Dendrimer conjugates of antidepressants and antipsychotics and methods of use thereof

Abstract

Formulations of dendrimers conjugated to one or more antidepressant or antipsychotic agents having higher selectivity to targeted cell types and reduced risk of side effects have been developed, as well as methods of using the same. Preferably, the dendrimer conjugated antidepressant or antipsychotic agent selectively binds to one or more receptors on or inside the surface of the target cell. The formulations are suitable for enteral or parenteral administration for the treatment of one or more diseases, disorders and injuries in the central and peripheral nervous system.

Description

Field of the Invention

[0001] The present invention relates generally to the field of antipsychotic / antidepressant drug formulations, and in particular to dendrimer-antipsychotic / antidepressant conjugates for selective delivery to the nervous system and peripheral sites of disease and to specific receptors on target cells.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Application No. 63 / 401,483, filed on August 26, 2022, entitled “DENDRIMER CONJUGATES OF ANTIDEPRESSANT AND ANTIPSYCHOTIC AGENTS AND THEIRMETHODS OF USE”, the applicant being The Johns Hopkins University, the inventors being Kunal Parikh, Kannan Rangaramanujam, Sujatha Kannan and Anjali Sharma, the entirety of which is hereby incorporated herein by reference.

[0004] Statement Regarding Federally Funded Research

[0005] none. Background of the Invention

[0007] Depression is a chronic and relapsing illness that may require lifelong treatment with different modalities. A significant proportion of patients with depression remain undertreated, especially in primary care settings. Nonadherence to treatment and premature discontinuation of treatment are important factors that may significantly contribute to poor treatment outcomes. Adverse effects associated with the use of antidepressant medications (ADs) are some of the most common factors leading to nonadherence and discontinuation of treatment. Studies have shown that up to 43% of patients with depression may discontinue their antidepressant medications due to treatment-emergent adverse effects. Existing first-line treatments have significant side effects, are ineffective in up to 33% of patients, and may take up to six weeks to achieve clinical efficacy in patients who do respond.

[0008] The introduction of tricyclic antidepressants (TCAs) and monoamine oxidase inhibitors in the 1950s revolutionized the treatment of depression. Since then, the search for more selective and potentially better tolerated ADs has continued. Rational drug development has led to the emergence of selective serotonin reuptake inhibitors (SSRIs). SSRIs have become the first-line drug for the treatment of depression and many other indications. With the successful launch of SSRIs, many newer generations of antidepressants have been approved for the treatment of depression, including serotonin and norepinephrine reuptake inhibitors (e.g., venlafaxine, desvenlafaxine, and duloxetine), bupropion (bupropion) (norepinephrine and dopamine reuptake inhibitor), mirtazapine (mirtazapine) (norepinephrine and selective serotonin antagonists), and trazodone (serotonin antagonists and reuptake inhibitors). In addition to agomelatine (with 5-HT 2c With the exception of melatonin receptor agonists with melatonin receptor antagonist properties, all other drugs work primarily by modulating monoaminergic neurotransmission. Recently, the U.S. Food and Drug Administration (FDA) has approved four additional antidepressants for the treatment of depression: vilazodone, levomilnacipran, vortioxetine, and the dextromethorphan / bupropion combination.

[0009] Over the years, efforts have been underway to develop more effective antidepressants with better safety and tolerability profiles. Tricyclic antidepressants (TCAs) increase levels of norepinephrine and serotonin (two neurotransmitters) and block the effects of acetylcholine (another neurotransmitter). These restore the balance of these neurotransmitters in the brain, thereby relieving depression. In addition to relieving depression, tricyclic antidepressants also cause sedation and, to some extent, block the effects of histamine.

[0010] There is no clear evidence to support clinically significant differences in efficacy and tolerability among the various newer antidepressants, and controversy remains. No clinically significant differences have been found in the efficacy of SSRIs and TCAs. Differences in tolerability between TCAs and SSRIs appear to be modest. In addition, concerns have been raised regarding the safety and tolerability of the newer antidepressants with long-term use.

[0011] The Center for Adverse Effects Monitoring (CARM) continues to receive reports of sexual dysfunction associated with the use of antidepressants and antipsychotics. The most commonly reported drugs since 1965 are fluoxetine, citalopram, paroxetine, venlafaxine, risperidone, and clozapine.

[0012] Sexual dysfunction can be divided into four categories:

[0013] Sexual desire disorder, which includes a partial or complete lack of sexual desire.

[0014] Sexual arousal disorders, including erectile dysfunction and insufficient vaginal lubrication.

[0015] Orgasmic disorders, including premature, delayed, or absent orgasm (anorgasm); and ejaculation failure.

[0016] Sexual pain disorders, including dyspareunia and vaginismus.

[0017] The prevalence of sexual dysfunction in major depression can be as high as 70%. This most commonly manifests as a lack of sexual desire. Sexual desire often improves with treatment of depression; however, successful antidepressant treatment often has other deleterious effects on sexual function. Antidepressant-associated sexual dysfunction may also result from inadequately treated depression, concurrent alcohol abuse, concurrent physical illness, relationship problems, or a combination of these factors.

[0018] Management of sexual dysfunction during treatment with antidepressants can be difficult, in part because of the often multifactorial etiology. Options for patients who experience problems include dose reduction (may increase risk of relapse), medication holidays (may jeopardize treatment adherence; do not allow spontaneity; may experience withdrawal symptoms in patients using antidepressants with short half-lives), switching antidepressants (may increase risk of relapse or different side effects), continuing treatment ("wait and see"), or adding a reversal agent (eg, phosphodiesterase inhibitors, which may cause additional side effects).

[0019] Newer antidepressants (ADs) are widely used as first-line treatment for major depression and are considered safer than tricyclics. Some side effects are transient and disappear after a few weeks after the start of treatment, but potentially serious adverse events may persist or occur later. These include gastrointestinal symptoms (nausea, diarrhea, gastric bleeding, dyspepsia), hepatotoxicity, weight gain and metabolic abnormalities, cardiovascular disturbances (heart rate, QT interval prolongation, hypertension, orthostatic hypotension), genitourinary symptoms (urinary retention, incontinence), sexual dysfunction, hyponatremia, osteoporosis and fracture risk, bleeding, central nervous system disturbances (lowered seizure threshold, extrapyramidal side effects, cognitive impairment), sweating, sleep disturbances, affective disturbances (apathy, switching, paradoxical effects), ocular manifestations (glaucoma, cataracts), and hyperprolactinemia. Sometimes, such adverse events may persist after discontinuation of the drug, leading to iatrogenic complications. Other areas of concern include suicidal tendencies, safety of overdose, withdrawal syndrome, risks during pregnancy and lactation, and risk of malignancy. Therefore, rational selection of an AD should consider potential benefits and risks, likelihood of response to treatment options, and vulnerability to adverse events. The findings of this review should remind physicians to carefully review AD prescriptions on an individual basis and consider alternative treatments, if available.

[0020] A significant proportion of patients often fail to achieve a complete response and remission. Antipsychotics are often used as adjunctive therapy in adults. To date, the U.S. Food and Drug Administration has approved four atypical antipsychotics (aripiprazole, quetiapine, brexpiprazole, and olanzapine) for this purpose. However, augmentation with antipsychotics is associated with higher discontinuation rates and more adverse events (AEs) compared with antidepressant monotherapy. In children and adolescents with depression, adjunctive antipsychotics along with antidepressants are often associated with movement disorders and seizures compared with antidepressant monotherapy.

[0021] Individuals with attention deficit disorder and its variants (including those diagnosed with attention deficit disorder and those with symptoms thereof) ("ADHD") are another group of individuals for whom current treatments have limited efficacy and many side effects. ADHD medications include methylphenidate, amphetamine, atomoxetine, clonidine, guanfacine, viloxazine, and their analogs / modifications. These medications are often used for patients with depression, post-traumatic stress disorder, and bipolar disorder. See https: / / www.webmd.com / add-adhd / adhd-medication-chart and https: / / www.additudemag.com / adhd-medication-for-adults-and-children.

[0022] Some of these work by mechanisms similar to antidepressants / antipsychotics, such as atomoxetine (SNRI), viloxazine (SNRI), and methylphenidate, which blocks the reuptake of dopamine and norepinephrine. Current antidepressants used to treat ADHD symptoms include bupropion, desipramine, imipramine, and nortriptyline.

[0023] ADHD drugs are usually divided into stimulants and non-stimulants. Both are usually associated with significant side effects related to systemic, non-targeted delivery. Although non-stimulants are typically not as effective as stimulants, they are used for up to 30% of patients who cannot tolerate or benefit from stimulants. There is an urgent need for alternatives with reduced side effects and improved selectivity.

[0024] It is therefore an object of the present invention to provide formulations and methods of use thereof to increase specificity of delivery to particular cells and receptors, reduce side effects of antidepressants and antipsychotics, and improve therapeutic efficacy (including time to onset). SUMMARY OF THE INVENTION

[0026] Formulations of dendrimers conjugated to one or more antidepressants or antipsychotics and methods of use thereof have been developed that have greater selectivity for targeted cell types and reduced risk of side effects. Preferably, the dendrimer-conjugated antidepressants or antipsychotics selectively bind to one or more receptors on or within the target cells. These formulations are suitable for enteral or parenteral administration for the treatment of one or more diseases, disorders and injuries of the central and peripheral nervous systems.

[0027] Representative antidepressants and antipsychotics include selective serotonin reuptake inhibitors (SSRIs), serotonin and norepinephrine reuptake inhibitors (SNRIs), norepinephrine-dopamine reuptake inhibitors (NDRIs), tricyclic antidepressants (TCAs), monoamine oxidase inhibitors (MAOIs), benzodiazepines, GABA modulators (e.g., neurosteroids), antipsychotics, atypical antipsychotics, or analogs thereof. Preferred dendrimers include glucose dendrimers and PAMAM dendrimers, which may be modified by functional group hydroxylation, pegylation, or other means to alter uptake. Preferred glucose dendrimers include G1, G2, and G3 glucose dendrimers, while preferred PAMAM dendrimers include G3, G4, G5, and G6 hydroxyl-terminated PAMAM dendrimers. In a preferred embodiment, the antidepressant or antipsychotic is conjugated to the dendrimer via a linker, which is preferably cleavable by hydrolysis. In some forms, the linker contains a triazole moiety.

[0028] The formulations may be administered orally, applied to a mucosal surface, or by injection.

[0029] The formulations can be used to prevent, treat, or manage symptoms of disorders such as major depressive disorder, treatment-resistant depression, postpartum depression, post-traumatic stress disorder, panic disorder, social anxiety disorder, anorexia nervosa, suicidal ideation, obsessive-compulsive disorder, premenstrual dysphoric disorder, anorexia, substance abuse disorders, epilepsy, bipolar disorder, autism spectrum disorder, attention deficit hyperactivity disorder, schizophrenia, cluster headaches, migraines, seizures, fibromyalgia, narcolepsy, obesity, Alzheimer's disease, Tourette's syndrome, pain (e.g., neuropathic pain and chronic pain), phobias, and cardiovascular disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1A and 1B Schematic diagram showing an exemplary synthetic route for a dendrimer-fluoxetine conjugate with a non-cleavable bond using click chemistry. Fluoxetine is first conjugated to a linker having an azide moiety ( Figure 1A ), and then conjugated to the dendrimer modified with surface alkyne groups via an azide-alkyne click reaction ( Figure 1B ).

[0032] Figure 2A and 2Bis a schematic diagram showing an exemplary synthetic route for a dendrimer-paroxetine conjugate having a non-cleavable bond. Paroxetine is first conjugated to a linker having an azide moiety ( Figure 2A ), and then conjugated to the dendrimer modified with surface alkyne groups via an azide-alkyne click reaction ( Figure 2B ).

[0033] Figure 3A and 3B is a schematic diagram showing an exemplary synthetic route for a dendrimer-venlafaxine conjugate having an enzyme-cleavable ester bond. Venlafaxine is first conjugated to a linker having an azide moiety ( Figure 3A ), and then conjugated to the dendrimer modified with surface alkyne groups via an azide-alkyne click reaction ( Figure 3B ).

[0034] Figure 4A and 4B Schematic diagram showing an exemplary synthetic route for a dendrimer-venlafaxine analog conjugate having a non-cleavable amide bond. The venlafaxine analog is first conjugated to a linker having an azide moiety ( Figure 4A ), and then conjugated to the dendrimer modified with surface alkyne groups via an azide-alkyne click reaction ( Figure 4B ).

[0035] Figure 5A and 5B Schematic diagram showing an exemplary synthetic route for a dendrimer-citalopram analog conjugate having a non-cleavable amide bond. The citalopram analog is first conjugated to a linker having an azide moiety ( Figure 5A ), and then conjugated to the dendrimer modified with surface alkyne groups via an azide-alkyne click reaction ( Figure 5B ).

[0036] Figure 6 is a schematic diagram showing the synthetic route for the glucose dendrimer-fluoxetine conjugate with a non-cleavable bond.

[0037] Figure 7 is a schematic diagram showing the synthetic route for glucose dendrimer-paroxetine conjugates with non-cleavable bonds.

[0038] Figure 8 is a schematic diagram showing the synthetic route for glucose dendrimer-venlafaxine conjugates having an enzymatically cleavable ester bond.

[0039] Fig. 9is a schematic diagram showing the synthetic route for glucose dendrimer-venlafaxine analog conjugates having non-cleavable amide bonds.

[0040] Fig.10 is a schematic diagram showing the synthetic route for glucose dendrimer-citalopram analog conjugates having non-cleavable amide bonds.

[0041] FIG. 11 is a schematic diagram showing a stepwise synthetic route for glucose dendrimer-DMT analog conjugates having non-cleavable amino-alkyl bonds.

[0042] FIG. 12 is a schematic diagram showing a step-by-step synthetic route for a glucose dendrimer-lysergic acid diethylamide (LSD) conjugate having a non-cleavable amino-alkyl bond.

[0043] Fig.13 Schematic overview of the major pharmacological targets of LSD, psilocybin, DMT, MDMA, and ketamine, the signaling cascades involved, hormonal regulation, and the main behavioral consequences following their administration in animals and humans.

[0044] Fig.14A Schematic diagram of the synthesis of PAMAM dendrimer-norketamine conjugate. Fig. 14B is a schematic diagram of the synthesis of glucose dendrimer-norketamine conjugate.

[0045] Fig.15A Graph of the NMDAR 1A / 2B antagonist assay for glucose dendrimer-ketamine (IC50 = 4.54 μM), hydroxy dendrimer-ketamine (IC50>100), and norketamine (IC50 = 6.96 μM). Fig. 15B is the % binding potency of the log concentration (μM) of the compound in the D2L human dopamine GPCR cell-based agonist cAMP assay. Norketamine (closed circles), glucose dendrimer-ketamine EC50 = 13.08 μM (open circles), and hydroxy dendrimer-ketamine EC50 = 4.263 μM (triangles). Fig. 15C is the % binding potency of the log concentration (μM) of ketamine in the TA1 human microamine GPCR cell-based agonist cAMP assay. Norketamine (closed circles), glucose dendrimer-ketamine EC50 = 13.08 μM (open circles), and hydroxy dendrimer-ketamine EC50 = 4.263 μM (triangles).

[0046] Fig.16A and 16Bis the composite neurobehavioral score of wild-type, knockout saline (control) and knockout mice treated with dendrimer-ketamine conjugate ( Fig.16A ) and the probability of survival within one day after birth ( Fig. 16B ) in the table below. Fig. 16C is a graph of distance travelled (m); Fig.16D is a graph of the speed at which mice travel; Fig.16E is a graph of the time spent in the corners. DETAILED DESCRIPTION OF THE INVENTION

[0048] I. Definitions

[0049] The term "antidepressant" refers to a compound that modulates the uptake of one or more monoamines (e.g., serotonin, noradrenaline, norepinephrine, dopamine) into presynaptic cells. In some forms, "antidepressants" inhibit the uptake of monoamines (e.g., serotonin, noradrenaline, norepinephrine, dopamine) into presynaptic cells; the continued presence of these monoamines in the synaptic cleft leads to increased postsynaptic receptor stimulation and, therefore, increased postsynaptic transmission. In some forms, these actions (presumably) correct or compensate for physiological defects that may lead to depression. Examples of drugs that inhibit monoamine reuptake include tricyclic antidepressants (TCAs), selective serotonin reuptake inhibitors (SSRIs), serotonin-norepinephrine reuptake inhibitors (SNRIs), etc. In some forms, "antidepressants" refer to compounds that inhibit the breakdown of monoamines (e.g., serotonin, norepinephrine, and dopamine) in the storage vesicles of presynaptic cells. The preservation of these monoamine substances is presumed to improve the efficiency of synaptic transmission. This can correct or compensate for the physiological defects that cause depression. Drugs that inhibit the decomposition of monoamines include monoamine oxidase inhibitors (MAOI). In other forms, "antidepressants" refer to compounds that increase serotonin reuptake (tianeptine), increase serotonin and / or norepinephrine release (mirtazapine), act directly on serotonin and melatonin receptors (agomelatine) or otherwise affect synaptic neurotransmission. These drugs (including ADHD drugs, such as methylphenidate, amphetamine (amphetamine), atomoxetine, clonidine, guanfacine, viloxazine and gabapentin (gabapentin) and lithium / lithium salts) are conjugated or compounded with dendritic macromolecules to selectively deliver to neurons and activated microglia for the prevention, treatment or control of a variety of diseases. In some forms, "antidepressants" can also improve mental health or neurological diseases via anti-inflammatory effects.

[0050] The term "antipsychotic" refers to a psychotropic compound that is administered to manage one or more symptoms of a psychotic disorder (e.g., including delusions, hallucinations, paranoia, or disorganized thought), such as in psychosis, schizophrenia, and bipolar disorder, as well as in a range of other conditions, such as those associated with mood disorders, anxiety disorders, or non-neurological disorders. Antipsychotic drugs inhibit dopaminergic transmission or dopaminergic and serotonergic transmission, and may also exert noradrenergic, cholinergic, and / or histaminergic blocking effects.

[0051] The terms "active agent" or "bioactive agent" are used interchangeably and refer to a chemical or biological compound that induces a desired pharmacological and / or physiological effect, which may be preventive, therapeutic or diagnostic. These may be nucleic acids, nucleic acid analogs, small molecules having a molecular weight of less than 2 kD and more typically less than 1 kD, peptide mimetics, proteins or peptides, carbohydrates or sugars, lipids or combinations thereof. These terms also include pharmaceutically acceptable, pharmacologically active drug derivatives, including but not limited to salts, esters, amides, prodrugs, active metabolites and analogs. The term "therapeutic agent" refers to an agent that can be administered to treat one or more symptoms of a disease or disorder. The term "diagnostic agent" generally refers to an agent that can be administered to reveal, precisely locate and define the location of a pathological process. A diagnostic agent can label target cells so that these labeled target cells can be subsequently detected or imaged.

[0052] "Analogs" are related to a given compound and refer to another compound that is structurally similar, functionally similar, or both to a specified compound. Structural similarity can be determined using any standard known in the art, such as the Tanimoto coefficient, which provides a quantitative measure of the similarity between the two compounds based on their molecular descriptors. Preferably, the molecular descriptors are 2D properties, such as fingerprints, topological indices, and maximum common substructures, or 3D properties, such as overall shape and molecular fields. For different and identical molecular pairs, the range of the Tanimoto coefficient is between 0 and 1 (inclusive). If the Tanimoto coefficient of the compound and the specified compound is between 0.5 and 1.0 (inclusive), preferably between 0.7 and 1.0 (inclusive), and most preferably between 0.85 and 1.0 (inclusive), then the compound can be considered to be an analog of the specified compound. If the pharmacological effect, physiological effect, or both induced by the compound are the same as those of a specific compound, the compound is functionally similar to a specific compound. "Analogs" can also refer to modifications of compounds, including but not limited to hydrolysis, reduction, or oxidation products. Hydrolysis, reduction, and oxidation reactions are known in the art.

[0053] The term "therapeutically effective amount" refers to an amount of a therapeutic agent that, when incorporated into and / or onto a dendrimer, produces some desired effect at a reasonable benefit / risk ratio applicable to any medical treatment. The effective amount may vary depending on factors such as the disease or condition being treated, the particular targeting construct being administered, the size of the subject, or the severity of the disease or condition. One of ordinary skill in the art can empirically determine the effective amount of a particular compound without undue experimentation. In some embodiments, the term "effective amount" refers to an amount of a therapeutic or prophylactic agent that reduces or alleviates the symptoms of one or more diseases.

[0054] In the context of inhibition, the terms "inhibit" or "reduce" refer to reducing or decreasing activity and number. This can be a complete inhibition or reduction of activity or number, or a partial inhibition or reduction. The inhibition or reduction can be compared to a control or standard level. The inhibition can be 5%, 10%, 25%, 50%, 75%, 80%, 85%, 90%, 95%, 99% or 100%. For example, a dendrimer composition comprising one or more inhibitors can inhibit or reduce the activity and / or number of diseased neurons by about 10%, 20%, 30%, 40%, 50%, 75%, 85%, 90%, 95% or 99% compared to the activity and / or number of the same cells in an equivalent tissue of a subject that has not received or been treated with the dendrimer composition. In some embodiments, inhibition and reduction are compared at the mRNA, protein, cell, tissue and organ levels. For example, inhibition and reduction of the rate of neuronal loss, rate of brain weight loss or rate of hippocampal volume loss compared to an untreated control subject.

[0055] The terms "treat" or "prevent" refer to ameliorating, alleviating or otherwise preventing a disease, disorder or condition from occurring or developing in an animal that may be susceptible to the disease, disorder and / or condition but has not yet been diagnosed as having the disease, disorder or condition; inhibiting the disease, disorder or condition, e.g., preventing its progression; and relieving the disease, disorder or condition, e.g., causing the disease, disorder and / or condition to regress. Treating the disease or condition includes ameliorating at least one symptom of the particular disease or condition, even if the underlying pathophysiology is not affected, such as treating pain in a subject by administering an analgesic, even if such agent does not treat the cause of the pain. Desired effects of treatment include reducing the rate of disease progression, improving or alleviating the disease state, and alleviating or improving prognosis. For example, an individual is successfully "treated" if one or more symptoms associated with depression are alleviated or eliminated, including but not limited to reducing the degree of anxiety, agitation or restlessness, improving feelings of sadness, tearfulness, emptiness or helplessness, improving the patient's quality of life, reducing the dose of other drugs required to treat the disease, and delaying the progression of the disease.

[0056] The phrases "pharmaceutically acceptable" or "biocompatible" refer to compositions, polymers and other materials and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications, and at a reasonable benefit / risk ratio. The phrase "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, solvent or encapsulating material, used to deliver or transport any subject composition from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the subject composition and not injurious to the patient.

[0057] The term "biodegradable" generally refers to materials that, under physiological conditions, will degrade or erode into smaller units or chemicals that can be metabolized, eliminated, or excreted from the body. Degradation time depends on composition and form.

[0058] The term "dendrimer" includes, but is not limited to, a molecular architecture having an inner core, inner layers or "generations" of repeating units regularly attached to the initiating core, and an outer surface attached to the terminal groups of the outermost generations.

[0059] The term "functionalization" refers to modifying a compound or molecule in some way to attach a functional group or moiety. For example, a molecule can be functionalized by introducing a molecule that makes it a strong nucleophile or a strong electrophile.

[0060] The term "targeting moiety" is a moiety that is designated to be located at or away from a particular location. The moiety can be, for example, a protein, a nucleic acid, a nucleic acid analog, a carbohydrate, or a small molecule. The location can be a tissue, a particular cell type, a subcellular compartment, or a molecule such as a receptor.

[0061] The term "extended residence time" refers to an increase in the time required for an agent to be cleared from a patient's body, or an organ or tissue of the patient. In certain embodiments, "extended residence time" refers to an agent whose clearance half-life is 10%, 20%, 50%, or 75% longer than a control standard (e.g., a comparable agent not bound to a delivery vehicle (e.g., a dendrimer). In certain embodiments, "extended residence time" refers to an agent whose clearance half-life is 2, 5, 10, 20, 50, 100, 200, 500, 1000, 2000, 5000, or 10000 times longer than a control standard (e.g., a comparable agent not bound to a delivery vehicle (e.g., a dendrimer)) that is specifically targeted to a particular cell type.

[0062] The terms "binding" and "encapsulation" refer to binding, formulating or otherwise including an agent into and / or onto a composition so as to allow release (e.g., sustained release) of the agent in a desired application. The agent or other material can be bound to the dendrimer by binding to one or more surface functional groups of the dendrimer (through covalent, ionic or other binding interactions), by physical admixture, by encapsulating the agent within the dendritic structure, and / or by encapsulating the agent within the dendritic structure.

[0063] As used herein, the central nervous system ("CNS") includes the brain and spinal cord. As used herein, the peripheral nervous system ("PNS") refers to the nerves outside the brain and spinal cord.

[0064] "Hydroxyl terminated" with respect to dendrimers refers to dendrimers having hydroxyl groups on their surface. These hydroxyl groups are not attached to the termini of the dendrimer via a sugar moiety (eg, a saccharide moiety).

[0065] "Sugar-capped" with respect to dendrimers means containing sugar moieties (eg, carbohydrate moieties) on the surface of the dendrimer rather than in its core.

[0066] "Sugar-based" with respect to dendrimers refers to dendrimers that contain sugar moieties (eg, saccharide moieties) in the core of the dendrimer or in both its core and its surface.

[0067] II. Composition

[0068] Dendrimer-active agent conjugates have been developed that are useful for delivering one or more antidepressants or antipsychotics to one or more target cells that express their receptors (e.g., neural cells, glial cells, and / or peripheral cells activated by the dendrimer), and further to specific receptors on these cells that are targeted by the drug. Typically, the antidepressant or antipsychotic binds to receptors on the surface of the target cell and / or to receptors within the target cell. Exemplary target cells include, but are not limited to, brain cells, such as microglia, astrocytes, and / or neurons, e.g., cells located within a pathological site in the brain or CNS; cells in the peripheral nervous system, such as peripheral neurons and glial cells, and / or peripheral cells, such as gastrointestinal cells, cardiovascular cells, and immune system cells. The microglia and / or astrocytes to which the antidepressant or antipsychotic is delivered can be active or inactive microglia and / or astrocytes.

[0069] The antidepressant or antipsychotic of the dendrimer-active agent conjugate binds to a target receptor on the surface of a target cell or inside a target cell. In some embodiments, the agent remains conjugated to the dendrimer while the antidepressant or antipsychotic binds to the target receptor. In these embodiments, after binding, the agent may be released from the dendrimer or may remain conjugated to the dendrimer. In some embodiments, the antidepressant or antipsychotic is released from the dendrimer near the target receptor and then binds to the target receptor on the target neural and / or glial cells.

[0070] A. Dendrimers

[0071] Dendrimers are three-dimensional, hyperbranched, monodisperse, globular and multivalent macromolecules that include surface end groups (Tomalia, DA, et al., Biochemical Society Transactions, 35, 61 (2007); and Sharma, A., et al., ACS Macro Letters, 3, 1079 (2014)).

[0072] The term "dendrimer" includes, but is not limited to, a molecular architecture having an inner core ("GO") and layers (or "generations") of repeating units attached to and extending from the inner core, each layer having one or more branch points, and an outer surface attached to the end groups of the outermost generation. In some embodiments, the dendrimer has a regular, tree-like or "starburst" molecular structure.

[0073] Typically, the diameter of the dendrimer herein is between about 1 nm and about 60 nm, more preferably between about 1 nm and about 50 nm, between about 1 nm and about 40 nm, between about 1 nm and about 30 nm, between about 1 nm and about 20 nm, between about 1 nm and about 10 nm, or between about 1 nm and about 5 nm. In some embodiments, the diameter is between about 1 nm and about 2 nm. In some embodiments, the diameter of the dendrimer is effective to penetrate the blood-brain barrier ("BBB") (less than 5 nm) and remain near or inside the target neural and / or glial cells to deliver the agent bound thereto. In some embodiments, the dendrimer has a diameter that effectively penetrates the BBB and is internalized into the target neural and / or glial cells (e.g., neurons, oligodendrocytes, astrocytes, microglia, and glial support cells) to deliver the agent bound thereto. In some embodiments, the dendrimer has a diameter that effectively penetrates a barrier interface (e.g., the blood-nerve barrier ("BNB")) and is internalized into neural and glial cells (e.g., neurons, Schwann cells, satellite cells, and glial support cells) of the peripheral nervous system to deliver the agent bound thereto. These typically have a diameter greater than 5 nm. In some embodiments, the dendrimer has a diameter that effectively remains in the peripheral circulation to deliver the agent bound thereto to target cells of the peripheral nervous system. In some embodiments, the dendrimer has a diameter that can be effectively retained in the peripheral circulation to deliver the drug conjugated thereto to target cells of the gastrointestinal system, cardiovascular system, and / or immune system.

[0074] In some embodiments, the molecular weight of the dendrimer is between about 500 Daltons and about 100,000 Daltons, between about 500 Daltons and about 50,000 Daltons, or between about 1,000 Daltons and about 20,000 Daltons, inclusive. Dendrimers with a size less than 30,000 Da are more suitable for transport across the BBB, while those with a size greater than 50,000 Da are more suitable for confinement in the periphery.

[0075] In some embodiments, the dendrimer has a supercore (e.g., dipentaerythritol) and one or more monosaccharide branching units. In some embodiments, the monosaccharide branching units are bound to the core or previous monomer layers by a linker such as a polyethylene glycol chain. In a preferred embodiment, the supercore is dipentaerythritol and the monosaccharide branching units are glucosyl branching units, as shown in structures II-IV. In a most preferred embodiment, the dendrimer is made entirely of glucose building blocks. PAMAM dendrimers modified with sugars can also work, but dendrimers made of sugars, especially glucose, are most preferred. Particularly preferred glucose dendrimers are G1 to G3 glucose dendrimers, such as G1, G2 and / or G3 glucose dendrimers.

[0076] The preferred dendrimer is a glucose dendrimer, but other dendrimers may also be used. Suitable dendrimer scaffolds include, but are not limited to, polyamidoamine (also known as PAMAM) or STARBURST TM Dendrimers; polypropylamine (POPAM), polyethyleneimine, polylysine, polyester, polystyrene, aliphatic polyethers, aromatic polyether dendrimers, dendrimers of sugars (e.g., glucose, galactose, mannose, fructose, etc.) and copolymers thereof, such as copolymers of sugars and alkylene glycols (e.g., dendrimers formed from glucose and ethylene glycol building blocks). Dendrimers can have multiple surface functional groups, such as carboxylic acids, amines, hydroxyls and / or acetamides. The terms "surface functional groups" and "terminal groups" are used interchangeably herein. Preferred dendrimers have surface hydroxyls to ensure selective uptake in neurons and activated microglia. In some embodiments, one or more of these surface functional groups are further modified with other molecules, such as sugars (e.g., glucose, galactose, mannose, fructose, etc.) and / or polyalkylene glycols (e.g., polyethylene glycol), thereby having sugar molecules and / or polyalkylene glycols as terminal moieties / molecules. Preferred PAMAM dendrimers include hydroxyl terminated PAMAM dendrimers, particularly G3 to G6 hydroxyl terminated PAMAM dendrimers, such as G3, G4, G5 and G6 hydroxyl terminated PAMAM dendrimers.

[0077] The dendrimer can be of any generation, including but not limited to 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, or 10th generation.

[0078] In some embodiments, by using higher generation dendrimers (e.g., 4th, 5th, or 6th generation PAMAM dendrimers, or 2nd, 3rd, or higher generation glucosyl dendrimers), the dendrimer-active agent conjugates can be confined to the peripheral circulation and specifically targeted to specific tissue regions and / or cell types, such as peripheral neural and glial cells and / or the gastrointestinal, cardiovascular, and / or immune systems. Alternatively, the dendrimer-active agent conjugates can be confined to the peripheral circulation by appropriate functionalization of the dendrimers (e.g., PEGylation).

[0079] In some embodiments, by using a dendrimer of a certain generation, e.g., PAMAM dendrimers and / or glucose dendrimers of generation 2 (G2), G3, G4, and G5, the dendrimers can be specifically targeted to specific tissue regions and / or cell types of the central nervous system (CNS), peripheral nervous system (PNS), and / or periphery, e.g., neurons and glial cells of the CNS, neurons and / or glial cells of the PNS, and / or peripheral cells such as gastrointestinal cells, cardiovascular cells, and / or immune system cells.

[0080] The term "PAMAM dendrimer" refers to poly(amidoamine) dendrimers that may contain different cores (with amidoamine building blocks) and may have any number of carboxylic acid, amine, acetamide and / or hydroxyl termini. In some embodiments, the dendrimer is a 4th, 5th or 6th generation ("G") dendrimer. In the most preferred embodiments, the dendrimer is made entirely of glucose building blocks. PAMAM dendrimers modified with sugars may also work, but dendrimers made of sugars, especially glucose, are most preferred.

[0081] Monosaccharide-based dendrimers

[0082] In some embodiments, the branching units include monosaccharides. In some embodiments, the monosaccharide branching units are attached to the core or a previous monomer layer via a linker such as a polyethylene glycol chain. In some embodiments, the monosaccharide branching units are glucose-based branching units. In some embodiments, the branching units may include PEG and / or alkyl chain linkers between different dendrimer generations. For example, the glucose layers are connected via a PEG linker and a triazole ring. In some embodiments, the branching units are the same for each generation of dendrimers generated from the core. Thus, for example, for the generation of the 1st generation of dendrimers, for the generation of the 2nd generation of dendrimers, and for the generation of the 3rd generation of dendrimers, the branching units are glucose-based branching units.

[0083] In some embodiments, the dendrimer has a supercore (e.g., dipentaerythritol) and one or more monosaccharide branching units. In some embodiments, the supercore is dipentaerythritol and the monosaccharide branching units are glucosyl branching units. In further embodiments, the spacer molecule may also be an alkyl CH2) n - Hydrocarbon-like unit.

[0084] In some embodiments, dendrimers synthesized using glucose building blocks, whose surfaces are composed primarily of glucose moieties, allow for specific targeting of cells, including damaged neurons, ganglion cells, and other neuronal cells in the brain, eye, and / or peripheral nervous system. In some embodiments, glucose-based dendrimers selectively target or are enriched within target neural and / or glial cells. In some embodiments, glucose-based dendrimers selectively target or are enriched within target neural and / or glial cells. In some embodiments, glucose-based dendrimers selectively target or are enriched on the surface of target neural and / or glial cells. In some embodiments, glucose-based dendrimers selectively target or are enriched within and / or on the surface of damaged, diseased, and / or overactive neurons and / or glial cells.

[0085] In some cases, dendrimers include an effective number of sugar molecules and terminal groups, such as glucose and / or hydroxyl groups, for targeting one or more neurons and / or glial cells of the CNS, PNS, and / or eye. The terminal hydroxyl groups of these dendrimers can be part of a terminal glucose molecule or additional hydroxyl groups that are not part of a glucose molecule, or a combination thereof. In some embodiments, all terminal hydroxyl groups are part of a terminal glucose molecule. In some embodiments, the number of sugar molecules at the end of the dendrimer is determined by the number of generations.

[0086] In some embodiments, the dendrimer is composed of glucose and oligoethylene glycol building blocks. Exemplary glucose dendrimers are shown in structures V and VII.

[0087] Some exemplary glucose dendrimers include a first generation glucose dendrimer with 24 hydroxyl (-OH) end groups, a second generation glucose dendrimer with 96 hydroxyl (-OH) end groups, a third generation glucose dendrimer with 396 hydroxyl (-OH) end groups, and a fourth generation glucose dendrimer with 1584 hydroxyl (-OH) end groups. For example, a glucose dendrimer is a second generation glucose-based dendrimer with 24 glucose molecules on the periphery and 6 glucose molecules embedded in the main chain, held together by PEG segments.

[0088] Dendrimer compositions have been developed that can selectively accumulate within neurons, particularly in the nuclei of injured and / or overactive neurons, termed "glucose dendrimers." These dendrimers can also accumulate in large quantities within activated microglia. However, in contrast to hydroxy dendrimers that accumulate primarily in microglia, these dendrimers primarily enter neurons. Johns Hopkins University describes glucose dendrimers in USSN 63 / 327,610, "Dendrimer Compositions for Targeted Delivery of Therapeutic Agents to Neurons," with inventors KannanRangaramanujam, Rishi Sharma, Anjali Sharma, Sujatha Kannan, NirnathSah, Mira Sachdeva, and Siva P Kambhampati, filed April 5, 2022.

[0089] Glucose dendrimers include (a) a central core, (b) one or more branching units, wherein the branching units are monosaccharide glucosyl branching units, optionally with linkers attached thereto; and optionally (c) one or more therapeutic, prophylactic and / or diagnostic agents. Typically, one or more branching units are attached to the central core, and the surface groups of the dendrimer are monosaccharide glucose molecules. In some embodiments, the central core is dipentaerythritol, or a hexapropargylated derivative thereof. In some embodiments, the branching units are attached to the central core via linkers such as hydrocarbon or oligoethylene glycol chains. In a preferred embodiment, the branching unit is α-D-glucopyranoside tetraethylene glycol azide having the following structure,

[0090]

[0091] or its peracetylated derivatives.

[0092] In some embodiments, the glucose dendrimer is a generation 1, 2, 3, 4, 5, or 6 dendrimer. In one embodiment, the dendrimer is a generation 1 dendrimer having the following structure:

[0093]

[0094] In a preferred embodiment, the dendrimer is a second generation dendrimer having the following structure:

[0095]

[0096] In some embodiments, one or more therapeutic, prophylactic and / or diagnostic agents are encapsulated, bound and / or incorporated into the dendrimer at a concentration of about 0.01% to about 30%, preferably about 1% to about 20%, more preferably about 5% to about 20% by weight. In some embodiments, the dendrimer is conjugated to a small molecule, an antibody or antigen-binding fragment thereof, a nucleic acid or a polypeptide. In some embodiments, the therapeutic agent conjugated to the dendrimer is an anti-inflammatory agent, an antioxidant, or an immunomodulatory agent. In other embodiments, the dendrimer is conjugated to one or more diagnostic agents, such as fluorescent dyes, near-infrared dyes, SPECT imaging agents, PET imaging agents, and radioisotopes.

[0097] In some embodiments, the dendrimer and one or more therapeutic, prophylactic or diagnostic agents are linked via one or more linkers or coupling agents (e.g., one or more hydrocarbon or oligoethylene glycol chains). Exemplary linkages are disulfide bonds, ester bonds, ether bonds, thioester bonds, and amide bonds.

[0098] PAMAM dendrimers

[0099] The term "PAMAM dendrimer" refers to poly(amidoamine) dendrimers that may contain different cores with amidoamine building blocks and may have carboxylic acid, amine, acetamide and / or hydroxyl termini of any generation, including but not limited to generation 1 to 10 PAMAM dendrimers. In some embodiments, the dendrimer is a generation 4, 5, or 6 ("G") dendrimer. In some embodiments, the PAMAM dendrimer has hydroxyl termini or is surface modified with monosaccharides.

[0100] In general, the complete structure of a dendrimer can be distinguished into a core part followed by radially linked branching units (ie generations), which are further modified with chemical functionalities carrying desired terminal groups on the outer surface of the dendrimer.

[0101] In some embodiments, the dendrimer is in the form of nanoparticles, as described in US 2011 / 0034422, US 2012 / 0003155, and US 2013 / 0136697. For example, the molecular weight of the dendrimer can be varied to prepare polymer nanoparticles that form particles with properties (e.g., drug release rate) optimized for a particular application.

[0102] In some embodiments, different types of dendritic macromolecules can be used as delivery vehicles to bind and deliver one or more active agents, including but not limited to dendrons and tectodendrimers. Dendrons are branch-like wedges with a core containing one function (functional group, f=1) and another function (f=8, 16, 32, etc.) at the periphery. Dendritic macromolecules are usually composed of a central dendritic macromolecule and multiple dendritic macromolecules attached to the periphery.

[0103] 1. Core

[0104] In some embodiments, dendrimers are prepared using a process that assembles from a multifunctional core and extends outward through a series of reactions. The multifunctional core moiety allows for the stepwise addition of branching units (ie, generations) around the core.

[0105] Exemplary chemical structures suitable as core moieties include dipentaerythritol, pentaerythritol, 2-(aminomethyl)-2-(hydroxymethyl)propane-1,3-diol, 2-ethyl-2-(hydroxymethyl)propane-1,3-diol, 3,3′,3″,3″′-silanetetrayltetrakis(propane-1-thiol), 3,3-divinylpenta-1,4-diene, 3,3′,3″-nitrilotripropionic acid, 3,3′,3″-nitrilotripropionic acid, tris(N-(2-aminoethyl)propionamide), 3,3′,3″,3″-(ethane-1,2-diylbis(azatriyl))tetrapropionamide, 3-(carboxymethyl)-3-hydroxyglutaric acid, 2,2′-((2,2-bis((2-hydroxyethoxy)methyl)propane-1,3-diyl)bis(oxy))bis(ethanol-1-ol), tetrakis(3-(trichlorosilyl)propyl)silane, 1-thioglycerol, 2,2,4 , 4,6,6-hexachloro-1,3,5,215,415,615-triazatriphosphine, 3-(hydroxymethyl)-5,5-dimethylhexane-2,4-diol, 4,4′,4″-(ethane-1,1,1-triyl)triphenol, 2,4,6-trichloro-1,3,5-triazine, 5-(hydroxymethyl)benzene-1,2,3-triol, 5-(hydroxymethyl)benzene-1,3-diol, 1,3,5-tri(dimethyl) (vinyl) silyl) benzene, carbosiloxane core, nitrilotrimethanol, ethylenediamine, propane-1,3-diamine, butane-1,4-diamine, 2,2′,2″-nitrilotri(ethane-1-ol), α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, cucurbituril, benzene-1,2,3,4,5,6-hexathiol, monosaccharides, disaccharides, trisaccharides, oligosaccharides or their azide, alkyne-modified parts. In some embodiments, the core part is chitosan. Therefore, azide-modified chitosan or alkyne-modified chitosan is suitable for binding to the branch unit using click chemistry.

[0106] In some embodiments, the core moiety is ethylenediamine or tetraethylene oxide. In some embodiments, the core moiety is dipentaerythritol. Examples of chemical structures suitable for use as core moieties are shown in Table 1 below.

[0107] Table 1. Structural representation of various building blocks (core, branching units, surface functional groups, monomers) used for dendrimer synthesis.

[0108]

[0109]

[0110]

[0111]

[0112]

[0113] Exemplary chemical structures suitable as branching units include monosaccharides. In some embodiments, the monosaccharide branching units are conjugated to the monomer core or the previous layer through a linker such as a polyethylene glycol chain. In a preferred embodiment, the monosaccharide branching units are glucose-based branching units. Exemplary glucose-based branching units are shown in structures II-IV. These are spacer molecules and therefore can also be alkyl (CH2)n-type hydrocarbon units.

[0114] The branching units are PEG or alkyl chain linkers between different generations of dendrimers, for example, glucose layers are connected via PEG linkers and triazole rings.

[0115] In a preferred embodiment, the branching units are identical for each generation of dendrimers generated from the core. Thus, in one embodiment, the branching units are glucose-based branching units used to generate the first generation dendrimers shown in structures V-VII.

[0116] In some embodiments, the branching unit is a supermonomer, i.e., AB n Building blocks. Exemplary supermonomers include AB4, AB5, AB6, AB7, AB8 building blocks. The supermonomer strategy greatly increases the number of available end groups. An exemplary AB4 supermonomer is peracetylated β-D-glucopyranoside tetraethylene glycol azide, as shown in structure III.

[0117] The chemical structures listed in Table 1 are also suitable as building blocks for forming branch units of dendritic macromolecules. For example, branch units of dendritic macromolecules are formed from dipentaerythritol, pentaerythritol, 2-(aminomethyl)-2-(hydroxymethyl)propane-1,3-diol, 2-ethyl-2-(hydroxymethyl)propane-1,3-diol, 3,3′,3″,3″′-silanetetrayltetra(propane-1-thiol), 3,3-divinylpenta-1,4-diene, 3,3′,3″-nitrilotripropionic acid, 3,3′ ...′,3″′- -nitrilotris(N-(2-aminoethyl)propionamide), 3,3′,3″,3″-(ethane-1,2-diylbis(azatriyl))tetrapropionamide, 3-(carboxymethyl)-3-hydroxyglutaric acid, 2,2′-((2,2-bis((2-hydroxyethoxy)methyl)propane-1,3-diyl)bis(oxy))bis(ethanol-1-ol), tetrakis(3-(trichlorosilyl)propyl)silane, 1-thioglycerol, 2 ,2,4,4,6,6-hexachloro-1,3,5,215,415,615-triazatriphosphine, 3-(hydroxymethyl)-5,5-dimethylhexane-2,4-diol, 4,4′,4"-(ethane-1,1,1-triyl)triphenol, 2,4,6-trichloro-1,3,5-triazine, 5-(hydroxymethyl)benzene-1,2,3-triol, 5-(hydroxymethyl)benzene-1,3-diol, 1,3,5 -tri(dimethyl(vinyl)silyl)benzene, carbosiloxane core, nitrilotrimethanol, ethylenediamine, propane-1,3-diamine, butane-1,4-diamine, 2,2′,2″-nitrilotri(ethanol-1-ol), α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, cucurbituril, benzene-1,2,3,4,5,6-hexathiol, monosaccharides, disaccharides, trisaccharides, oligosaccharides, or their azide, alkyne-modified parts, or combinations thereof.

[0118] Other examples of chemical structures suitable for forming branching units of dendrimers disclosed herein include, but are not limited to, sugar moieties such as glucose, galactose, mannose, and fructose, and alkylene glycols such as ethylene glycol, and combinations thereof. In some embodiments, the branching unit is chitosan. Thus, azide-modified chitosan or alkyne-modified chitosan is suitable for conjugation to a core moiety or other identical or different branching units using click chemistry. In some embodiments, the branching unit is methyl acrylate or ethylenediamine, or a combination thereof. In some embodiments, the branching unit is a linear or branched polyglycerol. In some embodiments, the branching unit is a copolymer of an alkylene glycol (e.g., ethylene glycol) and a sugar moiety (e.g., glucose, galactose, mannose, and / or fructose).

[0119] 2. Surface functional groups

[0120] The surface functional groups / molecules of the dendrimers are not limited to primary amine end groups, hydroxyl end groups, carboxylic acid end groups, acetamide end groups, sugar molecules, oligo- or polyalkylene glycols and / or thiol end groups. In some embodiments, the desired terminal functional groups can be added by one of the conjugation methods of the core and branch units.

[0121] In some embodiments, the surface functional groups are hydroxyl groups, such as hydroxyl groups of PAMAM dendrimers, hydroxyl groups of 2nd generation PEG dendrimers as shown in structure I, or hydroxyl groups of terminal glucose of dendrimers prepared with glucosyl branching units as shown in structures V and VII. In some embodiments, the desired surface functional groups can be modified or added by one of the conjugation methods of the core and branching units. Exemplary surface functional groups include hydroxyl end groups, amine end groups, carboxylic acid end groups, acetamide end groups, and thiol end groups, and combinations thereof.

[0122] In some embodiments, dendrimers can be specifically targeted to specific tissue regions and / or cell types, such as cells and tissues of the central nervous system (CNS), peripheral nervous system (PNS), and / or eye. In some embodiments, dendrimers specifically target neurons and / or glial cells of the CNS. In some embodiments, dendrimers specifically target neurons and / or glial cells of the PNS. In some embodiments, the glucose dendrimers are those of generation 1 (G1), G2, G3, G4, and G5, preferably G1, G2, and / or G3.

[0123] In some embodiments, the dendrimers contain an effective number of terminal glucose and / or hydroxyl groups for targeting one or more neurons and / or glial cells of the CNS, PNS, and / or eye.

[0124] Glucose dendrimers are preferred. In some embodiments, the dendrimers are composed of glucose and oligoethylene glycol building blocks. An exemplary first generation glucose dendrimer is shown in Structure VI, while a second generation glucose dendrimer is shown in Structure VIII.

[0125] In some embodiments, the dendrimer has a plurality of surface functional groups, such as hydroxyl (-OH) groups, amine groups, acetamide groups, and / or carboxyl groups (also referred to herein as surface functional groups or peripheral functional groups) at the periphery of the dendrimer. In some embodiments, the surface density of such peripheral functional groups is at least 1 group / nm 2 (Number of surface functional groups / nm 2 For example, in some embodiments, the surface density of surface functional groups (e.g., hydroxyl groups) is greater than 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 OH groups / nm 2, for example at least 10, 15, 20, 25, 30, 35, 40, 45, 50 or more 50 OH groups / nm 2 In some embodiments, the volume density of surface functional groups (eg, hydroxyl groups) is from about 1 to about 50 groups / nm 3 Between about 5 and about 30 groups / nm 3 Between or about 10 to about 20 groups / nm 3 In further embodiments, the surface density of surface functional groups (e.g., hydroxyl groups) is from about 1 to about 50 groups / nm 2 Between, preferably 5-20 groups / nm 2 (Number of surface functional groups / nm 2 The molecular weight of each surface functional portion is between about 100 Da and about 10 kDa, preferably between about 100 Da and 1000 Da.

[0126] In some embodiments, the amount of surface functional groups (eg, any of the above, eg, hydroxyl groups) of the dendrimer is at least 40%, at least 50%, greater than 40%, greater than 50%, or in the range of greater than 40% to 100%.

[0127] In some embodiments, one or more surface functional groups (e.g., any of the above) at the periphery of the dendrimer are further modified by conjugation with one or more carbohydrate molecules and / or one or more polyalkylene glycols (e.g., polyethylene glycol). In these embodiments, the surface density of the terminal carbohydrate moieties / molecules and / or polyalkylene glycols can be within any of the above hydroxyl ranges. Hydroxyl terminated PAMAM dendrimers, PAMAM dendrimers with surfaces modified with sugar moieties (where >10% of the surface groups are modified with sugars (especially glucose)), and glucose dendrimers (where the dendrimers are made of glucose building blocks) are preferred. For delivery to the brain, constructs with a total molecular weight of <30,000 Da are preferred. For primary confinement to the peripheral circulation, constructs with a total molecular weight of >50,000 Da are preferred. When dendrimers are formed from sugar moieties / molecules (e.g., glucose) or include sugar moieties / molecules at the termini, the terminal hydroxyl groups of these dendrimers can be part of the terminal sugar moiety / molecule or additional hydroxyl groups that are not part of the sugar moiety / molecule, or a combination thereof. In some embodiments, all terminal hydroxyl groups are part of a terminal sugar moiety / molecule.

[0128] a. Hydroxyl-terminated dendrimers

[0129] In some embodiments, the dendrimer comprises a plurality of hydroxyl groups. Some exemplary high density hydroxyl-containing dendrimers include commercially available polyester dendrimers, such as hyperbranched 2,2-bis(hydroxy-methyl)propionic acid polyester polymers (e.g., hyperbranched bis-MPA polyester-64-hydroxy, 4th generation), dendritic polyglycerols. In some embodiments, the hydroxyl-terminated dendrimers include hydroxyl-terminated PAMAM dendrimers, particularly G3 to G6 hydroxyl-terminated PAMAM dendrimers, such as G3, G4, G5, and G6 hydroxyl-terminated PAMAM dendrimers.

[0130] In some embodiments, the high-density hydroxyl-containing dendrimer is an oligoethylene glycol (OEG)-based dendrimer. For example, the second-generation OEG dendrimer (D2-OH-60) shown in Structure I can be synthesized using efficient, robust, and atom-economical chemical reactions, such as Cu(I)-catalyzed alkyne-azide click and photocatalyzed thiol-ene click chemistry. Orthogonal supermonomers and supercore strategies can be used to achieve very low-generation high-density polyol dendrimers with minimal reaction steps, for example as described in WO2019094952. In some embodiments, the dendrimer backbone has non-cleavable polyether bonds throughout the structure to avoid disintegration of the dendrimer in vivo and allow such dendrimers to be eliminated from the body as a single entity (non-biodegradable).

[0131]

[0132] Structure I. Second generation (G2) oligoethylene glycol dendrimers

[0133] In some embodiments, the dendrimer has a plurality of hydroxyl (-OH) groups at the periphery of the dendrimer. In some embodiments, the surface density of hydroxyl (-OH) groups is at least 1 OH group / nm 2 (Number of surface hydroxyl groups / nm 2 For example, in some embodiments, per nm 2 The surface density of hydroxyl groups is greater than 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 OH groups / nm 2 , such as at least 10, 15, 20, 25, 30, 35, 40, 45, 50, or greater than 50. In some embodiments, the volume density of hydroxyl groups is between about 1 and about 50 groups / nm 3 Between about 5 and about 30 groups / nm 3 Between or between about 10 and about 20 groups / nm 3In further embodiments, the surface density of hydroxyl (-OH) groups is from about 1 to about 50, or from 5 to 20 OH groups / nm 2 (number of surface hydroxyl groups / surface area in nm2), and a molecular weight of about 100 Da to about 10 kDa, preferably about 100 Da to 1000 Da. In some embodiments, the amount of surface hydroxyl groups of the dendrimer is preferably greater than 35%, at least 40%, at least 50%, greater than 40%, greater than 50%, or in the range of greater than 40% to 100%. In some embodiments, the dendrimer may have a small portion of the hydroxyl groups exposed on the outer surface, while the remaining hydroxyl groups are located in the inner core of the dendrimer.

[0134] In some embodiments, the dendrimers specifically target specific tissue regions and / or cell types after administration into the body. In some embodiments, the dendrimers specifically target specific tissue regions and / or cell types without a targeting moiety. In some embodiments, the dendrimers include an effective number of hydroxyl groups for targeting CNS cells and / or PNS cells, such as microglia, astrocytes, and / or neurons associated with a disease, disorder, or injury of the central nervous system or peripheral nervous system. In some embodiments, the dendrimers specifically target specific tissue regions and / or cell types without a targeting moiety, and the active agent bound thereto binds directly to receptors on the surface and / or inside the target neural and / or glial cells.

[0135] In some embodiments, the dendrimers are able to specifically target specific tissue regions and / or cell types, preferably cells and tissues of the central nervous system (CNS) and the eye. In some embodiments, the dendrimers specifically target neurons of the CNS and the eye. Unmodified PAMAM dendrimers with hydroxyl end groups are not as enriched in neurons of the brain and / or retinal ganglion cells (RGCs) of the eye as these glucose dendrimers. Glucose dendrimers with terminal glucose monosaccharides and a high density of hydroxyl functional groups effectively target neurons in a generation-dependent manner. The embodiments show that generation 2 (G2) is effective, and G3 and G4 should be effective. G5 and above are more difficult to use.

[0136] In a preferred embodiment, the dendrimer contains an effective number of terminal glucose and / or hydroxyl groups for targeting one or more neurons of the CNS or eye. The hydroxyl groups on the surface of the dendrimer are part of a glucose molecule. There are no additional hydroxyl groups other than the glucose molecules on the surface. The number of sugar molecules on the surface is determined by the generation. All generations are expected to target neurons.

[0137] In some embodiments, the dendrimers are made of glucose and oligoethylene glycol building blocks. Exemplary glucose dendrimers are shown in the embodiments, for example, the first generation dendrimers shown in structures IV-VI and the Figure 1A and 1B Some exemplary glucose dendrimers include a first generation glucose dendrimer with 24 hydroxyl (-OH) end groups, a second generation glucose dendrimer with 96 hydroxyl (-OH) end groups, a third generation glucose dendrimer with 396 hydroxyl (-OH) end groups, and a fourth generation glucose dendrimer with 1584 hydroxyl (-OH) end groups. In a preferred embodiment, the glucose dendrimer is a second generation glucose based dendrimer with 24 glucose molecules at the periphery and 6 embedded glucose molecules in the backbone, which are held together by PEG segments.

[0138] b. Carbohydrate-modified dendrimers

[0139] In some embodiments, the dendrimer comprises one or more carbohydrate molecules at the termini. These terminal carbohydrate molecules can be prepared by conjugating one or more surface functional groups (e.g., amine, carboxyl, or hydroxyl) of the dendrimer to one or more carbohydrate molecules. In a preferred embodiment, prior to carbohydrate conjugation, the dendrimer is a hydroxyl-terminated dendrimer, such as a hydroxyl-terminated PAMAM dendrimer, and one or more hydroxyl groups are conjugated to one or more carbohydrate molecules.

[0140] In some embodiments, hydroxy-terminated dendrimers modified with surface glucose molecules selectively target central and / or peripheral nerves and / or glial cells in vitro and in vivo; and / or selectively accumulate on the surface and / or inside these target nerve cells, glial cells and / or macrophages, allowing one or more active agents conjugated thereto to bind to one or more receptors on / in the target nerve cells and / or glial cells.

[0141] In some embodiments, the carbohydrate moiety used to modify one or more surface functional groups of the dendrimer is a monosaccharide. Exemplary monosaccharides suitable for modifying the dendrimer include glucose, glucosamine, galactose, mannose, fructose, dehydroascorbic acid, uric acid, inositol. In some embodiments, the dendrimer is conjugated to glucose and thus comprises glucose as a terminal moiety / molecule. In some embodiments, the hydroxyl-terminated dendrimer is modified with one or more glucose moieties to the dendrimer ("D-Glu"). In some embodiments, the dendrimer is conjugated to galactose. In some embodiments, the dendrimer is conjugated to mannose. In some embodiments, the dendrimer is conjugated to fructose. In some embodiments, the dendrimer is conjugated to one or more monosaccharides other than glucose (e.g., galactose, mannose, and / or fructose). For example, the carbohydrate moiety is an oligosaccharide, the terminal of which is one or more monosaccharides, including glucose, glucosamine, mannose, fructose, thereby exposing these sugar moieties on the surface for conjugation.

[0142] Glucose dendrimers or glucose modified dendrimers can be used to obtain selective uptake by target cells. Drugs bound to dendrimers can bind to receptors or other sites of action. In preferred embodiments, dendrimers (e.g., glucose or hydroxyl terminated PAMAM dendrimers) or carbohydrate functionalized dendrimers are conjugated to one or more carbohydrate moieties active agents that have affinity for and are suitable for binding to one or more of the following: serotonin receptors, such as 5HT-1A, 5HT-2B, 5HT-2A, 5HT-2B, 5HT-2C, 5HT-3, 5HT-4. In some embodiments, the dendrimer is conjugated to one or more carbohydrate moieties having affinity for and suitable for binding to one or more of the following: norepinephrine (NE) receptors, e.g., α1A adrenergic receptor, α1B-adrenergic receptor, α1D-adrenergic receptor, α2A-adrenergic receptor, α2B-adrenergic receptor, α2C-adrenergic receptor, β1-adrenergic receptor, and β2-adrenergic receptor. In some embodiments, the dendrimer is conjugated to one or more carbohydrate moieties having affinity for and suitable for binding directly or indirectly to dopamine D1 and D2 receptors. In some embodiments, the dendrimer is conjugated to one or more carbohydrate moieties having affinity and suitable for binding to one or more non-cannabinoid receptors, such as: G protein-coupled receptors, such as GPR55, GPR18, GPR3, GPR6, GPR12, GPR40, GPR43, GPR41, GPR120, GPR23, GPR92, GPR84, GPR119, or GPR35; adenosine receptors, such as adenosine A3; muscarinic acetylcholine receptors, such as M1 and M4; serotonin receptors, such as 5-HT1A, 5-HT2A; opioid receptors, such as μ- and δ-opioid receptors; and tachykinin NK2 receptors. In some embodiments, the dendrimer is conjugated to one or more carbohydrate moieties, or is made of sugar moieties that have affinity and are suitable for transport through one or more of GLUT1, GLUT2, GLUT3, GLUT4, GLUT5, GLUT6, GLUT7, GLUT8, GLUT9, GLUT10, GLUT11, GLUT12, GLUT13, and GLUT14. In further embodiments, the dendrimer is conjugated to one or more glucose and / or glucosamine moieties. In other embodiments, the dendrimer is conjugated to one or more oligosaccharides terminated with glucose and / or glucosamine moieties, i.e., the glucose and / or glucosamine moieties are exposed on the surface of the dendrimer conjugate, suitable for binding to one or more GLUT, GLUT, 5HT receptor, DA receptor, NE receptor, and / or transporter.

[0143] In some embodiments, the dendrimer has multiple carbohydrate moieties / molecules, e.g., monosaccharides, e.g., glucose, or sugar building blocks for the dendrimer at its periphery. In some embodiments, the surface density of carbohydrate molecules, e.g., monosaccharides, e.g., glucose, is at least 1 carbohydrate molecule / nm 2 (Number of surface carbohydrate groups / nm 2 In some embodiments, the surface area per nm 2 The surface density of carbohydrate molecules is greater than 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 OH groups / nm 2 , such as at least 10, 15, 20, 25, 30, 35, 40, 45, 50 or more than 50. For example, per nm 2 The surface density of carbohydrate molecules is greater than 10. In some embodiments, the volume density of surface carbohydrate molecules is from about 1 to about 50 groups / nm 3 , about 5 to about 30 groups / nm 3 or about 10 to about 20 groups / nm 3 In a further embodiment, the surface density of carbohydrate molecules is 2 From about 1 to about 50, from about 5 to about 20 (surface carbohydrate molecules / nm 2 In some embodiments, the surface area of ​​the dendrimer is about 100 Da (about 1000 Da) and the molecular weight of each carbohydrate moiety is about 100 Da to about 1000 Da. In these embodiments, i.e., one or more surface functional groups of the dendrimer are modified to introduce one or more sugar moieties / molecules at the termini, the terminal hydroxyl group can be part of the terminal sugar moiety / molecule or an additional hydroxyl group that is not modified by the sugar moiety / molecule and is therefore not part of the sugar moiety / molecule, or a combination thereof.

[0144] In some embodiments, the weight content of carbohydrate molecules (e.g., monosaccharides, such as glucose) is between about 1% and 40% of the total weight of the glycosylated dendrimer, for example, between about 2% and 20%, between about 5% and 15%, or between 9% and 12% of the total weight of the glycosylated dendrimer. For example, in some embodiments, after conjugation, the carbohydrate moiety is present in an amount of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% of the total weight of the glycosylated dendrimer. In some embodiments, the conjugation of carbohydrate molecules through one or more surface functional groups occurs through about 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20% or 25% of the total available surface functional groups (preferably hydroxyl groups) of the dendrimer before conjugation. In other embodiments, conjugation of carbohydrate molecules occurs on less than 5%, less than 10%, less than 15%, less than 20%, less than 25%, less than 30%, less than 35%, less than 40% of the total available surface functional groups of the dendrimer prior to conjugation.

[0145] c. Dendrimers modified with polyalkylene glycol

[0146] In some embodiments, the dendrimer contains one or more polyalkylene glycols at the termini. These terminal polyalkylene glycols can be prepared by conjugating one or more surface functional groups (e.g., hydroxyl groups) of the dendrimer with a polyalkylene glycol (e.g., PEG). In some embodiments, prior to conjugation, the dendrimer is a hydroxyl-terminated dendrimer (e.g., a hydroxyl-terminated PAMAM dendrimer), and at least a portion of the surface hydroxyl groups are conjugated with PEG.

[0147] In some embodiments, the dendrimer has a plurality of polyalkylene glycols, such as PEG, at the periphery of the dendrimer. In some embodiments, the surface density of the polyalkylene glycol, such as PEG, is at least 1 polyalkylene glycol / nm 2 (Number of polyalkylene glycols on the surface / nm 2 In some embodiments, the surface area per nm 2 The surface density of the polyalkylene glycol is greater than 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, such as at least 10, 15, 20, 25, 30, 35, 40, 45, 50 or greater than 50. For example, per nm 2 The surface density of the polyalkylene glycol is greater than 10. In some embodiments, the volume density of the surface polyalkylene glycol is from about 1 to about 50 groups / nm 3 , about 5 to about 30 groups / nm 3or about 10 to about 20 groups / nm 3 In a further embodiment, the surface density of the polyalkylene glycol (eg, PEG) is 2 From about 1 to about 50, from about 5 to about 20 (number of surface polyalkylene glycols / nm 2 , and has a molecular weight of about 100 Da to about 10 kDa.

[0148] In some embodiments, the weight content of the polyalkylene glycol molecules (e.g., PEG) is about 1% to 40% of the total weight of the PEGylated dendrimer, such as about 2% to 20%, about 5% to 15%, or 9% to 12% of the total weight of the PEGylated dendrimer. For example, in some embodiments, the weight content of the polyalkylene glycol molecules (e.g., PEG) is about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% of the total weight of the PEGylated dendrimer after conjugation.

[0149] In some embodiments, the conjugation of polyalkylene glycol molecules (e.g., PEG) through one or more surface functional groups of the dendrimer occurs through about 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, or 25% of the total available surface functional groups (preferably hydroxyl groups) of the dendrimer before conjugation. In other embodiments, the conjugation of polyalkylene glycol molecules (e.g., PEG) occurs on less than 5%, less than 10%, less than 15%, less than 20%, less than 25%, less than 30%, less than 35%, less than 40% of the total available surface functional groups of the dendrimer before conjugation.

[0150] B. Antidepressants and antipsychotics

[0151] The dendrimer is complexed or conjugated with one or more antidepressants and antipsychotics. Exemplary antidepressants and antipsychotics include, but are not limited to, a range of drug classes including selective serotonin reuptake inhibitors (SSRIs), serotonin-norepinephrine reuptake inhibitors (SNRIs), antipsychotics, atypical antipsychotics, tricyclic antidepressants (TCAs), monoamine oxidase inhibitors (MAOIs), benzodiazepines, Antidepressants, antipsychotics and their derivatives typically bind to one or more receptors to regulate the conduction of neurotransmitter signaling in the central nervous system. Antidepressants typically inhibit the reuptake of neurotransmitters (particularly serotonin, dopamine and norepinephrine) through selective receptors, thereby increasing the concentration of these specific neurotransmitters in the synaptic cleft. Antipsychotics typically reduce neurotransmission in the dopamine pathway through D2 antagonism or partial D2 receptor agonism. Therefore, partial antagonism, functional selectivity and inverse agonism all play an important role in determining the cellular response to specific neurotransmitter receptor ligands.

[0152] 1- Antidepressants

[0153] a. Selective serotonin reuptake inhibitors (SSRIs)

[0154] The composition may include a dendrimer complexed with one or more SSRIs. "SSRIs" or "selective serotonin reuptake inhibitors" are a class of drugs typically used as antidepressants to treat major depressive disorder and anxiety. SSRIs are thought to increase extracellular levels of the neurotransmitter serotonin by limiting its reabsorption (reuptake) into presynaptic cells, thereby increasing the level of serotonin available in the synaptic cleft to bind to postsynaptic receptors.

[0155] SSRIs are known to selectively inhibit serotonin transport by inhibiting serotonin reuptake, particularly by binding to the serotonin transporter (SERT). This action of SSRIs results in a sharp increase in serotonin in the cell body dendrite region of serotonergic neurons, resulting in desensitization of the cell body dendrite serotonin-1A autoreceptors. Therefore, neuronal pulse flow increases. It causes an increase in the release of serotonin from axon terminals, which ultimately leads to desensitization of postsynaptic serotonin receptors. The desensitization of these receptors may contribute to the therapeutic effects of SSRIs, or may explain the reasons for tolerance to the acute side effects of SSRIs. SSRIs cause a strong but slow disinhibition of 5-HT neurotransmission in the central nervous system (CNS) and peripheral nervous system (PNS). In this case, the action of antidepressants is mediated by a pathway from the midbrain raphe nuclei to the prefrontal cortex. Since most of the serotonin in the human body is produced in the intestine, SSRIs affect enteric neurons, thereby increasing the availability of serotonin, increasing enteric neurogenesis, and accelerating in vitro GI motility. In fact, serotonin receptors are widely expressed in the GI tract, and five of the seven known families, namely 5-HT1, 5-HT2, 5-HT3, 5-HT4 and 5-HT7 receptors, are expressed in the intestine and can affect intestinal function (Mawe and Hoffman (2013), Nat Rev Gastroenterol Hepatol. 2013 Oct; 10(10): 564).

[0156] In some forms, one or more SSRIs may be conjugated to a dendrimer, including but not limited to fluoxetine (Prozac TM ), Paxil TM ), sertraline (Zoloft TM ), Citalopram (Celexa TM ), fluvoxamine (Luvox TM ), escitalopram (Lexapro TMIn other forms, SSRI-like drugs that may be included in the composition include, but are not limited to, one or more of Dapoxetine (Priligy), R-fluoxetine, Indalpine (Upstène), Zimelidine (Zelmid), Alaproclate (GEA-654), Centpropazine, Cericlamine (JO-1017), Femoxetine (Malexil: FG-4963), Ifoxetine (CGP-15210), Omiloxetine, Panuramine (WY-26002), Pirandamine (AY-23713), and Seproxetine ((S)-norfluoxetine).

[0157] In some forms, the composition may include one or more SSRI-related compounds in an amount effective to deliver a low dose of the SSRI-related compound. For example, although described as SNRIs, duloxetine (Cymbalta), venlafaxine (Effexor), and desvenlafaxine (Pristiq) can selectively act as serotonin reuptake inhibitors (SRIs). They are at least 10 times more selective for serotonin reuptake inhibition than for norepinephrine reuptake inhibition. The selectivity ratio for venlafaxine is about 1:30, for duloxetine is about 1:10, and for desvenlafaxine is about 1:14. At low doses, these SNRIs act as SSRIs; only at higher doses do they also significantly inhibit norepinephrine reuptake. In another example, Milnacipran (Ixel, Savella) and its stereoisomer, levomilnacipran (Fetzima), are the only widely marketed SNRIs that inhibit serotonin and norepinephrine to a similar degree, with the ratio of the two approaching 1:1. In a third example, Vilazodone (Viibryd) and vortioxetine (Trintellix) are SRIs that also act as modulators of serotonin receptors and are described as serotonin modulators and stimulators (SMS). Vilazodone is a 5-HT1A receptor partial agonist, while vortioxetine is a 5-HT1A receptor agonist and a 5-HT3 and 5-HT7 receptor antagonist. Litoxetine (SL 81-0385) and lubazodone (YM-992, YM-35995) are SRIs, of which ritoxetine is a 5-HT3 receptor antagonist and lubazodone is also a 5-HT2A receptor antagonist.

[0158] In a preferred embodiment, the dendrimer is conjugated to one or more SSRIs or SSRI derivatives as shown below in structures I-VI.

[0159]

[0160]

[0161] b. Serotonin-norepinephrine reuptake inhibitors (SNRIs)

[0162] The composition may include a dendrimer complexed with one or more SNRIs. "SNRI" refers to any of a class of compounds that act on and increase the levels of two monoamine neurotransmitters in the brain, namely serotonin and norepinephrine (Arroll B et al., Annals of Family Medicine, 2005; 3(5): 449-456). SNRIs exert their effects by: (i) inhibiting the presynaptic reuptake of serotonin from the synaptic cleft by binding to the SERT receptor; and (ii) inhibiting the presynaptic reuptake of norepinephrine from the synaptic cleft by binding to the norepinephrine (NAT) transporter. Norepinephrine (NE) is present in cell bodies in the pons and medulla oblongata. These cell bodies project neurons to the hypothalamus, thalamus, limbic system, locus coeruleus, lateral tegmental area, and cerebral cortex. In the peripheral nervous system, norepinephrine is used as a neurotransmitter by sympathetic ganglia located near the spinal cord or in the abdomen and Merkel cells located in the skin. NE is also released directly into the blood by contractions of the adrenal glands and the sphincters of the abdominal viscera and GI tract and bladder. Preventing reuptake can prolong the persistence of these monoamines in the synaptic clefts in the central nervous system (CNS) and the peripheral nervous system (PNS). Therefore, this can lead to increased postsynaptic receptor stimulation and additional postsynaptic neuron transmission.

[0163] SNRIs are often considered to have "dual action" (because of their mechanism of action) and are sometimes referred to as non-tricyclic serotonin and norepinephrine reuptake inhibitors. However, the specific extent of norepinephrine and serotonin reuptake inhibition depends on the dose and the drug. Suitable SNRIs that can be conjugated to the dendrimer composition include, but are not limited to, venlafaxine (Effexor XR TM ), desvenlafaxine (Pristiq and Khedezla TM ), duloxetine (Cymbalta, Irenka), milnacipran (Savella), levomilnacipran (Fetzima), tramadol hydrochloride (Tramadol HCl), sibutramine (Meridia), atomoxetine (Strattera) and bicifadine (DOV-220,075) and their derivatives.

[0164] In some forms, the SNRI conjugated to the dendrimer has a stronger binding affinity for the serotonin receptor than the norepinephrine receptor. For example, duloxetine and venlafaxine preferentially bind to the serotonin receptor than the norepinephrine receptor, thereby exerting greater efficacy on serotonin reuptake relative to norepinephrine reuptake. In some forms, the SNRI conjugated to the dendrimer has a stronger binding affinity for the norepinephrine receptor. In some forms, the SNRI conjugated to the dendrimer has equal binding affinity for the serotonin receptor and the norepinephrine receptor.

[0165] In some forms, SNRIs conjugated to dendrimers indirectly modulate signaling at non-serotonin and non-adrenergic receptors. In an exemplary embodiment, the dendrimer can be conjugated to venlafaxine, a synthetic bicyclic derivative of phenylethylamine and a substrate for P-glycoprotein (P-gp), which pumps it out of the brain. At high doses, venlafaxine decreases presynaptic reuptake of dopamine, thereby enhancing dopamine levels, particularly in the prefrontal cortex. The mechanism of action for the increase in dopamine levels involves inhibition of norepinephrine transporters. These transporters have a significant affinity for dopamine, resulting in the transporter being able to act on both dopamine and norepinephrine. Therefore, inhibition of norepinephrine transporters can lead to an increase in dopamine. This increase in dopamine occurs particularly in the prefrontal cortex, where dopamine transporters are scarce and reuptake is more heavily dependent on norepinephrine transporters. Venlafaxine also indirectly modulates opioid receptors, muscarinic acetylcholine receptors (mAChRs), histaminergic receptors, and α1 and α2-adrenergic receptors.

[0166] In some embodiments, the dendrimer is conjugated to one or more SNRIs or derivatives thereof in an amount effective to provide about 40 mg / day to about 60 mg / day, about 60 mg / day to about 80 mg / day, about 40 mg / day to about 100 mg / day, about 100 mg / day to about 150 mg / day, about 150 mg / day to about 200 mg / day, about 200 mg / day to about 250 mg / day, 250-300 mg / day, about 350 mg / day to about 450 mg / day of one or more SNRIs or derivatives thereof to the cell.

[0167] In some embodiments, dendrimers conjugated to SSRIs can exert anti-inflammatory effects on certain cells, which may improve the efficacy of treatments for mental health and neurological disorders.

[0168] In some embodiments, conjugation of SSRIs to dendrimers resulting in improved pharmacokinetics and receptor targeting may also significantly reduce the time required for onset of effect.

[0169] In a preferred embodiment, the dendrimer is conjugated to one or more SSRIs or SSRI derivatives as shown in Formula VIII-XI below.

[0170]

[0171]

[0172] c. Atypical antidepressants

[0173] The composition may include a dendrimer complexed with one or more atypical antidepressants. "Atypical antidepressants" refers to any antidepressant that works in a different manner than most other antidepressants. Atypical antidepressants are often used in patients with major depression who have had an inadequate response or intolerable side effects during first-line treatment with SSRIs. Suitable atypical antidepressants that can be conjugated to the dendrimer include, but are not limited to, agomelatine, bupropion, mianserin, mirtazapine, nefazodone, opipramol, tianeptine, and trazodone.

[0174] In an exemplary embodiment, the dendrimer can be compounded with agomelatine. Agomelatine is a receptor agonist for melatonin MT1 and MT2 receptors, and a receptor antagonist for serotonin 5-HT2C and 5-HT2B receptors. By antagonizing 5-HT2C, it specifically inhibits / increases norepinephrine and dopamine release in the frontal cortex. Therefore, it is sometimes classified as a norepinephrine-dopamine de-inhibitor. Antagonism of 5-HT2B is presumably the antidepressant property shared by agomelatine and several atypical antipsychotics such as aripiprazole.

[0175] In a second exemplary embodiment, the dendrimer can be complexed with mirtazapine. Mirtazapine has a dual mode of action. It is a noradrenergic and specific serotonergic antidepressant (NaSSA) that acts by antagonizing adrenergic α2-autoreceptors and α2-isoreceptors and blocking 5-HT2 and 5-HT3 receptors. Therefore, mirtazapine can enhance the release of norepinephrine and 5-HT1A-mediated serotonin transmission. Mirtazapine is extensively metabolized in the liver by cytochrome (CYP) P450 isozymes CYP1A2, CYP2D6, and CYP3A4.

[0176] In a third exemplary embodiment, the dendrimer can be complexed with nefazodone. Nefazodone is a phenylpiperazine compound that is structurally related to trazodone and is described as a serotonin antagonist and reuptake inhibitor (SARI). Nefazodone primarily acts as a potent antagonist of the serotonin 5-HT2A receptor, and to a lesser extent, a potent antagonist of the serotonin 5-HT2C receptor. It also has high affinity for α1-adrenergic receptors and serotonin 5-HT1A receptors, and has relatively low affinity for α2-adrenergic receptors and dopamine D2 receptors. Nefazodone also has significant affinity for serotonin, norepinephrine, and dopamine transporters, and therefore acts as a weak serotonin-norepinephrine-dopamine reuptake inhibitor (SNDRI).

[0177] In a fourth exemplary embodiment, the dendrimer can be complexed with trazodone. Trazodone is a mixed agonist and antagonist of various serotonin receptors, an antagonist of adrenergic receptors, a weak histamine H1 receptor antagonist, and a weak serotonin reuptake inhibitor. In some forms, trazodone is an antagonist of 5-HT2A and 5-HT2B receptors, a partial agonist of 5-HT1A receptors, and an antagonist of α1 and α2 adrenergic receptors.

[0178] In another exemplary embodiment, the dendrimer can be complexed with bupropion, a dopamine and norepinephrine reuptake inhibitor. Bupropion can reduce the presynaptic reuptake of dopamine and norepinephrine and acts as an antagonist of multiple nicotinic acetylcholine receptors. Bupropion is generally metabolized to three known active metabolites, hydroxybupropion (R, R-hydroxybupropion and S, S-hydroxybupropion) and diastereomers, namely threohydrobupropion and erythrohydrobupropion.

[0179] In yet another exemplary embodiment, the dendrimer can be complexed with mianserin. Mianserin is a tetracyclic antidepressant that has antihistamine and hypnotic-sedative effects but little anticholinergic effects. Mianserin appears to exert its effects via antagonism of histamine and serotonin receptors and inhibition of norepinephrine reuptake. More specifically, mianserin is a histamine H1 receptor, serotonin 5-HT 1D , 5-HT 1F , 5-HT 2A , 5-HT 2B , 5-HT 2CMianserin is an antagonist / inverse agonist at most or all sites of the 5-HT, 5-HT3, 5-HT6 and 5-HT7 receptors and adrenergic α1 and α2 adrenergic receptors, and is also a norepinephrine reuptake inhibitor. As a high-affinity H1 receptor inverse agonist, mianserin has a strong antihistamine effect (e.g., sedative effect). In contrast, it has a lower affinity for muscarinic acetylcholine receptors and therefore lacks anticholinergic properties. Mianserin has been found to be a low-affinity but potentially significant partial agonist of the K-opioid receptor.

[0180] d. Tricyclic antidepressants (TCAs)

[0181] The composition may include a dendrimer complexed with one or more tricyclic antidepressants. "Cyclic antidepressants" refers to any antidepressant designated as tricyclic or tetracyclic, depending on the number of rings in its chemical structure - three (tri) or four (tetra). Tricyclic antidepressants act on approximately five different neurotransmitter pathways to achieve their effects (Stahl et al., Prim. Care Companion J. Clin. Psychiatry 2004; 6(4): 159). Tricyclic antidepressants block the reuptake of serotonin and norepinephrine in the presynaptic terminal, which results in an increased concentration of these neurotransmitters in the synaptic cleft. Increased concentrations of norepinephrine and serotonin in the synapse may contribute to the antidepressant effects of tricyclic antidepressants. In addition, in some forms, they can act as competitive antagonists of postsynaptic alpha cholinergic receptors (α1 and α2), muscarinic and histaminergic receptors (H1). The structure of the receptor greatly affects the binding affinity of TCAs.

[0182] The chemical structure of TCAs consists of a tricyclic structure with attached secondary or tertiary amines. Secondary amines include desipramine, nortriptyline, and protriptyline, while tertiary amines include amitryptiline, clomipramine, doxepin, imipramine, and trimipramine. Tertiary amines tend to have a greater blocking effect on serotonin reuptake, while secondary amines have a greater blocking effect on norepinephrine uptake. The combination of different amine structures and the variation in chemical composition contribute to the variety of adverse reactions seen with TCA use, as these factors affect the affinity and binding of the TCA receptors.

[0183] Suitable tricyclic antidepressants that can be conjugated to the dendrimer include, but are not limited to, amitriptyline (e.g., Elavil, Endep), amitriptylinoxide (e.g., Amioxy, Ambivalon, Equilibrin), clomipramine (e.g., Anafranil), desipramine (e.g., Norpramin, Pertofrane), dibenzepin (e.g., Noveril, Victoril), dimetacrine (e.g., Istonil), dosulepin (e.g., Prothiaden), doxepin (e.g., Adapin, Sinequan), imipramine (e.g., Tofranil), lofepramine (e.g., Lomont, Gamanil), amoxapine, melitracen (e.g., Dixeran, Melixeran, Trausabun), nitro-nitropropene (e.g., nitropropene), nitro-propene (e.g., nitro- ... (Nitroxazepine) (e.g., Sintamil), Nortriptyline (e.g., Pamelor, Aventyl), Noxiptiline (e.g., Ageedal, Elronon, Nogedal), Opipramol (e.g., Insidon), Pipofezine (e.g., Azafen / Azaphen), Protriptyline (e.g., Vivactil), and / or Trimipramine (e.g., Surmontil).

[0184] Suitable tetracyclic antidepressants that can be conjugated to the dendrimer include, but are not limited to, Maprotiline (Ludiomil; also classified as a TCA and grouped with secondary amines), Mianserin (Tolvon), Mirtazapine (Remeron), Setiptiline (Tecipul), Amoxapine (Asendin: usually classified as a TCA and grouped with secondary amines), Quetiapine (Seroquel; an atypical antipsychotic sometimes used as an adjunctive antidepressant). Other tetracyclic antidepressants that can be conjugated to dendrimers include, but are not limited to, Benzoctamine (e.g., Tacitin), Loxapine (e.g., Adasuve, Loxitane), Mazindol (e.g., Mazanor, Sanorex), Aptazapine (CGS-7525A: a close analog of mirtazapine), Esmirtazapine (ORG-50,081; the (S)-(+) enantiomer of mirtazapine), Oxaprotiline (C 49-802BDA; a close analog of maprotiline), and Ciclazindol (WY-23,409; a close analog of mazindol). Benzoctamine (e.g., Tacitin) is a tetracyclic compound and is closely related to maprotiline, with the two compounds differing only in the length of the side chain, but Benzoctamine is typically used as an anxiolytic. Loxapine (e.g., Adasuve, Loxitane) is a typical antipsychotic that produces amoxapine as a major metabolite and is said to have antidepressant effects, but is not generally considered a tetracyclic antidepressant. Drugs that contain four rings that are not all fused together but can still be classified as tetracyclics include mazindol. Mazindol (Mazanor, Sanorex) is a monoamine reuptake inhibitor that is generally used as an appetite suppressant and has antidepressant effects.

[0185] e. Monoamine oxidase inhibitors (MAOIs)

[0186] The compositions may include a dendrimer complexed with one or more monoamine oxidase inhibitors (MAOIs).

[0187] MAOIs are responsible for blocking the enzyme monoamine oxidase (Shulman KI et al., CNS drugs. 2013; 27(10): 789-797). MAOIs break down different types of neurotransmitters from the brain: norepinephrine, serotonin, dopamine, and tyramine. MAOIs inhibit the breakdown of these neurotransmitters, thereby increasing their levels and allowing them to continue to affect cells already affected by depression.

[0188] There are two types of monoamine oxidase, A and B. MAO A is mainly found in the placenta, intestines, and liver, but MAO B is found in the brain, liver, and platelets. The ratio of MAO-A to MAO-B varies throughout the body. In the human brain, the ratio of MAO-A to MAO-B is 25% to 75%, while in the liver, the ratio is 50% to 50%, in the intestine, the ratio is 80% to 20%, and in peripheral adrenergic neurons, the ratio is 90% to 10%.

[0189] Serotonin and norepinephrine are substrates for MAO A, but phenylethylamine, methylhistamine, and tryptamine are substrates for MAO B. Dopamine and tyramine are metabolized by MAO A and B. Selegiline and rasagiline are irreversible and selective inhibitors of MAO type B, but safinamide is a reversible and selective MAO B inhibitor.

[0190] In some forms, the dendrimer is complexed with one or more reversible and / or irreversible MAOIs. Reversible inhibitors of monoamine oxidase A (RIMA) are a subclass of MAOIs that selectively and reversibly inhibit the MAO-A enzyme. Suitable reversible MAOIs include, but are not limited to, befloxatone (MD-370,503), brofaromine (Consonar), furazolidone (Furoxone), linezolid (Zyvox), moclobemide (Aurorix), and / or toloxatone (Humoryl). Some MAOIs covalently bind to monoamine oxidases, thereby irreversibly inhibiting them; the bound enzyme cannot function and thus enzyme activity is blocked until the cell produces new enzymes. Suitable irreversible MAOIs that can be conjugated to the dendrimer include, but are not limited to, clorgyline, iproniazid (marsilid), isocarboxazid (Marplan), Nialamide (Niamid), Pargyline (Eutonyl), Phenelzine (Nardil), Procarbazine (Matulane), Rasagiline (Azilect), Selegiline (Selegiline), l-deprenyl (Eldepryl and Emsam), and Tranylcypromine (parnate).

[0191] f. Benzodiazepine Drugs

[0192] The composition may include one or more benzodiazepines Dendrimers of drug-like complexes. BZD, BDZ, BZ are a class of psychoactive drugs whose core chemical structure is a benzene ring and diazepam. Benzodiazepines act on the central nervous system (CNS) Receptor (BZ-R). The receptor is a protein containing five transmembrane subunits that form a chloride channel in the center, the GABA-A receptor. The five subunits consist of two α, two β, and one γ subunit. The extracellular parts of the α and β subunit proteins form the receptor site for gamma-aminobutyric acid (GABA), an inhibitory neurotransmitter. The extracellular parts of the α and γ subunit proteins form the benzodiazepine The binding site for drugs of this class. Activation of BZ-R causes a conformational change in the central pore, which allows chloride ions to enter the neuron. The influx of chloride anions hyperpolarizes the neuron, resulting in a decrease in the firing of action potentials in that neuron.

[0193] In some forms, the benzodiazepine conjugated to the dendrimer Ultra-short-acting benzodiazepines Ultra-short-acting benzodiazepines Non-limiting examples of the class include, but are not limited to, flunitrazepam, diazepam, and midazolam. In some forms, the benzodiazepine conjugated to the dendrimer Benzodiazepines Short-acting benzodiazepines Non-limiting examples of the class include, but are not limited to, alprazolam, chlordiazepoxide In some forms, benzodiazepines conjugated to dendrimers Benzodiazepines Long-acting benzodiazepines Non-limiting examples of the class include, but are not limited to, clonazepam, oxazepam, and prazepam.

[0194] g. Other antidepressants

[0195] The composition may include a dendrimer complexed with one or more beta-blockers, neurosteroids and / or anticonvulsants (Wang M., Neurosteroids and GABA-A Receptor Function. 2011; 2).

[0196] "Beta blockers," also called beta-adrenergic blockers, block norepinephrine and epinephrine (adrenaline) from binding to beta receptors on nerves. Blocking these transmitters helps control some of the physical symptoms of anxiety. It also helps to lower heart rate and lower blood pressure. Non-limiting examples of beta blockers include fluoxetine, paroxetine, duloxetine, and bupropion.

[0197] As used herein, "neurosteroids" refer to steroids that are produced in the brain or transported to the brain that alter neuronal excitability. Suitable neurosteroids include, but are not limited to, brexanolone and SGE-217, and include GABA modulators such as ganaxolone, minaxolone, and zuranolone.

[0198] As used herein, "anticonvulsant" refers to drugs used to prevent, reduce or control seizures, including antidepressants, steroids and other drugs.

[0199] 2. Antipsychotics

[0200] a. Typical antipsychotic drugs

[0201] The composition may include a dendrimer complexed with one or more typical antipsychotics (also known as first generation antipsychotics). Typical antipsychotics generally work by inhibiting dopaminergic neurotransmission; they work best when they block activity at D2 dopamine receptors in the brain (Wang Shanghai archives of psychiatry. 2013; 25(3): 134-140). In some forms, typical antipsychotics may also have noradrenergic, cholinergic, and histaminergic blocking effects. The one or more typical antipsychotics conjugated to the dendrimer may belong to one of the following subclasses: phenothiazines, butyrophenones, thioxanthenes, indolinones, diphenylpiperidines, and / or diphenylbutylpiperidines.

[0202] i. Phenothiazines

[0203] The composition may include a dendrimer compounded with one or more phenothiazines. Phenothiazines are the largest chemical group of typical antipsychotic drugs, containing more than 40 compounds (divided into three subtypes). Phenothiazines have the same three-ring structure, in which different side chains are attached to the nitrogen atom of the middle ring. The activity of this group may be affected by substitutions at the 2 or 10 positions. Phenothiazines are divided into three subclasses based on substitutions at the 10 position: aliphatic phenothiazines, piperidine phenothiazines and piperazine phenothiazines.

[0204] In some forms, the one or more phenothiazines conjugated to the dendrimer are aliphatic (low / medium potency) phenothiazines. Suitable aliphatic phenothiazines include, but are not limited to, Chlorpromazine, Levomepromazine, Promazine, and Triflupromazine.

[0205] In some forms, the one or more phenothiazines conjugated to the dendrimer are piperidine phenothiazines. Piperidine phenothiazines are low or medium potency agents. Suitable piperidine phenothiazines include, but are not limited to, mesoridazine, pericyazine, pipotiazine, and thioridazine.

[0206] In some forms, the one or more phenothiazines conjugated to the dendrimer are piperazine phenothiazines. Piperazine phenothiazines are medium-acting or high-acting agents. Suitable piperazine phenothiazines include, but are not limited to, Perphenazine, Fluphenazine, and Trifluoperazine.

[0207] In an exemplary embodiment, the phenothiazines conjugated to the dendrimer have the structural formula shown below as Formula X1.

[0208]

[0209] ii. Non-phenothiazine drugs

[0210] The composition may include a dendrimer complexed with one or more non-phenothiazine drugs. The non-phenothiazine drugs that may be conjugated to the dendrimer may be one or more of butyrophenones, thioxanthenes, indolinones, diphenylpiperidines, and / or diphenylbutylpiperidines.

[0211] In some forms, the dendrimer is conjugated to one or more butyrophenones and / or diphenylbutylpiperidines. Butyrophenones and diphenylbutylpiperidines are highly potent agents. Non-limiting examples of butyrophenones include, but are not limited to, Benperidol, Droperidol, and Haloperidol. Non-limiting examples of diphenylbutylpiperidines include, but are not limited to, Fluspirilen and Pimozide.

[0212] In some forms, the dendrimer is conjugated to one or more thioxanthenes, dibenzepines, and / or dihydroindolones. Thioxanthenes, dibenzepines, and dihydroindolones are low or medium potency agents. Non-limiting examples of thioxanthenes include, but are not limited to, clopentixol, flupenthixol, thiothixene, and zuclopenthixol. Non-limiting examples of dihydroindolones include, but are not limited to, molindone. Non-limiting examples of dibenzepines include, but are not limited to, clotiapine and loxapine.

[0213] b. Atypical antipsychotics

[0214] The composition may include a dendrimer complexed with one or more atypical antipsychotics. The term "atypical" refers to antipsychotics that produce minimal extrapyramidal side effects (EPS) at clinically effective antipsychotic doses, have a low tendency to cause tardive dyskinesia (TD) after long-term treatment, and treat both positive and negative signs and symptoms of mental health disorders (e.g., schizophrenia). Suitable atypical antipsychotics that can be conjugated to the dendrimer compositions include, but are not limited to, clozapine (Clozaril), risperidone (Risperdal), olanzapine (Zyprexa), quetiapine (Seroquel), ziprasidone (Geodon), aripiprazole (Abilify), zotepine, cariprazine, buxipirazole, asenapine (Saphris), iloperidone (Fanapt), amisulpride, blondanserin, melperone, perospirone, remoxipride, sertindole, sulpiride, lurasidone (Latuda), and paliperidone (Invega), which is the active metabolite of risperidone.

[0215] Typically, the dendrimer is conjugated to one or more atypical antipsychotics that are highly selective for serotonin (5-HT) and dopamine (D) receptors. More preferably, the one or more atypical antipsychotics bind as antagonists for 5-HT2A receptors and D2 receptors. For example, the dendrimer is conjugated to one or more of clozapine (Clozaril), risperidone (Risperdal), olanzapine (Zyprexa), quetiapine (Seroquel), ziprasidone (Geodon), aripiprazole (Abilify), zotepine, cariprazine, brexipiprazole, asenapine (Saphris), iloperidone (Fanapt), amisulpride, blonanserin, melperone, perospirone, remopride, sertindole, sulpiride, lurasidone (Latuda), and paliperidone (Invega), which is an active metabolite of risperidone.

[0216] The dendrimers may be conjugated to one or more atypical antipsychotics, which are selective for one or more other receptors. For example, some atypical antipsychotics are also potent serotonin-1A (5-HT1A; aripiprazole), serotonin-1C (5-HT1C; clozapine, olanzapine, risperidone), histamine-1 (H1; olanzapine, quetiapine) and α1- (aripiprazole, clozapine, olanzapine, paliperidone, quetiapine) and α2-adrenergic (clozapine, olanzapine, paliperidone, quetiapine, risperidone) receptor blockers. In another example, some atypical antipsychotics, such as clozapine, olanzapine and quetiapine, bind to histamine-1 (H1) receptors with comparable affinity to D2, 5-HT2A / 2C and α2-adrenergic receptors.

[0217] In some embodiments, the dendrimers may be conjugated to one or more atypical antipsychotics that have different effects on the central nervous system and nervous system in a tissue-specific manner. For example, risperidone and aripiprazole (despite their inhibitory effects on the firing activity of 5-HT neurons) each increase cortical 5-HT levels on their own and enhance the increase in 5-HT concentrations in the frontal cortex induced by escitalopram. In addition, risperidone enhances the anxiolytic and antidepressant-like behavioral effects of escitalopram.

[0218] Drugs that are particularly useful for treating ADHD and its symptoms can be conjugated to dendrimers for use in treating individuals. These include ADHD drugs such as methylphenidate, amphetamine, atomoxetine, clonidine, guanfacine, viloxazine and their analogs / modifications. These drugs are often used in patients with depression, PTSD and bipolar disorder. See https: / / www.webmd.com / add-adhd / adhd-medication-chart and https: / / www.additudemag.com / adhd-medication-for-adults-and-children / . In some forms, the efficacy of non-stimulant ADHD drugs can be significantly improved via conjugation to dendrimers.

[0219] Some of these have a mechanism of action similar to that of antidepressants / antipsychotics, such as atomoxetine (SNRI), viloxazine (SNRI), and methylphenidate, which blocks the reuptake of dopamine and norepinephrine. Current antidepressants used to treat symptoms of ADHD include bupropion, desipramine, imipramine, and nortriptyline.

[0220] C. Coupling agents and spacers

[0221] Dendrimer-active agent conjugates can be formed by covalent conjugation or non-covalent attachment of one or more active agents to a dendrimer. In a preferred embodiment, one or more active agents are covalently conjugated to a dendrimer.

[0222] Optionally, one or more active agents are conjugated to the dendrimer via one or more spacers. The term "spacer" includes chemical moieties and functional groups used to connect the active agent to the dendrimer. The spacer can be a single chemical entity or two or more chemical entities linked together. The spacer can include any small chemical entity, a peptide, or a polymer with a thiol, thiopyridine, succinimidyl, maleimide, vinyl sulfone, carbonate, etc.

[0223] In some embodiments, the spacer for conjugating the active agent to the dendrimer comprises different bonds, such as disulfide bonds, ester bonds, carbonate bonds, carbamate bonds, thioester bonds, hydrazine bonds, hydrazide bonds, ether bonds, and amide bonds. The spacer between the dendrimer and the active agent can be designed to provide a releasable or non-releasable form of the dendrimer conjugate in vivo. In some embodiments, the conjugation between the active agent and the dendrimer is carried out through an appropriate spacer, which comprises an ester bond between the active agent and the dendrimer. In some embodiments, one or more spacers between the dendrimer and the active agent can provide the desired and effective release kinetics in vivo. These spacers can contain cleavable linkages (e.g., esters, disulfide bonds, phosphodiesters, triglycyl peptides, and hydrazines) or non-cleavable linkages (e.g., amides, ethers, and aminoalkyls). The conjugation between the active agent and the dendrimer can be carried out using reactions known in the art, such as click chemistry, acid amine coupling, Steglich esterification, and the like.

[0224] In some embodiments, the conjugation between the active agent and the dendrimer is through a spacer comprising a disulfide bond, an ester bond, an ether bond, a thioester bond, a carbamate bond, a carbonate bond, a hydrazine bond, an ether bond, or an amide bond, or a combination thereof. In some embodiments, the conjugation between the active agent and the dendrimer is through an appropriate spacer comprising an ester bond or an amide bond between the agent and the dendrimer, depending on the desired release kinetics of the agent.

[0225] The spacer can be selected from compounds terminated by thiol, thiopyridine, succinimidyl, maleimide, vinyl sulfone and carbonate. The spacer can include compounds terminated by thiopyridine, such as dithiodipyridine, N-succinimidyl 3-(2-pyridyldithio)-propionate (SPDP), succinimidyl 6-(3-[2-pyridyldithio]-propionamido)hexanoate LC-SPDP or sulfo-LC-SPDP. The spacer may also include peptides, wherein the peptide is linear or cyclic, essentially having a thiol group, such as glutathione, homocysteine, cysteine ​​and its derivatives, arg-gly-asp-cys (RGDC), cyclo(Arg-Gly-Asp-d-Phe-Cys)(c(RGDfC)), cyclo(Arg-Gly-Asp-D-Tyr-Cys), cyclo(Arg-Ala-Asp-d-Tyr-Cys). The spacer may be a thiol acid derivative, such as 3-mercaptopropionic acid, thioglycolic acid, 4-mercaptobutyric acid, thiolane-2-one, 6-mercaptohexanoic acid, 5-mercaptopentanoic acid and other thiol derivatives, such as 2-mercaptoethanol and 2-mercaptoethylamine. The spacer can be thiosalicylic acid and its derivatives, (4-succinimidyloxycarbonyl-methyl-α-2-pyridylthio) toluene, (3-[2-pyridylthio] propionyl hydrazide, the spacer can have a maleimide terminal group, wherein the spacer includes a polymer or a small chemical entity, such as bismaleimido diethylene glycol and bismaleimido triethylene glycol, bismaleimido ethane, bismaleimido hexane. The spacer can include vinyl sulfones, such as 1,6-hexane-bisvinyl sulfone. The spacer can include a thioglycoside, such as thioglucose. The spacer can be a reduced protein, such as bovine serum albumin and human serum albumin, any thiol-terminated compound capable of forming a disulfide bond. The spacer can include polyethylene glycol with maleimide, succinimidyl and / or thiol terminal groups.

[0226] D. Dendrimer-Drug Conjugates

[0227] Dendrimer-active agent conjugates can be formed by covalently conjugating or non-covalently attaching an antidepressant and / or antipsychotic to a dendrimer, dendrimer or hyperbranched polymer. Methods for conjugating one or more active agents to dendrimers are known, such as those described in U.S. Publication Nos. 2011 / 0034422, 2012 / 0003155 and 2013 / 0136697.

[0228] In some embodiments, one or more active agents are covalently conjugated to one or more terminal groups (e.g., terminal hydroxyl groups) of the dendrimer. In some embodiments, the dendrimer conjugates include one or more active agents conjugated to the dendrimer via one or more spacers. The spacer between the dendrimer and the active agent can be designed to provide a releasable or non-releasable form of the dendrimer conjugate (in vivo). For example, the spacer can be cleavable or contain a cleavable chemical bond, for example, cleaved by exposure to an intracellular compartment of a target nerve and / or glial cell or binding to a receptor on the surface or inside a target nerve and / or glial cell in vivo. Examples of cleavable bonds that can be used for the spacer of the dendrimer-active agent conjugate include esterase-sensitive ester bonds, glutathione-sensitive disulfide bonds, phosphatase-sensitive phosphodiester bonds, oligopeptides (e.g., triglycyl peptide linkers capable of lysosomal release), acid-cleavable hydrazine bonds, and the like. In some embodiments, the spacer between the dendrimer and the active agent can provide the desired and effective release kinetics in vivo. In some embodiments, the spacer between the dendrimer and the active agent may be non-cleavable or comprise a non-cleavable chemical bond, such as an amide bond, an ether bond, and an aminoalkyl bond.

[0229] Typically, the spacer between the dendrimer and the active agent is of sufficient length to enable the active agent conjugated thereto to reach and bind to a target receptor on the surface and / or inside the target cell. The length of the spacer can vary, depending on the location of the target receptor (e.g., on the cell surface, in the cytoplasm of the cell, or in the intercellular compartment of the cell) and / or the density of the receptor when located on the cell surface.

[0230] Dendrimers can be of generation 2, 3, 4, 5, 6, up to generation 10. In some embodiments, the dendrimer is conjugated to one or more active agents via a spacer containing a cleavable (ester, disulfide, phosphodiester, triglycine, and hydrazine) or non-cleavable (amide, ether, and aminoalkyl) bond.

[0231] The density of active agents covalently bound or non-covalently attached to the dendrimer can be adjusted depending on the specific antidepressant and / or antipsychotic drug being delivered, the target receptor, the target nerve and / or glial cell, the location of the target nerve and / or glial cell, etc. For example, multiple active agents conjugated to the dendrimer are located at the periphery of the dendrimer and the surface density of the active agents is at least 1 active agent / nm 2 (Number of conjugated active agents / nm 2 For example, in some embodiments, per nm 2The surface density of the active agent is greater than 2, 3, 4, 5, 6, 7, 8, 9, 10, such as at least 10, 15, 20, 25, 30, 35, 40, 45, 50 or greater than 50. In some embodiments, the volume density of the active agent is from about 4 to about 50 groups / nm 3 , about 5 to about 30 groups / nm 3 or about 10 to about 20 groups / nm 3 .

[0232] Typically, the dendrimer-active agent conjugate has a hydrodynamic volume in the nanometer range. For example, in some embodiments, the diameter of the glucose dendrimer-active agent conjugate comprising one or more antidepressants and / or antipsychotics conjugated to the dendrimer is from about 2 nm to about 100 nm, or greater than 100 nm, up to 500 nm, depending on the generation of the dendrimer, the chemical composition and amount of the active agent conjugated thereto. In some embodiments, the dendrimer-active agent conjugate comprising one or more antidepressants and / or antipsychotics conjugated to the dendrimer has a diameter that is effective to penetrate brain tissue and remain on the surface and / or in the target neural and / or glial cells for a period of time sufficient for the active agent to bind to a targeted receptor on the surface and / or in the target neural and / or glial cells. In some embodiments, the dendrimer-active agent conjugates comprising one or more antidepressant and / or antipsychotic drugs bound to the dendrimer have a diameter effective to remain in the peripheral circulation and to remain on the surface and / or in target neural and / or glial cells for a sufficient period of time to allow the active agent to bind to a targeted receptor on the surface and / or in target neural and / or glial cells (such as, for example, neural and / or glial cells of the gastrointestinal system).

[0233] The dendrimer-active agent conjugate can be neutral, have a positive charge or a negative charge. In some embodiments, the dendrimer-active agent conjugate is neutral. The presence of an antidepressant and / or antipsychotic can affect the surface charge of the dendrimer-active agent conjugate. In some embodiments, the surface charge of the dendrimer conjugated to an antidepressant and / or antipsychotic is between -100 mV and 100 mV, between -50 mV and 50 mV, between -25 mV and 25 mV, between -20 mV and 20 mV, between -10 mV and 10 mV, between -10 mV and 5 mV, between -5 mV and 5 mV, or between -2 mV and 2 mmV. The above ranges include all values ​​from -100 mV to 100 mV. In preferred embodiments, the surface charge of the dendrimer-active agent conjugate is neutral or nearly neutral, ie, from about -10 mV to about 10 mV, inclusive.

[0234] An exemplary dendrimer-active agent conjugate is represented by formula (I). The dendrimer of the exemplary conjugate contains surface hydroxyl groups, wherein one or more surface hydroxyl groups are conjugated to one or more active agents via one or more spacers as shown in formula (I) below.

[0235]

[0236] wherein D can be a 1st to 10th generation or a 2nd to 10th generation dendrimer, such as any of the above, such as PAMAM (e.g., a hydroxyl-terminated PAMAM dendrimer) or a glucosyl dendrimer; each occurrence of L can be any suitable chemical moiety suitable for providing tailored drug release and receptor binding, preferably containing a triazole moiety; Y can be a bond or selected from a secondary amide (-CONH-), a tertiary amide (-CONR-), a sulfonamide (-S(O)2-NR-), a secondary carbamate (-OCONH-; -NHCOO-), a tertiary carbamate (-OCONR-; -NRCOO-), a carbonate (-OC(O)-O-), a urea (-NHCONH-; -NRCONH-; -NHCONR-, -NRCONR-), a carbinol (-CHOH-, -CROH-), a disulfide bond, a phosphodiester group

[0237]

[0238] A linkage of hydrazine, hydrazone, hydrazide, ester (-C(O)-O-), ether (-O-) and oligopeptide (e.g., triglycyl peptide), wherein R is an alkyl, aryl or heterocyclic group; each occurrence of X may be an antidepressant and / or antipsychotic, wherein the functional group of X (e.g., an amino group, including a primary, secondary or tertiary amino group; a carboxyl group; or a hydroxyl group) forms part of the linkage Y; n may be an integer from 1 to 100; and m may be an integer from 16 to 4096.

[0239] The oxygen atoms in formula (I) are derived from surface functional groups of the dendrimer, such as surface hydroxyl groups, wherein the surface hydroxyl groups may or may not be part of a terminal sugar moiety / molecule (e.g., glucose). Although not illustrated in formula (I), one or more hydroxyl groups in the dendrimer that are not conjugated to an active agent may be modified with one or more carbohydrates and / or polyalkylene glycols (e.g., PEG).

[0240] When administered to a subject in need thereof, the antidepressant and / or antipsychotic X of formula (I) can bind to a target receptor on the surface of a target cell or inside a target cell. In some embodiments, when the antidepressant and / or antipsychotic X binds to the target receptor, the agent X remains conjugated to the dendrimer. In these embodiments, after binding, the agent X can be released from the dendrimer or remain conjugated to the dendrimer as a complete dendrimer-active agent conjugate. In some embodiments, the antidepressant and / or antipsychotic X is released from the dendrimer at a location close to the target receptor and then binds to the target receptor.

[0241] In some embodiments, each occurrence of L can be represented by -A'-L1-B'-L2-, wherein A' can be a carbonyl (-C(O)-) or a bond (including single bonds, double bonds and triple bonds, such as single bonds); B' can be a bond (including single bonds, double bonds and triple bonds, such as single bonds), amide, ester, ether, thiol, dithiol, aryl, heteroaryl, polyaryl, heteropolyaryl or heterocycle; and L1 and L2 can independently be a bond, alkylene, heteroalkylene, aryl, aralkyl, ether, polyether, thiol, dithiol, thiol ether, polythioether, oligopeptide, polypeptide, oligo(alkylene glycol) or polyalkylene glycol, or L1 and L2 can independently be composed of a combination of these groups, such as a combination of alkylene and polyether, a combination of alkylene and thiol or dithiol, a combination of alkylene and oligopeptide, a combination of alkylene, polyether, thiol or dithiol, or a combination of polyether and thiol or dithiol. In some forms, L1-B'-L2- together form a chemical moiety selected from -alkylene-triazole-di(alkylene glycol)-, -di(alkylene glycol)-triazole-alkylene-, -alkylene-triazole-oligo(alkylene glycol)-, -oligo(alkylene glycol)-triazole-alkylene-, -alkylene-triazole-poly(alkylene glycol)-, -poly(alkylene glycol)-triazole-alkylene-, -alkylene-triazole-ether-, -alkylene-triazole-alkylene-, -alkylene-amide-alkylene-, and combinations thereof.

[0242] In some embodiments, B' can be a bond (including single, double, and triple bonds, such as a single bond), an amide group, or a heterocyclic group, such as a triazole group.

[0243] In some embodiments, L1 can be a bond; an alkylene group, such as C1-C 10 alkylene, C1-C8 alkylene, C1-C6 alkylene, C1-C5 alkylene, C1-C4 alkylene or C1-C3 alkylene; or oligo- or poly-(alkylene glycol), for example

[0244]

[0245] 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.

[0246] In some embodiments, L2 can be a bond; an alkylene group, such as C1-C 10 alkylene, C1-C8 alkylene, C1-C6 alkylene, C1-C5 alkylene, C1-C4 alkylene or C1-C3 alkylene; oligo- or poly-(alkylene glycol), for example

[0247]

[0248] wherein p is an integer from 1 to 20, 1 to 18, 1 to 16, 1 to 14, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 or 2; an oligopeptide or polypeptide, such as a triglycyl peptide; a thiol; or a dithiol; or L2 is composed of a combination of two or more of an alkylene, an oligopeptide or poly(alkylene glycol), an oligopeptide or polypeptide, a thiol, and a dithiol. For example, L2 is represented by

[0249]

[0250] wherein p, q, r, s, t and u are independently integers from 0 to 10, 0 to 8, 0 to 6, 0 to 5, 0 to 4, 0 to 3 or 0 to 2, such as 0, 1 or 2; and G' is a thiol, a dithiol, an oligopeptide (e.g., a triglycyl peptide) or a polypeptide.

[0251] In some embodiments, Y is the least cleavable linkage in vivo. In some embodiments, Y is a cleavable linkage in vivo. In some embodiments, Y is an amide (-CONH-), an ester (-C(O)-O-), an ether (-O-), a phosphodiester, or a disulfide.

[0252] In some embodiments, L and Y are both single bonds, and D is directly conjugated to X (active agent or analog thereof) through an ether bond.

[0253] In some embodiments, D is a 2nd generation PAMAM dendrimer, a 3rd generation PAMAM dendrimer, a 4th generation PAMAM dendrimer, a 5th generation PAMAM dendrimer, a 6th generation PAMAM dendrimer, a 1st generation glucose dendrimer, a 2nd generation glucose dendrimer, a 3rd generation glucose dendrimer, a 4th generation glucose dendrimer, a 5th generation glucose dendrimer, or a 6th generation glucose dendrimer.

[0254] More specific exemplary dendrimer-active agent conjugates are shown in the following embodiments.

[0255] Diagnostic agents

[0256] In some cases, the agent delivered to the target neural and / or glial cells or tissues via the dendrimer is a diagnostic agent. Examples of diagnostic agents that can be delivered to the brain via the glucose dendrimer conjugate include paramagnetic molecules, fluorescent compounds, magnetic molecules and radionuclides, x-ray imaging agents and contrast agents. The dendrimer conjugate may include an agent that can be used to determine the location of the administered composition. Agents that can be used for this purpose include fluorescent labels, radionuclides and contrast agents.

[0257] Exemplary diagnostic agents include dyes, fluorescent dyes, near infrared dyes, SPECT imaging agents, PET imaging agents, and radioisotopes. Representative dyes include carbocyanine, indocarbocyanine, oxacarbocyanine, thüicarbocyanine and merocyanine, polymethine, coumarin, rhodamine, xanthan gum, fluorescein, boron-dipyrromethene (BODIPY), Cy5, Cy5.5, Cy7, VivoTag-680, VivoTag-S680, VivoTag-S750, AlexaFluor660, AlexaFluor680, AlexaFluor700, AlexaFluor750, AlexaFluor790, Dy677, Dy676, Dy682, Dy752, Dy780, DyLight547, Dylight647, HiLyte Fluor 647, HiLyteFluor 680, HiLyte Fluor 750, IRDye 800CW, IRDye 800RS, IRDye 700DX, ADS780WS, ADS830WS and ADS832WS.

[0258] Exemplary SPECT or PET imaging agents include chelators such as diethylenetriaminepentaacetic acid (DTPA), 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), diaminedithiol, activated mercaptoacetyl-glycyl-glycyl-glycine (MAG3), and hydrazinonicotinamide (HYNIC).

[0259] Exemplary isotopes include Tc-94m, Tc-99m, In-111, Ga-67, Ga-68, Gd3+, Y-86, Y-90, Lu-177, Re-186, Re-188, Cu-64, Cu-67, Co-55, Co-57, F-18, Sc-47, Ac-225, Bi-213, Bi-212, Pb-212, Sm-153, Ho-166, and Dy-166.

[0260] In an exemplary embodiment, the dendrimer composition includes one or more radioisotopes suitable for positron emission tomography (PET) imaging. Exemplary positron-emitting radioisotopes include carbon-11 ( 11 C), copper-64 ( 64 Cu), nitrogen-13( 13 N), oxygen-15( 15 O), gallium-68( 68 Ga) and fluorine-18 ( 18 F), such as 2-deoxy-2- 18 F-fluoro-β-D-glucose ( 18 F-FDG).

[0261] In further embodiments, a single dendrimer conjugate composition can simultaneously treat and / or diagnose a disease or condition at one or more locations in the body.

[0262] E. Exemplary Dendrimer-Agent Conjugates

[0263] In a preferred embodiment, the dendrimer is conjugated to fluoxetine as shown in structures I and II below, wherein n is an integer from 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0264]

[0265] In other embodiments, the dendrimer is conjugated to paroxetine as shown in Structure III below.

[0266]

[0267] In other embodiments, the dendrimer is conjugated to venlafaxine or a venlafaxine analog as shown in the following structures IV-VI, wherein n is an integer from 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0268]

[0269]

[0270] In other embodiments, the dendrimer is conjugated to citalopram or a citalopram analog as shown below in structures VII and VIII, wherein n is an integer from 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0271]

[0272] III. Methods for preparing dendrimers and conjugates thereof

[0273] Methods of synthesizing dendrimers and preparing dendrimer nanoparticles are also described.

[0274] A. Methods of Preparing Dendrimers

[0275] Dendrimers can be prepared by a variety of chemical reaction steps. Dendrimers are usually synthesized according to methods that allow control of their structure at every stage of their construction. Dendritic structures are mainly synthesized by two main different methods: divergent or convergent.

[0276] In some embodiments, dendrimers are prepared using a different approach in which the dendrimer is assembled from a multifunctional core that is extended outward through a series of reactions, typically Michael reactions. This strategy involves coupling monomer molecules with reactive and protective groups to a multifunctional core moiety, which results in a stepwise increase in the number of generations around the core, followed by removal of the protecting groups. For example, PAMAM-NH2 dendrimers are first synthesized by coupling N-(2-aminoethyl)acrylamide monomers to an amine core.

[0277] In other embodiments, dendrimers are prepared using a convergent approach, where the dendrimer is built from small molecules that eventually reach the surface of the sphere, and the reactions proceed inward, building inward and ultimately attaching to the core.

[0278] There are many other synthetic routes for the preparation of dendrimers, such as the orthogonal method, the accelerated method, the two-stage convergence method or the supercore method, the supermonomer method or the branched monomer method, the double exponential method; the orthogonal coupling method or the two-step method, the double monomer method, and the AB2-CD2 method.

[0279] In some embodiments, the core, one or more branching units, one or more spacers, and / or one or more surface functional groups of the dendrimer may be modified to allow conjugation with further functional groups (branching units, spacers, surface functional groups, etc.), monomers, and / or agents by click chemistry using one or more copper-assisted azide-alkyne cycloadditions (CuAAC), Diels-Alder reactions, thiol-ene and thiol-alkyne reactions, and azide-alkyne reactions (Arseneault M et al., Molecules. 2015 May 20; 20(5): 9263-94). In some embodiments, preformed dendrimers are clicked onto high density hydroxyl polymers. "Click chemistry" involves, for example, the coupling of two different moieties (e.g., a core group and a branching unit; or a branching unit and a surface functional group) through a 1,3-dipolar cycloaddition reaction between an alkyne moiety (or its equivalent) on the surface of the first moiety and an azide moiety (e.g., present on a triazine composition or its equivalent) or any reactive end group (e.g., a primary amine end group, a hydroxyl end group, a carboxylic acid end group, a thiol end group, etc.) on the second moiety. In some embodiments, one or more hydroxyl groups on the surface of a dendrimer (a hydroxyl-terminated PAMAM dendrimer or a glucose dendrimer) are modified to contain an alkyl group, and the drug is modified to contain an azide group. Alternatively, one or more hydroxyl groups on the surface of a dendrimer (a hydroxyl-terminated PAMAM dendrimer or a glucose dendrimer) are modified to contain an azide group, and the drug is modified to contain an alkynyl group. The azide and alkynyl groups are then reacted by a 1,3-dipolar cycloaddition reaction to form a triazole moiety.

[0280] In some embodiments, the synthesis of dendrimers relies on one or more reactions such as thiol-ene click reaction, thiol-alkyne click reaction, CuAAC, Diels-Alder click reaction, azide-alkyne click reaction, Michael addition, epoxy ring opening, esterification, silane chemistry, and combinations thereof.

[0281] In some embodiments, the method involves one or more steps of protecting and deprotecting functional groups (e.g., hydroxyl groups) on the central core, branching units, and / or therapeutic, prophylactic, or diagnostic agents to facilitate the addition of branching units to generate the desired dendrimer, or the addition of therapeutic, prophylactic, or diagnostic agents to generate the desired dendrimer conjugate. In the case of hydroxyl groups, they can be protected by forming ethers, esters, or acetals. Other exemplary protecting groups include Boc and Fmoc.

[0282] Any existing dendritic platform can be used to prepare dendrimers with desired functionality, i.e., dendrimers with a high density of surface hydroxyls by conjugating high hydroxyl content moieties such as 1-thioglycerol or pentaerythritol. Exemplary dendritic platforms such as polyamidoamine (PAMAM), poly(propyleneimine) (PPI), poly-L-lysine, melamine, poly(ether hydroxylamine) (PEHAM), poly(esteramine) (PEA), and polyglycerol can be synthesized and explored.

[0283] Dendrimers can also be prepared by combining two or more dendrimers. A dendrimer is a wedge-shaped portion of a dendrimer with a reactive focal functional group. Many dendrimer scaffolds are commercially available. They are available in 1, 2, 3, 4, 5, and 6 generations, with 2, 4, 8, 16, 32, and 64 reactive groups, respectively. In certain embodiments, one type of agent is linked to one type of dendrimer, and a different type of agent is linked to another type of dendrimer. The two dendrimers are then linked to form a dendrimer. Two dendrimers can be linked by click chemistry, i.e., a 1,3-dipolar cycloaddition reaction between an azide moiety on one dendrimer and an alkyne moiety on another dendrimer to form a triazole linker.

[0284] Exemplary methods for preparing dendrimers are described in detail in International Patent Publication Nos. WO2009 / 046446, WO2015168347, WO2016025745, WO20160.25741, WO2019094952, and U.S. Pat. No. 8,889,101.

[0285] 1. Method for preparing glucose dendrimers

[0286] In some embodiments, the glucose-based dendrimers are prepared using a different approach, where the dendrimers are assembled from a multifunctional core that is extended outward through a series of reactions. This strategy involves coupling monomer molecules with reactive and protective groups to the multifunctional core moiety, resulting in a stepwise increase in the number of generations around the core, followed by removal of the protecting groups.

[0287] In some embodiments, glucose dendrimers are synthesized by coupling AB4 peracetylated α-D glucose-PEG4-azide monomers to hexapropargylated cores. In a preferred embodiment, the supercore is prepared from dipentaerythritol, for example, by propargylating dipentaerythritol to obtain a hexapropargylated core. An exemplary scheme for preparing such glucose dendrimers is shown in Scheme 1.

[0288]

[0289] Scheme 1. Synthesis of hypernuclei

[0290] In some embodiments, the branching unit is a supermonomer, i.e., AB n Building units. Exemplary supermonomers include AB3, AB4, AB5, AB6, AB7, AB8 building units. The supermonomer strategy greatly increases the number of available end groups. An exemplary supermonomer is the AB4 orthogonal supermonomer, which includes an azide functional group and four allyl groups and is prepared by reacting dipentaerythritol having five allyl groups with monotosylated triethylene glycol azide.

[0291] In some embodiments, the branching unit is a linear or branched polyglycerol, for example, as shown in Formula III. Other monomers include disaccharides and oligosaccharides, and sugars such as fructose, lactose, and sucrose.

[0292] a. Synthesis of AB4 building blocks

[0293] Some exemplary synthesis methods of supermonomers AB4 are described below. In some embodiments, supermonomers AB4 are based on glucose molecules. In preferred embodiments, supermonomers AB4 are conjugated with polyethylene glycerol (e.g., tetraethylene glycol (PEG4)). In one embodiment, supermonomers AB4 are peracetylated β-D-glucopyranoside tetraethylene glycol azide.

[0294] In some embodiments, the synthesis of glucose-OAc-TEG-OTs involves the following steps: a solution of peracetylated β-D-glucopyranoside (10 g, 25.6 mmol) is dissolved in 50 mL of anhydrous dichloromethane (DCM), followed by the addition of 2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (6.2 g, 17.9 mmol), and the reaction mixture is cooled to 0°C. Boron trifluoride etherate (2.5 eq.) is added and the reaction is allowed to return to room temperature. The reaction is monitored by TLC and quenched after 5 hours by the addition of saturated sodium bicarbonate solution at 0°C. After stirring for 10 minutes, DCM (300 mL) is added and the organic layer is washed 3 times with saturated sodium bicarbonate solution until the effervescence is quenched. The reaction mixture is dried over sodium sulfate, filtered, and evaporated under reduced pressure. The crude product is purified by combiflash chromatography using an ethyl acetate / hexane (70:30) mixture as the eluent. The desired compound is obtained in a yield of 60%. The structure of glucose-OAc-TEG-OT is shown below:

[0295]

[0296] In some embodiments, the synthesis of glucose-OAc-TEG-N3 involves the following steps: a solution of glucose-OAc-TEG-OT (6 g, 8.8 mmol) is dissolved in 40 mL of anhydrous DMF, and then sodium azide (2 eq) is added and the reaction mixture is heated to 50 ° C overnight. After completion, the reaction mixture is filtered and the DMF is evaporated. After drying, the crude reaction mixture is passed through combiflash using ethyl acetate: hexane (70: 30) as the eluent. The structure of glucose-OAc-TEG-N3 is as follows:

[0297]

[0298] In some embodiments, the synthesis of glucose-OH-TEG-N3 comprises the following steps: dissolving peracetylated β-D-glucopyranoside tetraethylene glycol azide in anhydrous methanol, and adding sodium methoxide to adjust the pH to about 8.5-9. The reaction is stirred at room temperature overnight, then diluted with methanol, and the pH is adjusted to about 6-7 with Amberlist IR-120+. The reaction mixture is separated by filtration, and the solvent is removed by rotary evaporation. The structure of glucose-OH-TEG-N3 is shown below.

[0299]

[0300] b. Synthesis of Glucose Dendrimers

[0301] In some embodiments, glucose dendrimers are synthesized by coupling AB4 peracetylated β-D-glucose-PEG4-azide monomers to a hexapropargylated core. In a preferred embodiment, the hexapropargylated core is linked to the AB4 β-D-glucose-PEG4-azide building unit (2) via a click reaction to obtain a first generation dendrimer.

[0302] In some embodiments, the first generation dendrimer D1-Glu6-OAc24 is prepared according to the following steps: The hexapropargylated compound (0.5 g, 1 mmol) and the azido derivative ((4.1 g, 7.4 mmol), 1.2 equivalents / acetylene) are suspended in a 1:1 mixture of DMF and water in a 20 mL microwave vial equipped with a magnetic stir bar. CuSO4·5H2O (5 mol% / acetylene, 75 mg) and sodium ascorbate (5 mol% / acetylene, 60 mg) dissolved in a minimum amount of water are added. The reactants are irradiated in a microwave at 50°C for 6 hours. The reaction mixture is dialyzed against DMF and then dialyzed against water containing EDTA. EDTA is further removed by dialysis against a large amount of water. The product is lyophilized to obtain D1-Glu6-OAc24. The structure of D1-Glu6-OAc24 is shown below.

[0303]

[0304] In some embodiments, the first generation dendrimer D1-Glu6-OH 24 The preparation of the first generation peracetylated glucose dendrimer (1 g, 0.26 mmol) was as follows: peracetylated first generation glucose dendrimer (1 g, 0.26 mmol) was dissolved in anhydrous methanol and sodium methoxide was added to adjust the pH to about 8.5-9. The reaction was stirred at room temperature overnight and then diluted with methanol and IR-120+ adjusts the pH to about 6-7. The reaction mixture is separated by filtration, the solvent is removed by rotary evaporation, and then dialyzed against water. The structure of the first generation glucose dendrimer D1-Glu6-OH24 is shown below.

[0305]

[0306] In some embodiments, the first generation glucose dendrimer D1-Glu6-OH24 is propargylated to provide D1-Acetylene24 as follows: D1-GLu6-OH24 (2 g, 0.721 mmol) was dissolved in anhydrous dimethylformamide (DMF, 50 mL) by ultrasound. Sodium hydride [60% dispersion in mineral oil] (951 mg, 39.65 mmol) was slowly added to the solution in portions under stirring at 0°C. The solution was stirred for another 15 minutes at 0°C. Propargyl bromide (3.85 mL, 34.608 mmol, 80% w / w toluene solution) was then added at 0°C and stirring was continued for another 6 hours at room temperature. The reaction mixture was quenched with ice and water, filtered, dialyzed with DMF, and then dialyzed with water to obtain D1-acetylene 24. The structure of D1-acetylene 24 is shown below.

[0307]

[0308] In some embodiments, the first generation dendrimer D1-acetylene 24 is further reacted with AB4 β-D-glucose-PEG4-azide to yield a second generation dendrimer with 24 glucose molecules and 96 surface hydroxyl groups.

[0309] The preparation method of the exemplary second generation dendrimer D2-Glu24-OAc96 is as follows: D1-acetylene dendrimer 24 (0.5 g, 0.13 mmol) and glucose-OAc-TEG-azide (2.2 g, 4 mmol) were suspended in a 1:1 mixture of DMF and water in a 20 mL microwave vial equipped with a magnetic stir bar. CuSO4·5H2O (5 mol% / acetylene, 5 mg) and sodium ascorbate (5 mol% / acetylene, 10 mg) dissolved in a minimum amount of water were added thereto. The reactants were irradiated in a microwave at 50°C for 8 hours. Upon completion, the reaction mixture was dialyzed against DMF and then against water containing EDTA. EDTA was further removed by dialysis against a large amount of water. The product was lyophilized to obtain D2-G1u24-OAc96.

[0310] In some embodiments, the preparation method of the second-generation dendritic macromolecule D2-Glu24-OH96 is as follows: dissolve the peracetylated second-generation glucose dendritic macromolecule D2-Glu24-OH96 in anhydrous methanol, add sodium methoxide to adjust the pH value to about 8.5-9.0, stir the reaction at room temperature overnight, then dilute with methanol, IR-120+ adjusts the pH value to about 6-7. The reaction mixture is filtered to remove the resin, and the filtrate is rotary evaporated and dialyzed against water to obtain an off-white solid product.

[0311] The structure of the second generation glucose dendrimer D2-Glu24-OH96 is shown below.

[0312]

[0313] In some embodiments, the second generation dendrimer D2-Glu24-OH96 is propargylated at one or more terminal hydroxyl groups and is suitable for further conjugation with one or more therapeutic, prophylactic or diagnostic agents. In some embodiments, one or more terminal hydroxyl groups of the second generation dendrimer D2-G1u24-OH96 are propargylated as follows: D2-Glu24-OH96 (5b) (200 mg, 0.016 mmol) was dissolved in anhydrous dimethylformamide (DMF, 10 mL) by ultrasound. Sodium hydride [60% dispersion in mineral oil] (22 mg, 0.934 mmol) was slowly added in portions to the stirred solution at 0°C. The solution was stirred for an additional 15 minutes at 0°C. Propargyl bromide (18.0 μL, 80% weight / weight solution in toluene) was then added at 0°C and stirring was continued for 6 hours at room temperature. The solvent was evaporated using a V10 evaporator system and the mixture was filtered through a PD10 The crude product was purified by G25 M column. The aqueous solution was lyophilized to obtain an off-white solid product.

[0314] In some embodiments, one or more fluorescent dyes (e.g., infrared fluorescent Cy5 dye) are conjugated to the second generation dendrimer D2-Glu24-OH96. In one embodiment, Cy5-D2-Glu24-OH96 ( Figure 1B Compound 7 in (200 mg, 0.016 mmol) was prepared as follows: Compound 6 (200 mg, 0.016 mmol) and Cy5 azide (20.7 mg, 0.02 mmol) were suspended in a 1:1 mixture of DMF and water in a 25 mL round bottom flask equipped with a magnetic stir bar. CuSO4·5H2O (5 mol% / acetylene, 0.3 mg) and sodium ascorbate (10 mol% / acetylene, 0.5 mg) dissolved in a minimum amount of water were added thereto. The reaction was stirred at room temperature for 24 hours. After the reaction was completed, DMF was evaporated with V10, purified with a PD10 Sephadex G25 M column, and the aqueous solution was freeze-dried to obtain a blue solid product.

[0315] In some embodiments, the total number of hydroxyl groups available for further conjugation to active agents (including therapeutic agents and / or diagnostic agents) is about 1-30, 2-20, or 5-10 of the 96 total available hydroxyl groups of an exemplary second generation dendrimer, wherein 24 glucose molecules contain 96 surface hydroxyl groups.

[0316] B. Methods of Preparing Dendrimer-Agent Conjugates

[0317] Methods of conjugating pharmaceutical agents to dendrimers are generally known in the art, for example as described in US Published Application Nos. US2011 / 0034422, US2012 / 0003155, and US2013 / 0136697.

[0318] In some embodiments, one or more pharmaceutical agents are covalently attached to the dendrimer. In some embodiments, the pharmaceutical agent is attached to the dendrimer via a spacer designed to be non-cleavable in vivo. In some embodiments, the pharmaceutical agent is 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, enzymatically, or a combination of both, thereby providing sustained release of the pharmaceutical agent in vivo. In some embodiments, the chemical structure of the spacer and its point of attachment to the pharmaceutical agent can be selected so that cleavage of the spacer releases the pharmaceutical agent or a suitable prodrug thereof. The chemical structure of the spacer can also be selected based on the desired release rate of the pharmaceutical agent.

[0319] In some embodiments, the conjugation between the agent and the dendrimer is achieved through one or more of a disulfide bond, an ester bond, an ether bond, a phosphodiester bond, a triglycyl peptide bond, a hydrazine bond, an amide bond, or an aminoalkyl bond. In some embodiments, the conjugation between the agent and the dendrimer is through an appropriate spacer that provides an ester bond or an amide bond between the agent and the dendrimer depending on the desired release kinetics of the agent. In some cases, an ester bond or a disulfide bond is introduced to form a releasable form of the drug. In other cases, an amide bond or an aminoalkyl bond is introduced to form a non-releasable form of the drug.

[0320] The spacer generally contains one or more organic functional groups. Examples of suitable organic functional groups contained in the spacer include secondary amides (-CONH-), tertiary amides (-CONR-), sulfonamides (-S(O)2-NR-), secondary carbamates (-OCONH-; -NHCOO-), tertiary carbamates (-OCONR-; -NRCOO-), carbonates (-OC(O)-O-), ureas (-NHCONH-; -NRCONH-; -NHCONR-, -NRCONR-), methanols (-CHOH-, -CROH-), disulfides, hydrazones, hydrazides, ethers (-O-) and esters (-COO-, -CH2O2C-, CHRO2C-), wherein R is an alkyl, aryl or heterocyclic group. In general, the identity of the one or more organic functional groups in the spacer is selected based on the desired release rate of the agent. In addition, the one or more organic functional groups can be selected to facilitate covalent conjugation of the agent to the dendrimer. In some embodiments, the conjugation between the agent and the dendrimer is carried out through an appropriate spacer that provides a disulfide bond between the agent and the dendrimer. In some embodiments, the dendrimer-active agent conjugate is capable of rapidly releasing the agent through a thiol exchange reaction under reducing conditions in vivo.

[0321] In certain embodiments, the spacer comprises a combination of one or more of the above-mentioned organic functional groups and a linking group. The linking group can be composed of any atomic combination, including oligomeric chains and polymer chains; For example, the total number of atoms in the linking group is 3 to 200 atoms, 3 to 150 atoms, 3 to 100 atoms, or 3 to 50 atoms. Examples of suitable linking groups include alkyl, heteroalkyl, alkylaryl, oligo and polyethylene glycol chains and oligo and poly (amino acid) chains. The change of the linking group provides additional control of the release of the agent in vivo. In the embodiment where the spacer includes a linking group, one or more organic functional groups are generally used to connect the linking group to the anti-inflammatory agent and the dendritic macromolecule.

[0322] Reactions and strategies for covalently attaching agents to dendrimers are known in the art. See, for example, March, Advanced Organic Chemistry, 5th edition, 2001, Wiley-Interscience Publication, New York) and Hermanson, Bioconjugate Techniques, 1996, Elsevier Academic Press, USA. The appropriate covalent conjugation method for a given agent can be selected based on the desired linking moiety and the structure of the agent and dendrimer, as this is related to the compatibility of the functional groups, the protection group strategy, and the presence of labile bonds.

[0323] The amount of active agent (drug load) in the dendrimer-active agent conjugate depends on many factors, including the choice of active agent, the structure and size of the dendrimer, and the tissue to be treated. In some embodiments, one or more antidepressants and / or antipsychotics are conjugated to the dendrimer at a concentration of between about 0.01% and about 45%, including between about 0.1% and about 30%, including between about 0.1% and about 20%, including between about 0.1% and about 10%, including between about 1% and about 10%, including between about 1% and about 5%, including between about 3% and about 20%, including between about 3% and about 10%. However, the specific drug load for any given active agent, dendrimer, and target site can be identified by conventional methods, such as those described.

[0324] In some embodiments, conjugation of the agent / spacer occurs through about 1%, 2%, 3%, 4%, or 5% of the total available surface functional groups (e.g., hydroxyl groups) of the dendrimer prior to conjugation. In other embodiments, conjugation of the agent / spacer occurs on less than 5%, less than 10%, less than 15%, less than 20%, less than 25%, less than 30%, less than 35%, less than 40%, less than 45%, less than 50%, less than 55%, less than 60%, less than 65%, less than 70%, less than 75% of the total available surface functional groups of the dendrimer prior to conjugation with the active agent. In some embodiments, the dendrimer-active agent conjugate retains an effective amount of surface functional groups for targeting neural and / or glial cells while being conjugated to an effective amount of the agent for treating, preventing, and / or imaging a disease or condition. In some embodiments, the dendrimer-active agent conjugate retains an effective amount of the active agent for targeting to target neural and / or glial cells and binding to target receptors on the surface or within target neural and / or glial cells.

[0325] The following embodiments describe more specific methods for preparing exemplary dendrimer-active agent conjugates.

[0326] IV. Pharmaceutical Preparations

[0327] Pharmaceutical compositions comprising the dendrimer-active agent conjugates may be formulated in conventional manner using one or more physiologically acceptable carriers, optionally including excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically for oral, mucosal (intranasal, buccal, sublingual, vaginal, rectal or pulmonary), transdermal or injectable (intravenous, subcutaneous, intraperitoneal, intramuscular or intrathecal administration).

[0328] Representative excipients include aqueous buffers, wetting agents, viscosity modifiers, tonicity agents, stabilizers, and combinations thereof. Suitable pharmaceutically acceptable excipients are preferably selected from materials that are generally recognized as safe (GRAS) and can be administered to an individual without causing adverse biological side effects or unwanted interactions.

[0329] Typically, the pharmaceutically acceptable salt of an active agent can be prepared by reacting the free acid or base form of the drug with a stoichiometric amount of an appropriate base or acid in water or an organic solvent or a mixture of the two; typically, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred. Pharmaceutically acceptable salts include drug salts derived from inorganic acids, organic acids, alkali metal salts and alkaline earth metal salts, and salts (e.g., quaternary ammonium salts) formed by reacting a drug with a suitable organic ligand. A list of suitable salts can be found in, for example, Remington's Pharmaceutical Sciences, 20th edition, Lippincott Williams & Wilkins, Baltimore, Maryland, 2000, p. 704. Examples of ophthalmic drugs sometimes administered in the form of pharmaceutically acceptable salts include timolol maleate, brinzolamide tartrate, and diclofenac sodium.

[0330] The composition is preferably formulated in dosage unit form for ease of administration and uniformity of dosage. The phrase "dosage unit form" refers to physically discrete units of the conjugate suitable for the patient to be treated. However, it should be understood that the total single dose of the composition will be determined by the attending physician within the scope of reasonable medical judgment. The therapeutically effective dose can be initially estimated in cell culture assays or animal models (usually mice, rabbits, dogs or pigs), or inferred from human data. Animal models are also used to achieve the desired concentration range and route of administration. Then, this information should help determine the effective dose and route of administration for humans. The therapeutic effect and toxicity of the conjugate can be determined by standard pharmaceutical procedures in cell culture or experimental animals, for example, ED50 (the dose has a therapeutic effect on 50% of the population) and LD50 (the dose is lethal to 50% of the population). The dose ratio of toxicity to therapeutic effect is the therapeutic index, expressed as the LD50 / ED50 ratio. Pharmaceutical compositions with large therapeutic indices are preferred. Data obtained from cell culture tests and animal studies can be used to formulate dosage ranges for human use.

[0331] In certain embodiments, the composition is administered topically, for example, by direct injection into the site to be treated or via an implant or pump. In some embodiments, the composition is injected, topically applied, or otherwise administered directly into the vascular system, which is located on vascular tissue at or near the site of injury, surgery, or implantation. For example, in embodiments, the composition is topically applied to exposed vascular tissue during surgery. Typically, topical administration results in an increase in the local concentration of the composition, which is greater than the concentration that can be achieved with systemic administration.

[0332] Pharmaceutical compositions for administration by parenteral (intramuscular, intraperitoneal, intravenous or subcutaneous injection) and enteral routes of administration are described. These include administration to mucosal surfaces (nasal, buccal, sublingual, pulmonary, vaginal and rectal).

[0333] A. Parenteral Administration

[0334] The compositions of the dendrimer-active agent conjugates can be administered parenterally. The phrases "parenteral administration" and "intestinal administration" are art-recognized terms that include modes of administration other than enteral and topical administration. The dendrimers can be administered orally, intranasally, subcutaneously, intraperitoneally, intravenously, intrathecally or intramuscularly. For liquid formulations, pharmaceutically acceptable carriers can be, for example, aqueous or non-aqueous solutions, suspensions, emulsions or oils. Parenteral carriers (for subcutaneous, intravenous, arterial or intramuscular injection) include, for example, sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's solution, and fixed oils. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, and injectable organic esters, such as ethyl oleate. Aqueous carriers include, for example, water, alcohol / aqueous solutions, cyclodextrins, emulsions or suspensions, including saline and buffered media. The dendrimers can also be administered in the form of an emulsion, such as water-in-oil. The example of oil is petroleum, animal oil, vegetable oil or synthetic oil, for example peanut oil, soybean oil, mineral oil, olive oil, sunflower seed oil, cod liver oil, sesame oil, cottonseed oil, corn oil, olive oil, vaseline and mineral oil. Suitable fatty acids for parenteral preparations include for example oleic acid, stearic acid and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters.

[0335] Preparations suitable for parenteral preparations may include antioxidants, buffers, antibacterial agents, and solutes that make the preparation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions that may include suspending agents, solubilizing agents, thickening agents, stabilizers, and preservatives. Intravenous carriers may include fluid and nutrient supplements, electrolyte supplements, such as supplements based on Ringer's dextrose. In general, water, saline, aqueous glucose and related sugar solutions and glycols, such as propylene glycol or polyethylene glycol are preferred liquid carriers, particularly for injectable solutions.

[0336] Injectable pharmaceutical carriers for injectable compositions are well known to those of ordinary skill in the art (see, for example, Pharmaceutics and Pharmacy Practice, JB Lippincott Company, Philadelphia, Pennsylvania, Banker and Chalmers, eds., pp. 238-250 (1982), and ASHP Handbook on Injectable Drugs, Trissel, 15th edition, pp. 622-630 (2009)).

[0337] B. Enteral administration

[0338] The dendrimer-active agent conjugate composition can be administered enterally (orally, sublingually, vaginally, rectally, buccally, intranasally, pulmonary or transdermally). The carrier or diluent can be a solid carrier such as a capsule or tablet or a diluent for a solid formulation, a liquid carrier or a diluent for a liquid formulation, or a mixture thereof.

[0339] For liquid preparations, pharmaceutically acceptable carriers can be, for example, aqueous or non-aqueous solutions, suspensions, emulsions or oils. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol and injectable organic esters, such as ethyl oleate. Aqueous carriers include, for example, water, alcohol / aqueous solutions, cyclodextrins, emulsions or suspensions, including saline and buffered media.

[0340] The example of oil comprises petroleum, animal oil, vegetable oil or synthetic oil, for example peanut oil, soybean oil, mineral oil, olive oil, sunflower seed oil, cod liver oil, sesame oil, cottonseed oil, corn oil, olive, vaseline and mineral oil. The fatty acid suitable for parenteral preparation comprises for example oleic acid, stearic acid and isostearic acid. Ethyl oleate and isopropyl myristate are the examples of suitable fatty acid esters.

[0341] The carrier includes for example sodium chloride solution, Ringer's dextrose, glucose and sodium chloride, lactated Ringer's solution and fixed oil.Preparation includes for example aqueous and non-aqueous isotonic sterile injection solution, it can contain antioxidant, buffer, antibacterial agent and make the preparation and the solute of the blood isotonic of the expected recipient, and aqueous and non-aqueous sterile suspension, it can include suspending agent, solubilizing agent, thickening agent, stabilizer and preservative.Carrier can include for example fluid and nutritional supplement, electrolyte supplement, for example supplement based on Ringer's dextrose.In general, water, saline, aqueous glucose and relevant sugar solution are preferred liquid carriers.These also can be prepared together with protein, fat, carbohydrate and other components of infant formula.

[0342] Oral preparations can be in the form of chewing gum, gel strips, tablets, capsules or lozenges. Encapsulating materials used to prepare enteric coated oral preparations include cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate and methacrylate copolymers. Solid oral preparations such as capsules or tablets are preferred. Elixirs and syrups are also well-known oral preparations.

[0343] Formulations for administration to mucosal surfaces (e.g., the nasal cavity, oral surfaces, or the lungs) typically contain pharmaceutically acceptable excipients (e.g., excipients for parenteral administration), alone or in combination with various surfactants, penetration enhancers, and the like.

[0344] The compositions can also be formulated as aerosols (i.e., they can be "nebulized") for administration by inhalation. Aerosols can be placed in pressurized acceptable propellants, such as dichlorodifluoromethane, propane, nitrogen and air. For administration by inhalation, the compound is delivered in the form of an aerosol spray from a pressurized pack or nebulizer, using a suitable propellant.

[0345] In some embodiments, the dendrimer conjugates can improve the effectiveness and durability of treatment, thereby reducing the need for repeated dosing to once a week, once a month, once every six months, once a year, or other long-term dosing schedules. Some embodiments can be incorporated into drug delivery systems (e.g., implants, pumps, patches, creams, etc.) to provide controlled, sustained delivery in a manner that reduces compliance and potential for abuse.

[0346] How to use

[0347] In preferred embodiments, the dendrimer compositions cross the barrier interface of the central and peripheral nervous systems and selectively target specific cells and specific receptors on cells to address a variety of diseases, disorders, injuries and conditions. These methods include administering to a subject in need thereof an amount of the composition effective to increase the permeability of the antidepressant and / or antipsychotic drug across the barrier interface of the central and peripheral nervous systems and / or to increase the binding of the antidepressant and / or antipsychotic drug to specific receptors in specific cells, particularly to serotonin receptors, norepinephrine receptors, adrenaline receptors and dopamine receptors in the central nervous system, peripheral nervous system, and / or cells in the peripheral circulation, such as neural cells, glial cells, cardiovascular cells, gastrointestinal cells, immune cells.

[0348] A. Treatment methods

[0349] The compositions can be administered to treat, prevent or control symptoms of various disorders, diseases and conditions, including but not limited to anxiety, mood disorders, eating disorders, personality disorders, stress disorders and / or psychiatric disorders as well as ADHD and individuals with symptoms of ADHD. In some forms, when the dendrimer is complexed with one or more antidepressants, the compositions can be administered to treat one or more neurological disorders, such as mental health disorders, such as mood disorders, anxiety disorders, eating disorders, substance-related disorders and post-traumatic disorders; degenerative diseases, such as Parkinson's disease and Alzheimer's disease; pain disorders, such as neuropathic pain, gastrointestinal diseases and / or cardiovascular diseases. In some forms, when the dendrimer is complexed with one or more antipsychotics, the compositions can be administered to treat psychosis occurring in bipolar disorder, schizophrenia and / or degenerative diseases. In other forms, the dendrimer-antipsychotic conjugates can be administered to subjects in need of mood stabilization, for example, patients with bipolar disorder, to reduce anxiety in patients with anxiety disorders, to reduce tics in patients with Tourette syndrome. In other forms, the dendrimers are complexed with one or more antidepressants and / or antipsychotics to treat non-neurological disorders, such as gastrointestinal and cardiovascular diseases.

[0350] Typically, an effective amount of a dendrimer complex comprising a dendrimer in combination with one or more therapeutic, prophylactic and / or diagnostic agents is administered to an individual in need thereof. Dendrimers may also include targeting agents, but as demonstrated in the examples, these are not required for delivery to damaged tissues in the spinal cord, brain and related areas.

[0351] In some embodiments, the dendrimer complex includes an agent attached or conjugated to the dendrimer that is capable of preferentially releasing the drug at a target receptor. The agent may be covalently attached or dispersed or encapsulated within the molecule. The amount of the dendrimer complex administered to the subject is selected to deliver an effective amount to reduce, prevent or otherwise alleviate one or more clinical or molecular symptoms of the disease or condition being treated as compared to a control (e.g., a subject treated with the active agent without the dendrimer).

[0352] 1. Site-specific targeting

[0353] The compositions and methods are intended to circumvent existing challenges of selective drug delivery to the central and peripheral nervous systems. The compositions and methods can increase the bioavailability of drugs in the central and peripheral nervous systems by one or more of the following: (i) increasing the density of drugs across the brain barrier, particularly the blood-brain barrier and the blood-cerebrospinal fluid barrier, (ii) increasing drug solubility, (iii) promoting target binding, i.e. increasing site-specific binding, (iv) improving drug pharmacokinetics, and (v) improving brain distribution. For example, in some forms, the compositions and methods allow for the selective delivery of compounds to the peripheral nervous system, thereby increasing the potential of the compositions for selective treatment of peripheral specific diseases and conditions, including but not limited to neuropathic pain, local anesthesia, traumatic nerve injury, and hereditary and inflammatory neuropathies.

[0354] a. Improve the permeability of drugs across barrier interfaces

[0355] Dendrimer compositions and methods can improve the delivery of antidepressants and / or antipsychotics across one or more barrier interfaces in the brain and nervous system, particularly the blood-brain barrier (BBB), CSF-blood barrier, and blood-nerve barrier. These barrier interfaces generally protect neurons from blood-borne substances and help maintain water homeostasis and an environment suitable for neuronal function in the blood. Due to the clinical significance of antidepressants and / or antipsychotics, dendrimer compositions can be used to deliver antidepressants and / or antipsychotics with improved permeability across these barrier interfaces to achieve site-specific targeting.

[0356] i. Blood-brain barrier

[0357] The "blood-brain barrier" (BBB) ​​is a continuous endothelial membrane that, together with pericytes and other components of the neurovascular unit, restricts the entry of toxins, pathogens, and blood cells into the brain. However, the BBB also impedes drug delivery to the central nervous system (CNS), in part because (1) systemic delivery of drugs intended for the brain may result in excessive peripheral drug concentrations and (2) the complex interactions between cells and molecules that make up the structure and function of the BBB make it difficult to determine drug permeability across the BBB, drug distribution in the brain, and target engagement in the brain.

[0358] Brain microvascular endothelial cells, pericytes, astrocytes, tight junctions, neurons, and basement membranes constitute physically tight brain capillaries within the BBB. Brain capillary endothelial cells have no pores, which restricts the diffusion of small molecules and proteins. Interendothelial junctions connect endothelial cells to a continuous barrier, severely limiting the permeation of water-soluble substances. Pericytes, astrocytes, and basement membranes surround endothelial cells, ultimately forming an impermeable BBB. In addition, efflux transporters are located in brain capillary endothelial cells, which further prevents substances from entering the brain. The permeability of the BBB is mainly controlled by interendothelial junctions, which are protein complexes such as adherens junctions, tight junctions, and gap junctions. Adherens junctions mainly regulate the permeability of the endothelial barrier. Tight junctions play a crucial role in maintaining the permeability barrier of epithelial and endothelial cells, which controls tissue homeostasis. Gap junctions are composed of six connexin molecules and allow direct electrical and chemical communication between endothelial cells. Finally, the composition of the BBB does not have a static structure, but is constantly adjusted in response to various physiological changes in the brain. The dendrimer compositions and methods of the present application overcome the above challenges and are suitable for delivering antidepressants and / or antipsychotics across the blood-brain barrier via one or more of the above transport mechanisms.

[0359] Molecules cross the BBB via the paracellular pathway (between adjacent cells) or the transcellular pathway (across cells). For the paracellular pathway, ions and solutes cross the BBB by passive diffusion using concentration gradients. The transcellular pathway includes different mechanisms such as passive diffusion, receptor-mediated transport, and transcytosis.

[0360] Physicochemical factors that affect BBB permeability include molecular weight, charge, lipid solubility, surface activity, and relative size of molecules. BBB permeability may also be affected by physiological factors, such as efflux transporters, such as P-glycoprotein (P-gp), enzyme activity, plasma protein binding, and cerebral blood flow. Hydrophilic molecules (such as proteins and peptides) enter the brain through specific and saturable receptor-mediated transport mechanisms (such as glucose transporter-1 (GLUT-1), insulin transporter, and transferrin transporter). These endogenous transporters are expressed on the luminal and extraluminal endothelial cell membranes. Among these transport mechanisms, receptor-mediated transcytosis has been widely studied for drug delivery to the brain. The dendrimer compositions and methods of the present application are suitable for delivering antidepressants and / or antipsychotics across the blood-brain barrier through one or more of the above-mentioned mechanistic pathways.

[0361] ii. Blood-Nerve Barrier (BNB)

[0362] The blood-nerve barrier (BNB) defines the physiological space in which the axons, Schwann cells, and other related cells of the peripheral nerve function, thereby ensuring the normal function of the peripheral nerve and maintaining the homeostasis of the endoneurial environment. The BNB is composed of the endoneurial microvessels and perineurium within the nerve bundle. Tight junctions between endothelial cells and pericytes in the endoneurial vessels isolate the endoneurium from the blood, thereby preventing the uncontrolled leakage of molecules and ions from the circulatory system into the peripheral nerve. In addition, there is a diffusion barrier within the perineurium, which is formed by tight junctions between adjacent perineurial cells and the basement membrane surrounding each perineurial cell layer. Endoneurial capillaries and perineurial channels are restrictive barriers that separate the endoneurial extracellular environment of peripheral nerves from the space around the epineurial bundle and the systemic circulation, thereby protecting the endoneurial microenvironment from drastic concentration changes in blood vessels and other extracellular spaces.

[0363] For drug targets located in peripheral nerves, BNBs may be problematic because they may limit or prevent the drug from reaching its site of action, thereby negatively affecting drug efficacy. In addition, the expression profiles of transporters in peripheral nerves may be very different from those in the central nervous system. The dendrimer compositions of the present application can be used to improve the permeability of antidepressants and / or antipsychotics across BNBs, thereby improving the delivery of antidepressants and / or antipsychotics to peripheral nerve targets.

[0364] iii. Blood-CSF Barrier

[0365] The composition can be used to improve the delivery of antidepressants and / or antipsychotics to target sites through the blood-cerebrospinal fluid barrier (blood-CSF barrier) and the ventricles. The choroid plexus is a vascular tissue present in all ventricles. The functional unit of the choroid plexus is composed of capillaries wrapped by a layer of differentiated ependymal epithelium. Unlike the capillaries that form the blood-brain barrier, the choroid plexus capillaries have holes and no tight junctions. Therefore, the endothelium does not form a barrier to the movement of small molecules. In contrast, the blood-CSF barrier of the choroid plexus is formed by epithelial cells and the tight junctions connecting them. Another part of the blood-CSF barrier is the arachnoid membrane that wraps the brain. The cells of this membrane are also connected by tight junctions.

[0366] The CSF space and brain structures near the CSF compartment are pharmacological targets for CNS diseases. For example, the subarachnoid space, perivascular or periventricular spaces are areas where pathogenic lymphocytes, monocytes and neutrophils accumulate in neuroinflammatory diseases (such as multiple sclerosis and related experimental autoimmune encephalitis) or virus-induced neurological diseases (including neuro-AIDS and CMV infection). B cell foci producing potentially harmful antibodies detected in different CNS autoimmune diseases are believed to be mainly located in the pia mater. Therefore, in some forms, dendrimer compositions can be used to deliver antidepressants and / or antipsychotics to target areas via the blood-CSF space connected to the deep cervical lymph nodes to improve or treat symptoms associated with neuroinflammatory diseases.

[0367] In some forms, dendrimer compositions can be used to deliver antidepressants and / or antipsychotics to target sites to improve or treat symptoms and conditions associated with vascular degeneration. For example, cerebral amyloid angiopathy induces degenerative vascular changes caused by amyloid beta peptide (Aβ), cystatin c, transthyretin, or gelsolin deposits that infiltrate perivascular areas. These deposits are accessible through the interconnected CSF / perivascular spaces.

[0368] In some forms, dendrimer compositions can be used to deliver antidepressants and / or antipsychotics to target sites to improve or treat symptoms and conditions associated with tumor development. For example, periventricular tumors (including meningiomas, drug-resistant ependymomas, and leptomeningeal metastases from peripheral primary tumors) are in direct contact with the CSF. The blood-tumor barrier is generally considered to be porous due to the reduced efficacy of tight junctions (which allow contrast enhancement in magnetic resonance imaging). However, many antidepressants and / or antipsychotics are lipophilic and are prevented from crossing the BBB by multidrug resistance (MDR) efflux proteins that control transcellular pathways. In many periventricular tumors (e.g., ependymomas), MDR proteins are still well expressed on the blood-tumor barrier. Therefore, in some forms, dendrimer compositions can be used to utilize pharmacological pressure from the CSF to achieve therapeutic concentrations of antidepressants and / or antipsychotics within tumor tissue.

[0369] b. Improve target specific binding

[0370] The compositions can be used to deliver antidepressant and / or antipsychotic drugs with increased binding affinity and specificity for one or more receptors to modulate serotonin (5HT) receptors, such as 5HT-1A, 5HT-2B, 5HT-2A, 5HT-2B, 5HT-2C, 5HT-3, 5HT-4; one or more norepinephrine (NE) receptors, such as α 1A Adrenergic receptors, alpha 1B-Adrenergic receptors, alpha 1D -Adrenergic receptors, alpha 2A -Adrenergic receptors, alpha 2B -Adrenergic receptors, alpha 2C -adrenergic receptors, β1-adrenergic receptors and β2-adrenergic receptors; monoamine transporters, such as serotonin reuptake transporter (SERT) and / or norepinephrine transporter (NET). Dendrimer compositions can be used to deliver antidepressant and / or antipsychotic drugs with increased binding affinity and specificity for one or more receptors to modulate one or more of the GLUT1, GLUT2, GLUT3, GLUT4, GLUT5, GLUT6, GLUT7, GLUT8, GLUT9, GLUT10, GLUT11, GLUT12, GLUT13 and GLUT14 transporters.

[0371] i. Improve serotonin receptor binding

[0372] Dendrimer compositions can improve binding to one or both serotonin receptors, thereby modulating signaling in a cell-specific and tissue-specific manner.

[0373] In the central nervous system (CNS), serotonin is almost entirely produced by neurons derived from the raphe nuclei located in the midline of the brainstem. These neurons that produce serotonin form the largest and most complex efferent system in the human brain. The caudal raphe nuclei dominate the spinal cord, while the more anterior raphe nuclei, dorsal raphe nuclei, and medial raphe nuclei dominate most of the rest of the CNS through diffuse projections. Almost every cell in the brain is close to serotonin fibers, and almost all behaviors and many other brain functions are regulated by serotonin.

[0374] Both within the CNS and throughout the body, serotonin plays a variety of roles in vascular biology, ranging from the control of vascular resistance and blood pressure to the control of hemostasis and platelet function. Serotonin causes vasoconstriction or vasodilation in different vascular beds, depending on the specific receptors expressed in each vessel wall and surrounding smooth muscle tissue. Thus, in some embodiments, dendrimer-antidepressant compositions can be used to activate 5-HT1B receptors on cerebral blood vessels to promote vasodilation, resulting in analgesic effects on migraine.

[0375] Platelets have large vesicular serotonin stores but lack the enzymes to synthesize serotonin; instead, they absorb serotonin from plasma via the serotonin transporter. Serotonin is then secreted through platelet dense granules during platelet activation and plays a role in promoting platelet aggregation and vasoconstriction of surrounding blood vessels, thereby facilitating hemostasis. Intracellular serotonin also promotes platelet activation by covalently linking to small G proteins via tissue transglutaminase. This modification constitutively activates G protein-dependent signaling pathways and stimulates platelet aggregation. In addition, serotonin covalently cross-links a variety of adhesion proteins and coagulation factors on the platelet cell surface, a process that is essential for activating a portion of platelets. Therefore, serotonin also acts through non-covalent interactions with membrane-bound receptors.

[0376] Serotonin regulates many different aspects of cardiac function, ranging from electrical conduction to valve closure to post-MI remodeling. Thus, in some forms, the dendrimer-SSRI composition can be administered to reduce the risk of myocardial infarction. For example, in some forms, the composition can be used as a 5-HT4 antagonist to help improve cardiac function and prevent pathological remodeling in congestive heart failure. In other forms, the composition can be used as a 5-HT2A antagonist to treat vasospastic angina and ischemic heart disease, and / or as a 5-HT3 antagonist to treat post-MI pain.

[0377] ii. Improved noradrenergic receptor binding

[0378] Dendrimer compositions can improve binding to one or both noradrenergic receptors, thereby modulating signaling in a cell-specific and tissue-specific manner.

[0379] Norepinephrine (also known as norepinephrine) is a neurotransmitter of the brain that plays a crucial role in regulating arousal, attention, cognitive function, and stress responses. It also functions as a peripheral hormone in the "fight or flight" response as part of the sympathetic nervous system. In states of stress or anxiety, norepinephrine and epinephrine are released and bind to adrenergic receptors throughout the body, exerting effects such as dilation of the pupils and bronchioles, increase in heart rate and vasoconstriction, increase in renin secretion from the kidneys, and inhibition of peristalsis. The noradrenergic system plays a role in the pathogenesis of several major neuropsychiatric disorders and has emerged as an important pharmacological target in a variety of psychiatric, neurological, and cardiopulmonary diseases.

[0380] The central noradrenergic system is composed of two main ascending projections derived from the brainstem: the dorsal noradrenergic bundle (DNB) and the ventral noradrenergic bundle (VNB). The DNB is derived from the A6 locus coeruleus located dorsal to the pons, and is mainly composed of noradrenergic neurons. It works as the main site of noradrenaline production in the central nervous system. It sends out projections to specifically control the cerebral cortex, hippocampus and cerebellum, and has a projection that overlaps with the projection from the VNB, to control the area of ​​the amygdala, hypothalamus and spinal cord. The VNB is derived from the nucleus in the pons and medulla oblongata, and sends out projections to control the area of ​​the amygdala, hypothalamus and midbrain and medulla oblongata.

[0381] The sympathetic nervous system and neuroendocrine chromaffin cells (located in the adrenal medulla) are primarily responsible for the synthesis and exocytosis of norepinephrine and other catecholamines into the blood circulation. These hormones act on alpha and beta adrenergic receptors on smooth muscle cells and adipose tissue throughout the body.

[0382] After an action potential enters the presynaptic terminal, voltage-gated calcium channels are stimulated and bring calcium from the extracellular space into the intracellular space. This influx causes norepinephrine (stored in vesicles) to bind to the cell membrane and be released into the synaptic cleft via exocytosis. Norepinephrine can then go on to bind to three major receptors: alpha 1 (α-1), alpha-2, and beta receptors. These receptors are classified as G-protein coupled receptors, have either inhibitory or excitatory effects, and have different binding affinities for norepinephrine.

[0383] The alpha-1 receptor is further subdivided into alpha-1a, alpha-1b, and alpha-1d receptors. These receptors are located in postsynaptic areas of the brain, including the locus coeruleus, olfactory bulb, cerebral cortex, dentate gyrus, amygdala, and thalamus. The alpha-1 receptor has a moderate binding affinity for norepinephrine and is coupled to the Gq protein signaling pathway. In this pathway, phospholipase C (PLC) is activated, which converts phosphatidylinositol 4,5-bisphosphate (PIP2) to inositol 1,4,5-triphosphate (IP3) and diacylglycerol (DAG) on the cell membrane. IP3 is released into the cytosol and binds to transmembrane IP3 receptors located on the endoplasmic reticulum (ER), which functions as a calcium channel. Upon binding, the receptor undergoes a conformational change that results in the release of calcium from the ER into the cytosol. DAG is retained in the cell membrane and actively regulates protein kinase C (PKC), which functions to phosphorylate other proteins. These combined actions produce excitatory cellular effects.

[0384] The alpha-2 receptors are subdivided into alpha-2a, alpha-2b, and alpha-2c receptors. These receptors are located in presynaptic and postsynaptic areas of the brain, including the locus coeruleus, amygdala, and hypothalamus. These receptors have the highest binding affinity for norepinephrine and are coupled to the Gi / o protein signaling pathway. In this pathway, cAMP levels are reduced, resulting in decreased adenylate cyclase activity, producing an inhibitory cellular effect. Presynaptic noradrenergic terminals contain alpha-2 autoreceptors, which prevent further release of norepinephrine.

[0385] Beta receptors are subdivided into beta-1, beta-2, and beta-3 receptors. These receptors are located in different areas of the brain, with beta-1 and beta-2 receptors being most prevalent in the cerebral cortex. These receptors have the lowest binding affinity for norepinephrine and are coupled to the Gs protein signaling pathway. In this pathway, cAMP levels increase, leading to activation of protein kinase A (PKA), which in turn phosphorylates other proteins within the cell and leads to excitatory cellular effects. Beta-2 receptors are also coupled to the Gi protein signaling pathway. Beta-3 receptors are found in adipose tissue.

[0386] In the adrenal medulla, acetylcholine stimulates the release of epinephrine and norepinephrine. Acetylcholine binds to nicotinic receptors located on adrenal chromaffin cells, which generates an action potential maintained by voltage-gated sodium and potassium channels. This action potential triggers an influx of calcium into the cytosol, which causes norepinephrine vesicles to bind to the cell membrane, resulting in the release of norepinephrine into the circulation, where it travels to bind to alpha and beta receptors on smooth muscle and fat cells.

[0387] Norepinephrine can be degraded intracellularly or in the synaptic cleft by monoamine oxidase (MAO) or catechol-O-methyltransferase (COMT). MAO oxidizes norepinephrine, while COMT deaminates norepinephrine via O-methylation metabolism. MAO and COMT are found in adrenal chromaffin cells, while sympathetic nerves contain only MAO. COMT is found in all organs. The liver is responsible for the complete degradation of norepinephrine to vanillylmandelic acid (VMA).

[0388] B. Disease to be treated

[0389] The compositions are useful for treating one or more diseases, disorders and injuries in the central and peripheral nervous systems. The compositions may also be used to treat a variety of diseases, disorders and injuries, including mental health disorders, gastrointestinal diseases, cardiovascular diseases and / or to treat other tissues in which nerves play a role in the disease or disorder. The compositions and methods are also useful for preventive uses. For example, the compositions may be administered to a patient in need to improve, treat or prevent symptoms associated with a variety of disorders, diseases and conditions, including but not limited to anxiety, mood disorders, eating disorders, ADHD, personality disorders, stress disorders and / or psychiatric disorders. In some forms, when the dendrimer is compounded with one or more antidepressants, the compositions may be administered to treat one or more nervous system disorders, such as mental health disorders, such as mood disorders, anxiety disorders, eating disorders, substance-related disorders and post-traumatic disorders; degenerative diseases, such as Parkinson's disease and Alzheimer's disease; pain disorders, such as neuropathic pain, gastrointestinal diseases and / or cardiovascular diseases. In some forms, when the dendrimer is compounded with one or more antipsychotics, the compositions may be administered to treat bipolar disorder, schizophrenia and / or psychosis occurring in degenerative diseases. In other forms, the compositions can be administered to subjects in need of mood stabilization (e.g., bipolar disorder), to reduce anxiety in anxiety disorders, and to reduce tics in Tourette syndrome. In other forms, the dendrimers are compounded with one or more antidepressants and / or antipsychotics to treat non-neurological disorders, such as gastrointestinal disorders and cardiovascular diseases.

[0390] The dendrimer complex is administered in a dosage unit amount effective to treat or alleviate a condition associated with a pathological condition of the central and peripheral nervous systems. For example, the dendrimer complex is used in an effective dosage unit amount to treat or alleviate a condition associated with a pathological condition affecting neurons, microglia, and astrocytes. In general, by targeting these cells, the dendrimer can specifically deliver an agent to treat neuroinflammation.

[0391] In preferred embodiments, the composition may include glucose or hydroxy dendrimers having a diameter of 5 nm or less and conjugated with antidepressants, antipsychotics, or other drugs that act by modulating monoaminergic neurotransmission for use in the treatment of mental health and CNS diseases. In other preferred embodiments, the composition may include glucose or hydroxy dendrimers having a diameter greater than 5 nm and conjugated with antidepressants, antipsychotics, or other drugs that act by modulating monoaminergic neurotransmission for use in the treatment of peripheral nervous system diseases.

[0392] 1. Mental health disorders and conditions

[0393] These compositions and methods are useful for treating a variety of mental health disorders and conditions, including, but not limited to, affective or mood disorders, anxiety disorders, childhood disorders, eating disorders, personality disorders, schizophrenia and other psychotic disorders, and substance-related disorders.

[0394] a. Affective or emotional disorders

[0395] Affective or mood disorders are marked disturbances of mood (low moods are called depression, and high moods are called hypomania or mania). These disorders include bipolar disorder, cyclothymia, hypomania, major depressive disorder, disruptive mood dysregulation disorder, treatment-resistant depression, persistent depressive disorder, premenstrual dysphoric disorder, seasonal affective disorder, medical condition-related depression, and depression caused by substance or medication use.

[0396] In some forms, the compositions can be administered to a subject in need of treatment, prevention, or management of symptoms of major depressive disorder, including, but not limited to, persistent low mood or depression, anhedonia or decreased interest in pleasurable activities, feelings of guilt or worthlessness, low energy, difficulty concentrating, changes in appetite, psychomotor retardation or agitation, sleep disturbances, and / or suicidal thoughts.

[0397] b. Anxiety disorder

[0398] Anxiety disorders are different from normal nervousness or anxiety, which involve excessive fear or worry. Anxiety disorders include generalized anxiety disorder, panic disorder, social anxiety disorder, and various phobia-related disorders.

[0399] Generalized anxiety disorder (GAD) generally involves persistent anxiety or fear, which interferes with daily life. It is different from occasionally worrying about things or feeling anxious because of stressful life events. People with GAD often feel anxious, which lasts for months or even years. These compositions and methods are suitable for treating one or more symptoms of GAD, including but not limited to restlessness, fatigue, difficulty concentrating, irritability, headache, muscle pain, stomach pain or unexplained pain, excessive worry, sleep problems (such as difficulty falling asleep or difficulty staying asleep).

[0400] In some forms, the compositions can be administered to patients in need to treat, prevent or control one or more symptoms associated with panic disorder. Panic disorder is an anxiety disorder characterized by sudden, recurrent attacks of intense fear accompanied by physical symptoms, including chest pain, palpitations, shortness of breath, dizziness or abdominal discomfort, or a feeling of loss of control even when there is no apparent danger or trigger. People with panic disorder often worry about when the next attack will occur and actively try to prevent future attacks by avoiding places, situations or behaviors associated with panic attacks. Panic attacks may occur as frequently as several times a day or as rarely as several times a year. These compositions and methods are suitable for treating one or more panic attack symptoms, including but not limited to palpitations, excessive sweating, shaking or tingling, chest pain, and feelings of being out of control, such as feelings of impending doom and feelings of being out of control.

[0401] Social anxiety disorder is an intense and persistent fear of being watched and judged by others. For people with social anxiety disorder, the fear of social situations can be so intense that it seems beyond their control. For some people, this fear can prevent them from going to work, school, or doing everyday things. The compositions and methods are useful for treating one or more symptoms of social anxiety disorder, including, but not limited to, excessive blushing, sweating or shaking, heart palpitations, stomach aches, stiff body posture or soft voice, and self-consciousness or fear of negative judgment.

[0402] A phobia is an intense fear or aversion to a particular object or situation. Although the anxiety felt is real in certain situations, the fear felt by a person with a phobia is disproportionate to the actual danger posed by the situation or object. These compositions and methods are useful for treating one or more phobia symptoms, including, but not limited to, unreasonable or excessive worry about encountering the phobic object or situation, immediate intense anxiety upon encountering the phobic object or situation, and tolerating the unavoidable objects and situations with intense anxiety.

[0403] c. Eating disorders

[0404] Eating disorders are serious and often fatal conditions involving severe disturbances in a person's eating behaviours and related thoughts and emotions. Eating disorders involve an obsessive focus on food, weight and body shape. Common eating disorders include anorexia nervosa, bulimia nervosa and binge eating disorder.

[0405] Anorexia nervosa is a disorder in which people refuse to eat, severely restrict food, or eat only very small amounts of certain foods. They may also weigh themselves repeatedly. They may think they are overweight even when they are severely underweight. There are two subtypes of anorexia nervosa: restricting and binge-purge. People with restricting anorexia nervosa severely restrict the amount and types of food they eat. People with binge-purge anorexia nervosa also greatly restrict the amount and types of food they eat. In addition, they may have binge eating and purging episodes - eating large amounts of food in a short period of time and then vomiting or using laxatives or diuretics to get rid of the food they ate.

[0406] Symptoms of anorexia nervosa include, but are not limited to, thinning bones (osteopenia or osteoporosis), mild anemia and muscle weakness, brittle hair and nails, dry, yellow skin, growth of fine hair all over the body (lanugo), severe constipation, low blood pressure, slowed breathing and pulse, impaired heart structure and function, brain damage, multiple organ failure, a drop in internal body temperature causing a person to feel cold all the time, lethargy, lack of energy or tired all the time, and infertility.

[0407] Bulimia nervosa is a disorder in which the patient repeatedly and frequently eats large amounts of food and feels unable to control these eating. After a binge, the patient engages in behaviors to compensate for the overeating, such as forced vomiting, excessive use of laxatives or diuretics, fasting, excessive exercise, or a combination of these behaviors. People with bulimia nervosa may be slightly underweight, normal weight, or overweight. Symptoms of bulimia nervosa include chronic inflammation and pain in the throat, swelling of the salivary glands in the neck and jaw area, wear of tooth enamel and tooth sensitivity and decay due to exposure to stomach acid, acid reflux and other gastrointestinal problems, intestinal discomfort and irritation from laxative abuse, severe dehydration due to purging of fluids, and electrolyte imbalances (too low or too high levels of sodium, calcium, potassium and other minerals), which can lead to a stroke or heart attack.

[0408] d. Schizophrenia

[0409] The compositions and methods are useful for treating symptoms associated with schizophrenia.

[0410] Schizophrenia is characterized by a marked impairment in the way reality is perceived and behavioral changes associated with persistent delusions, where the individual persists in believing that something is real despite evidence to the contrary; persistent hallucinations, where the individual may hear, smell, see, touch, or feel things that are not there; experiences of influence, control, or passivity, where one experiences that one's feelings, impulses, actions, or thoughts are not generated by oneself but are placed in or withdrawn from one's mind by others, or that one's thoughts are being directed to others; disorganized thinking, often manifested as garbled or irrelevant speech; highly disorganized behavior, such as when the individual does things that seem strange or purposeless, or when the individual has unpredictable or inappropriate emotional responses that interfere with their ability to organize their behavior; "negative symptoms" such as very limited speech, restricted experience and expression of emotions, inability to experience interests or pleasure, and social withdrawal; and / or extreme agitation or slowed movements, with unusual postures.

[0411] Antipsychotic drugs are the mainstay of drug treatment for schizophrenia. Therefore, the composition is suitable for treating one or more of the above-mentioned positive, negative, cognitive, confusional and emotional symptoms in a subject in need thereof.

[0412] 2. Neurological diseases and neurodegenerative diseases

[0413] The compositions and methods are useful for treating symptoms associated with neurological and neurodegenerative diseases.

[0414] Neurodegenerative diseases are chronic progressive neurological diseases that affect neurological and behavioral functions, involving biochemical changes that lead to different histopathological and clinical syndromes (Hardy H et al., Science. 1998; 282: 1075-9). Abnormal proteins that are resistant to cellular degradation mechanisms accumulate within cells. The pattern of neuronal loss is selective, that is, one group of neurons is affected while other neurons remain intact. Usually, there is no clear inciting event for the disease. Classic neurodegenerative diseases include Alzheimer's disease, Huntington's disease and Parkinson's disease.

[0415] Neuroinflammation mediated by activated microglia and astrocytes is a major hallmark of various neurological diseases, making them potential therapeutic targets (Hagberg, H et al., Annals of Neurology 2012, 71, 444; Vargas, DL et al., Annals of Neurology 2005, 57, 67; Pardo, CA et al., International Review of Psychiatry 2005, 17, 485). Several scientific reports have shown that reducing early neuroinflammation by targeting these cells can delay the onset of the disease, thereby providing a longer therapeutic window for treatment (Dommergues, MA et al., Neuroscience 2003, 121, 619; Perry, VH et al., Nat Rev Neurol 2010, 6, 193; Kannan, S et al., Sci. Transl. Med. 2012, 4, 130ra46; and Block, ML et al., Nat Rev Neurosci 2007, 8, 57). It is a challenging task to deliver therapeutic agents across the blood-brain barrier. Neuroinflammation can lead to damage of the blood-brain barrier (BBB). The damaged BBB in neuroinflammatory diseases can be used to deliver drug-loaded nanoparticles to various parts of the brain (Stolp, HB et al., Cardiovascular Psychiatry and Neurology 2011, 2011, 10; and Ahishali, B et al., International Journal of Neuroscience 2005, 115, 151).

[0416] The compositions and methods can also be used to deliver active agents to treat neurological or neurodegenerative diseases or disorders or central nervous system disorders. In preferred embodiments, the compositions and methods can effectively treat and / or alleviate neuroinflammation associated with neurological or neurodegenerative diseases or disorders or central nervous system disorders, or peripheral symptoms caused by nervous system or neurodegenerative diseases or disorders. The method generally includes administering to the subject an effective amount of the composition to increase cognition or reduce cognitive decline, increase cognitive function or reduce cognitive function decline, increase memory or reduce memory decline, increase learning ability or talent or reduce learning ability or talent decline, or a combination thereof.

[0417] Neurodegeneration refers to the gradual loss of neuronal structure or function, including neuronal death. For example, the compositions and methods can be used to treat subjects with diseases or conditions, such as Parkinson's disease (PD) and PD-related conditions, Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD) and other dementias, multiple sclerosis (MS), post-encephalitic dementia, cancer and chemotherapy-related cognitive impairment and dementia, and dementia and pseudodementia caused by depression.

[0418] 3. Pain

[0419] The compositions and methods are useful for treating neuropathic and / or non-neuropathic pain associated with various diseases, such as, for example, complex regional pain syndrome, peripheral neuropathy, multiple sclerosis, and sciatica. For example, tricyclic antidepressants (TCAs) are effective for treating neuropathic (chronic) neuralgia. Chronic neuralgia, also known as neuropathic pain, is caused by nerve damage or other nerve-related problems and is generally unresponsive to conventional analgesics (e.g., paracetamol). In some forms, the compositions can be used to treat chronic pain that does not involve nerves (non-neuropathic pain). For example, the dendrimer compositions can be compounded with TCAs, SSRIs, and / or serotonin-norepinephrine reuptake inhibitors (SNRIs) and used to treat chronic non-neuropathic pain. Conditions that cause non-neuropathic pain that may benefit from treatment with the compositions include, but are not limited to, fibromyalgia, chronic back pain, and chronic neck pain.

[0420] 4. Gastrointestinal diseases

[0421] The compositions and methods are suitable for treating gastrointestinal diseases and / or digestive system diseases.

[0422] Functional gastrointestinal (GI) and motility disorders are the most common GI diseases in the general population. Functional GI diseases are diseases of gut-brain interaction. It is a group of diseases classified according to GI symptoms associated with any combination of the following: motility disorders, visceral hypersensitivity, changes in mucosal and immune function, changes in intestinal microbiota and changes in central nervous system (CNS) processing, such as difficulties in the ability of the brain to regulate pain signals from the GI tract. The term "functional" is generally applicable to diseases in which the body's normal activities in terms of intestinal motility, intestinal nerve sensitivity, or the way the brain controls some of these functions are impaired. In some forms, the composition can be administered to treat intestinal diseases and / or abdominal pain, for example, irritable bowel syndrome (IBS), functional bloating / distension, constipation, diarrhea, and / or constipation caused by opioids. People with IBS may have abnormal awareness and interpretation of these activities (abnormal perception). In some forms, the composition can be administered to treat centrally mediated gastrointestinal pain diseases, such as centrally mediated abdominal pain syndrome (CAPS) and / or narcotic bowel syndrome (NBS) / GI hyperalgesia caused by opioids.

[0423] 5. Irritable Bowel Syndrome

[0424] IBS is characterized by abnormal changes in intestinal muscle movements (dysmotility), heightened sensations from intestinal activity (increased visceral sensitivity), and brain-gut dysfunction, particularly difficulties with the brain's ability to regulate pain signals from the GI tract. Instead of normal muscle activity (motility) for digestion, people with IBS may experience spasms and cramps. If the movements are too fast, diarrhea may result, and if the movements are too slow, constipation may result. Both of these conditions may also cause abdominal discomfort or pain in people with IBS. Dysmotility may also be associated with abdominal cramping, belching, urinary urgency, or other unpleasant GI symptoms.

[0425] For people with IBS, there may also be increased sensitivity of the nerves in the GI tract. This can occur after a GI infection or surgery that causes damage to the nerves in the intestines. This can lead to a lowered threshold for experiencing gut sensations, which can cause abdominal discomfort or pain. For people with visceral hypersensitivity, even eating small amounts of food can cause stretching on the intestines, which can cause discomfort.

[0426] When nerve impulses from the gut reach the brain, they may feel more or less severe depending on the regulatory activity of the brain-gut axis. Signals of pain or discomfort travel from the gut to the brain. The brain can "reduce" pain by sending signals that block nerve impulses generated in the GI tract. This ability to reduce pain is impaired in people with IBS. Additionally, pain may become more severe when the individual is experiencing psychological distress. This can often occur due to stress in life or even the stress and frustration of GI symptoms. This brain-gut dysfunction can be treated with psychotherapy or antidepressants or a combination of both.

[0427] Thus, the dendrimer-antidepressant compositions can be used as effective analgesics (drugs that relieve pain) to treat the symptoms of IBS and other functional GI disorders. In some forms, the dendrimer-antidepressant compositions are effective for treating abdominal pain and relieving other IBS symptoms, such as diarrhea, constipation, bloating, nausea, or urinary urgency. For example, the dendrimer-antidepressant conjugates can help regulate abnormal intestinal function, such as diarrhea and constipation, and other IBS symptoms. In other forms, tricyclic antidepressants (TCAs) can be included in the dendrimer-antidepressant conjugates to help treat diarrhea, such as amitriptyline (Elavil), imipramine (Tofranil), desipramine (Norpramin), and nortriptyline (Pamelor). In another example, the dendrimer-antidepressant conjugates can include serotonin reuptake inhibitors (SSRIs) to help treat constipation. In an exemplary embodiment, the dendrimer-antidepressant conjugates can help treat other problems, such as anxiety and depression, which are often associated with chronic pain disorders.

[0428] 6. Sleep disorders

[0429] The compositions and methods are useful for treating or managing symptoms associated with sleep disorders and / or sleep disruption, including, but not limited to, insomnia, restless legs syndrome, narcolepsy, and sleep apnea.

[0430] In some forms, the compositions and methods are also applicable to the treatment of symptoms associated with narcolepsy. Narcolepsy is a rapid onset REM sleep disorder characterized by excessive daytime sleepiness (EDS), frequent uncontrollable sleep attacks, and sleep fragmentation, and may be associated with cataplexy, sleep paralysis, and hypnagogic hallucinations. There are two types: narcolepsy type 1 (formerly known as narcolepsy with cataplexy) and narcolepsy type 2 (formerly known as narcolepsy without cataplexy). In some forms, these compositions and methods may be applied to patients in need to treat or manage one or more symptoms of cataplexy, including epileptic seizures (especially asthenic seizures), periodic paralysis, cardiogenic syncope, orthostatic syncope, neurogenic syncope, and / or psychotic symptoms. In some forms, these compositions and methods may be applied to patients in need to treat or control excessive daytime sleepiness in narcolepsy patients. In some forms, these compositions and methods can be administered to a patient in need thereof to treat or manage depression and / or anxiety symptoms in a patient with narcolepsy.

[0431] 7. Neurodevelopmental and attention deficit disorders

[0432] The compositions and methods are useful for treating or managing symptoms associated with attention deficit disorders, such as attention deficit hyperactivity disorder, autism spectrum disorder, Tourette syndrome, sensory integration disorder, auditory processing disorder, and other specific learning difficulties.

[0433] In some forms, the composition can be applied to patients in need to treat or manage symptoms associated with developmental disorders (e.g., autism spectrum disorders). Autism spectrum disorders (ASD) include a range of neurodevelopmental disorders. This spectrum is characterized by repetitive behavior patterns, interest patterns, activity patterns, and social interaction problems. ASD is a complex neurodevelopmental disorder characterized by behavioral and psychological problems in children. These children feel distressed when their surroundings change because their ability to adapt is extremely low. These symptoms begin to appear in early childhood and affect daily function. Children with ASD have a higher rate of language problems, intellectual disabilities, and epilepsy than the general population.

[0434] In some forms, the composition can be administered to a patient in need to treat or manage symptoms associated with attention deficit / hyperactivity disorder (ADHD). ADHD is one of the most common neurodevelopmental disorders, usually first diagnosed in childhood and often persisting into adulthood. Individuals with ADHD may have difficulty concentrating, controlling impulsive behavior (may act without considering the consequences), or being overactive. Symptoms of ADHD include, but are not limited to, inattention, e.g., confusion, problems with being unable to focus on tasks, frequent daydreaming, not paying attention when speaking in person; impulsivity, e.g., making impromptu decisions without considering the possibility of harm or long-term effects, often acting quickly to obtain immediate rewards, and often interrupting others; and hyperactivity, e.g., twisting, fidgeting, tapping, talking, and constantly moving, especially in inappropriate situations.

[0435] Tourette syndrome (also known as Tourette disorder) is a common neurodevelopmental disorder that affects up to 1% of the population. It is characterized by multiple motor and vocal tics and begins in childhood. In some forms, the composition can be administered to patients in need to treat or manage symptoms associated with Tourette syndrome, including but not limited to simple tics, such as sudden, brief, repetitive movements involving a limited number of muscle groups, such as blinking and other eye movements, facial expressions of pain, shrugging, and head or shoulder twitching, repeated throat clearing, sniffing, barking or grunting, and / or complex tics, such as unique coordinated movement patterns involving multiple muscle groups (e.g., facial expressions of pain combined with head twisting and shrugging), sniffing or touching objects, jumping, bending or twisting, repeating one's own words or phrases, repeating the words or phrases of others (called echolalia), or, more rarely, using vulgar, obscene or dirty words (called profanity).

[0436] C. Dosage and Effective Amount

[0437] In some in vivo methods, the dendrimer complex is administered to a subject in a therapeutically effective amount. The term "effective amount" or "therapeutically effective amount" refers to a dose sufficient to treat, inhibit or alleviate one or more symptoms of the disease being treated or otherwise provide the desired pharmacological and / or physiological effect. The exact dose will vary depending on a variety of factors, such as subject-related variables (e.g., age, immune system health, etc.), the disease or condition, and the treatment affected.

[0438] Typically, the dosage of the composition can be from about 0.001 to about 100 mg / kg of the body weight of the subject being treated, from about 0.01 to about 100 mg / kg of body weight, from about 0.1 mg / kg to about 10 mg / kg, and from about 0.5 mg to about 5 mg / kg of body weight. The subject is typically a mammal, most preferably a human.

[0439] For example, the amount of the dendrimer complex composition is effective to deliver one or more active agents to cells at or near the site of inflammation, particularly central nervous system inflammation or ocular inflammation. Thus, in some embodiments, the amount of the dendrimer complex composition comprising one or more active agents is effective to improve inflammation in a subject. In a preferred embodiment, the effective amount of the dendrimer complex composition does not induce significant cytotoxicity in cells of the subject compared to an untreated control subject. Preferably, the amount of the dendrimer complex composition is effective to prevent or reduce inflammation and / or further associated symptoms of a disease or condition in a subject compared to an untreated control.

[0440] In general, the time and frequency of administration will be adjusted to balance the efficacy of a given therapeutic or diagnostic regimen with the side effects of a given delivery system. Exemplary dosing frequencies include continuous infusion, single and multiple administrations, such as hourly, daily or weekly administrations.

[0441] In some embodiments, the dosage is administered once, twice, or three times a day, or once every other day, two days, three days, four days, five days, or six days. In some embodiments, the dosage is administered once or twice a week, once or twice every two weeks, or once every three weeks. In some embodiments, the dosage is administered once or twice a month, once or twice every two months, once or twice every three months, once or twice every four months, once or twice every five months, or once or twice every six months.

[0442] Those of ordinary skill in the art will appreciate that the dosing regimen can be any length of time sufficient to treat the subject's disease. The term "chronic" means that the length of the dosing regimen can be hours, days, weeks, months, or possibly years.

[0443] In some embodiments, the regimen includes one or more cycles of treatment followed by a drug holiday (e.g., no drug). The treatment cycle can be, for example, the dosing cycle described above. Likewise, the drug holiday can be 1, 2, 3, 4, 5, 6, or 7 days: or 1, 2, 3, 4 weeks, or 1, 2, 3, 4, 5, or 6 months.

[0444] The dendrimer complexes may be administered in combination with one or more additional therapeutically active agents known to treat the above-mentioned conditions or diseases.

[0445] D. Control

[0446] The effect of the dendrimer complex composition can be compared to a control. Suitable controls are known in the art and include, for example, untreated cells or untreated subjects. In some embodiments, the control is untreated tissue from a treated subject or from an untreated subject. Preferably, the control cell or tissue is derived from the same tissue as the treated cell or tissue. In some embodiments, the untreated control subject suffers from or is at risk for the same disease or condition as the treated subject.

[0447] E. Combination therapy

[0448] The dendrimer complex composition can be administered alone or in combination with one or more other active agents as part of a therapeutic or prophylactic treatment regimen, including other antidepressants or psychedelics, cannabinoids, or hallucinogens. 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 including the dendrimer complex composition can be administered on the first, second, third, or fourth day, or a combination of these days.

[0449] The term "combination" or "combination" is used to refer to the concomitant, simultaneous or sequential administration of two or more drugs. Thus, the combination can be administered simultaneously (e.g., as a mixture), separately but simultaneously (e.g., through different intravenous lines into the same subject), or sequentially (e.g., one compound or drug is administered first and then the second is administered).

[0450] VI. Kit

[0451] The composition can be packaged in a kit. The kit can include a single or multiple doses of a composition comprising one or more antidepressants and antipsychotics bound or conjugated to a dendrimer (e.g., one or more hydroxyl-terminated PAMAM dendrimers or glucose dendrimers as described in the Examples), and instructions for administering the composition. Specifically, the instructions direct the administration of an effective amount of the dendrimer composition to an individual suffering from a particular disease / disorder. The composition can be formulated with reference to a particular treatment method as described above, and can be packaged in any convenient manner.

[0452] The invention will be further understood with reference to the following non-limiting examples. Example

[0453] Example 1: Synthesis of Hydroxyl-Polyamidoamine (PAMAM-OH) Dendrimers Conjugated with Antipsychotic / Antidepressant Drugs

[0454] The conjugation of PAMAM-OH to one or more antipsychotics or antidepressants is described below. Selective serotonin reuptake inhibitor (SSRI) drugs are used as exemplary drugs. Exemplary SSRIs include fluoxetine Paroxetine Venlafaxine, Citalopram wait.

[0455] The synthesis of PAMAM-OH-SSRI conjugates is achieved using a combination of linking chemistry and linkers (cleavable and non-cleavable). In short, the surface hydroxyl groups on PAMAM-OH are modified with linkers to bring complementary groups on the surface, which can further react with complementary groups on the drug linker. On the other hand, the drug is modified by a linker to bring complementary functional groups so as to react with the dendrimer-linker. The linker on the drug is connected by a cleavable or non-cleavable bond. Examples of cleavable bonds include ester bonds sensitive to esterases, glutathione sensitive disulfide bonds, phosphatase sensitive phosphodiester bonds, triglycyl peptide linkers (CX) capable of lysosomal release, acid cleavable hydrazine bonds, etc. Examples of non-cleavable bonds include ether bonds, aminoalkyl bonds, or amide bonds. The linker can be an amino acid, a peptide, polyethylene glycol (n=2-15) or a hydrocarbon chain.

[0456] 1. Synthesis of hydroxyl-terminated PAMAM dendrimers using unbreakable bonds - Fluoxetine Conjugate

[0457] according to Figure 1A and 1B The reaction shown utilizes a non-cleavable bond to prepare a PAMAM dendrimer-fluoxetine (Prozac) conjugate. Figure 1A and 1B is a schematic diagram showing an exemplary synthetic route for conjugating fluoxetine to a dendrimer using click chemistry.

[0458] 2. Synthesis of hydroxyl-terminated PAMAM dendrimers using unbreakable bonds - paroxetine Conjugate

[0459] according to Figure 2A and 2B The reaction shown utilizes a non-cleavable bond to prepare a PAMAM dendrimer-paroxetine (Paxil) conjugate. Figure 2A and 2B is a schematic diagram showing an exemplary synthetic route for conjugating paroxetine to a dendrimer using click chemistry.

[0460] 3. Synthesis of hydroxyl-terminated PAMAM dendrimer-venlafaxine conjugates using enzyme-cleavable bonds

[0461] according to Figure 3A and 3B The reaction shown utilizes an enzyme-cleavable ester bond to prepare a PAMAM dendrimer-venlafaxine conjugate. Figure 3A and 3B is a schematic diagram showing an exemplary synthetic route for conjugating venlafaxine to a dendrimer using click chemistry.

[0462] 4. Synthesis of Hydroxyl-Terminated PAMAM Dendrimer-Venlafaxine Analog Conjugates Using Non-Broken Bonds

[0463] according to Figure 4A and 4B The reaction shown utilizes a non-cleavable amide bond to prepare a PAMAM dendrimer-venlafaxine analog conjugate. Figure 4A and 4B is a schematic diagram showing an exemplary synthetic route for conjugating venlafaxine analogs to dendrimers using click chemistry.

[0464] 5. Synthesis of Hydroxyl-Terminated PAMAM Dendrimers-Citalopram Using Unbreakable Bonds Analog conjugate

[0465] according to Figure 5A and 5B The reaction shown utilizes a non-cleavable amide bond to prepare a PAMAM dendrimer-Citalopram (Celexa) conjugate. Figure 5A and 5B is a schematic diagram showing an exemplary synthetic route for conjugating citalopram to a dendrimer using click chemistry.

[0466] Example 2: Synthesis of Glucose Dendrimers (GD) Conjugated with Antipsychotic / Antidepressant Drugs

[0467] The following describes the conjugation of glucose dendrimers (GD) to one or more antipsychotics or antidepressants. Selective serotonin reuptake inhibitor (SSRI) drugs are used as exemplary agents. Exemplary SSRIs include fluoxetine Paroxetine Venlafaxine, Citalopram wait.

[0468] The synthesis of GD-SSRI conjugates is achieved using a combination of various linking chemistries and linkers (cleavable and non-cleavable). In short, the surface hydroxyl groups on GD are modified with linkers to bring complementary groups on the surface, which can further react with complementary groups on the drug linker. On the other hand, the drug is modified by a linker to produce complementary functional groups in order to react with the dendrimer-linker. The linker on the drug is connected by a cleavable or non-cleavable bond. Examples of cleavable bonds include ester bonds sensitive to esterases, glutathione sensitive disulfide bonds, phosphatase sensitive phosphodiester bonds, triglycyl peptide linkers (CX) capable of lysosomal release, acid cleavable hydrazine bonds, etc. Examples of non-cleavable bonds include ether bonds or amide bonds. The linker can be an amino acid, a peptide, polyethylene glycol (n=2-15) or a hydrocarbon chain.

[0469] 1. Synthesis of glucose dendrimers-fluoxetine using non-breakable bonds Conjugate

[0470] according to Figure 6 The reaction shown in Figure 1 utilizes a non-cleavable bond to prepare a glucose dendrimer-fluoxetine conjugate. The synthesis of fluoxetine-azide is as follows: Figure 1A The synthesis of the glucose dendrimer and fluoxetine conjugate was achieved by partially modifying the OH groups of the glucose dendrimer with complementary groups on the surface of the glucose dendrimer, which reacted with the complementary linker containing an azide attached to fluoxetine to generate a glucose dendrimer-fluoxetine conjugate ( Figure 6 ).

[0471] 2. Synthesis of Glucose Dendrimers-Paroxetine Using Unbreakable Bonds Conjugate

[0472] according to Figure 7 The reaction shown in Figure 1 utilizes a non-cleavable bond to prepare a glucose dendrimer-paroxetine conjugate. Paroxetine-azide synthesis is shown in Figure 2 Figure 2A The synthesis of the glucose dendrimer and paroxetine conjugate was achieved by partially modifying the OH groups of the glucose dendrimer with complementary groups on the surface of the glucose dendrimer, which reacted with the complementary linker containing an azide attached to paroxetine to generate the glucose dendrimer-paroxetine conjugate ( Figure 7 ).

[0473] 3. Synthesis of glucose dendrimer-venlafaxine conjugates using enzyme-cleavable bonds

[0474] according to Figure 8The reaction shown in Figure 1 is used to prepare glucose dendrimer-venlafaxine conjugates using an enzyme that can cleave the ester bond. Venlafaxine is first modified with an azide, such as Figure 3A An exemplary synthetic route of glucose dendrimer and venlafaxine conjugate is shown in Figure 8 shown.

[0475] 4. Synthesis of Glucose Dendrimer-Venlafaxine Analog Conjugates Using Non-Broken Bonds

[0476] according to Fig. 9 The reaction shown in Figure 1 utilizes a non-cleavable amide bond to prepare a glucose dendrimer-venlafaxine conjugate. Venlafaxine is first modified with an azide, such as Figure 4A An exemplary synthetic route of glucose dendrimer and venlafaxine conjugate is shown in Fig. 9 shown.

[0477] 5. Synthesis of Glucose Dendrimer-Citalopram Analog Conjugates Using Non-Broken Bonds

[0478] according to Fig.10 The reaction shown in Figure 1 utilizes a non-cleavable bond to prepare a glucose dendrimer-citalopram analog conjugate. First, the citalopram analog is modified with an azide, such as Figure 5A An exemplary synthetic route for the conjugate of glucose dendrimer and citalopram analog is shown in Fig.10 shown.

[0479] Unless otherwise stated, reactions were performed in flame-dried glassware under a positive pressure of nitrogen using dry solvents. Commercial grade reagents and anhydrous solvents were purchased from chemical suppliers and used without further purification. 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC.HCl), N,N-diisopropylethylamine (DIPEA), 4-(dimethylamino)pyridine (DMAP) trifluoroacetic acid (TFA), anhydrous dichloromethane (DCM), and N,N′-dimethylformamide (DMF) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cyanine 5 (Cy5)-monoNHS ester was purchased from Amersham Bioscience-GE Healthcare. Deuterated solvents dimethyl sulfoxide (DMSO-d6), water (D2O), and chloroform (CDCl3) were purchased from Cambridge Isotope Laboratories Inc. (Andover, MA). Ethylenediamine core polyamidoamine (PAMAM) dendrimers, generation 4.0, hydroxyl surface (G4-OH; diagnostic grade; consisting of 64 hydroxyl end groups), methanol solution (13.75% w / w) were purchased from Dendritech Inc. (Midland, MI, USA). Dialysis membranes were purchased from Spectrum Laboratories Inc. (Rancho Dominguez, CA, USA). Hu308 tryptamine 1-(2-amino-1-(4-methoxyphenyl)ethyl)cyclohexanol, norketamine, serotonin, psilocybin analogs, psilocybin analogs, and cannabidiol drugs were purchased from Cayman Chemicals.

[0480] Synthesis of hydroxy-PAMAM dendrimer drug conjugates.

[0481] PAMAM-G4-OH (D4-OH) dendrimers consisting of approximately 64 terminal hydroxyl groups were used in the synthesis. After each synthetic step, the product was purified by dialysis in DMF for 24 h to eliminate small molecule impurities, followed by dialysis against water to remove DMF. 1 H NMR (in DMSO-d6 and D2O) and analytical HPLC were used to confirm the formation and purity of the intermediates and final products. The monofunctional D4-OH was functionalized with alkyne groups to afford the D-hexyne difunctional dendrimer by treating 5-hexynoic acid with EDC.HCl and 4-DMAP in DMF at room temperature under standard esterification conditions for 36 hours. The number of alkyne groups on the surface of the dendrimer was selected to be maintained at ~10-15 to maintain the overall water solubility of the conjugate. The crude product was dialyzed against ultrapure water using a 1 kDa membrane for 24 hours to remove low molecular weight impurities by selective diffusion across the semipermeable dialysis membrane. 1H NMR and analytical HPLC confirmed product formation and the purity of the intermediate and final products.

[0482] Synthesis of D-hexyne

[0483] A solution of PAMAM G4-OH 1 (10.00 g, 0.7 mmol) in DMF (50 mL) was treated with 5-hexynoic acid (1.40 g, 12.6 mmol), DMAP (2.41 g, 12.6 mmol) and stirred at room temperature for 5 minutes. Then EDC.HCl (1.54 g, 12.6 mmol) was added to the reaction mixture in batches over 5 minutes. The reaction mixture was stirred at room temperature for 36 hours. The crude product was transferred to a 1 kD MW cut-off cellulose dialysis tubing, dialyzed against DMF for 12 hours, and then dialyzed against water for 24 hours. The aqueous layer was frozen and lyophilized to give D-hexyne as a hygroscopic white solid (yield of 75%). 1 H NMR (500 MHz, DMSO-d6) 8.21-7.57 (m, internal amide H), 4.71 (s, GABA amide H, 50H), 4.01 (t, 22-24 CH2), 3.5-2.1 (m, dendrimer CH2) 1.71-1.59 (m, 22-24 CH2). HPLC C18 retention time 4 minutes: purity -99%.

[0484] Synthesis of dendrimer-drug conjugates.

[0485] A solution of D-hexyne and drug azide in DMF (5 mL) was treated with copper sulfate pentahydrate (CuSO4.5H2O) and sodium ascorbate aqueous solution. The reaction mixture was stirred in a microwave synthesizer and heated at 50°C for 10 hours. Upon completion, the reaction mixture was dialyzed against DMF in a 1 KDa cutoff cellulose dialysis tubing. To this solution was added EDTA (50 μL, 0.5 M) solution to remove copper by chelation. Dialysis against DMF was followed by dialysis against water overnight. Drug loading was calculated by proton integration, where the peaks corresponding to the dendrimer and drug were compared.

[0486] G2-glucose dendrimer (GD2)-drug conjugate

[0487] The second generation of glucose dendrimers (GD2) consist of 24 glucose molecules (96 surface hydroxyl groups) for conjugation. Glucose dendrimers are primarily composed of glucose moieties consisting of a central core of dipentaerythritol and one or more branching units of monosaccharide glucose molecules. Unlike hydroxyl-terminated PAMAM dendrimers, glucose dendrimers are primarily taken up by damaged neurons and were found to specifically target overexcitable neurons in culture and in vivo mouse models.

[0488] Synthesis and characterization of glucose dendrimers (GD2).

[0489] The GD synthesis begins with the reaction of the hexapropargylated core with AB4 and β-D-glucose-PEG4-azide to generate the first generation of glucose dendrimers (GD1) via a click reaction. The OH groups on GD1 are propargylated to give GD1-acetylene 24, which reacts with β-D-glucose-PEG4-azide to give the second generation (GD2) with 24 glucose moieties, providing 96 surface hydroxyl groups. In addition, a Cy5 fluorescent tag is attached to GD2 by propargylation of ~2-3 hydroxyl groups to give the alkyne-containing GD2 dendrimer. The GD intermediates and final products are purified using dialysis and used 1 The physicochemical properties of GD2 dendrimers were also evaluated (Table 2).

[0490] Table 2: Physicochemical properties of GD2

[0491] size 4.4nm Zeta potential -6.0 Glucose quantity 24 HPLC purity ＞99％ Molecular weight 12800Da

[0492] Synthesis and characterization of GD2-drug conjugates

[0493] A click chemistry strategy was used to conjugate the drugs of norketamine, tryptamine, venlafaxine, and Hu308 to GD2-hexynoic acid dendrimers. The linker-linked drug moiety was conjugated to the glucose dendrimer using a Cu(I)-catalyzed click (CuAAC) reaction in the presence of catalytic amounts of CuSO4.5H2O and sodium ascorbate to afford the GD2-drug conjugates. Trace amounts of copper were removed by dialysis against ethylenediaminetetraacetic acid (EDTA). The final GD2-drug conjugates were characterized by NMR and HPLC. Fig.14A Schematic diagram of the synthesis of PAMAM dendrimer-norketamine conjugate. Fig. 14B Schematic diagram of the synthesis of glucose dendrimer-norketamine conjugate.

[0494] Characterization of dendrimer-drug conjugates with venlafaxine and fluoxetine showed that these conjugates were pure and significantly more soluble in water than the free drug, >200-fold and >2900-fold, respectively. This improved solubility allowed them to be formulated in saline without the use of relatively toxic formulations.

[0495] Table 3: Properties of the conjugated compounds

[0496]

[0497] Table 3: Drugs and their conjugates

[0498]

[0499]

[0500] Table 4. Physical properties of dendrimer-drug conjugates.

[0501]

[0502] *https: / / go.drugbank.com / metabolites / DBME T 00 1 89

[0503] Example 4: Binding assay of dendrimer-ketamine conjugates

[0504] Materials and methods

[0505] Human Serotonin 5-HT2A Receptor (Agonist Radioligand):

[0506] Objective: To evaluate the affinity of compounds for the human 5-HT2A receptor in transfected HEK-293 cells by determining the affinity in a radioligand binding assay.

[0507] Experimental protocol: Cell membrane homogenates (30 μg protein) were incubated with 0.1 nM [125I]DOI in a buffer containing 50 mM Tris-HCl (pH 7.4), 5 mM MgCl2, 10 μM pargyline and 0.1% ascorbic acid at 22°C for 60 min in the presence or absence of test compounds.

[0508] Non-specific binding was determined in the presence of 1 μM DOI.

[0509] After incubation, the samples were quickly filtered under vacuum through a glass fiber filter (GF / B, Packard) pre-soaked with 0.3% PEI and rinsed several times with ice-cold 50 mM Tris-HCl using a 96-sample cell collector (Unifilter, Packard). The filters were dried and radioactivity was counted in a scintillation counter (Topcount, Packard) using a scintillation cocktail (Microscint 0, Packard).

[0510] Results are expressed as percent inhibition of specific binding of control radioligand.

[0511] The standard reference compound is DOI, which is tested in each experiment at multiple concentrations to obtain a competition curve from which its IC50 is calculated.

[0512] See Bryant et al. (1996), A novel class of 5-HT2A receptor antagonists: aryl aminoguanidines, Life Sci., 15: 1259.

[0513] Δ(DOP) Human Opioid GPCR Cell-Based Antagonist cAMP Assay:

[0514] Purpose: To evaluate the potency (IC50) and efficacy (maximal response) of compounds against the human Δ(DOP) receptor in stably transfected CHO-K1 cells. The assay principle is a cAMP cell-based assay.

[0515] Experimental protocol: Cells were seeded into white-walled 384-well microplates in a total volume of 20 μL and incubated overnight at 37° C. Prior to testing, the cell plating medium was exchanged with 10 uL of assay buffer (HBSS+10 mM HEPES).

[0516] Briefly, intermediate dilutions were made to the sample stock to generate 4X samples in assay buffer. 5 μL of 4X sample was added to the cells and incubated at 37°C for 30 minutes. 5 uL of 4X EC80 in 4X forskolin reagent was added and the cells were incubated at 37°C for 30 minutes. The final assay vehicle concentration was 1%. Results are expressed as percent inhibition of the control ligand.

[0517] General information

[0518] Assay volume and format: 10 μl in 384-well plate

[0519] Compound addition: 5ul 4X compound.

[0520] Maximum Tolerable DMSO Concentration: Client compound (in DMSO) diluted in solvent to a [100x] solution. 1%

[0523] Detection temperature: 37℃

[0524] Incubation time: 30 minutes

[0525] Forskolin concentration: 20uM

[0527]

[0528] TA1 Human Trace Amine GPCR Cell-Based Agonist cAMP Assay

[0529] Objective: To determine the evaluation of the potency (EC50) and efficacy (maximal response) of compounds at the human TA1 receptor in stably transfected CHO-K1 cells in a GPCR cell-based cAMP assay.

[0530] Experimental Protocol: Cells were seeded into white-walled 384-well microplates in a total volume of 20 μL and incubated overnight at 37°C prior to testing.

[0531] Prior to testing, the cell plating medium was exchanged with 15 μL of assay buffer (HBSS + 10 mM HEPES). In brief, intermediate dilutions were made to the sample stock solution to generate 4X samples in assay buffer. 5 μL of 4X was added to the cells and incubated at 37°C for 30 minutes. The final assay vehicle concentration was 1%.

[0532] Results are expressed as percent efficacy relative to the maximal response of the control ligand.

[0533] General information

[0534]

[0535] result

[0536] Table 5: Results of binding assays

[0537]

[0538] Fig.15A Graph of NMDAR 1A / 2B antagonist assays of glucose dendrimer-ketamine (IC50 = 4.54 μM), hydroxy dendrimer-ketamine (IC50>100), and norketamine (IC50 = 6.96 μM). Fig. 15B is the % binding potency of the compound log concentration (μM) in the D2L human dopamine GPCR cell based agonist cAMP assay. Norketamine (closed circles), glucose dendrimer-ketamine EC50 = 13.08 μM (open circles) and hydroxy dendrimer-ketamine EC50 = 4.263 μM (triangles). Fig. 15C is the % binding potency of the log concentration (μM) of ketamine in the TA1 human microamine GPCR cell-based agonist cAMP assay. Norketamine (closed circles), glucose dendrimer-ketamine EC50 = 13.08 μM (open circles), and hydroxy dendrimer-ketamine EC50 = 4.263 μM (triangles).

[0539] Significant efficacy data have been obtained with clinically important drugs that are derivatives of ketamine (norketamine). Norketamine has been tested for depression and addiction. Hydroxydendrimer-drugs target these receptors in microglia / macrophages, while glucose dendrimer-drugs target these receptors in neurons and microglia.

[0540] The conjugate can be active with or without releasing the drug. Binding affinity measures the activity of the intact conjugate. Typically, conjugates of psychedelic drugs are prepared in a non-releasing form (e.g., tryptamine, pseudoergotamine, psilocybin, ketamine) and are intended to be active in the intact form. This allows the intact conjugate to be released through the kidney without the toxicity from drug release. Alternatively, they can be designed to be released through analogs.

[0541] Both GD-ketamine and HD-ketamine exhibit unique and beneficial effects on neuropsychiatric disorders. Both dendrimer-ketamine conjugates have greater binding to opiate μ, opiate k, and σ2 receptors than norketamine, suggesting that dendrimer conjugation provides a positive benefit for binding to these receptors. Norketamine does not bind to opioid μ receptors (Ki of 53 μM for GD-ket and 40 μM for HD-ket). Norketamine has a Ki of 381 uM for opioid k, which is about 4 times better for GD-ket (Ki=82 μM) and about 7 times better for HD-Ket (Ki=556 μm). Ketamine's antidepressant effects are thought to be mediated through opioid k receptors, and dendrimer binding increases affinity for these receptors.

[0542] Improved binding was also seen in the case of the σ2 receptor after dendrimer conjugation. The greater ability of HD ketamine to bind to the σ2 receptor suggests a greater neuroprotective effect. The mechanism of neuroprotection may be related to increased NGF and BDNF production. σ2 receptor activation may also be beneficial in neuropsychiatric disorders such as schizophrenia and psychosis. Reduced anxiety and greater antidepressant effects were seen in the case of σ2 receptor targeting. Ketamine binds to the σ2 receptor at mM concentrations (Pergolizi, 2023; Bonaventura, 2021).

[0543] The ability to bind to dopamine 2 receptors was only seen with dendrimer-conjugated ketamine. Free norketamine does not appear to bind to dopamine 2 receptors. Similarly, ketamine also did not show any affinity for dopamine 2 receptors. The dopaminergic effects of ketamine are thought to be indirect. However, it has been shown that glucose dendrimer-ketamine has an RC50 of 13.07 μM for D2 receptors, while hydroxy dendrimer-ketamine is more effective with an RC50 of 4.3 μM. The antidepressant and neuroprotective effects of ketamine are primarily mediated through these and dopamine receptors. Ketamine, norketamine, and ketamine metabolites do not appear to have a direct effect on dopamine receptors and do not bind to dopamine receptors. The increased activity seen at D2 receptors in functional assays suggests that dendrimer-ketamine will be more effective as an antidepressant.

[0544] GD-ketamine and HD-ketamine exhibited different profiles. Binding to hydroxy dendrimers (HD) appeared to alter ketamine function compared to glucose dendrimers (GD), which was highly anticipated. HD-ketamine did not exhibit NMDAR 1A / 2B ion channel blockade, whereas GD-ketamine did show that the type of dendrimer bound to ketamine was critical for the difference in function between them. GD-ketamine also exhibited agonist activity against 5HT1A receptors, which was not seen in the case of HD-ketamine.

[0545] These results suggest that dendrimer conjugation yields unexpected benefits over free drug in terms of receptor binding and is dependent on the dendrimer structure. Microglial targeting of hydroxy dendrimers and additional neuronal targeting of glucose dendrimers yield unexpected results with clinical significance.

[0546] The conjugates target specific receptors on any cell they may be located on. This includes neurons, microglia, macrophages, and other cells. These receptors can be located anywhere in the brain. Examples include serotonin receptors such as 5HT1A, 5HT2A, NMDA, etc. Serotonin receptors are also found in microglia. These compounds bind to and act on these receptors. The "intrinsic cellular targeting" of these dendrimers (hydroxy dendrimers - microglia / macrophages, glucose dendrimers - neurons) is somewhat secondary to the effects on specific receptors because these compositions target specific receptors. Important receptors include serotonin receptors [5HT1A, 5HT2A (agonism, antagonism, inverse or inverse agonism)] and NMDA receptors. In many cases, using dendrimers to bind to specific receptors and not to other receptors can enhance the efficacy of binding and reduce the side effects of these drugs.

[0547] When combined with the cell targeting capabilities of glucose dendrimers, e.g., targeting damaged neurons (primary), microglia / macrophages (secondary), or hydroxy dendrimers (microglia / macrophages), which exhibit >5-200-fold increased water solubility in the case of dendrimer conjugation, these conjugates offer distinct benefits: altered binding, increased targeting selectivity, improved ease of formulation and delivery, and reduced side effects.

[0548] Using tryptamine as an example, it was demonstrated that the binding affinity of drugs conjugated to these dendrimers has unexpected properties.

[0549] Both the hydroxy and GD dendrimers have OH surface groups. When tryptamines were bound to these dendrimers with the same attachment chemistry, the affinity for serotonin and other receptors varied greatly. In cell-based binding assays (indicative of in vivo efficacy), the conjugates exhibited lower affinity than the free drug. The lower affinity may make the binding less tight and may allow us to modulate the drug to have an undesirably strong effect on this receptor. Second, the hydroxy dendrimer conjugates were not active, but the glucose dendrimer conjugates were. This was unexpected and may be due to the different internal structures of glucose and hydroxy dendrimers. The drug may fold into the hydrophobic core of the hydroxy dendrimer, but may open outward in the hydrophilic interior of the glucose dendrimer.

[0550] Example 5: Treatment of Rett syndrome animal model with ketamine and ketamine-dendrimer conjugates

[0551] Rett syndrome (RTT) is an inherited neurodevelopmental disorder of females that affects 1 in 10,000-15,000 births. Affected females develop normally for 6-18 months, but then lose voluntary motor skills, including speech and hand skills. Most people with RTT are heterozygous for mutations in the X-linked gene MECP2, which encodes a protein that binds to methylation sites in genomic DNA and promotes gene silencing. The symptoms, progression, and severity of Rett syndrome can vary significantly from person to person. A variety of disabilities may be associated with Rett syndrome. Symptoms typically develop in stages. RTT is typically characterized by a period of normal development after birth, followed by regression of speech and hand movements, abnormal gait, irregular hand movements, and slowed head growth. Other diagnostic criteria for RTT include irregular breathing, gastrointestinal and musculoskeletal problems, seizures, poor sleep, decreased response to physical pain, and behavioral problems.

[0552] Ketamine is a well-established anesthetic drug that causes "dissociative anesthesia" and exerts central and peripheral effects, including hypnotic, analgesic, and sympathomimetic effects, leading to hypertension and tachycardia. The primary mechanism is thought to be due to its action as an antagonist at the N-methyl-d-aspartate (NMDA) receptor. This can lead to the rapid action and response seen in the treatment of refractory depression, MDD, and suicidal ideation, which is different from SSRIs and SNRIs that only show delayed effects in controlling depression. Ketamine is also an effective treatment for chronic pain, again due to its effects on NMDAR inhibition. Increased dopamine release and its effects. However, ketamine also exerts neuroprotective effects through non-NMDAR-mediated mechanisms, such as increasing BDNF and mTOR. Ketamine also acts on opioid receptors and can increase dopamine. These effects are beneficial for chronic pain, refractory depression, MDD, and refractory epilepsy / seizures.

[0553] However, ketamine has several short-term and long-term side effects. Ketamine can cause respiratory depression at high doses and can lead to systemic side effects such as increased heart rate, hypertension, hyperthermia, loss of coordination, dizziness, nausea, vomiting, sensory perceptual disturbances, and high rates of auditory and visual hallucinations. More than half of patients treated with ketamine experience a flare-up as the effects of ketamine wear off, characterized by euphoria, vivid dreams, hallucinations, illusions, distortions of body image and objects, and delirium, which can be very disturbing and may lead to self-harm. Some of these symptoms are related to symptoms of schizophrenia. Long-term use of ketamine can lead to impaired memory and decreased executive function. Ketamine can also lead to tolerance and addiction, which can lead to withdrawal and dependence. Due to these significant side effects, ketamine should only be used in a controlled environment.

[0554] Ketamine has been shown to be effective in a mouse model of Rett syndrome, with bolus doses of 8 mg / kg delivered IP daily for 40 consecutive days (total dose of 320 mg / kg) and increased survival by 50% at 80 days after birth (Patrizi et al., 2016).

[0555] Materials and methods

[0556] Dendrimer conjugates were prepared as described above.

[0557] As reported by Guy et al., a mouse Mecp2-null mutation causes neurological symptoms that mimic Rett syndrome. Nat Genet. 2001 Mar; 27(3): 322-6. doi: 10.1038 / 85899. PMID: 11242117. Both Mecp2-null mice and mice in which Mecp2 is knocked out in the brain show severe neurological symptoms at about six weeks of age.

[0558] Patrizi A et al., Chronic Administration of the N-Methyl-D-Aspartate Receptor Antagonist Ketamine Improves Rett Syndrome Phenotype. Biol Psychiatry. 2016 May 1;79(9):755-764. doi:10.1016 / j.biopsych.2015.08.018. Epub 2015 Aug 24. PMID:26410354; PMCID:PMC7410367, report a systematic, randomized preclinical trial of chronic administration of low-dose (8 mg / kg, intraperitoneal) ketamine, an NMDAR antagonist, in Mecp2-null mice starting early in development or at the onset of the RTT phenotype. Mice were treated with 8 mg ketamine / kg / day ip from day 15 to day 55 or from day 30 to day 55 (total dose of 320 mg / kg for 40 days or 200 mg / kg for 25 days). Treatment starting on day 30 was ineffective, while treatment starting on day 15 showed some efficacy at a total dose of 320 mg / kg.

[0559] Experimental Paradigm for MECP2 Knockout ("KO") Mice:

[0560] Treatment starts at 28 days of age (4 weeks of age, symptomatic). Untreated animals die around 55-60 days.

[0561] Mice were treated with ketamine or ketamine conjugated to dendrimers. Control groups were wild-type treated with saline and knockout treated with saline.

[0562] Animals were dosed ip twice weekly at 2.5 mg / kg / dose. The total dose at the time of evaluation (60 days of age) was approximately 22.5 mg / kg.

[0563] Dendrimer ketamine was tested IP in the Rett syndrome mouse model at a dose of 2.5 mg / kg (ketamine) every two weeks for 8 weeks (total ketamine dose of 40 mg / kg).

[0564] Mecp2 Kos (knockouts) at 4 weeks of age were randomized and treated with saline or 2.5 mg / Kg ip of ketamine or D-ketamine (WT-saline, KO-saline, KO-ketamine, KO D-ketamine groups) every two weeks for 8 weeks. Neurobehavior was evaluated weekly by recording their comprehensive neurobehavioral score (NBS), which is based on a scale that includes assessments of mobility, gait, paw clenching, tremor, and respiration, each with a score of 0-3; the higher the score, the worse the phenotype. The D-ketamine treated group showed a slowed progression of the disease phenotype with better post-treatment neurobehavioral scores, while the untreated KO did not.

[0565]

[0566]

[0567] Open Field Test: Long-term behavioral changes in saline (WT), ketamine, and D-ketamine groups versus KO-saline group at week 8.5 of treatment. Motor function was assessed by recording mice in an open arena (10.5" x 19" x 8"). The mice were recorded in the same room as they were housed to avoid stress and changes in testing procedures. Each mouse was placed in a clean open arena and allowed to explore for 10 minutes, and its activity was recorded. Animals were placed in the same manner in the open field 5 cm away from the longer walls.

[0568] result

[0569] Animals were assessed for survival, neurobehavioral scores, and activity (total distance traveled, speed, and time spent in a corner).

[0570] Fig.13 A and 13B are the composite neurobehavioral scores of wild-type, knockout saline (control), and knockout mice treated with dendrimer-ketamine conjugates ( Fig.13 A) and the probability of survival within one day after birth ( Fig.13 B) diagram. Fig.13 C is a graph of the combined neurobehavioral score; Fig.13 D is a graph of travel distance (m).

[0571] Dendrimers conjugated to ketamine resulted in increased efficacy with increased binding to NMDARs without associated side effects. The doses used were significantly lower than free ketamine, which should reduce side effects. This was tested in a mouse model of Rett syndrome as a proof of concept, as Rett syndrome is a disease with increased glutamate production and increased NMDAR expression / activation.

[0572] A significant improvement in survival was seen with D-ketamine up to 90 days of age (with 100% survival) compared to untreated animals and animals treated with free ketamine. Previous published data by others in a similar model showed only a 50% survival rate following treatment with 320 mg / kg of ketamine (an 8-fold higher dose).

[0573] In the case of D-ketamine, significant improvements were seen in motor function, with behavior similar to normal healthy controls.

[0574] In the open field, significant differences were seen in the total distance traveled by WT mice and KO mice over a duration of 10 minutes. Significant improvements in motor function were also observed, as evidenced by an increase in the total distance traveled by D-ketamine treated KO in the open field test. Significant improvements were observed in maximum speed and time spent in a corner by D-ketamine compared to the KO saline group.

[0575] The video shows a dramatic improvement in the phenotype, bringing them close to healthy mice.

Claims

1. A composition comprising a dendrimer, preferably a hydroxyl-terminated dendrimer, a sugar-terminated dendrimer and / or a sugar-based dendrimer, covalently conjugated to at least one antidepressant and / or antipsychotic compound, wherein the hallucinogen and psychedelic is not ketamine, mephedrone, a derivative of ketamine, a cannabinoid, ibogaine or a derivative of a cannabinoid or ibogaine.

2. The composition according to claim 1, wherein the covalent conjugation is a covalent connection between modified or unmodified surface groups or internal groups of the dendrimer, including amide bonds, ester bonds, disulfide bonds, ether bonds or phosphate bonds.

3. The composition according to any one of claims 1 or 2, wherein the covalent bond between the dendrimer and the antidepressant and / or antipsychotic compound is cleaved after administration.

4. A composition according to any one of claims 1 to 3, wherein the dendrimer is a glucose dendrimer or a polyamidoamine (PAMAM) dendrimer of generation 1 to 10 (e.g., generation 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10), preferably wherein greater than 40% to 100% of the surface groups are hydroxylated or conjugated to monosaccharides.

5. The composition according to any one of claims 1 to 4, wherein the dendrimer is a glucose dendrimer, preferably a 1st, 2nd or 3rd generation glucose dendrimer.

6. The composition of claim 1, wherein the dendrimer is a generation 2-7 PAMAM dendrimer modified with sugar moieties, wherein the sugar groups are selected from the group consisting of glucose, galactose, mannose and fructose.

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

8. The composition of claim 1, wherein the dendrimer is a 2nd to 10th generation PAMAM dendrimer (e.g., a hydroxyl-terminated PAMAM dendrimer), preferably wherein greater than 40% of the functional groups are in the form of hydroxyl (OH) or monosaccharide groups.

9. The composition of claim 1, wherein the dendrimer is a glucose dendrimer made of glucose and optionally ethylene glycol building blocks having greater than 10 surface glucose moieties.

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

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

12. The composition of any one of claims 1-11, wherein the compound is a selective serotonin reuptake inhibitor (SSRI), a serotonin-norepinephrine reuptake inhibitor (SNRI), an antipsychotic, an atypical antipsychotic, a tricyclic antidepressant (TCA), a monoamine oxidase inhibitor (MAOI), a benzodiazepine, a beta blocker (BB) and an anticonvulsant.

13. The composition according to claim 12, wherein the antidepressant is a serotonin receptor antagonist, preferably selected from citalopram, escitalopram, fluoxetine, paroxetine, paxparoxetine, vilazodone, sertraline, fluvoxamine, analogs or derivatives thereof.

14. The composition according to claim 12, wherein the antidepressant is a serotonin and norepinephrine receptor antagonist, preferably selected from imipramine, nortriptyline, amitriptyline, doxepin, desipramine, duloxetine, venlafaxine, desvenlafaxine, levomilnacipran, analogs or derivatives thereof.

15. The composition according to claim 12, wherein the antidepressant is a MAO-A and / or MAO-B receptor antagonist, wherein the antidepressant is selected from the group consisting of tranylcypromine, phenelzine, isocarboxazid, and selegiline.

16. The composition of claim 12, wherein the antidepressant alters neuronal excitability, preferably wherein the antidepressant is ganaxolone, minaxolone, zolanolone, an analog or derivative thereof.

17. The composition according to claim 12, wherein the antidepressant is a norepinephrine and dopamine receptor antagonist, preferably wherein the antidepressant is bupropion and / or mirtazapine, an analogue or derivative thereof.

18. The composition according to claim 12, wherein the antipsychotic compound is a typical antipsychotic and / or an atypical antipsychotic, preferably wherein the antipsychotic is quetiapine, haloperidol, risperidone, olanzapine, clozapine, an analogue or derivative thereof.

19. The composition of claim 12, wherein the compound is selected from the group consisting of methylphenidate, amphetamine, atomoxetine, clonidine, guanfacine, viloxazine, analogs or derivatives thereof.

20. The composition of any one of claims 1-19, wherein the drug solubility of the dendrimer-antidepressant and / or antipsychotic composition is increased by at least 10-fold compared to the free antidepressant and / or antipsychotic, or wherein the dendrimer-antidepressant and / or antipsychotic composition reduces the onset of action compared to the free antidepressant and / or antipsychotic.

21. The composition of any one of claims 1-19, wherein the dendrimer-antidepressant and / or antipsychotic drug composition reduces the time to drug onset by 50% or more.

22. The composition of any one of claims 1-19, wherein the dendrimer-antidepressant and / or antipsychotic drug combination reduces side effects of the free drug.

23. The composition of any one of claims 1-22, wherein the dendrimer-antidepressant and / or antipsychotic drug conjugate further comprises a pharmaceutically acceptable excipient.

24. The composition according to any one of claims 1 to 23, comprising higher generation dendrimers, preferably PAMAM dendrimers of generation 3, 4, 5 or 6 (e.g. hydroxyl terminated PAMAM dendrimers), glucose dendrimers of generation 1, 2, 3 or higher, or functionalized with PEG, wherein the conjugate is confined to peripheral circulation.

25. The composition according to any one of claims 1 to 24, wherein the at least one antidepressant and / or antipsychotic compound is conjugated to the dendrimer via a spacer comprising a hydrocarbon (e.g. an olefin), a diethylene glycol moiety, an oligoethylene glycol chain, a triazole moiety or a combination thereof, preferably wherein the spacer comprises a triazole moiety.

26. A method of administering a dendrimer conjugate according to any one of claims 1 to 23 to an individual in need thereof, preferably via a route selected from the group consisting of mucosal administration, enteral administration or injection, preferably intranasal, intravenous, oral, sublingual, subcutaneous, inhalation, transdermal, intraperitoneal or intrathecal administration.

27. The method of claim 26, comprising preventing or treating depression, such as major depressive disorder, refractory depression and postpartum depression, post-traumatic stress disorder, panic disorder, social anxiety disorder, anorexia nervosa, suicidal ideation, obsessive-compulsive disorder, ADHD or its symptoms, premenstrual dysphoric disorder, anorexia, substance abuse disorders, epilepsy, bipolar disorder, autism spectrum disorder, attention deficit hyperactivity disorder, schizophrenia, cluster headaches, migraines, seizures, fibromyalgia, narcolepsy, obesity, Alzheimer's disease, Tourette syndrome, pain, such as neuropathic pain and chronic pain, phobias and cardiovascular disease, wherein the composition comprises a dendrimer conjugated to or formulated with a therapeutic agent, such as a selective serotonin reuptake inhibitor (SSRI), a serotonin and norepinephrine reuptake inhibitor (SNRI), a norepinephrine-dopamine reuptake inhibitor (NDRI), a tricyclic antidepressant (TCA), a monoamine oxidase inhibitor (MAOI), a benzodiazepine A drug, a GABA modulator such as a neurosteroid, an antipsychotic, an atypical antipsychotic, or an analog thereof, wherein the composition is delivered via an oral, subcutaneous, transdermal, intranasal, intramuscular, intravenous, or intrathecal route.

28. The method of claim 27, comprising treating a neuroinflammatory mental health disorder by modulating activation of a receptor on microglia by administering the dendrimer-agent conjugate.

29. The method of claim 27, comprising treating a mental health disorder by modulating activation of a receptor on a neuron by administering the dendrimer-agent conjugate.

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