidiol by attaching it to an azide-terminated orthogonal linker via a cleavable ester bond (Figures 2A and 2B). The purpose of the azide group is to participate in a CuAAC reaction with alkyne functional groups on the surface of the dendrimer. The hydroxyl groups in cannabidiol (1) are coupled to N-(OCH2CH2) in the presence of N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC) and 4-(dimethylamino)pyridine (DMAP) as coupling agents. n —COOH(2). The crude product is purified using column chromatography to give cannabidiol-azide (Figure 2A).

[0310] Alternatively, dendrimers can be modified to attach linkers bearing complementary alkyne groups (Figure 2B). Hydroxyl-terminated PAMAM dendrimer (D-OH; 4) is reacted with hexynoic acid in the presence of coupling agents EDC and DMAP to yield partial alkyne-terminated dendrimer (5) with 10 linkers (Figure 2B). Finally, CuAAC click reaction is performed between dendrimer (5) and cannabidiol-azide (3) to yield a D-cannabidiol conjugate with 10 drug molecules attached. The structure and purity of all intermediates and the final conjugate were confirmed by 100% NMR (Figure 2B). H This is achieved by NMR and HPLC. Synthesis of dendrimer-cannabidiol conjugates with non-cleavable ester linkers via click chemistry

[0311] Dendrimer-cannabidiol can also be obtained by click chemistry using a non-cleavable ester linker (Figures 3A and 3B). The synthesis begins with the modification of cannabidiol by attaching it to an azide-terminated orthogonal linker via a non-cleavable ester bond (Figure 3A). The hydroxyl group in cannabidiol (6) is converted to N-(OCHCH) in the presence of potassium carbonate (KCO) and dimethylformamide (DMF) as coupling agents. n —Br (7). The product is purified using column chromatography to give cannabidiol-azide (8), shown in FIG. 3A.

[0312] The dendrimer is further modified to attach a linker bearing a complementary alkyne group (Figure 3B). The hydroxyl-terminated PAMAM dendrimer (D-OH; 9) is reacted with hexynoic acid in the presence of coupling agents EDC and DMAP to yield the partial alkyne-terminated dendrimer (10) with 10 linkers (Figure 3B). Finally, a CuAAC click reaction is performed between dendrimer (9) and cannabidiol-azide (3) to yield the D-cannabidiol conjugate with 10 drug molecules attached. The final conjugate is purified. The structure and purity of all intermediates and the final conjugate were confirmed by 100% NMR (Fig. 3B). H This is achieved by NMR and HPLC. Example 3 Synthesis of dendrimer-HU-308 conjugates Synthesis of dendrimer-HU-308 conjugates bearing phosphatase-cleavable phosphodiester linkers via amidation reaction

[0313] Amidation reactions can yield dendrimer-HU-308 conjugates with phosphatase-cleavable phosphodiester linkers (Figures 4A and 4B). The synthesis begins with the modification of HU-308 by attaching a phosphodiester linker. As shown in Figure 4A, the hydroxyl group in HU-308 (1) is reacted with diphosphoryl chloride (POCl) and tetrahydrofuran (THF) at -40 °C to generate compound 1, which is then reacted with FmocHN-CHCH-HPO in the presence of triethylamine (EtN), 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 yield compound 3 (Figure 4A).

[0314] As shown in Figure 4B, D-OH is reacted with glutaric anhydride in DMF and DMAP to produce compound 4, which is then reacted with compound 3 to produce a D-HU-308 conjugate containing a phosphatase-cleavable phosphodiester linker. Synthesis of dendrimer-HU-308 conjugates with ester linkers via click reaction

[0315] The synthesis of dendrimer-HU-308 can also be achieved by modifying HU-308 to attach an azide-terminated orthogonal linker via a cleavable ester bond (Figures 5A and 5B). The hydroxyl group in HU-308 is converted to N-(OCHCH) in the presence of EDC and DMAP as coupling agents. n D-OH can be reacted with -COOH to give HU-308-azide (Figure 5A). As shown in Figure 5B, D-OH is reacted with hexynoic acid in the presence of coupling agents EDC and DMAP to give a partial alkyne-terminated dendrimer with 10 linkers attached (Figure 5B). Finally, a click reaction is performed between the dendrimer and HU-308-azide to give the D-HU-308 conjugate with 10 drug molecules attached. The structure and purity of all intermediates and the final conjugate were confirmed by 100% NMR (Figure 5C).H This is achieved by NMR and HPLC. Figure 5C is a schematic representation of the conjugation of HU-308 to PAMAM-G4-OH dendrimer. Synthesis of dendrimer-HU-308 conjugates bearing glutathione-sensitive disulfide linkers via click reaction

[0316] Dendrimer-HU-308 with a glutathione-sensitive disulfide linker can also be synthesized by the click reaction (Figures 6A and 6B). The hydroxyl group in HU-308 can be reacted with N3-SS-PEG2-acid in the presence of EDC and DMAP as coupling agents to give HU-308-azide (Figure 6A).

[0317] Meanwhile, D-OH is reacted with hexynoic acid in the presence of coupling agents EDC and DMAP to obtain a partial alkyne-terminated dendrimer with 10 linkers attached (Figure 6B). Finally, a click reaction is performed between the dendrimer and HU-308-azide to obtain a D-HU-308 conjugate with 10 drug molecules attached (Figure 6B). The structure and purity of all intermediates and the final conjugate were confirmed by 100% NMR (Figure 6C). H This is achieved by NMR and HPLC. Example 4 Synthesis of dendrimer-tetrahydrocannabinol conjugates Synthesis of dendrimer-tetrahydrocannabinol conjugates with esterase-sensitive ester linkers via click reaction

[0318] The synthesis of dendrimer-tetrahydrocannabinol (D-THC) conjugates bearing esterase-sensitive ester linkers can be achieved by click reaction (Figures 7A and 7B). As shown in Figure 7A, the hydroxyl group in THC was converted to N3-(OCH2CH2) in the presence of EDC and DMAP as coupling agents. nThe D-OH is then reacted with hexynoic acid in the presence of coupling agents EDC and DMAP to yield a partial alkyne-terminated dendrimer with 10 linkers (Figure 7B). Finally, a click reaction is performed between the dendrimer and THC-azide to yield a D-THC conjugate with 10 drug molecules attached. The structure and purity of all intermediates and the final conjugate were confirmed by 100% NMR (Figure 7C). H This is achieved by NMR and HPLC. Synthesis of dendrimer-tetrahydrocannabinol conjugates bearing triglycyl peptide linking groups for lysosomal release via click reaction

[0319] The synthesis of dendrimer-tetrahydrocannabinol (D-THC) conjugates with triglycyl peptide linkers can also be achieved by click reaction (Figures 8A and 8B). The triglycyl peptide linker allows for lysosomal-mediated cleavage of the dendrimer-THC conjugate. The hydroxyl group in THC is reacted with triglycyl peptide azide in the presence of DMF and K2CO3 as coupling agents to yield THC-triglycyl peptide azide (Figure 8A). As shown in Figure 8B, the D-OH is reacted with hexynoic acid in the presence of coupling agents EDC and DMAP to yield a partial alkyne-terminated dendrimer with 10 linkers attached. Finally, a click reaction is performed between the dendrimer and THC-triglycyl peptide azide to yield a D-THC conjugate with a triglycyl peptide linker and 10 drug molecules attached. Structural confirmation and purity of all intermediates and the final conjugate were confirmed by 100% NMR (Figure 8B). H This is achieved by NMR and HPLC. Example 5 Synthesis of dendrimer-anandamide conjugates Synthesis of dendrimer-anandamide conjugates with esterase-sensitive ester linkers via click reaction

[0320] Dendrimer-anandamide (D-AEA) conjugates with esterase-sensitive ester linkers can be synthesized by click reaction (Figures 9A and 9B). The hydroxyl group in AEA is converted to N-(OCHCH) in the presence of EDC and DMAP as coupling agents. n -COOH to give AEA-azide (Figure 9A). The AEA-azide is then conjugated to a dendrimer (D-OH) via a click reaction to give the D-AEA conjugate, as shown in Figure 9B. Synthesis of dendrimer-anandamide conjugates with non-cleavable ether linkers

[0321] The synthesis of dendrimer-anandamide (D-AEA) conjugates with non-cleavable ether linkers can also be achieved by click reaction (Figures 10A and 10B). The hydroxyl group in AEA is converted to N-(OCHCH) in the presence of KCO and DMF as coupling agents. n -Br to give AEA-azide (Figure 10A). As shown in Figure 10B, AEA-azide is conjugated to a dendrimer (D-OH) via a click reaction to give a D-AEA conjugate with a non-cleavable linker. Example 6 Synthesis of dendrimer-2-arachidonoylglycerol (D-2-AG) conjugates bearing esterase-sensitive ester linkers via click reaction

[0322] Dendrimer-2-arachidonoylglycerol (D-2-AG) conjugates bearing a non-cleavable esterase-sensitive ester linking group can be synthesized by the click reaction (Figures 11A and 11B). The hydroxyl group in 2-AG is converted to N-(OCHCH) in the presence of EDC and DMAP as coupling agents. n-COOH to give 2-AG-azide (Figure 11A). As shown in Figure 11B, 2-AG-azide is conjugated to a dendrimer (D-OH) via a click reaction to give a D-2-AG conjugate with an esterase-sensitive ester linker. Example 7 Synthesis and conjugation of cannabinoid drugs with hydroxyl-terminated PAMAM, glucose dendrimers, and hydroxylated bisMPA dendrimers

[0323] This methodology targets specific receptors on the surface of cells, such as neurons, microglia, macrophages, and other cells. These receptors can be found anywhere in the body. For example, CB1 / CB2 receptors are present in various organs throughout the body, including the brain. The conjugates described herein bind to these receptors. Therefore, the "specific cellular targeting" of these dendrimers, for example, the targeting of hydroxyl dendrimers to microglia and / or macrophages and the targeting of glucose dendrimers to neurons, is secondary to binding to specific receptors and is an effect on these receptors.

[0324] Targeting specific receptors can be used to achieve desired results. In one embodiment, the conjugate is targeted to CB1 and can cause agonism, antagonism, reverse agonism, or inverse agonism. In another embodiment, the conjugate is targeted to CB2 and can cause agonism, antagonism, or reverse agonism, or a combination of activity against CB1 / CB2, TRPV1, and other receptors.

[0325] When combined with the cell targeting capabilities of glucose dendrimers (injured neurons (primary), microglia / macrophages (secondary) or hydroxyl dendrimers (microglia / macrophages)), the conjugates can be specifically targeted by specific cannabinoid receptors of specific cells in specific organs.

[0326] The dendrimer conjugates not only aid in targeting, but also improve aqueous solubility by over 200-fold compared to the non-dendrimer-bound drug, which greatly facilitates formulation and delivery.

[0327] The following examples, using tryptamine (as a non-limiting example), demonstrate the efficacy of the conjugates in terms of selective targeting, alteration of binding affinity, and Hu-308 in terms of aqueous solubility. Materials and Methods

[0328] Unless otherwise noted, reactions were performed in flame-dried glassware under a positive nitrogen pressure using dry solvents. Commercial-grade reagents and anhydrous solvents were purchased from chemical suppliers and used without further purification. 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC.HCl), N,N-diisopropylethylamine (DIPEA), 4-(dimethylamino)pyridine (DMAP), trifluoroacetic acid (TFA), anhydrous dichloromethane (DCM), and N,N'-dimethylformamide (DMF) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cyanine 5 (Cy5)-mono-NHS ester was purchased from Amersham Bioscience-GE Healthcare. Deuterated solvents dimethyl sulfoxide (DMSO-d6), water (DO), and chloroform (CDCl3) were purchased from Cambridge Isotope Laboratories Inc. (Andover, MA). Ethylenediamine-core polyamidoamine (PAMAM) dendrimer, generation 4.0, hydroxy surface (G4-OH; diagnostic grade; consisting of 64 hydroxyl end groups), methanol solution (13.75% w / w) was purchased from Dendritech Inc. (Midland, MI, USA). Dialysis membranes were purchased from Spectrum Laboratories Inc. (Rancho Dominguez, CA, USA). Hu308, tryptamine, 1-(2-amino-1-(4-methoxyphenyl)ethyl)cyclohexanol, nor-ketamine, 5-hydroxytryptamine, psilocybin analogs, psilocyn analogs, and cannabidiol drugs were purchased from Cayman Chemicals. Synthesis of hydroxyl-PAMAM dendrimer-drug conjugates

[0329] The PAMAM-G4-OH (D4-OH) dendrimer, consisting of approximately 64 terminal hydroxyl groups, was used in the synthesis. After each synthesis step, the product was purified by dialysis in DMF for 24 hours to remove small molecular weight impurities, followed by water dialysis to remove the DMF.1 H NMR (in DMSO-d6 and DO) and HPLC analysis were used to confirm the formation and purity of the intermediates and final products. Monofunctional D4-OH was functionalized with alkyne groups by treatment with 5-hexynoic acid under standard esterification conditions using EDC.HCl and 4-DMAP in DMF at room temperature for 36 hours to yield the D-hexyne bifunctional dendrimer. The number of alkyne groups on the dendrimer surface was selected to maintain approximately 10-15 to maintain the overall aqueous solubility of the conjugate. The crude product was dialyzed against ultrapure water for 24 hours using a 1 kDa membrane to remove low molecular weight impurities by selective diffusion through the semipermeable dialysis membrane. 1 H NMR and HPLC analyses were used to confirm product formation and purity of intermediates and final products, which are shown in Figures 12A-12C. Synthesis of D-hexyne

[0330] A solution of PAMAM G4-OH 1 (10.00 g, 0.7 mmol) in DMF (50 mL) was treated with 5-hexynoic acid (1.40 g, 12.6 mmol) and DMAP (2.41 g, 12.6 mmol) and stirred for 5 minutes at room temperature. EDC.HCl (1.54 g, 12.6 mmol) was then added portionwise to the reaction mixture over 5 minutes. The reaction mixture was stirred at room temperature for 36 hours. The crude product was transferred to cellulose dialysis tubing with a 1 kD MW cutoff and dialyzed against DMF for 12 hours and then against water for 24 hours. The aqueous layer was frozen and lyophilized to afford D-hexyne as a hygroscopic white solid (75% yield). 1 H NMR (500 MHz, DMSO-d6) 8.21-7.57 (m, internal amide H), 4.71 (s, GABA amide H, 50H), 4.01 (t, 22-24 CH2), 3.5-2.1 (m, dendrimer CH2) 1.71-1.59 (m, 22-24 CH2). HPLC C18 retention time 4 minutes: purity approximately 99%. Synthesis and purification of Hu308-azide

[0331] This method can be used for other cannabinoids. A solution of Hu308 in anhydrous dimethylformamide (DMF) was treated with hexanoic acid-PEG4-azide, DMAP, and EDC in anhydrous DMF. 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, extracted with DCM, and the organic layer was then washed successively with saturated sodium bicarbonate solution, followed by saturated ammonium chloride solution, and finally brine. The organic layer was then dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was dried on a rotary evaporator and purified by CombiFlash® chromatography on silica gel using a gradient of 40% ethyl acetate / hexane as the eluent. The desired Hu308-PEG4-azide was obtained as a colorless oil. Synthesis of dendrimer-drug conjugates

[0332] A solution of D-hexyne and drug-azide in DMF (5 mL) was treated with copper sulfate pentahydrate (CuSO4.5H2O) and sodium ascorbate in water. The reaction mixture was stirred and heated at 50 °C for 10 h in a microwave synthesizer. Upon completion, the reaction mixture was dialyzed against DMF in 1 KDa cutoff cellulose dialysis tubing. EDTA (50 μL, 0.5 M) solution was added to the solution to remove copper by chelation. DMF dialysis was followed by water dialysis overnight. Drug loading was calculated by proton integration comparing the peaks corresponding to the dendrimer and drug. [Table 2] G2-glucose dendrimer (GD2)-drug conjugate

[0333] The second-generation glucose dendrimer (GD2) consists of 24 glucose molecules (96 surface hydroxyl groups) used for conjugation. Glucose dendrimers are primarily made up of a central core of di-pentaerythritol and glucose moieties consisting of one or more branching units of monosaccharide glucose molecules. Unlike hydroxyl-terminated PAMAM dendrimers, glucose dendrimers are primarily taken up by injured neurons and have been found to specifically target hyperexcitable neurons in both culture and in vivo mouse models. Synthesis and characterization of glucose dendrimer (GD2)

[0334] GD synthesis began by reacting the hexapropargylated core with AB4, a β-D-glucose-PEG-azide building block, via a click reaction to yield the first-generation glucose dendrimer (GD1). Propargylation of the OH groups on GD1 yielded GD1-acetylene 24, which was reacted with β-D-glucose-PEG-azide to yield the second-generation GD2 with 24 glucose moieties, resulting in 96 surface hydroxyl groups. Furthermore, a Cy5 fluorescent tag was attached onto GD2 via propargylation of approximately 2–3 hydroxyl groups to yield an alkyne-containing GD2 dendrimer. GD intermediates and final products were purified using dialysis. 1 The physicochemical properties of the GD2 dendrimer were also evaluated (Table 3). [Table 3] Synthesis and characterization of GD2-drug conjugates

[0335] The Hu308 drug was conjugated to a GD2-hexynoic acid dendrimer using a click chemistry strategy. The linker-attached drug moiety was conjugated to a glucose dendrimer using a Cu(I)-catalyzed click (CuAAC) reaction in the presence of catalytic amounts of CuSO4.5HO and sodium ascorbate to yield the GD2-drug conjugate. Traces of copper were removed by dialysis against ethylenediaminetetraacetic acid (EDTA). The final GD2-drug conjugate was characterized by NMR and HPLC. [Table 4] result

[0336] These compounds are shown in Figure 12. Figures 13A-13B show the synthesis and construction. Figure 13C shows the binding and release of the compounds.

[0337] FIG. 14 is a graph of % binding efficiency versus log tryptamine (μM) in a 5-HT1A human serotonin GPCR cell-based agonist cAMP assay.

[0338] Both hydroxyl and GD dendrimers have OH surface groups. When tryptamine was conjugated to these dendrimers using the same linking chemistry, very different affinities were observed for serotonin and other receptors (Figure 13C). Binding assays showed that the conjugates had lower affinity than the free drug in cell-based binding assays (an indicator of in vivo efficacy). Lower affinity may not allow for stronger binding, potentially modulating the drug's undesirably strong effect on the receptor. Second, the hydroxyl dendrimer conjugates were not active, whereas the glucose dendrimer conjugates were. This was unexpected and may be due to differences in the internal structures of the glucose and hydroxyl dendrimers. The drug can fold within the hydrophobic core of the hydroxyl dendrimer but open outward in the hydrophilic interior of the glucose dendrimer.

[0339] The conjugates can be active with or without drug release, and can be engineered to release or not release the drug using the appropriate linking chemistry described herein. Binding affinity measures the activity of the intact conjugate. For example, the Hu-308 conjugate releases the drug over a period of time (FIG. 13C) and is active in both the conjugated and released forms. Cannabidiol (CBD) conjugates can be engineered to not release the drug and can be active in the conjugated form. Example 8 Localization of dendrimer-Hu308 in peripheral nerve injury / sciatic nerve injury: Materials and Methods

[0340] Ten days after sciatic nerve ligation, animals were intravenously injected with dendrimer-Hu308 tagged with the fluorescent dye Cy5. These were then perfused and imaged 24 hours later. Microglia on the injured side are known to overexpress CB2 receptors, which modulate neuropathic pain. result

[0341] In a rodent model of chronic neuropathic pain induced by sciatic nerve injury, dendrimers conjugated to the CB2 receptor agonist Hu308 bind to CB2 receptors overexpressed in microglia surrounding the injury site. Hu308 binds to the intracellular portion of the CB2R, and dendrimer-Hu308 was observed to localize at high concentrations within glia, allowing intracellular binding of CB2 receptors.

[0342] The hydroxyl-terminated dendrimer-Hu308-Cy5 localized only to microglia surrounding the area of ​​sciatic nerve injury. These microglia are known to overexpress CB2R after injury. In contrast, the glucose-dendrimer GD tagged with Cy5 localized to injured neurons along the nerve and to neurons and microglia in the dorsal root ganglion ipsilateral to the injury in the acute phase immediately after injury. GD2-Cy5 localized to neurons near the ligation site and to neurons and glia in the dorsal root ganglion. This indicates that GD-Hu308 can bind to CB2 receptors in injured neurons and glia immediately after peripheral nerve injury.

[0343] Modifications and variations of the present invention will be obvious to those skilled in the art and are intended to fall within the scope of the appended claims.

Claims

1. 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 said conjugation is covalent.

2. 10. The composition of claim 1, wherein the compound is covalently conjugated by a covalent linkage between a modified or unmodified surface group or an interior group of the dendrimer comprising an ester, amide, disulfide, or ether linking group.

3. 3. The composition of claim 1, wherein the covalent link between the dendrimer and the cannabinoid compound is cleaved after administration.

4. 4. The composition of any of claims 1 to 3, wherein the dendrimer is a 1st to 7th generation (such as 1st, 2nd, 3rd, 4th, 5th, 6th or 7th generation) polyamidoamine (PAMAM) dendrimer, preferably wherein between greater than 40% and 100% of the surface groups are hydroxylated.

5. The dendrimer is a first to seventh generation PAMAM dendrimer (e.g., first, second, third, fourth, fifth, sixth or seventh generation), and optionally has terminal groups such as COOH, NH ２ The composition of any one of claims 1 to 4, wherein the hydroxybenzoate is selected from the group consisting of acetamide and acetamide.

6. 2. The composition of claim 1, wherein the dendrimer is a 1st to 7th generation PAMAM dendrimer (such as 1st, 2nd, 3rd, 4th, 5th, 6th or 7th generation) 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 dendrimer is a dendritic polymer, optionally a hyperbranched polymer.

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

9. 2. The composition of claim 1, wherein the dendrimer is a glucose dendrimer, preferably a first, second or third generation glucose dendrimer, preferably made from glucose building blocks and optionally ethylene glycol building blocks, having more than 10 surface glucose moieties.

10. 2. The composition of claim 1, wherein the dendrimer is a galactose dendrimer made from galactose building blocks and optionally ethylene glycol building blocks, having more than 10 surface sugar moieties.

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

12. 12. The composition of any of claims 1 to 11, wherein the cannabinoid is cannabidiol (CBD) or a CBD-analogue.

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

14. The composition according to any one of claims 1 to 11, wherein the cannabinoid is a CB1 agonist, preferably selected from the group consisting of arachidonylcyclopropylamide (ACPA) and methanandamide.

15. The 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. 12. The composition of any of claims 1 to 11, wherein the cannabinoid is a CB2 agonist, preferably the CB2 agonist is selected from the group consisting of HU-308, JWH-133, AM1241, GW405833 and GW842166X.

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

18. 12. The 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 tetrahydrocannabivarin (THCV) and THCV-analogues.

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

21. 12. The composition of any of claims 1-11, wherein the dendrimer-cannabinoid composition has at least a 10-fold increase in drug solubility compared to the free cannabinoid.

22. 22. A composition according to any preceding claim, wherein the dendrimer-cannabinoid conjugate further comprises a pharmaceutically acceptable excipient and optionally a non-cannabinoid active agent.

23. 23. The composition of any of claims 1 to 22, wherein the conjugate is restricted to the peripheral circulation by the use of a higher generation dendrimer, preferably a 4th, 5th or 6th generation PAMAM dendrimer, a 2nd, 3rd or higher generation glucose dendrimer, or is functionalized with PEG.

24. 24. The composition of any one of claims 1 to 23, wherein the at least one natural cannabis derivative or synthetic cannabinoid compound is conjugated to the dendrimer via a spacer comprising a hydrocarbon (such as an alkylene), a diethylene glycol moiety, an oligoethylene glycol chain, a triazole moiety or a combination thereof, preferably 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. 26. The pharmaceutical composition of claim 25, in a form selected from a hydrogel, nanoparticles, microparticles, suspension, powder, tablet, capsule, cream, solution, or combinations thereof.

27. 25. 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. 28. The method of claim 27, wherein the individual has an ocular disorder and preferably the dendrimer-cannabinoid conjugate is administered intravitreally, suprachoroidally, subconjunctivally, topically or systemically.

29. 28. The method of claim 27, comprising treating an individual with peripheral nerve pain, such as peripheral neuropathy or chemotherapy-induced pain.

30. 28. 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-analogue.

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

32. 28. The method of claim 27, comprising treating an individual in need thereof for symptoms of arthritis, diabetic nephropathy, renal interstitial disease, fibrosis, multiple sclerosis, irritable bowel syndrome, an autoimmune disorder, traumatic brain injury, acute pain, chronic inflammatory pain, neuropathic pain, ocular pain, or an inflammatory disorder, 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. 28. The method of claim 27, comprising treating an individual in need thereof for symptoms of neuropathic pain, cancer-related pain, Alzheimer's disease, Parkinson's disease, HIV-associated neuropathy, wherein the conjugate is a dendrimer-CB1 / CB2 agonist conjugate, and preferably the cannabinoid is selected from the group consisting of anandamide, Δ9-THC, and 2-arachidonoylglycerol.

34. 28. The method of claim 27, comprising treating an individual in need of glaucoma treatment by IOP lowering or neuroprotection, wherein the cannabinoid is selected from plant cannabinoids, synthetic cannabinoids, endocannabinoids and combinations thereof.

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

36. 36. The method of any of claims 27 to 35, comprising targeting the composition to neurons or neurons in disease-affected areas, made possible by conjugating the CB1 agonist or antagonist to a glucose dendrimer that targets damaged neurons.

37. 36. A method according to any one of claims 27 to 35, comprising administering the cannabinoid via the high generation and / or surface functionalised dendrimer, thereby targeting peripheral neurons without appreciable brain uptake.

38. A method for targeting a therapeutic, prophylactic or diagnostic agent to neurons in a disease or disorder affected area of ​​an individual, the method comprising administering the agent having 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. 39. The method of claim 38, wherein the dendrimer is a PAMAM dendrimer.

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

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

42. 42. The method of any of claims 38 to 41, administered in combination with radiation therapy, exercise or other concomitant treatment.

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

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